NUCIS number 12. September 2004. 52 pages (full ... - IAMZ - ciheam

N U C I SN E W S L E T T E RInformation Bulletin of the Research Network on Nuts (FAO-CIHEAM)INIAINSTITUTO NACIONAL DE INVESTIGACIÓN Y TECNOLOGÍA AGRARIA Y ALIMENTARIANumber 12 September 2004CIHEAMIRTA - Mas Bové ● Coordination Centre of the Research Network on NutsFAO CIHEAMNut NetworkYoung almond orchard growing on the slopes of Sierra de María, Almería, Andalusia, SpainEDITORIALActivities 2003-2004The activities of the Nut Network havebeen affected by the assessment andevaluation phase of the European Systemof Cooperative Research Networks inAgriculture (ESCORENA). This systemwas established in 1974 by FAO and Europeanresearch institutions, and currentlyconsists of eleven active networks. Dueto budgetary reductions, FAO is no longerin a position to provide the ESCORENAsecretariat for servicing the Networks norof supporting activities. Nonetheless,most of the planned activities under theFAO-CIHEAM Interregional CooperativeResearch Network on Nuts could be implementedthrough the support providedby the FAO’s Regional Office for Europeand some FAO technical services(AGPS) and financial support providedby INIA of Spain, INRA of France andthe co-sponsor and partner CIHEAM permitsthe most planned activities to be finallydeveloped for exemple the publicationof this Newsletter and the Inventoryof Walnut Germplasm, Research and References.In addition, some researchersfrom developing or transition countrieshave been supported to participate in differentmeetings and congresses.During 2003 and part of 2004 a number ofother activities was implemented withinthe framework of the FAO-CIHEAM InterregionalCooperative Research Networkon Nuts. In June 2003, the XIII GREMPAAlmond and Pistachio Meeting was heldin Mirandela, Tras os Montes, Portugaland the proceedings will be published inthe Options Méditerranéennes Cahiers byNovember 2004. In June 2004, the VIFAO-CIHEAM - Nucis-Newsletter, Number 12 September 20041

ISHS International Congress on Hazelnutwas held in Tarragona, Spain and theproceedings will be published in ActaHorticulturae. During 2004 two moreSymposia are planned: (i) the III ISHS InternationalSymposium on Chestnut to beheld in October in Portugal, Chaves, VilaReal; and (ii) the V ISHS InternationalWalnut Symposium to be held in Sorrento,Italy in November.The Network’s Coordinator participated intwo Scientific Committee Meetings andthe Convention of the International TreeNut Council (INC) held in Berlin (Germany)and Las Vegas (USA) respectively.The importance of promoting the healthycomponent of nut consumption in the humandiet through advertisement wasstressed. This is considered essential forsuccessful marketing and to sustain demand.Worldwide nut production is increasingover the years and its supply istaken by the also increasing world consumption.It seems that nut markets canbe further developed. The INC expressedan interest to collaborate and work closelywith the Nut Network. The issues ofpreventing and reducing kernel’s aflatoxincontamination through better crop preharvestand postharvest management is ofparamount importance for the INC.Almond trees starting bloom at IRTA,E.E. Lleida, SpainESCORENA’s reviewThe ESCORENA networks can be classifiedinto three groups according to thefield of agricultural activity: crop production,animal production and environment.The 23 rd Regional Conference for Europe(Nicosia, Cyprus, May 2002) recognizedthe importance of ESCORENA as the researchnetworks brought added value notonly to European scientists and their researchinstitutions, but also to partnersand beneficiaries beyon European borders.Furthermore, the Conference endorsedthe recommendation of the 32 ndSession of the European Commission onAgriculture (ECA) that an assessmentand evaluation be undertaken for eachnetwork. During 2003, this assessmentwas carried out by M. Larbier, a visitingscientist seconded to FAO by INRA, andhis report and proposals for the futurewere submitted to the 33 rd Session ofECA held in Rome in March 2004. OurReseach Network on Nuts was consideredamong those ten to be retained andfor having good potential for continuity inthe future. This conclusion was reacheddue to dynamic coordination, adaptationof research towards the aim of sustainabledevelopment, importance of the programmeand quality of the scientific andtechnical communication means used.The ECA was also informed of CIHEAM’sintention to continue to cooperate and cosponsorthree of these ten networks, includingour Nut Network. The report submittedto the ECA included an estimate ofthe necessary funds needed to ensure futurefunctioning of ESCORENA. Consideringthe budgetary constraints faced byFAO, the proposal to establish a multi-donorTrust Fund was made. The ECA reportwas submitted to the 24 th FAO RegionalConference for Europe (Montpellier,May 2004) but no decission on this issuewas taken. It seems that the financialsupport towards our Network by themember countries under circumstancesof financial difficulties and uncertainty isessential to foster activities.The Nut Network’s futureNut tree production, trade and industry isan important economical activity relatedto sustainable agriculture, often in marginallands, in both European and NearEastern Regions. Nuts are of major importanceand are typical components ofthe traditional and healthy Mediterraneandiet. To accomplish sustainable developmentand food security, a future combinedeffort in R&D, environmental managementand communication is needed.Due to overall budgetary constraints, theNetwork on Nuts has in recent years, undergonea restructuring exercise which isnow fully implemented. Thus, the Networkhas reorganized and simplified its structureand is trying to focus more on developmentoriented issues rather than its previousstrong genetic resources collection,description and improvement. In addition,as a result of the assessment carried outin 2003 mentioned above, two workinggroups: Genetic Resources and Economicswere suggested for elimination.Considering the current and future relevanceof the Network, it is generally seenby its members as a useful tool for bettercollaboration, and the opportunities tocome together are viewed as very importantfor project development. Accordingly,recent Network developments are focusedon research issues which are of priorityimportance for developing countriesand for countries in transition, particularlyin the Balkans, Caucasus and North Africa,but also beyond, wherever researchresults are relevant to stimulate growthand alleviate poverty and hunger. The relevanceand potential usefulness of ourNut Network is based on the availabilityof considerable genetic resources andtechnical experience for the developmentof nut tree crops in Europe and the NearEast regions. In addition, useful contributionsto rural development and the globalgenetic resources understanding can bemade. Major research areas of broad interest,which should be addressed are: foodsafety, biodiversity, biotechnology and sustainablemanagement of natural resources,including low-input agriculture. To strengthenFAO’s support to our Network it is necessaryto adopt a “project approach” ofactivities. This should be carried out incollaboration with relevant FAO Units.Genetic resources inventoriesThree Inventories on Germplasm, Researchand References have been alreadypublished: Almond (1997, RTS 51),Hazelnut (2000, RTS 56) and Chestnut(2001, RTS 65). These inventories publishedin the REU Technical Series(http://www.fao.org/regional/europe/PUB),are important compilations of the currentlyavailable species genetic resourcesand information on ongoing research projectsand bibliography. In addition, twomore inventories are being compiled andare at different stages of completion. TheInventory on Walnut being compiled by E.Germain is to be published shortly andthe Inventory on Pistachio is being collatedby N. Kaska and B.E. Ak. All thesecatalogues are being supported by FAO’sRegional Office for Europe and the Seedand Plant Genetic Resources Service(AGPS) together with CIHEAM-IAMZ.In addition, a draft Descriptors List forHazelnut has been developed by membersof the Network and has now beensubmitted to IPGRI for assessment andeventual editing and publication.Changes in the FAO Regional Officefor EuropeAs mentioned above, Mr. Michel Larbier,a visiting scientist seconded from INRA,France, took the task to evaluate andsuggest a proposal to re-organize ESCO-RENA. The efforts made by M. Larbier inreviewing the whole ESCORENA systemare acknowledged. Ms. Jutta Krause, RegionalRepresentative for Europe (FAORegional Office for Europe in Rome) isthe FAO contact person for ESCORENA.2 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

The Nut Network on the webIn addition to the already existing informationon the Nut Network at http://www.iamz.ciheam.org/ingles/nuts.htm,and on the previous FAO web page createdin 2000, the FAO European System ofCooperative Research Networks in Agriculture(ESCORENA) and the Nut Networkweb page has now been closed dueto lack of commitment from coordinatorsto make use of the interactive system fordata entry and lack of resources fromFAO to take over this role. However, essentialinformation on network coordinators,major network publications (e.g.REU Technical Series), ESCORENA reportsand links to existing network websitessuch as the nut network is included inthe REU website (see http://www.fao.org/world/regional/REU/content/escorena/index_en.htm ).NUCIS on the webA short version of the Newsletter (editorial,contents and back page) from issuenumber 6 and onwards, is available onthe Internet web pages of both FAO (http://www.fao.org/regional/europe/public-e/nucis.htm) and CIHEAM (http://www.iamz.ciheam.org/ingles/nuts.htm# publications).A full electronic version of NU-CIS is available from issue number 9 andonwards (http://www.iamz.ciheam.org/ingles/nucis9.pdf).The contents of thisNewsletter can be browsed through andalso copied and printed.Contributions to NUCISAs in past NUCIS editorials, it is againstressed that this Newsletter must be aneffective vehicle of communication for allthe Network members. The pages of thisbulletin are open to all readers who wouldlike to suggest ideas or to express theiropinion about the work developed by theNetwork (activities carried out and planned)or to publish short articles and reportson relevant horticultural subjects ofgeneral interest. A sufficient number ofcontributions are received from the MediterraneanBasin and overseas for the articlesand reports section. However, thesections on news and notes and also oncongresses and meetings are usually difficultto cover due to the scarce informationreceived and thus, contributions aremost welcomed. Otherwise, the Editorhas to report on the issues he is aware of,but certainly there must be many more issueson-going throughout the year whichmerit reporting. Also, the place for ‘grey’bibliography (references and documentswhich are difficult to search like Mastersor Ph. Theses) is scarcely filled.The NUCIS Newsletter is distributedworldwide free of charge to 1.400 readersfrom over 60 countries. The disseminationof information originated by the Networkis of paramount importance and throughthis bulletin has been largely successful.The first NUCIS was published in 1993,this issue of the NUCIS Newsletter isnumber 12 and during these eleven yearsgreat editing effort has been made. I acknowledgeand gratefully thank all contributorsfor their efforts and interest to produceand send me valuable information.The exchange of information betweenNetwork members through the pages ofthis Newsletter is the basis for developingcollaboration. The editing task in the elevenNUCIS issues already published hasbeen huge (NUCIS 1, 9 pages; 2, 20 pages;3, 24 pages; 4, 28 pages; 5, 36 pages;6, 52 pages, 7, 44 pages, 8, 46 pages9, 68 pages, 10, 48 pages, 11, 48 pagesand 12, 52 pages). In order to reducethe time consuming formal editing contributorswho send articles, news, notes, bibliographicreferences, etc. to the differentsections are asked to provide themwell organized and elaborated. Informationshould be sent in satisfactory English.Contributions could be sent throughInternet using the Editor’s email. The alternativeis to provide them on disketteand also in printed format. This bulletin isreproduced in black and white only, includingpictures. Please send your contributionsfor the next issue, number 13 by theend of October 2005; however it cannotbe guarantied that the article will be publisheddue to the uncertainty of funding.We apologize to readers for the delay inprinting this issue due to late funding. Wethank all who have contributed to this issue.The EditorThe designations employed and thepresentation of material in this publicationdo not imply the expression of anyopinion whatsoever on the part of theFood and Agriculture Organization ofthe United Nations concerning the legalstatus of any country, territory, city orarea or of its authorities, or concerningthe delimitation of its frontiers or boundaries.This publication contains the collectiveviews of an international group of expertsand does not necessarily representthe decisions or the stated policyof the Food and Agriculture Organizationof the United Nations, the InternationalCentre for Advanced MediterraneanAgronomic Studies nor of the Organizationfor the Economic Cooperationand Development.Contributions should be written conciselyin English. Please send contributionson paper and diskette (Microsoft ®Word or Word Perfect ® ). Authors areresponsible for the content of their papers.Reproduction of the articles is authorized,provided that the original sourceis clearly stated.CONTENTSEDITORIAL .......................................... 1ARTICLES AND REPORTS• Consumers’ acceptance of industrial productsmade with hazelnuts and almonds .............................. 4• Differential flower and fruit damagesby spring frosts in almond .......................................... 5• Prunus webbii as a way to look intoself-compatibility in almond ....................................... 8• Application of molecular markers in almondbreeding programmes ................................................ 9• Fusicoccum canker in almond orchards –biotechnology tools for early selectionof tolerant genotypes ................................................ 12• Virus screening for Australian almonds .................... 15• The pistachio industry in Italy: current situationand prospects ........................................................... 16• Pistachio production in Tunisia ................................ 19• Breeding monoecious pistachio cultivars ................. 21• Chilling stress and injury in pistachiofemale flowers .......................................................... 23• Underground drip system: the new irrigation methodfor pistachio and almond orchard in Turkey ............. 24• Iranian agricultural system to develop higher qualityand healthier pistachio production ........................... 26• Pistacia vera L. 1753 in Iran ..................................... 26• Pistacia atlantica desf. 1800 in Iran .......................... 27• Pistacia khinjuk stocks 1852 in Iran ......................... 30• Germplasm collection of pistacia at the JacobBlaustein Institute for desert research. Phenotypictraits and molecular markers .................................... 31• Where in a pistachio tree is Xanthomonasand how did it get there? .......................................... 33• Chestnut production in Andalusia ............................. 33• The world macadamia industry and researchactivities ................................................................... 36• Carob tree development in China .............................. 39NOTES AND NEWS• In memorian: my professor Dale Kester ................... 41CONGRESSES AND MEETINGSPage• XIII GREMPA meeting on pistachios and almonds ... 41• VI International Congress on Hazelnut ...................... 42TO BE HELD ................................................................. 45BIBLIOGRAPHY ........................................................... 46BACKPAGE ................................................................... 52FAO-CIHEAM - Nucis-Newsletter, Number 12 September 20043

ARTICLESAND REPORTSCONSUMERS’ ACCEPTANCEOF INDUSTRIAL PRODUCTSMADE WITH HAZELNUTSAND ALMONDSINTRODUCTIONHazelnuts and almonds are used to makeindustrial products like snacks, turrons,marzipan or chocolates. Although there iswide information about their chemicalcomposition and the influence of manyprocesses on the products´ final characteristics,there is a lack of informationabout consumers’ acceptance of these typesof products when hazelnut and almondvarieties change.For these reasons IRTA started in 1999 aresearch programme in order to analyzethe importance of selecting different nutvarieties to make different industrial products,as a consequence of consumers’acceptance. This information could be relevantnot only to producers but also forthe confectionery industry. Results for differentnut products have been publishedin several scientific reports (see references).An overall analysis of hazelnut andalmond commodities is made in this article.Variety is used as synonym of cultivar.METHODOLOGYFrom 1999 to 2002 four hazelnut varieties(‘Negret’, ‘Pauetet’, ‘Tomboul’ and ‘Tondadi Giffoni’) and eight almond varieties(‘Desmayo Largueta’, ‘Francolí’, ‘Glorieta’,‘Marcona’, ‘Masbovera’, ‘Nonpareil’and ‘Planeta’) were used to make severalproducts: dark chocolate with nuts, whitechocolate with nuts, brown chocolate withnuts, bonbons made with roasted nuts enrobedwith burnt sugar and coated withdark chocolate, marzipan and hard andsoft turrons.Chocolates and bonbons were made incollaboration with the industry “L’Art de laXocolata” (El Vendrell, Tarragona), whilemarzipan and turrons were made in theindustry “Turrons i Mel Alemany SL” (Osde Balaguer, Lleida). Hazelnut and almondsamples came from IRTA´s collections,while ‘Tomboul’ hazelnuts and‘Nonpareil’ almonds were purchased onthe market.Each commodity was assessed for industrialfacility, final quality and consumers’acceptance. Commodity acceptance wasevaluated using the consumers panelFigure 1.- Chocolate with nuts.“L’Art de la Xocolata” industryfrom IRTA (table 1), including over 670people (almost 300 families), and distributedall around Catalonia (Spanish NE).Consumer’s answers were weighted byage and education level in order to extrapolateconclusions to Catalonian people.MAIN RESULTSResults show that there are statistical differencesin variety acceptance, dependingon the industrial product studied. Sometimesthese differences can be solvedchanging the technological process, althoughthis has been observed for roastedhazelnuts and not for other products.For example, dark chocolate (Figure 1)was the most accepted, mainly for ‘Pauetet’hazelnut variety and for ‘Masbovera’almond variety. ‘Negret’ and ‘Tomboul’hazelnuts and ‘Desmayo Largueta’ and‘Francolí’ almonds showed a very goodtechnological behaviour to make bonbons(figure 2). ‘Marcona’ almonds were thebest accepted to make marzipan (Figure3) and hard turron (Figure 4), while ‘Negret’variety was the most accepted forhard turron made with hazelnuts. Finally,soft turrons (Figure 5) were most accep-Table 1.- Consumers distribution by sex, age, education level and city size in boththe IRTA panel and the Catalonian people.IRTA’s panelFigure 2.- Chocolate coating to make bonbons.“L’Art de la Xocolata” industryFigure 3.- Cutting marzipan.“Turrons i mels Alemany” industryted when they were made with ‘Marcona’or ‘Desmayo Largueta’ almond varieties.The overall analysis of the results alsoshows that sensorial characteristics fromhazelnuts and almonds must be consideredto make an industrial product. Inmost of them, sensorial profile from roastednuts should be known while in a fewof them the most important is the profilefrom raw nuts. Even though, when a productincludes sugar in its recipe the differencesin taste are not relevant enough,but it is still important that nut taste canbe identified.CataloniaSex: % %Men 49.7 48.8Women 50.3 51.2Age:< 30 years old 23.5 36.730 to 65 years old 68.6 46.1> 65 years old 7.9 17.2City habitants:< 30.000 40.4 43.6> 30.000 59.6 56.4Education level:Basic 34.9 69.3Middle 22.9 20.7High 42.2 10.04 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

Figure 4.- Hard turron ‘Alicante’.Labelled and vacuum packaged tabletsFigure 5.- Soft turron ‘Jijona’.Labelled and vacuum packaged tabletsRaw hazelnuts and almonds homogeneityis very important. Even though, this pointshould be studied in more detail for suchkind of products where nuts are grounded,not roasted or sugar enriched. Maindefects to be considered in raw nut bulksare rancidness, bitterness or burnt kernels.Some hazelnut and almond varietiesseem to show serious difficulties to beadapted to some industrial processes,due to a very high fragility or strength, anexcess of splits generation, an inadequatesurface to volume ratio, bad roastingaptitude, etc.Finally, both hazelnuts and almondsseem to show the same acceptation atconsumers level for those particular productsstudied. This could confirm thatthey can substitute each other dependingon the marked price, as it is usually assumed.However, it should be further confirmedfor other products.ACKNOWLEDGMENTSResearch supported by Instituto Nacionalde Investigación y Tecnología Agraria yAlimentaria (INIA) of Spain, project numberSC99-002.REFERENCESRomero, A., Tous, J.; Guerrero, L.;Gou, P.; Guardia, Mª D., 1999. Aplicacionesdel análisis sensorial en el tostadode avellana en grano. FruticulturaProfesional (Especial Frutos Secos II),104: 71-77.Gou, P.; Díaz, I.; Guerrero, L.; Valero,A.; Arnau, J.; Romero, A. 2000. Physico-chemicaland sensory property changesin almonds of ‘Desmayo Largueta’variety during toasting. Food Sci. Tech.Int., 6 (1): 1-7.Romero, A.; Tous, J., 2000. Nutricionalvalue of hazelnut (Corylus avellana, L.).Nucis-Newsletter, 9 (12): 25-27.Tous, J.; Romero, A., 2002. Almondand hazelnuts products. Consumer´spreferences. Nucis-Newsletter, 10 (12):12-13.Romero, A.; Tous, J.; Plana, J.; Guardiola,MD.; Díaz, I., 2002. How varietychoice affects Spanish consumer´s acceptanceof marzipan and chocolatesmade with almonds. Ed. ISHS. ActaHorticulturae, 591: 117-123.Romero, A.; Tous, J.; Plana, J., 2003.Importancia de la variedad de almendraen la aceptación de turrones tipo ‘Alicante’y ‘Jijona’. II Congreso Nacionalde Ciencia y Tecnología de los Alimentos.Orihuela (Alicante). Ed. UniversidadMiguel Hernández (UMH). VolumenII: 611-614.Romero, A.; Tous, J.; Plana, J. 2004.How cultivar choice affects spanishconsumers’ acceptance of chocolates,bonbons and hard turron made with hazelnuts.6th Int. Congress on Hazelnut.Tarragona-Reus, 14-18 June.A.Romero and J.TousIRTA, Centre Mas Bové. Apartat 415, 43280-Reus (Tarragona, SpainE-mail: agusti.romero@irta.esDIFFERENTIAL FLOWER ANDFRUIT DAMAGES BY SPRINGFROSTS IN ALMONDINTRODUCTIONFlowers and young fruits of almost all deciduousfruit trees are vulnerable to frostdamage (Andrews et al., 1983). Consequentlyspring frost injury is a major limitingfactor in the production and the distributionof horticultural crops (Ashworth,1992). The anatomical, physiological andbiological aspects of this damage havebeen reviewed in fruit trees (Rodrigo,2000). In Prunus, fruit damage is detectableby observing ovule browning, ice formation(Saunier, 1960), and browning ofthe petals and pistils.Almond (Prunus amygdalus, Batsch) isan early blooming species. As a result, almondcultivation was restricted to regionswith low risk of spring frosts. However,the expansion of the culture into inlandMediterranean areas, where the occurrenceof spring frost is common and overlappingwith bloom of most almond cultivars,increased the risk of reducing oreven nullifying yield (Socias i Company etal., 1999). Thus, almond breeding aimedto create and select late blooming cultivars(Kester and Assay, 1975). Nevertheless,this solution is not enough to overcomespring frost damages, and frosthardness is also considered a selectionobjective in our breeding programme (Sociasi Company et al, 1998).In almond, little is known about the minimumthreshold temperature causing damagesin flowers and young fruits. However,differences among several commercialalmond cultivars were described byFelipe (1988), concluding that at –2.5ºCsome cultivars suffered heavy productionlosses. Our aim was to assess springfrost damage in the flower and youngfruits of several almond selections andtheir consequences on final fruit set.MATERIALS AND METHODSThe study was carried out during the winter-springof 2003 on 21 self-compatibleselections from CITA´s breeding programmeat Zaragoza. They were graftedonto the almond x peach hybrid rootstock‘Garnem’ (Gómez Aparisi et al., 2001).The situation of this orchard, in relation tospring frost, was considered of high risk,because of the frequent incidence of frost(Felipe, 1988).To evaluate the damages caused in flowersand young fruits, samples were collectedtwo days after frost occurrence,placed in polyethylene bags, and taken tothe laboratory. Damage evaluation wasbased on the morphological symptoms,such as brown discolouration of the stylebase in flowers or browning of the ovulein young fruits. To assess the effect offrost on fruit set, all bud flowers werecounted on two branches of each selectionbefore bloom, thus much before frostincidence. Fruit set was recorded in June.Bud density was also measured accordingto Socias i Company (1988).RESULTS AND DISCUSSIONA frost of –2.5°C during five hours tookplace in the early morning of March 18,2003. On that day, the almond selectionswere at different phenological stages andcan be separated in two groups (Fig. 1).The first group comprised 12 selectionswhose bloom was already over, thus thefrost affected young fruits. The secondgroup comprised 9 selections that on thatday were opening early flowers; consequently,the frost only affected buds andearly flowers. The behaviour of bothgroups was different and needs to beexamined differently for each group of selections.FAO-CIHEAM - Nucis-Newsletter, Number 12 September 20045

Early blooming selections: Frost damageon small fruits ranged from 20% (G-6-39) to 77% (H-3-39) and produced a heavyreduction of fruit set, which was practicallynil in some (G-1-38 and H-3-37) andeconomically insufficient in others (G-2-23, G-2-7, G-5-18, G-6-24, and H-3-39).These could be due to the fact that theseselections had a young developing fruit,which is considered the most vulnerablestage to frost (Proebsting and Mills,1978b).Figure 1: Blooming time and fruit set of the studied selectionsIn addition, some selections showed anintermediate fruit set, although not commerciallyacceptable (G-6-39, I-1-95, andI-2-12), and selection G-1-58 showedvery high fruit set (33.5%), of a commerciallevel (Kester and Griggs, 1959). Thesedifferences, observed in selectionsthat were at the same phenological stage,could be due to a differential resistance ofthese selections to frost damage. Thesame observation was reported for otheralmond cultivars previously studied (Felipe,1988), as spring frost damage on reproductiveorgans depends mainly on thecultivar (Proebsting and Mills, 1978a) andon the intensity and duration of the lowtemperature (Mazur, 1969). In fact, differencesin yield losses have been observedamong almond cultivars at differentcritical low temperatures (Felipe, 1988).This suggests that selection G-1-58 hassuffered lighter losses than the other selectionsof this group with a temperatureof –2.5°C.A high negative correlation was observedbetween the percentage of fruit damageand fruit set (-0.65), showing how theamount of damaged fruits significantly reducesfruit set. There was also a high negativecorrelation between the percentageof damaged fruits and bud density (-0.63), showing that the effect of springfrosts is higher in selections with a lowerbud density. Moreover, a significant positivecorrelation was found between buddensity and fruit set (0.42), thus, fruit setof selections with a medium to high buddensity is higher than that of selectionswith a low bud density (Fig 2). This couldbe due to the fact that a high potential toproduce bud flowers may compensate thespring frost damages, and, together witha good frost resistance, as in the case ofselection G-1-58, could maintain the fruitset at a commercially acceptable level.Late blooming selections: flower andbud frost damage ranged from 2% (G-2-2) to 47% (G-3-5), generally lower thanthe fruit damage on selections of the earlyflowering group, confirming that frostdamage is highly dependent on the stageof bud development, as selections of thelate flowering group (Fig 1) were in theearly blooming stages, C, D and a few E(Felipe, 1977) when the frost happened.Also, the flower hardness of each selection(genotype) reflects the differencesamong selections at the same phenologicalstage (Westwood, 1993).In general, fruit set decreased when theamount of damaged flowers increased(Figure 3), which explains the significantnegative correlation between fruit set andpercentage of damaged flowers (-0.37).In addition, in spite of the high percentageof damaged flowers, selection G-3-5shows a medium fruit set (Figure 3). Thiscould be due to the high bud density ofthis selection, which has compensatedthe loss of flowers and maintained a mediumfruit set.CONCLUSIONIn general, the amount of damages causedby frost in selections at the beginningof bloom (late flowering group) wereslight in comparison to the damages causedin early blooming selections (earlyflowering group), confirming that frost damageis highly dependent on the phenologicalstage of the bud/flower/fruit, theearly developing fruit being the most susceptiblestage (Proebsting and Mills,1978b). Furthermore, differences observedamong selections for the amount offlower and fruit losses show different frosthardness for each selection. Consequently,only tolerant or resistant genotypes(cultivars or selections) can produce acrop when frost takes place at or laterthan blooming time.In some cases frost produced the completeloss of the crop, and in others animportant reduction of fruit set and, consequently,of yield. Moreover, selectionshaving a high bud density gave a fruit setfrom medium to acceptable. Hence, ahigh bud density seems to compensatespring frost damages.These data were not enough to explainthe effect of spring frost damage on fruitset, because fruit set in almond dependson several factors, including other weatherconditions (such as rain), insect activityamong inter-compatible and overlappingblooming cultivars or the ability fornatural autogamy in self-compatible cultivars(Socias i Company and Felipe,1992). However, a wide variability of responsesto frost were observed in theseselections, showing that even with thisfrost some genotypes can reach a 25-40% fruit set, considered as the economicallyacceptable threshold for a commercialcrop (Kester and Griggs, 1959). Consequently,selection for frost resistancecan be efficiently undertaken to obtain6 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

Figure 2. Percentage of damaged young fruits, bud density and fruit setof selections of the early flowering group.Felipe, A. J., 1988. Observaciones sobrecomportamiento frente a heladastardías en almendro. Rap. EUR, 11557.145-148.Felipe, A.J., 1977. Almendro. Estadosfenológicos. Inf. Técn. Econ. Agrar., 27:8-9.Gómez Aparisi, J., Carrera, M., Felipe,A.J., Socias i Company, R., 2001. ‘Garnem’,‘Monegro’ y ‘Felinem’: nuevos patroneshíbridos almendro x melocotoneroresistentes a nematodos y de hojapara frutales de hueso. Inf. Técn. Econ.Agrar., 97V(3): 282-288.Kester, D.E., Asay, R.D., 1975. Almonds.In: Advances in Fruit Breeding.Janick, J. and Moor, J. N (eds). PurdueUniv. Press, West Lafayette (IN), pp.387-419.Kester, D.E., Griggs, W.H., 1959. Fruitsetting in almond: the effect of crosspollinatingvarious percentages of flowers.Proc. Amer. Soc. Hort. Sci. 74:214-219.Mazur, P., 1969. Freezing injury inplants. Annu. Rev. Plant Physiol. 20,419-448.Figure 3. Percentage of damaged flowers, bud density and fruit set of selectionsof the late flowering group.Proebsting, E.L., Mills, H.H., 1978a. Asynoptic analysis of peach and cherryflower bud hardiness. J. Amer. Soc.Hort. Sci. 103, 842-845.cultivars adapted to the harsh weatherconditions of many almond growing regionsin continental climates.ACKNOWLEDGMENTSThis work was supported by the projectAGL2001-1054-C03-02 of the SpanishCICYT. OK gratefully acknowledges agrant from Instituto Nacional de Investigacióny Tecnología Alimentaría (INIA).Technical assistance by J. Búbal, O.Frontera and meteorological data from A.Cabezas are highly appreciated.REFERENCESAndrews, P. K., Proebsting, E.L.,Gross, D.C., 1983. Differential thermalanalysis and freezing injury of deacclimatingpeach and sweet cherry tissues.J. Amer. Soc. Hort. Sci. 108, 755-759.Ashworth, E.N., 1992. Formation andspread of ice in plant tissues. Hort. Rev.13, 215-255.Proebsting, E.L., Mills, H.H., 1978b.Low temperature resistance of developingflower buds of six deciduous fruitspecies. J. Amer. Soc. Hort. Sci. 103,192-198.Rodrigo. J., 2000. Spring frosts in deciduousfruit trees- morphological damageand flower hardiness. Scientia Hort.85, 155-173.Saunier, R., 1960. La lutte contre lesgelées printanières chez les arbres fruitiers(I). Pomol. Fr. 2: 5-12.Socias i Company, R., 1988. La densitéflorale comme critère variétale chezl’amandier. Rap. EUR, 11557. 145-148.Socias i Company, R., Felipe, A.J.,1992. Self-compatibility and autogamyin ‘Guara’ almond. J. Hort. Sci. 67: 313-317.Socias i Company, R., Felipe, A.J. andGomez-Aparisi, J., 1999. A major genefor flowering time in almond. PlantBreed., 118: 443-448.Socias i Company, R., Felipe, A.J., GómezAparisi, J., García, J.E., Dicenta,F., 1998. The ideotype concept in almond.Acta Hort., 470: 51-56.Westwood, M.N., 1993. TemperatezonePomology: Physiology and Culture.Timber Press, Portland.O. Kodad and R. Socias i CompanyUnidad de Fruticultura CITA, DGAApartado 727, 50080 Zaragoza, SpainE-mail: okodad@aragob.esFAO-CIHEAM - Nucis-Newsletter, Number 12 September 20047

Prunus webbiiAS A WAY TO LOOK INTOSELF-COMPATIBILITYIN ALMONDINTRODUCTIONAlmond [Prunus amygdalus Batsch or P.dulcis Miller (D.A. Webb)], derives fromone or more wild almond species (suchas P. fenziliana, P. bucharica, P. kuramica,P. triloba) that evolved in the desertsof Central Asia. By 450 BC, almond startedspreading in the Mediterranean basinand countries like Italy, Spain, France,Portugal, Morocco, Tunisia became importantcentres of production (Kester andRoss, 1996).The fact that almond is mainly a self-incompatiblespecies accounts for the greatdiversity found and also for the differencesamong populations of different origins(with distinct histories of hybridization).Furthermore, as pointed out by Socias iCompany (2002) it is expected that hybridizationswith wild Mediterranean species(such as P. webbii) may have taken placeduring almond expansion, resulting in thealmond populations that can be currentlyfound in this region.SELF-INCOMPATIBILITYMany flowering plants display self-incompatibility(SI), a genetically determinedcharacteristic which prevents inbreedingby promoting outcrossing. Self-incompatibilityis determined by a single multialleliclocus, the S locus. Whenever there is amatch between the S alleles expressed inthe pistils and those of the pollen, fertilizationis hampered. In the families Solanaceae,Rosaceae and Schrophulariaceae,the products of the S locus in thepistil are S-RNases that are thought todestroy the RNA of incompatible pollen(Wang et al., 2003).Even though SI is believed to have playedan important role in the success ofAngiosperms, it represents a limitation toplant breeders when fruit production isimportant. In almond, SI is almost 100%effective, which means that unless compatiblecultivars are present in an orchard,nut production will never be achieved.It has long been the desire of almondbreeders to obtain self-compatible(SC) cultivars that are, at the same time,good almond producers.SELF-COMPATIBILITY IN ALMONDSome self-compatible almond varietieshave been identified and the self-compatibilitytrait is usually attributed to the presenceof a self-fertility allele S F(Raynaudand Grasselly, 1985). The Italian cultivar‘Tuono’ is the best characterized SC almondcultivar and analysis of stylar RNaseactivity indicate that S Fis inactive in‘Tuono’ styles and in its SC progeny(Boskovíc et al., 1999). The S Fgene sequencesegregating with SC was isolatedfrom ‘Tuono’ (Ma and Oliveira, 2000,2001; Channuntapipat et al., 2001)though so far it is not known why there isno corresponding RNase activity. Breedingtests confirmed segregation of S Fsequenceand lack of RNase activity withthe SC phenotype (Ortega and Dicenta,2003). Apparently, other SC almond cultivarsdisplay only one S-RNase in theirstyles (i.e. they lack one S-RNase) butthe corresponding genes have still notbeen isolated.THE ORIGIN OF SELF-COMPATIBILITYIN ALMOND‘Tuono’ is originally from Italy, from theregion of Apuglia (Herrero et al., 1977)where other Prunus such as P. webbii coexist.Because P. webbii is reported as aSC species, it has been assumed that SCtrait was introgressed into almond fromthis wild relative (Godini, 1979, 2000),probably through S f(Socias i Company,2002). A consequence of this assumptionis that P. webbii would have to carry theS fallele. Furthermore, if S fwas the onlycause of self-compatibility, all SC P. webbiiwould contain this (or similarly nonfunctional)allele(s). If, on the other hand,P. webbii would have functional S allelesstill being SC, then other loci should alsobe implicated in SI/SC in this species.THE S fALLELE IN P. webbiiWhen testing for the presence of S f(usingS fspecific primers) in six ecotypes of P.webbii (one from Spain and five fromApuglia) two were shown to carry the S fallele (Sanchez and Oliveira, unpublishedresults), which could account for theirself-compatibility. Since no RNase activityin the pistil can be attributed to S f, atleast half of the pollen grains would beable to grow in the pistil and set seed.Such Prunus webbii plants may have, atsome point, crossed with cultivated almondoriginating cultivars as ‘Tuono’ thatcarry S f. It must be noted that the hybridizationmust have occurred several generationsago since, as assessed by molecularmarkers, ‘Tuono’ is more closely relatedwith almond than with P. webbii(Martins et al., 2003), that is to say, ‘Tuono’is not an F 1hybrid and further crosseswith cultivated almond must have beentaken place in the origin of this cultivar.The fact that other ecotypes of P. webbiido not contain the S fallele but are still SCdismisses the notion that the presence ofS fis a sine qua non condition for selfcompatibility.OTHER S ALLELES IN P. webbiiBy using primers designed for the conservedregions of Rosaceae S alleles (Signalpeptide, C1 and C5) on P. webbii genomicDNA, it was possible to isolate otherS alleles besides S fin this species (Sanchezand Oliveira, 2003). Thus, for four ofthe six ecotypes tested, two S alleleswere isolated. As expected one of thesealleles was S fin the two ecotypes alreadyshown to carry S f. For the two remainingecotypes, only one S allele was found.The fact that P. webbii genome containsS-alleles does not constitute in itself asurprise. If SI was the original condition inPrunus and SC plants are derived from SIplants, it could be expected that all plantsof this genus may carry S-RNase genesthough some may be in a relic non-functionalform. Nevertheless, testing S-geneexpression on pistil RNA from four varietiesby RT-PCR, amplification was successful.This means that at least some P.webbii S-alleles are pistil-expressed suggestingthat these genes may be functional,a feature that could be confirmed bytesting the RNase activity of P. webbiipistils.S-ALLELESAND SELF-COMPATIBILITYHow plants carrying the S fallele could beself-compatible was already explained.Similarly plants carrying a single S allelemay be accounted for if we consider thatnot being able to isolate a second S-alleleis due to either its absence from the genomeor (more likely) an allele mutated tosuch an extreme that it is no longer recognisableby the primers and therefore itis most likely non-functional. The possibilitythat these individuals are homozygousfor the S-locus (since P. webbii is SC thiswould be possible) and therefore to identicalS-alleles are present which wouldnot be distinguishable by PCR. As for theecotypes that carry two distinct S-allelesthat as judged by their sequence seem tocode putatively functional S-RNases, thecause of SC may be due to a lack of (orreduced) expression of the S-genes (oneor both) or SC may be a result of the actionof other genes outside of the S-locus.It has long been known that the geneticbackground also affects the self-incompatibilityresponse.Self-fertile varieties of SI species areknown in other plants and in many casesmutations in the S-RNase coding regionor low expression levels of this proteincan explain self-fertility (Kowyama et al.,1994; Royo et al., 1994; Sassa et al.,1997). However other factors such asmutations on the putative S-pollen counterpartor at modifier loci can account forSC in the presence of fully functional S-RNases (Ai et al., 1991; Golz et al., 1998;Tao et al., 2002 and Tsukamoto et al.,2003). Similar situations are not to be discardedin Prunus webbii.8 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

ALMOND GENES IN P. webbiiAnalysis of P. webbii S-allele sequencesshowed that three ecotypes carry allelesidentical to almond functional S-alleles.The observation that self-compatible P.webbii carries alleles known to confer SIis perhaps unexpected but supports thehypothesis put forward by Socias i Company(2002) that at least to some extent,gene flow probably occurs (or has occurredin the past) between the cultivated almond(P.amygdalus or P. dulcis) andneighbouring P. webbii trees. Alternatively,but less likely, these S-alleles werepresent in a common ancestor of thesetwo species. It is interesting to note thatthe Spanish ecotype shares alleles withthe ecotypes collected in Italy. This couldreflect the ancient nature of the S-allelesthat would be present in the original populationof P. webbii or shows exchangeof plant material between these two Europeanregions.CONCLUSIONIn conclusion, S-RNase coding genescould be isolated from a self-compatiblespecies, Prunus webbii, suggesting thatthis species may have been self-incompatiblein the past. The fact that the S fallele known to confer self-compatibility inalmond was found in some ecotypes of P.webbii provides evidence to the suggestionof this species being the origin of theS fallele of ‘Tuono’. This gene introgressionmust be a result of ancient hybridizationevents. Furthermore, probably S-gene flow also occurred in the oppositedirection since almond S-alleles are presentin P. webbii. Finally, self-compatibleplants do not necessarily carry the S fallelepreviously identified, so other causesfor SC may exist that could be exploitedin breeding SC almonds.REFERENCESAi, Y., Kron, E. and Kao, T.-H. (1991).S-alleles are retained and expressed ina self-compatible cultivar of Petunia hybrida.Mol. General Genet. 230, 353-358.Boškovíc, R., Tobutt, K.R., Duval, H.and Batlle, I. (1999). A stylar ribonucleaseassay to detect self-compatibleseedlings in almond progenies. TheorAppl Genet 99: 800–810.Channuntapipat, C., Sedgley, M. andCollins G. (2001). Sequences of the cD-NAs and genomic DNAs encoding theS1, S7, S8, and S Falleles from almond,Prunus dulcis. Theor Appl Genet103:1115–1122.Godini A. (1979). Ipotesi sulla comparsadell’autocompatibilità nel mandorla.Sci Tec Agr 19: 1–10.Godini, A. (2000). About the possiblerelationships between Amygdalus webbiiSpach and Amygdalus communis L.Nucis Newsletter 9.Golz, J.F., Clarke, A.E., Newbigin, E.and Anderson, M. (1998). A relic S-RNase is expressed in the styles of selfcompatibleNicotiana sylvestris. Plant J.16, 591-599.Herrero, M., Cambra, M. and Felipe, A.J. (1977) lnterpolinización de variedadesde almendro. An. Inst. Nac. Invest.Agrar.,Ser. Prod. Veg. 7: 99-103.Kester, D.E.; Ross, N.W. (1996). HistoryIn Almond: Production Manual.W.C. Micke (ed) Publication 3364, Universityof California, Oakland.Kowyama, Y., Kunz, C., Lewis, I., Newbigin,E., Clarke, A.E. and Anderson,M.A. (1994). Self-compatibility in aLycopersicon peruvianum variant(LA2157) is associated with a lack ofstyle S-RNase activity. Theor. Appl. Genet.88, 859-864.Ortega, E. and Dicenta, F. (2003). Inheritanceof self-compatibility in almond:breeding strategies to assure self-compatibilityin the progeny. Theor. Appl.Genet. 106 (5): 904 – 911.Ma R.C. and Oliveira M.M. (2000). Molecularidentification of S-genotypes inalmond (Prunus dulcis). Proceedings ofthe ISHS symposium on molecular markersfor characterizing genotypes andidentifying cultivars in horticulture. ActaHorticulturae 546: 575-580.Ma R.C. and Oliveira M.M. (2001). Molecularcloning of the self-compatibilitygenes S1 and S3 from almond Prunusducis cv.Ferragnes. Sex Plant Reprod14:163-167.Tao, R., Habu, T., Namba, A., Yamane,H., Fuyuhiro F., Iwamoto, K. and Sugiura,A. (2002). Inheritance of S F-RNasein Japanese apricot (Prunus mume) andits relation to self-compatibility. TheorAppl Genet 105: 222–228.Crossa Raynaud, P and Grasselly, Ch.(1985). Existence de groupes d’interstérilitéchez l’amandier. Options MediterraneanCIMEAM/ IAM2 85/1: 43-45.Royo, J., Kunz, C., Kowyama, Y., Anderson,M.A., Clarke, A.E. and Newbigin,E. (1994) Loss of a histidine residueat the active site of S locus ribonucleaseis associated with self-compatibilityin Lycopersicon peruvianum. Proc.Natl Acad. Sci. USA 91, 6511-6514.Sanchez. A.M. and Oliveira, MM (2003).S-alleles in self-compatible Prunuswebbii. CIHEAM Cahiers Options Méditerranéenes,Proceedings of the XIIIGREMPA Meeting on Pistachios and Almonds(in press).Sassa, H., Hirano, H., Nishio, H. andKoba, T. (1997). Style-specific selfcompatiblemutation caused by deletionof the S-Rnase gene in Japanese pear(Pyrus serotina). Plant J. 12, 223-227.Socias i Company. (2002). The relationshipof Prunus webbii revisited. Nucisnewsletter 11: 17-19.Tsukamoto, T., Ando, T., Kokubun, H.,Watanabe, H., Sato, T., Masada, M.,Marchesi, E. and and Kao, T.- H.(2003). Breakdown of self-incompatibilityin a natural population of Petuniaaxillaris caused by a modifier locus thatsuppresses the expression of an S-RNase gene. Sexual Plant Reproduction15: 255–263.Wang, Y., Wang, X., Skirpan, A.L. andKao, T.-H. (2003). S-RNase-mediatedself-incompatibility Journal of ExperimentalBotany, 54 (380): 115-122.A. Sanchez 1 and M. Oliveira 1, 21. Laboratory of Plant Genetic Engineering,IBET/ITQB, Quinta do Marquês, 2784-505Oeiras, Portugal2. Departamento de Biologia Vegetal,Faculdade de Ciências da Universidade deLisboa, Campo Grande 1749-016 Lisboa,PortugalE-mail: asanchez@itqb.unl.ptAPPLICATION OF MOLECULARMARKERS IN ALMONDBREEDING PROGRAMMESINTRODUCTIONDeveloping new cultivars is a long and tediousprocess in almond [Prunus dulcis(Mill.) D.A. Webb], involving the generationof large populations of seedlings fromwhich the best genotypes are selected.Whereas the capacity of breeders to generatebig populations from crosses is almostunlimited, its management, screeningand selection of seedlings are themain limiting factors in the generation ofnew releases. The development of molecularmarkers to be used in early selectionof seedlings in the almond breedingprogrammes, is interesting for the selectionof these new cultivars.Most of the work to develop molecularmarkers in almond has been carried outin setting up the different techniques andmapping the genome of the species, neverthelessthe application in breedingprogrammes has been more reduced. Differenttypes of molecular markers, includingisoenzymes, RFLPs, RAPDs, andSSRs, have been applied for the geneticcharacterization of germplasm, the establishmentof genetic relationships betweencultivars and species, and the constructionof genetic maps. In addition, methodologiesfor the analysis of marker-assistedselection include the use of mappingpopulations segregating for desiredcharacters and bulk segregant analysis.FAO-CIHEAM - Nucis-Newsletter, Number 12 September 20049

Currently there is a good knowledgeabout the almond genome, which togetherwith the quantity and suitability ofplant material raised from almond breedingprogrammes, can allow the developmentof useful molecular tools to apply tothe selection of genitors and seedlings.In this work, a review of the application ofmolecular markers in almond breeding ispresented, with special emphasis on thedevelopment of marker assisted selectionstrategies.MOLECULAR CHARACTERIZATIONAND GENETIC RELATIONSHIPSTraditionally, the identification and characterizationof almond cultivars has beenbased on morphological and agronomicaltraits. However, such traits are not alwaysavailable for analysis and are affected bychanging environmental conditions. Molecularmarker technology offers severaladvantages over the use of conventionaltraits. Molecular markers also offer apowerful tool to study genome evolution,and for understanding genome structureand determinants of genetic diversity(Wünsch and Hormaza, 2002).Isoenzymes: Isoenzymes were amongthe first genetic markers to be widely utilized.They have been used for cultivaridentification in almond because of theirenvironmental stability, their codominantexpression, and their good reproducibility.Nevertheless, their utilization is limitedby the small number of loci that can beanalysed with conventional enzyme stainingmethods, as well as a low variationin some loci (Arulsekar et al., 1986;Hauagge et al., 1987; Cerezo et al., 1989;Sathe et al., 2001). More recently, isoenzymesin combination with DNA-basedmarkers were applied to develop geneticmaps for woody perennials (see “Geneticmapping” below) and for the genetic characterizationof multiple embryos in almond(Martínez-Gómez and Gradziel,2003).RFLPs: Restriction fragment length polymorphism(RFLP) markers are based onthe differential hybridization of clonedDNA to bulk DNA fragments from restriction-enzymedigestion. RFLP markers arecodominant. RFLPs can detect a virtuallyunlimited number of markers, thus providingan efficient method for discoveringlinkages among markers and for constructinggenetic maps. RFLP have beenassayed in almond for map-based selection(Viruel et al., 1995). However, RFLPanalysis has important limitations: it is laboriousand time-consuming and it ofteninvolves the use of radioisotopes.PCR amplification of random locations inthe genome. RAPDs are characterized byusing arbitrary primers. A single oligonucleotideis used for the amplification ofgenomic DNA. In contrast to isoenzymesand RFLPs, RAPDs are dominant markers.This feature, as well as their variabledegree of repeatability and problemsin transferring across populations, limitstheir use primarily to map construction.RAPD techniques have been successfullyused in almond for identifying cultivars,estimating genetic diversity and assessingpossible origins for selected genotypes(Bartolozzi et al., 1998; Martins et al.,2003).SSRs: PCR-based, simple sequence repeat(SSR) markers (microsatellites) arebecoming the marker of choice for fingerprintingand genetic diversity studies for awide range of plants. Because of theirhigh polymorphism, abundance, and codominantinheritance, they are well suitedfor the assessment of genetic variabilitywithin crop species, and for the geneticrelationships among species (Powell etal., 1996). In the case of Prunus species,primer pairs flanking SSRs have beencloned and sequenced in different speciesincluding peach, apricot, and cherry(Aranzana et al., 2003). These SSR markershave been used for the molecularcharacterization and identification of almondcultivars (Martínez-Gómez et al.,2003a) and related Prunus species (Fig.1) (Martínez-Gómez et al., 2003b); andelaboration of genetic maps (Joobeur etal., 2000; Bliss et al., 2002) (see “Geneticmapping” below).Electrophoresis in polyacrilamide with radioactiveand silver staining was the firstmethod used in the analysis of the PCRamplified fragment of DNA obtained fromthe SSR markers. Electrophoresis in Metaphor® agarose was a method used asalternative to the polyacrilamide due to itseasier application (Fig. 2). More recently,new methods for the PCR amplified DNAhave been developed including the use ofautomated sequencers.Metaphor ® AgarosePolyacrylamideFigure 2. Allelic segregation of UDP96-013 SSRmarker in almond cultivars using Metaphor ®agarose and Polyarcilamide. First and lastsamples correspond to a 1 kb DNA ladderRAPDs: Random amplified polymorphicDNA (RAPD) markers are based on theFigure 1. Genetic relationship among peach and almond cultivars and related Prunus species obtained bya study with SSR markers (Martínez-Gómez et al., 2003b). Unrooted dendogram obtained by NeighbourJoining cluster analysis10 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

GENETIC MAPPINGSeveral intraspecific and interspecific almondmaps have been developed usingdifferent types of molecular markers. Theuse of PCR-based markers made it possibleto map and tag a wide range of traits(Arús et al., 1994; 1999). The analysis ofcosegregation among markers eases linkageanalysis between markers and majoror quantitative loci controlling horticulturallyimportant traits.Different research groups have releasedlinkage maps using morphological traits,isoenzymes, RFLPs and RAPDs in almond(Viruel et al., 1995; Joobeur et al.,1998; 2000; Jáuregui et al., 2001; Bliss etal., 2002). In addition, SSRs have beenused for mapping in almond (Joobeur etal., 2000; Bliss et al., 2002; Aranzana etal., 2003). The similar arrangement ofmarkers observed in different Prunusmaps suggests a high level of syntenywithin the genus (Aranzana et al., 2003).This homology among Prunus speciespartly explains the low level of breedingbarriers to interspecific gene introgressionand supports the opportunity for successfulgene transfer between closely relatedspecies (Gradziel et al., 2001).MARKER-ASSISTED SELECTIONSelection by molecular markers is particularlyuseful in fruit, nut, and other treecrops with a long juvenile period, andwhen the expression of the gene is recessiveor the assessment of the character isdifficult. Marker-assisted selection (MAS)is emerging as a very promising strategyfor increasing selection gains. If sufficientmapping information is available, MAScan highly shorten the number of generationsrequired to “eliminate” the undesiredgenes of the donor in backcrossing programmes(Arús and Moreno-González,1993). Marker loci linked to major genescan be used for selection, which is moreefficient than direct selection for the targetgene (Arús and Moreno-González,1993).The principal approach for the analysis ofmarker-trait association in almond is theuse of mapping populations segregatingfor the characters of interest. The differentlinkage maps developed in almondinclude markers associated with severaltraits of horticultural value. MappingTable 1. Markers associated to main agronomic traits in almond.quantitative characters by identifyingquantitative trait loci (QTL) is also becomingan important tool in tree breeding.QTLs are generally recognized by comparingthe degree of covariation for polymorphicmolecular markers with phenotypictrait measurements. Important charactersand QTLs that are presently beingmapped in almond include bloom time,self-incompatibility, shell hardness, orkernel taste (Asins et al., 1994; Viruel etal., 1995; Ballester et al., 1998, 2001;Jobeur et al., 1998; 2002; Bliss et al.,2002) (Table 1). The high degree of genomesynteny observed among Prunusspecies should also facilitate the successfultransfer of sets of markers and codingsequence among species (Aranzanaet al., 2003) (Table 1).Bulk segregant analysis (BSA), wheretwo pooled DNA samples are formed fromplant sources that have similar geneticbackgrounds but differ in one particulartrait, is another promising approach forthe analysis of molecular marker-horticulturaltrait association. A strategy combiningdifferent markers with bulk segregantanalysis was used to identify markerslinked to loci of specific characters inpeach x almond crosses (Warburton etal., 1996), and three RAPD markers associatedwith a gene conferring delayedbloom in almond (Ballester et al., 2001)(Table 1).A very promising application of molecularmarker assisted selection is the manipulationof self-compatibility in almond. Thisspecies is predominantly self-incompatible.Self-incompatibility is of the gametophytictype and acts to prevent self-fertilization.This character is controlled by asingle locus with multiple codominantalleles (Dicenta and García, 1993), and isexpressed within the styles of flowers asS-RNases glycoproteins (Bošković et al.,1997; 2003) that are responsible for thesubsequent inactivation of self-pollentube growth. Almond self-incompatibilityalleles (S-alleles) were initially identifiedin the field through controlled crosseswith a series of known S-genotypes (Kesterand Gradziel, 1996). More recently,molecular methods have been developedin two areas: identification of stylar S-RNases by electrophoresis in verticalpolyacrilamide gels (Bošković et al.,Trait Symbol Marker ReferenceSelf-incompatibility SI RFLP Joobeur et al. 1998Self-compatibility Sf RFLP Arús et al. 1999Kernel taste Sw RFLP Bliss et al. 2002Shell hardiness D RFLP Arús et al. 1999Late blooming Lb RAPD Ballester et al. 20011997, 2003), and the amplification of specificS-alleles using appropriately designedprimers for PCR and electrophoresisin horizontal agarose gels (Tamura et al.,2000; Channuntapipat et al., 2003). Thistechnique is being routinely used for theidentification of cross-incompatibility groupingsfor current almond cultivars and forefficiently breeding self-compatibility intonew cultivars (Gradziel et al., 2001b; Ortegaand Dicenta, 2003).ACKNOWLEDGMENTSThe authors acknowledge the support ofSpanish Ministry of Education and Science.BIBLIOGRAPHYAranzana M.J., Pineda A., Cosson P.,Dirlewanger E., Ascasibar J., CiprianiG., Ryder C.D., Testolin R., Abbott A.,King G.J., Lezzoni A.F. and P. Arús.2003. A set of simple-sequence (SSR)markers covering the Prunus genome.Theor. Appl. Genet. 106: 819-825.Arulsekar S., Parfitt D.E. and D.E. Kester.1986. Comparison of isozyme variabilityin peach and almond cultivars. J.Hered. 77: 272-274.Arús P. and J. Moreno-González. 1993.Marker-assisted selection. In: Hayward,M.D., Bosemark, N.O., and Romagosa,I. (eds.). Plant Breeding. Principles andProspects. London, United Kingdom:Chapman & Hall. p. 314-331.Arús P., Messeguer R., Viruel M.A., TobuttK., Dirlewanger E., Santi F., QuartaR. and E. Ritter. 1994. The EuropeanPrunus mapping project. Euphytica 77:97-100.Arús P., Ballester J., Jauregui B., JoobeurT., Truco M.J. and M.C. de Vicente.1999. The European Prunus mappingproject: update of marker developmentin almond. Acta Hort. 484: 331-336.Asins M.J., Mestre, P., García, J.E., Dicenta,F., and E.A. Carbonell. 1994.Genotype ∞ environment interaction inQTL analysis of an intervarietal almondcross by means of genetic markers.Theor. Appl. Genet. 89: 358-364.Ballester J., Bošković R., Batlle I., ArúsP., Vargas F. and M.C. de Vicente.1998. Location of the self-incompatibilitygene on the almond linkage map.Plant Breed. 117: 69-72.Ballester J., Socias i Company R., ArúsP. and M.C. de Vicente. 2001. Geneticmapping of a major gene delayingblooming time in almond. Plant Breed.120: 268-270.Bartolozzi F., Warburton M.L., ArulsekarS. and T.M. Gradziel. 1998. Geneticcharacterization and relatedness amongCalifornia almond cultivars and breedinglines detected by randomly amplifiedpolymorphic DNA (RAPD) analysis.J. Am. Soc. Hort. Sci. 123: 381-387.FAO-CIHEAM - Nucis-Newsletter, Number 12 September 200411

Bliss F.A., Arulsekar S., Foolad M.R.,Becerra V., Gillen A., Warburton M.L.,Dandekar A.M., Kocsine G.M. and K.K.Mydin. 2002. An expanded geneticlinkage map of Prunus based on an interspecificcross between almond andpeach. Genome 45: 520-529.Bošković R., Tobutt K.R., Batlle I. andH. Duval. 1997. Correlation of ribonucleasezymograms and incompatibilitygenotypes in almond. Euphytica 97:167-176.Bošković R. , Tobutt K R, Batlle I., Duval.H, Martínez-Gómez P. and T.M.Gradziel. 2003. Stylar ribonucleases inalmond: correlation with and predictionof self-incompatibility genotypes. PlantBreed. 122: 70-76.Cerezo M., Socias i Company R. and P.Arús. 1989. Identification of almond cultivarsby pollen isoenzymes. J. Am.Soc. Hort. Sci. 114: 164-169.Channuntapipat C., Wirthensohn M.,Ramesh S.A., Batlle I., Arús P., SedgleyM. and G. Collins. 2003. Identification ofincompatibility genotypes in almondusing specific primers based on the intronsof the S-alleles. Plant Breed. 122:164-168.Dicenta F. and J.E. García. 1993. Inheritanceof self-compatibility in almond.Heredity 70: 313-317.Gradziel T.M., Martínez-Gómez P., DicentaF., and D.E. Kester. 2001a. Theutilization of related almond species foralmond variety improvement. J. Am.Pomolog. Soc. 55: 100-109.Gradziel T.M., Martínez-Gómez P. andA.M. Dandekar. 2001b. The use of S-allele specific PCR analysis to improvebreeding efficiency for self-fertility in almond.HortScience 36: 440-440.Hauagge R., Kester D.E. and R.A.Asay. 1987. Isozyme variation amongCalifornia almond cultivars: I. Inheritance.J. Am. Soc. Hort. Sci. 112: 687-693.Jáuregui B., de Vicente M.C., MesseguerR., Felipe A., Bonnet A., SalessesG. and P. Arús. 2001. A reciprocaltranslocation between ‘Garfi’ almondand ‘Nemared’ peach. Theor. Appl. Genet.102: 1169-1176.Joobeur T., Viruel M.A., de VicenteM.C., Jáuregui B., Ballester J., DettoriM.T., Verde I., Truco M.J., MesseguerR., Battle I., Quarta R., Dirlewanger E.,and P. Arús. 1998. Construction of asaturated linkage map for Prunus usingan almond x peach F 2progeny. Theor.Appl. Genet. 97: 1034-1041.Joobeur T., Periam N., de Vicente M.C.,King G.J. and P. Arús. 2000. Developmentof a second generation linkagemap for almond using RAPD and SSRmarkers. Genome 43: 649-655.Kester D.E. and T.M. Gradziel. 1996.Almonds (Prunus). In: Moore, J.N. andJanick, J. (eds.). Fruit Breeding. NewYork: Wiley. p. 1-97.Martínez-Gómez P. and T.M. Gradziel.2003. Sexual polyembryony in almond.Sex. Plant Reprod. 16: 135-139.Martínez-Gómez P., Arulsekar S., PotterD., T.M. and Gradziel T.M. 2003a.An extended interspecific gene poolavailable to peach and almond breedingas characterized using simple sequencerepeat (SSR) markers. Euphytica 131:313-322.Martínez-Gómez P., Arulsekar S., PotterD., and T.M. Gradziel. 2003b. Relationshipsamong peach and almond andrelated species as detected by SSRmarkers. J. Amer. Soc. Hort. Sci. 128:667-671.Martins M., Tenreiro R. and M.M. Oliveira.2003. Genetic relatedness of Portuguesealmond cultivars assessed byRAPD and ISSR markers. Plant CellRep. 22: 71-78.Ortega E. and F. Dicenta. 2003. Inheritanceof self-compatibility in almond:breeding strategies to assure self-compatibilityin the progeny. Theor. Appl.Genet. 106: 904-911.Powell W., Morgante M., Andre C., HanafeyM., Vogel J., Tingey S., and A.Rafalski. 1996. Comparison of RFLP,RAPD, AFLP and SSR (microsatellite)markers for germplasm analysis. Mol.Breed. 2: 225-238.Sathe S.K., Teuber S.S., Gradziel T.M.,and K.H. Roux. 2001. Electrophoreticand immunological analyses of almond(Prunus dulcis L.) genotypes and hybrids.J. Agr. Food Chem. 49: 2043-2052.Tamura M., Ushijima K., Sassa H., HiranoH., Tao R., Gradziel T.M. and A.M.Dandekar. 2000. Identification of selfincompatibilitygenotypes of almond byallele-specific PCR analysis. Theor.Appl. Genet. 101: 344-349.Viruel M.A., Messeguer R., de VicenteM.C., Garcia-Mas J., Puigdomènech P.,Vargas F. and P. Arús. 1995. A linkagemap with RFLP and isozyme markersfor almond. Theor. Appl. Genet. 91:964-971.Wünsch A. and J.I. Hormaza. 2002.Cultivar identification and genetic fingerprintingof temperate fruit tree speciesusing DNA markers. Euphytica125: 56-67.1R. Sánchez-Pérez, 1 F. Dicenta, 2 T.M.Gradziel, 3 P. Arús, and 1 P. Martínez-Gómez.1CEBAS-CSIC. Departamento de Mejoray Patología Vegetal, Apdo 4195, 30.080,Murcia, Spain.2University of California-Davis,Dept. of Pomology, Davis,CA-95616, USA.3IRTA. Departament de Genètica Vegetal.Crta. Cabrils s/n, E-08348 Cabrils,Barcelona, Spain.E-mail: pmartinez@cebas.csic.esFusicoccum CANKERIN ALMOND ORCHARDS –BIOTECHNOLOGY TOOLSFOR EARLY SELECTIONOF TOLERANT GENOTYPESFusicoccum CANKER ON ALMONDAlmond is an important crop in the Mediterraneanregion, reaching about 28% ofthe world production. However, its susceptibility,particularly to certain fungal diseasessuch as brown rot blossom blight,constriction canker and almond scab,which can cause severe damage on foliageand branches of the tree, can be a majorimpediment for almond production.Regarding Fusicoccum canker or constrictioncanker, this is a damaging andeconomically important fungal disease inalmond orchards in the Mediterranean region,caused by Phomopsis amygdali(Del.) (perfect stage) Tuset and Portilla,formerly classified as Fusicoccum amygdaliDelacr. (imperfect stage). Besides almond,this fungus also infects peachtrees. Symptoms of this disease usuallyappear in early summer and become increasinglyevident as more infectedshoots appear through late summer. In almondmainly two types of symptoms arevisible, on shoots and on May spurs. Infectedtwigs and shoots wilt, and eventuallydie, because of elongate, brown,sunken cankers, often with a well delimitedoval pattern (Grasselly and Duval,1997) (Fig. 1).The group of leaves that form May spurswilt due to the infection, caused on thatspring or on the previous autumn. It isalso possible to observe cankers on theinternodes of these spurs. Leaves in generalcan also be affected, showing necrosisthat usually appear as big brownspots with circular or irregular shape.Gumming is commonly associated withthe cankers, although it is not a gooddiagnostic characteristic since otheragents may also be responsible for thissign. Constrictions formed at the base ofinfected shoots and leaf symptoms producedwell beyond the infection site resultfrom a translocatable toxin, fusicoccin, aphytotoxic terpenoid glucoside. Fusicoccin,increases the influx of potassium onguard cells inducing the permanent openingof the stomata and the consequenthigh loss of water can eventually lead toplant death (De Boer, 1997).The development of Fusicoccum cankeris favoured by high nitrogen levels in thesoil and by wet weather, since this diseaseis transmitted by rain (Rosenberg,1996). Most infections occur during12 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

Figure 1. Infected shoots showing brown sunken cankers. (Images kindly provided by I. Batlle)spring (growing season of the plants) andautumn, when apparently almond treesas well as peach trees are more susceptibleto the disease. Pycnidia (darkly pigmented,flask-shaped, conidia-bearing fruiting)are produced over the cankered surfacearea. Under wet weather the pycnidiaexude conidia in white tendrils. Theseconidia disseminated by rain, will infectnew plants through scars on leaves, budsor stipules, or directly through youngshoots during their growing stage by latewinter and early spring. By the end ofsummer, the trees can be severely affectedexhibiting numerous terminal deadzones spread through the tree canopy(Grasselly and Duval, 1997). These effectscan lead to significant losses on fruit productionin almond cultivars.The management of infected orchardsshould involve the implementation of fungicideapplication programmes and pruning(elimination) of infected limbs to reduceinoculum. Winter pruning of cankeredshoots, followed by pruning of youngshoots showing symptoms of the diseasein spring, are important practices to helpFusicoccum canker control (Barbé, 1993).Fungicides, applied just before budbreak(February-March) and in autumn, may benecessary in orchards where the diseaseis present (Grasselly & Duval, 1997). Keepingnitrogen at low to moderate levelsin the soil contributes to minimize the severityof the infection (Rosenberg, 1996).Wide differences on Fusicoccum tolerancebetween cultivars have been observed(Grasselly and Crossa-Raynaud, 1980;Romero and Vargas, 1981) and shouldbe taken into consideration when new orchardsare planned.Almond populations are found in mostMediterranean countries, where weatherconditions mainly in coastal orchards areusually favourable to Fusicoccum development.Therefore, the use of toleranceto Fusicoccum as a selectable trait in almondbreeding programmes and in theestablishment of new orchards can be ofgreat importance.MARKER ASSISTED SELECTION (MAS)Genetic markers, as transmissible entitiesassociated to economically importanttraits, can be used by breeders as importantselection tools (Darvasi & Soller,1994). After a decade of marker assistedselection in vegetable crops, increasingavailability of genomics-derived informationis starting to allow the use of molecularmarkers as tools to support classicalbreeding programmes in fruit tree species.Identification of molecular markers closelylinked to genes controlling traits of interestare essential in marker assisted selection.The closer the linkage, less is theprobability of recombination between themarker and the gene(s) that control thetrait of interest, thus the higher will be theefficiency of selection.The best perspectives for MAS applicationmay rely on breeding programmes aimingfor disease resistance improvement,particularly in systems where several genesare involved, with complex interactions,and where gene pyramiding is desirable(Kelly, 1995). Markers for diseaseresistance offer the additional advantageof permitting selection for resistance inthe absence of the pathogen(s) (Mehlenbacher,1995). Another possible applicationcan be on recurrent selection ofthe cultivated genotype when the aim isgene introgression from wild individuals.The construction of a linkage map of acertain population allows the study of cosegregationof markers with genes responsiblefor the variability found in thatpopulation, making possible the identificationof major genes or loci of quantitativecharacters (QTLs) on the map (Arús etal., 1999). Genetic maps developed in almond(or almond x peach) have beenused to identify markers linked to geneswith important effects on the morphologyand physiology of the plant, for exampleself-incompatibility, shell hardness or almondbitterness. Integration in thesemaps of new markers closely linked togenes of interest and their routine use oncurrent or future breeding programmes, isof high interest. However, despite its potentialbenefits, MAS applied to almondbreeding will likely be restricted to a limitedset of traits and parents where datafor segregating populations are available.Examples of applied MAS reported in theliterature are still scarce, mainly becauseprivate companies are the main users ofMAS, therefore information is more difficultto be available. Moreover, the costsand elaborated technology required formost of the markers available make MASapplication of limited use in breeding programmes.MAS applied to woody plants such as almondwould be extremely important,mainly for allowing the selection of charactersof interest on an initial stage ofplant development, which would only beexpressed after 2 to 5 years in the field,hence representing a significant reductionon time and area used when comparedto species with a shorter life cycle.SEARCH AND IDENTIFICATIONOF MOLECULAR MARKERS –THE CASE OF Fusicoccum LINKEDMARKERS IN ALMONDPlant breeding programmes require relativelylow-cost methods that can be usedfor hundreds or thousands of individuals,with a reliability of at least 95% (Rafalski& Tingey, 1993), these criteria make PCRbased techniques very attractive. Due totheir low cost, simplicity and easy detection,RAPD (Randomly Amplified PolymorphicDNA, Williams et al., 1990) markersare frequently chosen for this kind ofstudy. However, a poor specificity andlack of reproducibility can sometimes beassociated to this technique. In order toovercome these deficiencies, RAPD fragmentscan be converted into SCAR markers(Sequence Characterized AmplifiedRegion, Paran & Michelmore, 1993), morereproducible and sometimes codominants,or to CAPS (Cleaved Amplified PolymorphicSequences, Konieczny & Ausubel,1993; Jarvis et al., 1994).Michelmore et al. (1991) showed thatRAPD bands linked to a trait of interestwould be easily identified through the useof two samples of bulked DNA from a populationsegregating for that trait: withineach pool, or bulk, the individuals areidentical for the trait or gene of interestbut are arbitrary for all other genes. Anypolymorphism between the two poolsFAO-CIHEAM - Nucis-Newsletter, Number 12 September 200413

should be linked to the trait being analysed.Mapping on a segregating populationmay confirm the markers found. Thistechnique, identified as “bulk segregantanalysis” (BSA), is now commonly usedto map simple traits with RAPD markers.A variation of this technique, known as“bulked DNA analysis”, uses also twosamples of DNA mixtures, one of individualsthat express the trait, and anotherof individuals that do not express it, butwithout using a segregating population.Once a marker is identified as being relatedto a trait of interest, the developmentof SCAR or CAPS markers results in ahigher specificity and reproducibility ofthe fragments obtained. RAPD analysisfollowed by SCAR or CAPS developmenthas been used to identify markers associatedto traits of interest in plants, suchas downy mildew resistance in lettuce(Paran & Michelmore, 1993), the locusthat controls flower doubleness in carnation(Scovel et al., 1998), a new resistancegene to leaf stripe in barley (Tacconiet al., 2001), leptine content in Solanumphureja (Bouarte-Medina et al., 2002),sex determination in kiwifruit plants (Gillet al., 1998), genotypes of vine rootstocks(Xu et al., 1995), colour of fruit epidermisin apple (Cheng et al., 1996), Ma1 rootknotnematode resistance gene in Myrobalanplum (Lecouls et al., 1999) and resistanceto root-knot nematodes in peach(Lu et al., 1999).In our study, the RAPD technique wascombined with bulked DNA analysis toselect markers associated to either toleranceor sensitivity to Fusicoccum in almond.Fourteen almond cultivars classifiedas tolerant (7) or sensitive (7) to Fusicoccumwere screened with 120 RAPDprimers. Polymorphic bands were assessedfor co-segregation with tolerant orsusceptible phenotypes using 30 seedlingsobtained from a cross of ‘Masbovera’(T) x ‘Lauranne’ (S). Three RAPDmarkers were identified, two linked to tolerance(OPD-19 300and OPA-08 900) andone linked to sensitivity (OPD-19 500) toFusicoccum (Martins et al., 2001; Martinset al., 2003). SCAR primers were developedand segregation of SCAR markerswas tested, however, polymorphism betweenthe two groups (tolerant versus susceptible)was lost in this process. The developmentof CAPS markers as a possibleway to recover the lost polymorphismwill be analysed.Cosegregation analysis using the adequatepopulations is essential to confirmthe genetic base of the markers linked toa certain trait. In the case of almond, anew segregating population of some 120seedlings derived from a cross ‘Primorskyi’(T) x ‘Lauranne’ (S), is currently beingevaluated in the field, at IRTA, CentreMas Bové (Tarragona, Spain), for toleranceor sensibility to Fusicoccum. This materialwill allow the confirmation of whichfragments correspond to markers associatedto the disease. In this case it wouldbe of high interest to evaluate the possibilityof inserting these markers in the referenceEuropean Prunus genetic map, almond‘Texas’ x peach ‘Earlygold’ (TxE)(Joobeur et al., 1998).The genetic nature of tolerance to Fusicoccumin almond (or related species likepeach) is not yet defined, and further studiesare necessary to determine the modelthat can be applied to almond. Althoughartificial inoculation with Fusicoccumis currently being tested in almond(Cabrita et al., 2003), any procedure hasbeen established, so far, to efficientlyevaluate tolerance or sensibility of almondplants to Fusicoccum canker. Therefore,the identification of molecular markerslinked to tolerance or sensibility toFusicoccum, is of particular importancesince, to our knowledge, this is the onlymethod that will allow the early selectionof tolerant plants from breeding programmes,as well as the certification of tolerantcultivars to be implemented in neworchards. Symptoms development of thedisease in plants grown in the field derivedfrom breeding programmes takenabout 4 to 5 years under natural conditionsof high atmospheric humidity as it happensin the Mediterranean coast (I. Batlle,personal communication). Finally, it isimportant to highlight that the balanceand coordination among the use of moleculartechnologies and the fieldbasedwork, are essential for the success ofbreeding programmes (Scorza, 1996).REFERENCESArús P, Joobeur T, Jaúregui B, BallesterJ, Viruel MA, de Vicente MC, 1999. Marcadoresmoleculares en especies frutales:Prunus. In: Marcide JMO (ed) IdentificaciónMolecular de Germoplasmade Vid. Fundación Premio ARCE, InstitutoMadrileño de Investigación AgrariaY Alimentaria (IMIA), Madrid pp 153-165.Barbé L (1993) Favoriser le renouveaude la culture de l’amandier. Arboriculturefruitière. 358: 21-31.Bouarte-Medina T, Fogelman E, ChaniE, Miller AR, Levin I, Levy D, VeilleuxRE, 2002. Identification of molecularmarkers associated with leptine in reciprocalbackcross families of diploid potato.Theor Appl Genet 105: 1010-1018.Cabrita L, Branco V, Neves MA, Leitão J,2003. Barrinho Grado a new source of resistanceto Phomopsis amygdali in almond.XIII G.R.E.M.P.A. Meeting on Pistachioand Almond, Mirandela p 73 (Abstract).Cheng FS, Weeden NF, Brown SK,1996. Identification of codominantRAPD markers linked to fruit skin colorin apple. Theor Appl Genet 93: 222-227.Darvasi A and Soller M, 1994. Optimumspacing of genetic markers for determininglinkage between marker loci andquantitative trait loci. Theor Appl Genet89: 351-357.De Boer B, 1997. Fusicoccum - a key tomultiple 14-3-3 locks? Trends in PlantScience 2: 60-66.Gill GP, Harvey CF, Gardner RC, FraserLG, 1998. Development of sexlinkedPCR markers for gender identificationin Actinidia. Theor Appl Genet97: 439-445.Grasselly C & Crossa-Raynaud P, 1980.L’amandier. Maisonneuve et Larose,Paris.Grasselly C & Duval H, 1997. L’Amandier.Centre Technique Interprofessionneldes Fruits et Légumes, Paris.Jarvis P, Lister C, Szabo V, Dean C,1994. Integration of CAPS markers intothe RFLP map, generated using recombinantinbred lines of Arabidopsis thaliana.Plant Mol Biol 24: 685-687.Joobeur T, Viruel MA, de Vicente MC,Jáuregui B, Ballester J, Dettori MT, VerdeI, Truco MJ, Messeguer R, Batlle I,Quarta R, Dirlewanger E, Arús P, 1998.Construction of a saturated linkage mapin Prunus using an almond x peach F 2progeny. Theor Appl Genet 97: 1034-1041.Kelly JD, 1995. Use of random amplifiedpolymorphic DNA markers in breedingfrom major gene resistance to plantpathogens. HortScience 30: 461-465.Konieczny A & Ausubel FM, 1993. Aprocedure for mapping Arabidopsis mutationsusing codominant ecotype-specificPCR-based markers. Plant J 4:403-410.Lecouls AC, Rubio-Cabetas MJ, MinotJC, Voisin R, Bonnet A, Salesses G,Dirlewanger E, Esmenjaud D, 1999.RAPD and SCAR markers linked to theMa1 root-knot nematode resistancegene in Myrobalan plum (Prunus cerasiferaEhr.). Theor Appl Genet 99:328-335.Lu Z-X, Sossey-Alaoui K, Reighard GL,Baird WV, Abbott AG, 1999. Developmentand characterization of a codominantmarker linked to root-knot nematoderesistance, and its application topeach rootstock breeding. Theor ApplGenet 99: 115-122.Martins M, Sarmento D, Batlle I, VargasF, Oliveira MM, 2003. Development ofSCAR/CAPS markers linked to tolerance/sensitivityto Fusicoccum in almond.Options Méditerranéennes CIHEAM/IAMZ (in press).Martins M, Sarmento D, Batlle I, VargasF, Oliveira MM, 2001. Search for molecularmarkers linked to Fusicoccum to-14 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

lerance in almond. Acta Horticulturae577: 87-90.Mehlenbacher SA, 1995. Classical andmolecular approaches to breeding fruitand nut crops for disease resistance.HortScience 30: 466-477.Michelmore RW, Paran I, Kesseli RV,1991. Identification of markers linked todisease resistance genes by bulked segregantanalysis: A rapid method to detectmarkers in specific genomic regionsby using segregating populations.Proc Natl Acad Sci 88: 9828-9832.Paran I & Michelmore RW, 1993. Developmentof reliable PCR-based DNAmarkers linked to downy mildew resistancegenes in lettuce. Theor Appl Genet85: 985-993Rafalski JA & Tingey SV, 1993. Geneticdiagnostics in plant breeding: RAPDs,microsatellites and machines. TrendsGenet 9: 275-280.Romero MA & Vargas FJ, 1981. Contributionà la connaissance de la sensibilitéde l’Amandier au Fusicoccum amygdaliDel. Options Mediterranéennes,IAMZ. 81/1:143-147.Rosenberg D, 1996. Update on pestmanagement and crop development –Peach constriction disease. ScaffoldsFruit Journal Vol. 5.Scorza R, 1996. Genome mapping ofhorticultural crops: introduction to thecolloquium. HortScience 31: 1098.Scovel G, Ben-Meir H, Ovadis M, ItzhakiH, Vainstein A, 1998. RAPD andRFLP markers tightly linked to the locuscontrolling carnation (Dianthus caryophyllus)flower type. Theor Appl Genet96: 117-122.Socias i Company R & Felipe AJ, 1992.Almond: a diverse germplasm. HortScience27: 717-718, 863.Tacconi G, Cattivelli L, Faccini N, PecchioniN, Stanca AM, Valé G, 2001.Identification and mapping of a new leafstripe resistance gene in barley (Hordeumvulgare L.). Theor Appl Genet102: 1286-1291.Williams JGK, Kubelik AR, Livak KJ,Rafalski JA, Tingey SV, 1990. DNA polymorphismsamplified by arbitrary primersare useful as genetic markers. NucleicAcids Res 18: 6531-6535.Xu H, Wilson DJ, Arulsekar S, BakalinskyAT, 1995. Sequence-specific polymerasechain reaction markers derivedfrom randomly amplified polymorphicDNA markers for fingerprinting grape(Vitis) rootstocks. J Am Soc Hortic Sci120: 714-720.VIRUS SCREENINGFOR AUSTRALIAN ALMONDSSUMMARYAs part of the Australian almond breedingprogram, work is conducted on breedingparents and progeny to screen for viralpathogens. Transmission of virus of up to23% has been recorded when infectedparents are used. As a service to industrythe University also test the Monash almondbudwood facility.INTRODUCTIONAn important aspect of the Australian almondbreeding programme is the detectionof plant viruses in the breeding stockand progenies (Sedgley & Collins, 2002).The two most important viruses present inAustralia in almond (Prunus dulcis Mill.)are Prunus necrotic ringspot virus (PNR-SV) (Calico) and prune dwarf virus (PDV).Infection of nursery stock results in poorbud-take and retarded growth, and in commercialorchards growth and crop yieldsare affected. Leaf symptoms of PNRSVinclude chlorotic and necrotic leaf spots,mosaics, ringspots, and line patterns.There are other viruses, plum pox virus(PPV) (Sharka); tomato black ring virus(TBRV) (Enation); and tomato ringspot virus(ToRSV) (Peach yellow bud mosaic),which are present in Europe and Americabut not in Australia, however imported almondmaterial is tested by AQIS for viruseson arrival. As a service to the Australianalmond industry the University alsoregularly tests the Monash almond budwoodrepository for viruses thus ensuringthat only tested material is used forbudding in nurseries. The Monash facilityconsists of 601 almond trees representingall major cultivars grown in Australiaincluding those newly introduced fromEurope and the USA.ELISA TESTING AND WOODYINDEXINGWhen screening began for PNRSV andPDV in 1997 the methods used were theantibody-based enzyme-linked immunosorbentassay (ELISA) technique and woodyindexing, where almond budwood isgrafted onto the indicator plant Prunusserrulata cv. Shirofugen, then monitoredfor diagnostic symptoms. The detection ofPNRSV by ELISA was shown by BertozziFigure 1. Leaf symptoms of PNRSV on almondleavesFigure 2. Photo showing gumming symptomsof PNRSV on Shirofugen indicator plant aftergrafting to infected almond budwoodFigure 3. Photo of ELISA sampling tray showingpositive (yellow) and negative (clear) results forPNRSV testinget al. (2002) to be most sensitive whenplant material was collected in spring.All breeding parents and progenies derivedfrom crosses made in the programmefor the first two years, almost 8,000 trees,were tested. A small number have produceda positive response for PNRSV, butnone tested positive for PDV using ELI-SA. Where both parents were positive forPNRSV, transmission rates were high,22.6% & 12.5% for 1997 and 1998 respectively(Table 1). Seed transmissionfrom infected mother trees is generallyhigher than from virus-free mother treespollinated by infected pollen. The low levelsof infected progeny from negativeparents could be due to infections in thefield. It can be seen from these data thatbreeding parents should be virus tested ifpossible, so that potential new cultivarsare clean.M. Martins 1,2 , M.M. Oliveira 1,2(1) IBET/ITQB, Quinta do Marquês, 2784-505Oeiras, Portugal(2) Dep. Biologia Vegetal, Fac. Ciênciasde Lisboa, Campo Grande1749-016 Lisboa, PortugalE-mail: mmartins@itqb.unl.ptTable 1. The percentage of progeny testing positive for PNRSV.Progeny Both parents Pollen parent Seed parent Both parents-ve -ve -ve +ve1997 0.4% 0 6% 22.6%1998 0.1% 1.4% 1.8% 12.5%FAO-CIHEAM - Nucis-Newsletter, Number 12 September 200415

Figure 4. Agarose gel showing some cultivarswith a positive reaction to PNRSV primers.Lane 1: molecular marker ladder,lane 2: Carmel(-ve), Lane 3: Fritz (+ve),lane 4: Johnston (-ve), lane 5: Mission (-ve),lane 6: Nonpareil (+ve), lane 7: Nonpareil (-ve),lane 8: Parkinson (+ve), lane 9: Peerless (+ve),lane 10: Price (+ve), lane 11: Price (-ve),Lane 12: negative controlMONASHThe ELISA technique was used to screenall the Monash trees for PNRSV and PDVover 1997, 1998, & 1999 and half thetrees in 2000. PDV was not detected inany of the trees, however PNRSV wasdetected in 34 trees in 1997, 33 trees in1998, and 36 trees in 1999. Where treestest positive one year and negative thenext, this may be due to uneven distributionof the virus in the plant, or seasonalfluctuations of viral concentrations due toextremes of temperature. The increasefrom 1997 to 1999 may be attributed to anincrease in the virus titre of these trees tolevels able to be detected by ELISA, ormay represent new infections.RT-PCR TECHNIQUEA DNA based technique for virus detectionwas developed by G. Collins. This techniqueis based on the reverse transcription-polymerasechain reaction (RT-PCR)and is more sensitive than ELISA. The techniqueinvolves extracting ribonucleicacid (RNA) from fresh leaves and usingspecially designed primers that can detectthe viral coat protein within the plantcells. This technique was used on trees atMonash in 2000, 2001, 2002 and 2003.Generally RT-PCR confirmed the resultsof ELISA, however some trees were positivefor PNRSV where ELISA gave negativeresults. This is due to the more sensitivenature of RT-PCR, and has theadded advantage that plant material canbe tested at any time throughout thegrowing season.ACKNOWLEDGEMENTSThanks to Wanli Ma for technical assistancewith the RT-PCR. Thanks to the AlmondBoard of Australia, HorticultureAustralia Limited and Australian ResearchCouncil for funding. Reproducedwith permission from The AustralianNutgrower Journal.REFERENCESBertozzi, T., Alberts, E., and Sedgley,M. 2002. Detection of Prunus necroticringspot virus in almond: effect of samplingtime on the efficiency of serologicaland biological indexing methodologies.Australian Journal of ExperimentalAgriculture, 42: 207-210.Sedgley, M., and Collins, G. G. 2002.The Australian almond breeding programme.Acta Horticulturae, 591: 241-244.M. Wirthensohn 1 , S. Ramesh 1 , T. Bertozzi 2 ,E. Alberts 3 , G. Mekuria 4 , C. Bennett 5 ,G. Collins 1 , and M. Sedgley 11School of Agriculture & Wine, Universityof Adelaide, Waite Campus, PMB 1 GlenOsmond, SA 5064 Australia.2Present Address: Evolutionary Biology Unit,South Australian Museum, North Terrace,Adelaide, SA 5000 Australia.3South Australian Research and DevelopmentInstitute, GPO Box 397, Adelaide, SA 5001Australia.4Present address: Department of NaturalResources and Environment (Victorian Institutefor Dryland Agriculture), Private Bag 260,Horsham, Vic 3401 Australia.5Almond Board of Australia, HorticultureHouse, PO Box 52 Berri, SA 5343 Australia.E-mail: michelle.wirthensohn@adelaide.edu.auTHE PISTACHIO INDUSTRYIN ITALY: CURRENT SITUATIONAND PROSPECTSINTRODUCTIONPistachio (Pistacia vera L.), is a dioeciousand deciduous species native to westernAsia and Asia Minor. It was introducedinto Italy from Syria in 30 A.D. by the RomanGovernor of that Province, Lucius Vitellio,at approximately the same time itwas introduced in Spain (Minà Palumbo,1882). Its introduction into Sicily, whichnowadays accounts for almost the entirePistachio industry in Italy, probably occurredsome time later, following an initialperiod of cultivation in Campania which atthat time represented the roman “countryside”.Its cultivation in Sicily began tospread during the Arab domination (827-1040 A.D.) together with other major fruitand vegetable species and in parallel withthe improvement of the agronomic techniques.As a matter of fact, the Sicilian dialectalterm for Pistachio tree (Fastuca) isof arab derivation (Fustuq). However,very little is known until recent timesabout its distribution over the region. Historicreferences onto Pistachio cultivationare documented by the XVIIth century,even if first official records of the economicimportance of Pistachio industry arereferred to the first decades of the XXcentury.Soon after the second world war the entirestructure of the Sicilian Pistachio industryunderwent significant changes.The relative importance of the traditionalcultivation areas (Agrigento, Caltanissettaand Palermo) strongly decreasedwith the abandonment of large part of thePistachio cultivation which was partiallycompensated by a parallel increase of theproduction in the Catania Province (Fig. 1).Currently more than 90% of the total ItalianPistachio area is concentrated in onlyfew territories in the eastern Sicily (westernslopes of Mount Etna volcano), mainlylocated in the districts of Bronte andAdrano (Catania Province), onto approximately4.500 hectares. Few residuals ofthe past cultivation can still be found inCaltanissetta and Agrigento Provinces, insmall and scattered surfaces (as a whole,130 ha in specialized cultivation and 210ha in mixed cultivation).Despite this ancient origin and the longperiod of cultivation, only few female varietiesof Pistachio are now grown in Italy(about ten) together with an even morerestricted number of unnamed male selections.Among the female cultivars,‘Bianca’ (synonym: Napoletana) is practicallythe only cultivar grown, other varieties,namely ‘Femminella’, ‘Natalora’,‘Agostana’, ‘Silvana’, ‘Insolia’, ‘Cerasola’,‘Cappuccia’, representing not more than3% of the total (Barone et al., 1985) canstill be found, mostly in scattered, abandonedsettlements. This genetic pool isprobably the result of a two step introductionprocess of genetic material (Baroneet al., 1996) coincident with the earliestintroduction from Syria of the reddish ‘Cerasola’by the Romans and the later introductioncarried out by the Arabs. The reasonsof this low number of varieties isthought to be the exclusive use by thegrowers of vegetative propagation sinceancient times due to the long juvenilityperiod of P. vera seedlings, the long lifespanof the trees and easy hybridizationswith other Pistacia species. Nevertheless,within the Sicilian Pistachio germplasm,some valuable fruit characteristics suchas colour, flavour and nut quality arehighly appreciated in trade, specially thegreenness of the kernels and the rich oilynut-like flavour (Woodroof, 1967). Most ofthese appreciated characteristics havebeen maintained also by cultivars obtainedin the United States from Sicilianseeds imported in the early 1900’s suchas ‘Bronte’ and ‘Trabonella’ (Joley, 1969).In spite of these valuable characteristicsof the Italian pistachios, the contributionof Italian production represents nowadaysonly less than 0.6% of the world production.Nevertheless, with respect to productdestination, it is remarcable that Ita-16 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

(000)Tons250020001500100050001923/28lian pistachios maintain a dominant marketposition for the uses other than directconsumption (snack), since they are almostentirely used, and highly appreciated,by the confectionery and ice creamindustry.Due to this fact Italian pistachio exports,between 1000 and 2000 tons per year(Anonymous, 2000), are representedmainly by shelled and peeled pistachios,whereas imports (about 9000 tons) consistmainly of in-shell pistachios, consumedas salted and roasted snacks for domesticmarkets and, partially, for re-exportation.Figure 1 - Pistachio production in Italy1953/581974/79Sicily1980/851986/87Bronte area1993/942002/2003THE BRONTE AREAThis area of the eastern Sicilian provinceof Catania currently represents the mainPistachio growing area in Italy. From theecological point of view this area (Fig. 2)is of great naturalistic importance due tothe proximity to the Etna Volcano and theinclusion in the Regional system of parksand naturalistic reserves (“Parco dell’Etna”,59.000 ha, established in 1987).Figure 2 - The “Parco dell’Etna” area. The Bronte Pistachio area is encircledPistachio production in Bronte representsone of the main economic resources ofthe entire territory. A total of 3.300 ha ofspecialized Pistachio orchards are locatedin this area and 1.500 ha in the neighbourhoodterritories (Adrano, Biancavilla).The altimetry is comprised between350 (along the Simeto river bank) and900 m a.s.l.. The climate is typically Mediterraneanwith fall and winter rainfall (annualaverage: ~550 mm). Average monthlytemperatures of the area are betweena minimum of 6.0 °C (February) and amaximum of 32.6 °C (July) (Fabbri andValenti, 1998). Most of these Pistachioorchards are defined as “natural Pistachioplantings” because they have long beenobtained by grafting in situ spontaneousTerebinth (Pistacia terebinthus) plantsspread in the particular soils of the area,consisting of rocky, volcanic soils derivedin the centuries from the Mount Etna activity,in steep slopes (Barone et al., 1985).This traditional planting system took advantageof the extraordinary characteristicsof environmental adaptation of Terebinthto poor, dry, shallow soils of thearea, where no valid cultural alternativesare available, except for few other fruitspecies such as Prickly Pear and Fig.Thus, “natural Pistachio plantings” arecharacterized by the absence of regularplant spacings and by a wide range ofplant density (50-500 trees per ha). Alsothe age of the trees is consequently highlyvariable. Successive interplanting ofnursery terebinth seedlings is a commonpractice to replace dead plants and to increaseorchard density. A few local smallnurseries are active in the area of Bronteand supply plant material mainly as seedlingP. terebinthus rootstocks to begrafted directly in the orchard. Smallquantities of grafted P. vera/P.terebinthuspot plants are also produced. In such kindof conditions cultural operations are necessarilycarried out by hand, since mechanizationis almost impossible also dueto the training system adopted that canbe defined as an “open vase” with threefourmain irregular branches very close tothe soil surface. Pollination is ensured byspontaneous male P. terebinthus plantsor by scattered male P. vera pollinizersand also by natural hybrids between thetwo above mentioned species. Fruitsetproblems are likely to occur, speciallywhen pollination largely relies onto spontaneoussource of pollen, since male/femaleratio generally adopted in the Brontearea is particularly low (1/20). Besidesnatural orchards, new plantations arealso present in the area. In this case rationalorchard design (6m x 8m or 8m x8m) and common cultural practices areadopted.‘Napoletana’ (syn. Bianca) is the only cultivarwidely utilized both in the “natural” orin the regular plantations and Terebinth isthe only rootstock. Following the descriptors’list for Pistachio (Barone et al.,1997), ‘Bianca’ cultivar can be defined oflow-intermediate vigour. Growth habit isFAO-CIHEAM - Nucis-Newsletter, Number 12 September 200417

Picture 2. - Cultivar ‘Bianca’ pistachio kernelsPicture 3. - Cultivar ‘Bianca’ peeled pistachiosPicture 1. - Electrical hulling machinespreading; branching habit is intermediate.Terminal leaflet length is 10.1 cm, terminalleaflet width is 7.7 cm and the terminalleaflet/width ratio is 1.3. Peakbloom date occurs in the last week ofApril. Nut size is small to medium; nutlength is 21.6 mm, nut width is 11.6 mm,nut thickness is 9.9 mm, nut shape is generallyelongated, split nuts is generallylow (

Picture 4. - Pistachio shelling machinethe Bronte area, very few new plantationshave been established in the recentyears. The major constraints are, perhaps,the long unproductive period of theculture (full production is normally reachedby the 12 th or 13 th year), togetherwith the lack of sufficient experimentationon new orchard design, alternative rootstocks,mechanization and availability ofless alternate bearing cultivars. The questionthat arises is also related to the aimof new plantations: whether for industrialtransformation, following the present specialization,or for direct consumption, asin the rest of the emerging producercountries. In the first case we have geneticmaterial, market opportunities andenough know-how to imagine an easy development.In the second case, other varietiesshould be tested for the Sicilianenvironmental conditions, together withthe best suited cultural techniques. Forthis last option, strong concurrence, especiallyfor the cost of production, has tobe faced. In both cases there is a strongneed for research. At the Dipartimento diColture Arboree of the Università di Palermoresearches on alternate bearingphenomenon, rootstocks and germplasmevaluation are underway, in some casesfrom the 80’s. Some results of these researchactivities have been recently presentedto the last GREMPA meeting heldin Mirandela, Portugal (Barone et al.,2003; Caruso et al., 2003).Although currently the pistachio industrysituation, as mentioned before, could beconsidered stable, great opportunitiesare expected from the diffusion of mechanization,specially for harvest and othercultural practices. This is more likely tobe implemented in areas of new plantationswhere there are not the environmentalconstraints of the Bronte area.For this last area a strong impulse is expectedby obtaining the trade mark DOP(protected origin denomination) for thepistachios of Bronte, acknowledgementthat is underway. Finally, even if cooperationhas recently become stronger thanin the past, a further development of cooperationand, possibly, the establishmentof an unique Pistachio associationamong the growers are highly advisable topromote the Italian pistachio industry, as alreadydone in other producing countries.REFERENCESAnonymous. 2000. Italian PistachioHarvest Nears Zero. Statpub. com.Agric. Commodity Market News.Barone, E., T. Caruso, L. Di Marco.1985. Il pistacchio in Sicilia: superficicoltivate e aspetti agronomici. L’InformatoreAgrario 40: 35-42.Barone, E., L. Di Marco, F.P. Marra, M.Sidari. 1996. Isozymes and canonicaldiscriminant analysis to identify pistachio(P. vera L.) germplasm. Hortscience,31 (1): 134-138.Barone E., Caruso T. 1996. Genetic diversitywithin Pistacia vera in Italy. Reportof the workshop on: Taxonomy,Distribution, Conservation and Uses ofPistacia Genetic Resources. 29-30june, 1995, Palermo. IPGRI, Rome,Italy: 20-28.Barone E., Padulosi S., Van Mele P.1997. Descriptors for Pistachio (Pistaciavera L.). International Plant GeneticResources Institute, Rome, Italy.Barone E., La Mantia M., Marra F.P.,Motisi A., Sottile F., 2003. Manipulationof the vegetative and reproductive cycleof Pistachio (Pistacia vera L.). ProceedingsXIII GREMPA Meeting on Almondsand Pistachios, 1-5 giugno2003, Mirandela (Portugal). In press.Caruso, T., L. Di Marco, D. Giovannini,A. Di Pisa. 1986. Caratteristiche carpologichedi 4 cultivar di pistacchio coltivatein Sicilia. Frutticoltura 9-10: 63-66.Caruso, T., L. Di Marco, A. Raimondo.1987. Caratteristiche carpometriche diquattro cultivar di pistacchio (Pistaciavera L.) individuate in Sicilia. AgricolturaRicerca 80: 65-68.Caruso, T., L. Di Marco, R. Lo Bianco,F. Sottile. 1993. The pistachio germplasmof Sicily: pomological traits. Proceedingsof the IX G.R.E.M.P.A. Meeting- Pistachio. 1993. Bronte-Sciacca,20-21 May.Caruso T., Barone E., Marra F.P., SottileF., La Mantia M., De Pasquale C.2003. Effect of rootstock on growth,yield and fruit characteristics in cv.Bianca Pistachio (Pistacia vera L.)trees. Proceedings XIII GREMPA Meetingon Almonds and Pistachios, 1-5giugno 2003, Mirandela (Portugal). Inpress.Fabbri A., Valenti C. 1997. The SicilianPistachio industry: an overview. ActaHort. 470: 43-49.Granata G., Sidoti A., Corazza L. 1993.New acquisitions on control of Septoriapistaciae Desm. and Cytospora terebinthiBres. Proceedings of the IXG.R.E.M.P.A. Meeting - Pistachio.Bronte-Sciacca, 20-21 May. (Caruso T.,E. Barone and F. Sottile Eds.): 64-67.Greco F. 1998. Le principali avversitàdel pistacchio nel territorio di Bronte.Amministrazione Comunale di Bronte.Joley, L.E. 1969. Pistachio. In R.A. Jaynes(ed.). Handbook of Northern Americannut trees: 352-355. The NorthernNut Growers Association.Minà Palumbo, F. 1882. Monografia botanicaed agraria sulla coltivazione deipistacchi in Sicilia. Lorsnaider GiovanniTipografo, Palermo.Spina P. 1982 - Il pistacchio. FrutticolturaModerna vol. VI - Edagricole.Woodroof, J.G. 1967. Pistachio nuts. InTree nuts: Production, Processing, Products,261-287. AVI Publishing, Westport,Conn.E. Barone and F.P. MarraDipartimento di Colture Arboree -Università di Palermo - 90128 ItalyE-mail: ebarone@unipa.itPISTACHIO PRODUCTIONIN TUNISIAINTRODUCTIONPistachio (Pistacia vera L.) is a dioeciousspecies, widely present in the Mediterraneanarea. Its introduction in Tunisia mayhave been made by Carthaginians,however other introductions were madeby Romans and Arabs (Jacquy, 1973).Several species are scattered in differentparts of Tunisia (Jacquy, 1973; Nabli,1989): Pistacia terebinthus and Pistacialentiscus grown naturally in the dorsal hillfrom the north-west (Ghardimaou) to thenorth-east (Zaghouan). Pistacia atlanticaspreading from the dorsal hill on thenorth to the Saharian Atlas in the southwith important frequency in the predesertarea of the center and south of the country(Sened, Meknassy, Guettar…). Pistaciavera, the edible pistachio is foundalso from the north to the south withsome trees four to five hundred years old(Guettar oasis) and others two hundredyears old (Sfax – Mahres regions). Despitethis long history, development of pistachioproduction in Tunisia starts onlyduring the last 30 years.CURRENT SITUATIONPistachio is a hardy tree resistant todrought which can valorize poor soils,and therefore a lot of resources were putinto its development. Some developmentprojects were carried out with the supportfrom international organizations like FAOduring the period of the seventies. Fromless than 100 ha of pistachio orchard fiftyyears ago, the growing area is now morethan 43.000 ha localized mainly in theFAO-CIHEAM - Nucis-Newsletter, Number 12 September 200419

Table 1. Pistachio production in Tunisia (2002).Surface Number Yield Yield Density(ha) of trees (t) (kg/tree) (trees/ha)Irrigated crop 1.065 135.500 148,3 1,1 125Rainfed crop 41.983 2.888.618 1.029,5 0,4 70(Source: DGPA, 2002)Figure 2. Repartition of Pistachio growing area in TunisiaFigure 1. Geographical distribution of Tunisianpistachio growing area706050% of total area% of total productioncenter and the south of the country (Fig.1). Gafsa, Sidi Bouzid, Kasserine andSfax are the most important producing regionswith respectively 19.800 ha, 9.188ha, 7.668 ha and 3.290 ha.Despite this acreage increase the productionis still very weak reaching only1.170 t with important differences betweenirrigated and rainfed orchard and alsoregions (table 1). The north region representsless than 1% of the growing surfaceproducing more than 16% of the nationalyield (fig. 2).To understand this weak yield and characterizethe current situation of pistachiocrop in Tunisia, a survey was carried outin the important Tunisian pistachio growingzones (Ghrab et al., 2002b). Themost important observations were:• Inappropriate distribution of male andfemale trees on the orchard. Different ratiosof male to female varying from 2% to90% were observed reducing the profitabilityof the orchard.• Inadequate planted material that is supposedto be ‘Mateur’ as female and malesA40 and A25 cultivars. However the observationsmade revealed significant phenotypicdifferences between trees.• A shift of the flowering periods betweenA25 and A40 male cultivars and ‘Mateur’inducing an inefficient pollination. Thosecultivars were selected on the north ofthe country for their production, qualityand overlapping flowering period howeverunder the central and southern climaticconditions the flowering period of themale cultivars does not overlap with theblooming period of Mateur.• Since pistachio tree needs more thanfive years to bear under the central andsouthern semi-arid conditions, some growersopted for other species and quittheir orchards without any management.• For some pistachio growers, managementoperations such as fertilization, pruningand pest and disease control were%403020100Northignored and considered without any benefitfor this crop.CULTIVARS AND ROOTSTOCKSThe most commonly cultivated variety inTunisia is ‘Mateur’ (Fig. 3), selected in theNorth as well as two corresponding pollinatorsA40 and A25 (Mlika, 1980). Almostall the development of the pistachio industryin Tunisia is based on this cultivar.Few other local varieties are cultivated insome specific localities from which it tookits names as ‘Meknassy’ cultivars in the regionof Meknassy (Sidi Bouzid), ‘El Guettar’from the region of El Guettar (Gafsa) and‘Nouri’ and ‘Thyna’ from Sfax region.As rootstock, P. vera is almost the onlyused by growers for its good germinationrate. P. atlantica and P. terebinthus, inspite of their good characteristics asrootstock are barely used because oftheir seed germination difficulty.CULTURAL CONSIDERATIONPropagationT-budding is the most common propagationmethod. The budding is performed inJuly at the nursery on seedling of P. vera.The following winter trees are planted inthe orchard.PruningThe training system used for pistachio isusually the open vase, however a lot oforchards were grown without any training.CentreGrowing AreaSouthThe annual pruning is also rarely made. Ifit is made it is mainly to remove dried, diseased,infected, pest damaged, andnon-fruiting old shoots.PollinationPistachio trees are dioecious, with maleand female flowers borne on separatetrees. The designed orchard should ensurean adequate pollination. Male trees areoften planted in the center of a 3m x 3msquare of females, yielding an 8:1 ratio(Jacquy, 1973). Additional male trees aresometimes planted in border rows. Naturally,wind transports pollen from maleflowers to female flowers. Efficient pollinationwas assured when the floweringperiods of males is long enough to overlapwith the female flowering. But underTunisian conditions, a high percent ofblack fruit is obtained, and natural windpollination is proved inefficient and artificialpollination was widely adopted(Mlika, 1974). The male panicle is harvestedand kept on a dry, shaded place.The pollen is collected and mixed withtalc (1/9 ratio) and sprayed on the femalepanicle early in the morning. Three to foursprays are made during the female floweringperiod to insure a good pollinationrate.Pest and diseasePest and disease decrease quality andquantity of pistachio crop through their directand indirect damages. Few important20 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

Figure 3. Tree and nuts of ‘Mateur’ cultivar,Mateur, Tunisiapests and diseases are observed for theTunisian pistachio orchards. Eurytomaplotnikovi and Megastigmus pistaciae:the most pistachio fruit damaging wasps.They damage pistachio by feeding onnuts resulting in fruit blackness and drop.The most important disease is the leafspots, which caused a pre-matured leafing.Fertilization and irrigationFew growers fertilize and irrigate their orchardsdespite that the important growingareas are located on semi-arid zones withannual rainfall ranging from 150 mm to300 mm. This could explain the low yieldobtained. Some growers irrigate their orchardsonly during the first years of plantation.HarvestingIn Tunisia all orchards are hand harvestedfrom late August to early October dependingon the climate and the region.CONCLUSIONIn Tunisia the culture of pistachio existssince antiquity. But, in spite of this, its developmentstarted only thirty years agowith large planting programmes. However,its production is still low and researchwork should be done for pistachioproduction improvement. Based on thisstatement, different researches on pistachioare going on in Tunisia such as:• Developing micropropagation techniquesfor Pistacia species for rootstocksuse (Chelli Chaabouni and Gouta, 2002;Chelli Chaabouni and Drira, 2002).• Selection of male trees for Mateur cultivarunder the central and southern conditions(Ghrab et al., 2002a).• Survey and characterization and conservationof Tunisian pistachio germplasm.• Evaluation of introduced cultivars underTunisian conditions.AKNOWLEDGMENTSI wish to thank Mrs. Cherif R., Rhouma A.and Triki M. A. for their great help to completethe pest and disease part.REFERENCESChelli Chaabouni and Drira N., 2002.Culture Media and Hormone Effects onthe micro-propagation of two Pistachiospecies. III Inter. Symposium on Pistachiosand Almonds. Acta Hort., 591:395-398.Chelli Chaabouni and Gouta H., 2002.Effect of chemical scarification and giberellicacid on in vitro germination ofPistacia atlantica seeds. III Inter. Symposiumon Pistachios and Almonds.Acta Hort., 591: 73-76.Ghrab M., Ben Mimoun M., Triki H. andGouta H., 2002a. Evaluation of the performancesof seventeen male pistachiotreespecimens. III Inter. Symposium onPistachios and Almonds. Acta Hort.,591: 473-478.Ghrab M., Gouta H., Triki H., Jabeur M.,and Laroussi K., 2002b. L’amandier etle pistachier en Tunisie centrale: étatactuel et perspectives d’amélioration.Document technique, Institut de l’Olivier,1/2002, 16p.Jacquy R., 1973. La culture de pistachieren Tunisie. FAO, 97p.Mlika M., 1974. Contribution à l’étudede la pollinisation chez le pistachier.DEA Faculté des Sciences de Tunisie.Mlika M., 1980. Contribution à l’étudedu pistachier en Tunisie : choix des variétésmâles et femelles à floraison synchrone- Anatomie des fleurs. Mémoirede fin d’études de spécialisation INATunisie: 77 p.Nabli M. A. 1989. Essai de synthèse surla végétation et la phyto-écologie tunisiennes:I- Eléments de botanique et dephyto-écologie. Faculté des Sciencesde Tunis. 247 p.Ghrab M. 1 , Ben Mimoun M. 2 and Gouta H. 31Station d’Arboriculture Fruitière, Institutde l’Olivier, Route Soukra km 1.5, 3003 Sfax,Tunisie2Institut National Agronomique de Tunisia,43 av. Charles Nicolle, 1082 Tunis, Tunisia3Institut de l’Olivier, BP 1087, Sfax 3016,TunisiaE-mail: ghrab_m@yahoo.comBREEDINGMONOECIOUS PISTACHIOCULTIVARSINTRODUCTIONAll Pistacia (Anacardiaceae) species aredioecious and wind is the pollinatingagent (Zohary, 1952). For commercialpistachio orchards, approximately onemale tree is necessary for 8 - 11 females(Maranto and Crane, 1982). As a result,approximately 10% of a typical pistachioorchard is not productive. In the literature,only three cases of exceptional sex typesin Pistacia were reported. Firstly, Özbekand Ayfer (1958) found two hermaphroditetrees in Gaziantep province of Turkey.They supposed that these trees were eitherseedlings of Pistacia vera L., or hybridsbetween P. vera and P. terebinthus.The second report by Crane (1974) describedthree trees with exceptional phenotypes:(I) a branch bearing staminateflowers on a P. atlantica female tree, (II) ahybrid between P. vera and P. atlanticabearing approximately equal numbers ofstaminate and pistillate inflorescencesthat are mostly on separate branches,and (III) a similar hybrid, predominantlystaminate, but several branches bear pistillateinflorescences. Recently, Kafkas etal. (2000) reported a few monoecious P.atlantica trees in the Yunt Mountains ofthe Manisa province of Turkey. The distributionof staminate and pistillate inflorescenceswas found variable betweentrees. They found nine monoecious treeswith three different types: (Type I) allbranches of one of the trees were ‘fullymonoecious’ i.e., they bore a mixture ofmale and female inflorescences. (Type II)three trees had several branches withonly staminate flowers, while the rest ofthe branches bore pistillate inflorescences.(Type III) Five other trees had inflorescencesof both sexes on several branches,and pistillate inflorescences on theremaining branches. The authors testedpollen germination of monoecious treesand used them for pollination with P.vera. The pollen germination rates weresimilar with normal male trees of P. atlanticaand the hybridization with P. vera resultednormal fruit set and viable seedproduction.By using the preliminary results on thesemonoecious trees, a project supported byThe Scientific and Technical ResearchCouncil of Turkey (TUBITAK) was startedin the spring of 2001 by planning of a largescale of crosses with the aim to developmonoecious pistachio cultivars and tounderstand sex mechanism in Pistacia. Inthis paper, current status of the project isreported.FAO-CIHEAM - Nucis-Newsletter, Number 12 September 200421

MATERIALS AND METHODSType I (all branches bore a mixture ofmale and female infloresences) and TypeII (a tree has several branches with onlystaminate flowers, while the rest of thebranches bore pistillate inflorescences)monoecious trees found by Kafkas et al(2000) are the main materials of this study.Figure 1 shows some pictures fromthe fully monoecious tree (Type I). Thesetrees were used both as female and maleparents in this study. Additionally, two femaleP. vera cultivars (cvs ‘Siirt’ and‘Ohadi’) with a dioecious female P. atlanticatree as maternal tree, and dioeciousmale P. vera and P. atlantica trees wereused as pollinators. Totally, twenty differentcrosses were performed betweenfive female and four male parents.Before the stigma becomes receptive, femaleinflorescences were closed with apaper bag (Figure 1D). To collect pollenfrom male parents for artificial pollination,staminate branches of four pollinatorswere cut and put in water-containing potsin the laboratory to perform pollen tests andcross-pollination. In the following two days,pollen was collected and stored at -20 0 Cuntil pollination. Wheat flour was used as amixing agent with pollen at a ratio of 1:1.The harvested nuts were stratified for 60days and then they were germinated inthe greenhouse. Germinated plantletswere transplanted into the five liter plasticbags. The plants were transplanted to theorchard in the autumn of 2003 and dripirrigation is to be set up in 2004.Besides artificial pollination, the parentsused in this study were described morphologically.The progenies between P.atlantica genotypes and four pollinatorswere also characterized. The distributionof male/female branches/inflorescencesin the monoecious trees was observedyear to year by labeling them within thetree as well. When the progenies will reachthe reproductive stage, they would betested for their sex type, and they will beused as breeding parents to try to obtainmonoecious cultivated pistachios withhigh quality nuts. The segregating populationswill also be used to understandsex mechanism in Pistacia and will beused for molecular studies to clonegene(s) that are responsible for sex determination.Figure 1. (A) The fully monoecious P. atlantica tree, (B) a view from branches of the fully monoecious tree(F: pistillate inflorescences, M: staminate inflorescences), (C) close up view of the staminate and pistillateinflorescences in a branch, (D) P. vera cv Siirt cultivar tree with bagged branchesRESULTS AND DISCUSSIONArtificial pollination was made duringthree years. In 2001, the crosses betweenP. atlantica and four pollinators, in2002 the crosses between P. vera andthe pollinators, and in 2003 additionalcrosses were performed to increase plantnumbers. The crosses between P. atlanticaand four pollinators were made in Manisaprovince, whereas the crossesbetween P. vera and the pollinators wereperformed in Gaziantep Pistachio ResearchInstitute placed 1200 km far fromManisa province. The pollens were transportedto Gaziantep province in dry ice.Table 1 shows targeted and current plantnumbers in this project. We aimed to obtain500-700 seedlings from the crossesbetween P. vera cvs and monoecious P.atlantica Type I, 100-200 seedlings fromthe crosses between P. vera cvs and monoeciousP. atlantica Type II, 50-100seedlings from rest of the crosses. Totallyit is targetted to have 2000-3400 seedlingsin the project. Currently, we completedall the combinations and we havetotally 3054 seedlings in plastic bags.However, we also did additional crossesin (2003) to increase plant numbers especiallybetween P. vera and monoeciousgenotypes.Table 1. Targeted and current plant numbers in the crosses.No Female Male Targeted plant Current plantnumbers numbers1 P. atlantica x *M-PA-Type I 50-100 1082 P. atlantica x M-PA-Type II 50-100 1083 P. atlantica x P. vera 50-100 1084 P. atlantica x P. atlantica 50-100 1085 M-PA-Type I x M-PA-Type I 50-100 1106 M-PA-Type I x M-PA-Type II 50-100 587 M-PA-Type I x P. vera 50-100 698 M-PA-Type I x P. atlantica 50-100 989 M-PA-Type II x M-PA-Type I 50-100 7010 M-PA-Type II x M-PA-Type II 50-100 5211 M-PA-Type II x P. vera 50-100 5412 M-PA-Type II x P. atlantica 50-100 9713 P. vera cv. Ohadi x M-PA-Type I 500-700 63014 P. vera cv. Ohadi x M-PA-Type II 100-200 10815 P. vera cv. Ohadi x P. vera 50-100 5816 P. vera cv. Ohadi x P. atlantica 50-100 5617 P. vera cv. Siirt x M-PA-Type I 500-700 65818 P. vera cv. Siirt x M-PA-Type II 100-200 27319 P. vera cv. Siirt x P. vera 50-100 12020 P. vera cv. Siirt x P. atlantica 50-100 111Total plant numbers 2000-3400 3054M-PA: Monoecious P. atlantica22 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

S. Kafkas 1 , I. Acar 2 , H. Gozel 2 and S. Eti 11Department of Horticulture, Faculty ofAgriculture, University of Cukurova01330, Adana-Turkey.2Pistachio Research Institute, GaziantepTurkey.E-mail: skafkas@mail.cu.edu.trFigure 2. Variation of hermaphrodite flowers in the fully monoecious tree (St:stamen, P: pistil, Sg: stigma)(A) An hermaphrodite flower with five stamen and one very small pistil (B) A hermaphrodite flower withone stamen and one pistil, stigma with two parts (C) A hermaphrodite flower with one stamen and onepistil but stamen has stigma as wellCHILLING STRESS AND INJURYIN PISTACHIO FEMALEFLOWERSDuring the last three years, we have observedalso hermaphrodite flowers insome inflorescences of the fully monoecioustree. But they were not stable, becausethe numbers of anthers in a flowerchanged from 1 to 6. Figure 2 shows variationsin the hermaphrodite flowers ofType I tree. In this tree, only several ofthe inflorescences were fully hermaphroditeand a few of the others showed notonly hermaphrodite flowers but also hadstaminate and pistillate flowers as well. Insome inflorescences, one side showedstaminate flowers and the other side hadpistillate flowers, and hermaphrodite flowerswere in the middle of them. Also severalstamens having stigma as in thepistils (Figure 2C) were found. In 2001,these inflorescences were baggedwithout pollination and obtained 6 nutsraising five P. atlantica seedlings. In2002, open pollinated nuts of these inflorescenceswere germinated yielding 50seedlings. In 2003, these inflorescenceswere bagged and pollinated with Type Ipollen and now we have 48 seeds.A pistachio cultivar that has both staminateand pistillate or hermaphrodite inflorescencesin a single tree, together with highyield of quality nuts, would eliminate theneed for male trees in the orchard and increasesthe yield per hectare by about10%. Furthermore, protandry is a verycommon problem in pistachio orchards inTurkey and also in some of the other pistachioproducer countries. Therefore, theimportance of a developing monoeciouspistachio cultivar in pistachio productionand cultivation is clear. In the first cycle,we are not expecting to have a monoeciousgenotype with high yield and qualitynuts because of the hybridization of P.vera with the wild species. But, havingsegregating monoecious populations inthe first cycle and isolating the related genesand transferring them or performingsecond cycle crosses, there may arise acultivated monoecious pistachio cultivarwith high yield and quality.Sex determination mechanisms in plantsare diverse, and may involve either sexchromosomes such as in Actinidia (Mcneilage,1991) or individual sex gene(s)as in Cucumis (Perl-Treves, 1999). InPistacia, the genetic mechanism of sexdetermination is still unknown. But, knowingthe segregation of sex types in theprogenies of this project may explain sexinheritance and mechanism in Pistacia.ACKNOWLEDGMENTSThe authors acknowledge the support ofS. Karaca during the cross pollination studyin the Manisa province of Turkey. Thisproject is supported by The Scientific andTechnical Research Council of Turkey(TUBITAK).REFERENCESCrane, J.C. 1974. Hermaphroditism inPistacia. California Agriculture, 28: 3-4.Kafkas, S., Perl-Treves, R. and Kaska,N., 2000. Unusual Pistacia atlanticaDesf. monoecious sex types in the YuntMountains of the Manisa province ofTurkey. Israel Journal of Plant Sciences,48 (4): 277-280.Maranto, J. and Crane, J.C. 1982. Pistachioproduction. Division of AgriculturalSciences, Univ. of California, Leaflet2279, p.18.Mcneilage, M.A., 1991. Gender Variationin Actinidia deliciosa, the kiwifruit.Sex. Plant Reprod., 4: 267-273.Ozbek, S. and Ayfer, M. 1958. A hermaphroditePistacia found in the vicinity ofAntep, Turkey. Proc. Am. Soc. Hort.Sci., 72: 240-241.Perl-Treves, R., 1999. Male and femaleconversion along the cucumber shoot:approaches to studying sex genes andfloral development in Cucumis sativus.(C.C. Ainsworth editor). Sex Determinationin Plants. BIOS Scientific Publishers,pp. 189-216.Zohary, M. 1952. A monographical studyof the genus Pistacia. PalestineJournal of Botany, Jerusalem Series, 5:187-228.Despite of pistachio resistance to differentwell-known environmental stresses(drought, salt, unfavourable soils), thistree suffers from some serious stresseslike spring chilling. This injury can evennegatively affect crop yield. This stressspecially affects many temperate-zoneorchards of pistachio every year in Iran,causing significant yield losses and generaltree weakness, and severing alternatebearing. Varying regarding on differenttemperatures; these damages occurredthrough decreasing fruit set rate as effectivepollination period (through stigmaand style deterioration, decreasing ovaryreception period and flower age, and increasingpollen tube growth period), femaleflower abscission, inflorescence dieback, and flower buds’ death and abscissioncould occur.A research project has been directed tostudy the effects caused by this stress,strains, and the level of damages in differenttemperatures. Here, some resultsare briefly presented.The project involves simulated chillingstress in laboratory, simultaneous tooutdoor temperature decrease, resultingin remarkable effects on pistachio floweringand pollination in early spring 2003,we could compare our lab results withfield observations and records.In order to study the stress, shoots containingflower bud from cultivar “Ohadi” indifferent phenological stages (bud, andblooming and full-bloomed inflorescence)were collected and transferred to a controlledlow temperature chamber. Temperaturewas reduced to 2 o C in one hour.Every hour, some bud and shoot sampleswere assessed looking for macro- and microscopicvisible injuries.Chilling could adversely affect the reproductivestructures in different thermal basedand morpho-anatomical levels. Themost severe injuries (from lower to highertemperatures) were complete flower buds,current spring shoot (containing inflores-FAO-CIHEAM - Nucis-Newsletter, Number 12 September 200423

season cultural practices may affect phenologyof female flowers, especially thetime of bud bloom, this model can also beuseful for reliable cultural programming.Picture 1. Healthy control (left) and necrotic bud (right), Immediately before bloomY. GholipourIranian Pistachio Research institute (IPRI)Qazvin Pistachio Research StationE-mail: ygholipour_trade@yahoo.comPicture 2. Control (left) and injured necrotic stigma and style (right)UNDERGROUND DRIP SYSTEM:THE NEW IRRIGATIONMETHOD FOR PISTACHIOAND ALMOND ORCHARDIN TURKEYPicture 3. Complete flower necrosis (left) and current shoot and inflorescence die back (right)cences), inflorescence, pistil, style, andstigma necrosis and deterioration, respectively.Injury level analysisFlower buds: their chilling resistance isthe highest among all reproductive structuresand organs. Their resistance, however,decreases as they finish their dormancyperiod, especially before bloom.Light injury (low to about -4 o C for about2h) could lead to their delayed bloom withsome blooming and flowering abnormalities.Deeper injury (about -4 to -6 o C forabout 6-8h) resulted in the necrosis anddeterioration of very young pistils in closedbuds (Picture 1).Inflorescences: in the first visible injury level,the tip vegetative growth (on the inflorescenceshoot) became darker green incolor. In this stage, the fertilization rateand subsequent fruit set can decrease,possibly because of negative effect of thelow temperature on the pollen tube growthand shortening physiologically effectiveage of the pistil. With more temperaturereduction (low to about -2 o C for about 2h),stigma, and then style showed morphologicalnecrosis and anatomical deterioration,which resulted in pollen germinationand tube growth inhibition (Picture 2).Complete necrosis of flowers, and then,inflorescence, caused by about -4 o C forabout 2h, was observed (Picture 3).Comparing this simulated pattern of injuryin laboratory, with injuries records fromorchards, showed that we can predict anddetermine injury level and subsequentevents in fruiting and abscissions basedon field temperature data. For example,due to temperature drop, down to 2 o C forabout 2 hours, it could be foreseen that adecrease in fruit set would happen as resultof delayed pollen tube growth,without visible symptoms. In some areas,lower temperatures caused visible injuriesof stigma, resulting in problemmes infertilization. We are developing a modelbased on environmental temperature(with other climatic parameters) data andfemales’ injuries, to make our cultivationsready for next years potencial spring chillingstress. By the way, since the earlyIt is well known that pistachio trees (Pistaciavera L.) are drought resistant. InTurkey the crop was not irrigated until recently.During the last decade seminarsand meetings on Pistachio productionand management were given by Prof.Kaska and Ak to farmers. They both triedto convince farmers to irrigate pistachiotrees, when water supply is available.Farmers believe that pistachio treesshould not be irrigated, because theywould die. This belief is only true when awrong irrigation method is applied.Commercial orchards are budded ontoPistacia vera rootstocks. This rootstock issusceptible to some diseases. We tellgrowers the usefulness of applying asound irrigation system. Some farmershave started to irrigate by tanker system.They have seen that irrigation is usefulfor pistachio trees. They recognized itseffects on reducing alternate bearing.When irrigating pistachio trees somechanges could occur as below: Treegrowth will be higher, leaf size will be bigger,the number of current year’s shootlength will be increased, yield will be increased,nut size will be higher, splittingpercentage will increase, blank nut ratewill be reduced, and less alternate bearing.24 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

B. E. Ak standing close to a young almond treeYoung almond orchards growing at Anatolia, TurkeyIrrigation by underground drip systemsare being used on olive plantations inSpain. This system has been started tobe used for pistachio and almond orchardsin Turkey. However farmers arenot confident with this method. The advantagesof underground drip irrigationsystem are mentioned as follows: lowerwater consumption, better water distribution,greater uniformity, waste water canbe used, less evaporation, greater transpiration,location of fertilizer, less calcification,less diseases, possibility of tillage,Tree planting in Turkeyreduction of labour, longer lasting, novandalism.All above advantages are valid for all kindof fruit trees. In fact, currently this systemis experimentally being used in pistachioorchards at Ceylanpinar State Farm(South Anatolia). The yield seems to becomemore regular, as there is strong alternatebearing, especially in the ‘Kirmizi’cultivar. This underground irrigation systemwas established in 200 areas wherethe adult trees are 30 years old, using‘Siirt’ and ‘Kirmizi’ cultivars. The depth ofthe watering line is at approximately 20-25 cm deep, under the soil surface.In Sanliurfa, a private investor, establisheda new almond orchard in 2002 byusing this underground drip irrigation system.There were planted 30 ha of almondtrees budded 6m x 6m onto bitter almondrootstocks. The cultivars were ‘Ferragnes’,‘Ferraduel’, ‘Lauranne’, ‘Bertina’and ‘Felisia’. The new orchard for pistachiocv ‘Siirt’ and old orchard cv. ‘Kirmizi’and ‘Ketengömlegi’ have been started toirrigate with an underground drip irrigationsystem.As a result, irrigation is a big revolutionfor pistachio industry in Turkey. In the futurethe beneficial effects of irrigation willbe clear and all possible trees will be irrigated.The researches on this subject arebeing developed at the Pistachio ResearchStation in Gaziantep, the Universityof Kahramanmaras and Pistachio Researchand Application Center, Universityof Harran in Sanliurfa. In addition, almondorchards are expanding at South-easternpart of Turkey. The ecology is very suitableand the farmers prefer almond treesbecause of their many advantages.B.E. AkHarran University, Faculty of AgricultureDepartment of Horticulture63200- Sanliurfa (Turkey)Tel: (414)247 26 97 Fax: (414) 247 44 80E-mail: beak@harran.edu.trSoil ready for plantingUnderground irrigation system placedFAO-CIHEAM - Nucis-Newsletter, Number 12 September 200425

IRANIAN AGRICULTURALSYSTEM TO DEVELOP HIGHERQUALITY AND HEALTHIERPISTACHIO PRODUCTIONINTRODUCTIONIn recent years, improvement and optimizationof different stages of pistachio production,processing and handling, hasbeen important and has affected most research,extension, and governmental programmesand activities. These significantefforts have been directed to increase thequality and health of pistachio product regardingdifferent aspects: harvest, postharvest,processing, transportation andmarketing. This is based on internationalstandards, and taken place in order toprotect public health of domestic and foreignpistachio consumers. Here, we summarizeIranian programmes and activitiesto reach this aim.GOVERNMENTALAND ADMINISTRATIVE WORKSThe government of Iran, as one of theeffective and active members of WHO, iscommitted to supply healthy and safeproducts to international markets, andtries to use modern technology in variousstages of production:• Organizing the pistachio committee inthe Ministry of Agriculture and by attendingthe representatives from related ministriesand organizations (Ministry ofHealth and Medical Education, Customs,Ports and Shipping, Iranian center for developmentof export, Ministry of foreignaffairs, Iranian institute of standards, Pistachioresearch institute etc.)• Organizing the pistachio staff and aflatoxincommittees• Preparation and distributing of manualsand technical publications plus 90.000sheets of posters, 85.000 sheets oftracts, more than 500 m wall writings, andseveral extensional films among the pistachiogrowers.• Holding training workshops in order toretraining horticultural exports and promoters.• Convening nationwide seminars on aflatoxinin pistachio• Convening executive and applied workshopsand seminars for growers to developmodern and standard cultivation (e.g.pruning, methods of fertilization e.g. microelementapplication and deep placementfertilizer), harvesting (to develop ontime harvest with proper equipment andproper transport), processing (speciallydrying process) etc.• Huge investment to develop establishment,mobilization and equipping currentprocessing terminals and 634 new linesof hygienic processing of pistachio.• Organizing product forewarning networkswith cooperation of research centers,Iranian Ministry of Health and MedicalEducation and Academic departmentsof plant protection, in pistachio cultivatingprovinces, in order to study and announcethe most proper time of harvest. Thisaction has lead to significant decrease ofpistachio early split and aflatoxin contents.• Implementing the national project of“pistachio production structural improvementin country” within the last few yearsin ten main pistachio cultivation provinces.This project covers applying differenttechnical methods to decrease and removethe risk of aflatoxicity of the crop.• Persuasion of pistachio growers tochange their traditional methods of allpistachio cultivation practices, specially adecrease of pesticides (which lead to reduceto 37.5% per surface unit).• Establishing the nationwide union of pistachioproduction cooperative companiesin order to organizing and activating thecooperative companies in pistachio cultivationareas.RESEARCH ACTIVITIESIn recent years many projects have beendeveloped and directed to find and adaptapproaches to improve pistachio quality.A huge effort has been spent to carry outmany research and research-extensionalprojects and activities aiming to developmost proper ways of production and processing.Their results have been appliedto obtain higher quality product, and nowwe can see the results of all the referredactivities.H. Hokmabadi 1 , B. Gheibi 2 , Y. Gholipour 11Scientific board members, Iranian PistachioResearch Institute,2Pistachio Affairs Dept. Ministry of JahadAgriculture,E-mail: hokmabadi@pri.irE-mail: ygholipour_trade@yahoo.comE-mail: b.gheibi@agri-jihad.orgPistacia vera L. 1753 IN IRANINTRODUCTIONThe history of pistachio production andbusiness in Iran is long. The reason isthat the origin of pistachio trees was theIranian plateau and the first pistachiotrees were first grown by Iranians. Thewidest natural pistachio forests havebeen found in Iran. Expanding of pistachiogardens and improved quality andquantity of Iranian pistachio productshave not acquired all at once. But it is theresult of nonstop efforts and creativity ofThe forest of P.vera in SarakhsIranian farmers and gardeners during thelast centuries.The wild species of Pistacia vera is theorigin of all domesticated pistachio cultivarsin Iran. Since thousands of yearsago, the nuts or scions of this wild specieshave primarily transferred to Khorasanand then to other parts of Iran. Abrishamiinformed that at Safaviehe times thepistachio nuts and scions were introducedto Kerman province for the first time(Abrishami 1994). Today, Kerman andRafsanjan are the main pistachio producingregions in Iran.The study of the ecological distributionand genetic diversity of Iranian wild pistachiosmight be a functional mean to selectconvenient and resistant rootstocksfor Iranian domestic pistachios (Behboodi2003).BOTANICAL CHARACTERISTICSOF P. veraThe genus Pistacia belongs to theAnacardiaceae family. There are two synonymsfor P. vera, which are P. reticulataWilld 1797 and P. trifolia L. 1753. P.vera is a diocious plant and the only distinguishingtrait between female and maletrees are their flowers at flowering time.Trees 3-8 (-10) m high, cortex brownish,becoming gray. Leaves deciduous, coriaceous,imparipinate. Leaflets 1-2 (3)pairs, sometimes with only one leaflet,glabrous or along the mid rid puberulent,upper surface smooth, lower surface opaque,ovate or orbiculate, apex abruptlyacuminate, acute, rarely obtuse, 5-10(-12) cm x 3-6 (-8) cm; rachis terete, puberulent.Fruiting panicle upright straight.Drupe 10-20 cm x 6-12 mm, oblong-linearor ovate, apiculate (Rechinger 1969).DISTRIBUTION OF P. VERA FORESTSNative P. vera forests are placed in northeasternpart of Iran, particularly in Sarakhs,Zoor-Abad, Jam and Tayebad.26 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

Leaves of P.veraThese forests extend to Bokhara andSamarghand in Turkmenistan from thenorth, to Balkh, Thaleghan, and Harat inAfghanistan from the east. In Sarakhs region,more intense forests are seen atTchahtchahe, Khabeh, and Ameneh Morad-Tappeh.These jungles have sporadicsparse of trees with diffuse spread of pistachios.Male and female pistachios aremixed through these forests.The extend of P. vera forests have beenestimated about 75.000 ha and the mainforest of Sarakhs is about 17.500 ha. Inthe last century, these forests have facedintensive human damages. The grassedNuts of P.veragrounds of P. vera forests have encounteredovergrazing of livestock belongingto the different Iranian tribes who live inthis region and the wood of pistachiotrees has been used for their thermal requirements.For these reasons, the extensionof P. vera forests have dramaticallyreduced and the distance betweenpistachio trees has enlarged.P. vera is not found as forest in other regionsof Iran. Thus it seems that thesetrees have been adapted to the ecologiccondition of this region. P. vera is the originof all cultivated Iranian domestic cultivars.The Gazvini cultivars, regarding nutappearance, are similar to wild P. vera, inwhich both of them have small nuts. Inaddition, the nuts of P. vera are similar tothose of small Badamy cultivars.DISTRIBUTION OF P. veraACCORDING TO ECOLOGICALFACTORSP. vera species is extended from northwestto south-east of Sarakhs region at300-1500 m high. The average yearlyrainfall in these regions is about 200-300mm and the average daily temperature isbetween 10-20 ºC. Considering resistanceof P. vera trees regarding the changeof day and night temperatures, its lowwater requirement, and the absence ofGummous disease for wild P. vera, it ispossible to use it as resistant rootstockfor pistachio or to cross it for hybridizationto improve the quality of cultivated pistachiosin the future.REFERENCESAbrishami M., 1995. Persian Pistachio,a comparative history. University press,Tehran, Iran.Behboodi B. Sh., 2003. Ecological distributionstudy of wild pistachios for selectionof rootstocks. XIII GREMPAMeeting on Pistachios and Almond, 1-5June, Mirandela, Portugal.Rechinger K. H. (1969) Flora Des IranischenHochlandes Und Der UmaramendenGebirge. No. 63. AkademischeDruck-u Verlagsanstalt, Graz (Austria).Behrooz Sh. BehboodiDepartment of Biology, Faculty of Science,University of Tehran, Tehran, Iran.E-mail: behboodi@khayam.ut.ac.irPistacia atlanticaDESF. 1800 IN IRANINTRODUCTIONThree important Pistacia species P. vera,P. khinjuk and P. atlantica are found inIran. For this reason Iran is regarded aspistachio origin and rising. The synonymspecies of P. atlantica is P. mutica Fischand C.A. May 1838. This species hasthree subspecies in Iran, which are differentin their botanical appearance.Distribution of Pistacia vera in IranBOTANICAL CHARACTERISTICSOF P. atlanticaTrees up to 7 m high. Leaves deciduous,imparipinnate, leaflets 2-4 (-5)pairs, form and size very variable, rotundate-ovate,oblong, or lanceolate, (2-)FAO-CIHEAM - Nucis-Newsletter, Number 12 September 200427

P. atlantica sub-sp. mutica trees P. atlantica sub-sp. kurdica treesForest of P. atlantica sub-sp. cabulicaLeaves of P. atlantica sub-sp. muticaLeaves of P. atlantica sub-sp. kurdicaDistribution of P. atlantica sub-sp. muticaLeaves of P. atlantica sub-sp. cabulica2.5-8 cm x 0.7-2 (-2.5) cm, obtuse orblunt at apex, never mucronate, at marginciliate, becoming glabrous, nerves inconspicuousor slightly conspicuous. Rachismarginate or narrowly winged. Maleinflorescence paniculate, 2-8.5 cm x 6-9.5cm, different in subspecies. Floweringtime early spring. Fruit drupe, with aromaticpericarp, seed oily (Rechinger 1969).WORLD GEOGRAPHIC DISTRIBUTIONOF P. atlanticaP. atlantica is located in the Atlantic region,Turkey, Iran, Caucus, Afghanistan,Pakistan, Iraq, Syria, north Africa, and Algeria.However its origin is Iran, but it hasexpanded from east to southwest of Europeand from southeast to North of Africaand the Canary islands.P. atlantica subspecies in IranRechinger classified P. atlantica intothree subspecies: mutica, kurdica, cabulica(Rechinger 1969). However non-Iranianbotanists have not accepted thisclassification.P. atlantica subspecies mutica (Fishand May) Rech. 1969Rachis winged up to terminal leaflets,leaflets narrowly oblong, 3-8 cm x 1.5-3cm, at margin ciliate. Drupe slightlybroader than long. This subspecies is locatedin Turkey, Iran, and the Caucus. InIran, it is found in northwest, northeast,center, east and south including Azerbaijan,Esfahan, Kohkiloyeh Va Boyer-Ahmad,Fars, Kerman, Khorasan, Yazd,Semnan and Tehran provinces.28 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

Nuts of P. atlantica sub-sp. mutica Nuts of P. atlantica sub-sp. kurdica Nuts of P. atlantica sub-sp. cabulicaDistribution of P. atlantica sub-sp. kurdicaDistribution of P. atlantica sub-sp. cabulicaP. atlantica subspecies kurdica (Zohary)Rech. 1969Rachises winged up to terminal leaflets,leaflets ovate, 4-9 x 2-5 cm, glabrous orrarely ciliate at margin. Fruits depressedglobouse,5-8 mm x 8-10 mm. This subspecies,apart from Iran, is distributed inIraq, Syria, and Turkey. In Iran it is nativein Azarbaygan, Kurdestan, Kermanshahan,Kohkiloyeh Va Boyer-Ahmad, andTehran provinces.P. atlantica subspecies cabulica (Stocks)Rech. 1969Leaflets (2) 3-4 pairs, which are lanceolate,attenuated at the ends, 3-4 times longerthan broad. Drupe globous, withequal long and broad. This subspecies isfound in Afghanistan, Pakistan, and Iran.Its distribution in Iran is mainly limited tosoutheast and central parts of the countrywhere Hormozgan, Kerman and Balotchestanprovinces are placed.Ecological distribution of P. atlanticasubspeciesP. atlantica subspecies mutica is foundbetween 900-2800 m of hight. The meanyearly rainfall is between 200-400 mm inthese regions and the mean of yearly daytemperature is between 15-20 ºC.P. atlantica subspecies kurdica is distributedin hights between 900-2800 mwhere the yearly rain mean is about 500-600 mm and the average of yearly daytemperature is flanked by 5-15 ºC. Thedistribution of P. atlantica subspecies cabulicais essentially restricted to Iransoutheast where the hight of lands altersfrom 50 m to 2500 m. The subspecies cabulicais able to grow in poor soils. Themean rainfall in its growing places isyearly lower than 100 mm and increasesto 200 mm as maximum. The averageday temperatures are 15-25 ºC yearly. Itis the most tolerant subspecies regardinglow water requirements and standing hightemperatures (Behboodi 2003).CONCLUSIONSThe subspecies cabulica’s tolerance ishigher than in other P. atlantica subspeciesregarding enduring of xerothermicand warm weather. Thus to improve domesticpistachios we suggest to usesubspecies cabulica rootstocks for breedingwith cultivated rootstocks or to producenew rootstocks using P. vera x P.atlantica subspecies cabulica or P. khinjukx P. atlantica subspecies cabulica hybrids(Behboodi 2003).REFERENCESBehboodi B. Sh., 2003. Ecological distributionstudy of wild pistachios for selectionof rootstocks. CHEAM JournalOptions Mediterraneennes, in press.Behboodi B. Sh., 2003. Ecological distributionstudy of wild pistachios for selectionof rootstocks. XIII GREMPAMeeting on Pistachios and Almond, 1-5June, Mirandela, Portugal, AbstractBook.Rechinger K. H. (1969) Flora Des IranischenHochlandes Und Der UmaramendenGebirge. No.63. AkademischeDruck-u Verlagsanstalt, Graz (Austria)Behrooz Sh. BehboodiDepartment of Biology, Faculty of Science,University of Tehran, Tehran, Iran.E-mail: behboodi@khayam.ut.ac.irFAO-CIHEAM - Nucis-Newsletter, Number 12 September 200429

Pistacia khinjuk STOCKS 1852.IN IRANINTRODUCTIONThe scientific name of Pistacia khinjukderives from the Persian word “Khenjok”and the origin of this word is the Korasanprovince of Iran. This species is the synonymof the species P. acuminata Boissand Bushe 1860. P. khinjuk is describedas “Lentisque de Bombay” in French language,as “Östindisher Mastix” in Germany,and in English as “East IndianMastiche”. The P. khinjuk trees are widelydistributed in several mountainous regionsof Iran. It is resistant against xerothermicconditions and hot and cold weather.These properties suit it for graftingof pistachio trees (Behboodi, 2003).BOTANICAL CHARACTERISTICSOF P. khinjukTrees are dioecious, 3-7 m high, cortexsmooth, grey coloured. Leaves deciduous,imparipinnate, sometimes (var. populifoliaBoiss.) reduced only to one terminal leaflet,pubescent, velvety, becoming glabrousin age, petiole terete, leaflets 1-2(3) pairs, 3-10 cm x 1.5-7 (-8) cm, broadovate, oblique, narrowed suddenly into a± acuminate apex, on the upper surfaceshiny. Fruiting panicle straight. Drupe 4-6mm x 4-5 mm, globose, subcompressed,suboblique, apiculate (Rechinger 1969).DISTRIBUTION OF P. khinjukFORESTSP. khinjuk is reported to spread in Iran,Turkey, Pakistan, Iraq, Syria, North Africa,and Palestine.DISTRIBUTION OF P. khinjuk IN IRANACCORDING TO ECOLOGICALFACTORSP. khinjuk is distributed in west, center,east, south, and east-south of Iran, inKurdestan, Lorestan, Eshfahan, KohkiloyehVa Boyer-Ahmad, Farse, Hormozegan,Kerman, Balotchestan, Khorasan,Semnan and Tehran provinces. Only athin north band in Gillan and Mazandaranprovinces, on the coast of Caspian sea isblank of this species. P. khinjuk spread at700-2000 m heights on hills, flats and onmedium heights of Alborze and Zagrosemountains. This species withstands tomost trouble weather situations. The averagerainfall of its growing areas altersfrom 100 mm to 600 mm and the meantemperature varied from 10 ºC to 25 ºC.P. khinjuk is the most interesting pistachiospecies in Iran as it grows in diffe-P. khinjuk treerent regions and diverse climates. It is exclusivelydistributed as continuos populationsor accompanying with subspecies ofP. atlantica. Distribution of this species isnot affected by wide ranging changes inday and night temperatures, height, andlow rain. It is adapted to xerothermic weathers,soil sorts, and difficult environmentalsituations (Behboodi 2003).Using P. khinjuk rootstocks alone, or utilizingits hybrids such as P. khinjuk x P. atlanticasubspecies mutica or P. khinjuk xLeaves of P. khinjukThe forest of P. khinjuk tree30 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

GERMPLASM COLLECTIONOF Pistacia AT THE JACOBBLAUSTEIN INSTITUTEFOR DESERT RESEARCH.PHENOTYPIC TRAITSAND MOLECULAR MARKERSP. atlantica subspecies cabulica or P.khinjuk x P. vera may assist breeding programmesto obtain resistant pistachiorootstocks.REFERENCESBehboodi B. Sh., 2003. Ecological distributionstudy of wild pistachios for selectionof rootstocks. CHEAM JournalOptions Mediterraneennes, in press.Distribution of P. khinjukNuts of P. khinjukBehboodi B. Sh., 2003. Ecological distributionstudy of wild pistachios for selectionof rootstocks. XIII GREMPAMeeting on Pistachios and Almond, 1-5June, Mirandela, Portugal, Abstract Book.Rechinger K. H. 1969, Flora Des IranischenHochlandes Und Der UmaramendenGebirge. No.63. AkademischeDruck-u Verlagsanstalt, Graz (Austria)Behrooz Sh. BehboodiDepartment of Biology, Faculty of Science,University of Tehran, Tehran, Iran.E-mail: behboodi@khayam.ut.ac.irNatural habitats are shrinking in theworld, causing reduction in genetic variabilityof plants with important economicalvalue. Shift from traditional agriculture ofPistacia vera L. (pistachio nut tree, Anacardiaceae)to modern agriculture, ‘disturb’the genetic pool of the existing naturalpopulation of this species by introductionof selected cultivars. The lengthyprocess of cultivars selection leads onone hand to yield producing crops, but onthe other hand to a loss of importanttraits. Wild relatives of crop species arethus useful as potential genotypes inbreeding programmes. Gene banks andlive germplasm collections are importantin the conservation of the genetic variability,which still exists at a rather limitedscale in the wild.At the Jacob Blaustein Institute for DesertResearch (BIDR), Ben-Gurion Universityof the Negev, Israel, a live germplasm collectionof Pistacia species was established(Golan-Goldhirsh and Kostiukovsky,1998). The collection consists of ca. 600accessions of P. vera, P. atlantica, P.khinjuk, P. chinensis, P. lentiscus, P. terebinthus,and P. palaestina. All plantsgrowing in the field collection developedfrom seeds collected from natural populationsof Pistacia spp. around the Mediterraneanbasin, and Asia (Table 1). TheBIDR germplasm collection at its currentstatus serves as a small-scale gene bank,representing the genetic variability thatoccurs in nature. Thus, offering an opportunityto study the genetic relations amongand within Pistacia species. Young trees (6to 8 years old) are growing under similargrowth conditions and therefore, any phenotypicvariability can be related to geneticdifference (see Fig. 1). During the lastfew years we characterized the germplasmby assessing phenotypic traits, followingplant growth rate and related theseto molecular markers.Molecular marker techniques, amplifiedfragment length polymorphism (AFLP)and random amplified polymorphic DNA(RAPD) as well as morphological descriptorsand plant growth parameters (followingIPGRI descriptors for P. vera, 1997)were used to assess the relationshipsamong Pistacia species and amongaccessions of P. vera (Barazani et al.,2003; Golan-Goldhirsh et al., 2004).FAO-CIHEAM - Nucis-Newsletter, Number 12 September 200431

Table 1. Pistacia species growing inthe germplasm collection at BIDR andtheir geographical origin.Pistacia speciesOriginP. atlantica CyprusIsraelSpainSyriaP. chinensis USAP. khinjuk SyriaP. lentiscus CyprusIsraelTunisiaSpainP. palaestina IsraelSyriaP. terebinthus SyriaP. vera TurkmenistanSyriaFig.1. View of accessions of Pistacia atlantica(front) and Pistacia lentiscus (back) growing inthe germplasm collection at BIDRZohary (1952), in his famous monographon Pistacia spp. divided the eleven speciesof the genus into four different sections(Fig. 2). The three evergreen speciesof the genus: P. lentiscus, P. weinmanifoliaand P. saportae were includedin the section EU Lentiscus; the two Americanspecies were clustered in the sectionLenticella; in the section EU Terebinthushe included the five species P. terebinthus,P. palaestina, P. vera, P. khinjuk,and P. chinesis, while P. atlantica wasseparated into a single section Butmela(Zohary, 1952). Studies conducted atBIDR germplasm collection on the relationshipamong Pistacia species usinggenetic marker techniques (Golan-Goldhirshet al., 2004) and morphological descriptors,support Zohary’s division of thegenus (Fig. 2). However, while parsimonyanalysis of the morphological descriptorsclustered the six studied species accordingto Zohary’s division, molecular techniquesanalyses (AFLP and RAPD) includedP. atlantica, P. vera and P. khinjuk inthe same genetic subgroup. The geneticclustering indicated that the genetic distancebetween P. atlantica and P. vera-P.khinjuk was smaller than that of the lattertwo species to P. terebinthus-P. palaestina(Fig. 2). We assume that the sameFig. 2. Genetic markers and morphological descriptors clustering of six Pistacia species (bottom) incomparison to the classical division of the genus as was proposed in 1952 by Zohary (top)center of distribution of P. vera, P. khinjukand P. atlantica in central Asia is thecause to the close genetic relationsamong these species. However, a muchdeeper study, such as alignment of genomicnucleotide sequences of conservedgenes, might assist in deciphering thephylogenetic tree of Pistacia sp.The germplasm collection at BIDR includesaccessions of P. vera that were developedfrom seeds collected from Badghiz,Turkmenistan. There, natural standsof P. vera still exist in two populations,Kepele and Agachli, close to the Iranianand Afghan borders. Plant growth parameters(plant height and trunk diameter)over two years of measurements at thecollection site indicated that accessionsfrom Agachli had higher growth rate thanthese of Kepele. Statistical analysisshowed that these differences were significantat 96% and 94% levels for plantheight and trunk diameter, respectively(Barazani et al., 2003). Agachli accessionscan therefore be considered for futurebreeding programmes of P. vera.Moreover, significant statistical relationwas found between plant growth rate andthe genetic matrix as was assessed byRAPD (Barazani et al., 2003). Trees inthe two populations, Kepele and Agachli,although geographically close, grow indifferent habitats, which probably led tothe genotypical differences.The pistachio tree is an important nutcrop in many countries. However, P. atlantica,P. khinjuk, P. palaestina and P.terebinthus are used by local farmers inthe Middle East as rootstocks for graftingof P. vera. Therefore, wild relatives ofhorticultural cultivars may be useful stockmaterial for breeding programmes (Zohary,1995), pointing to the significance ofpreservation of the genetic variability.Increasing concern on loss of genetictraits has led, over the last decades, toconservation plans and more effort togene banks management, as well as establishmentof germplasm collections ofwild relatives of economically importantcrops. We destined the Pistacia spp. livegermplasm for this purpose and for selectionof suitable genotypes for future breedingprogrammes.ACKNOWLEDGMENTSThis research was supported in part underGrant No. TA-MOU-98-CA17-028 fundedby the U.S. – Israel Cooperative DevelopmentResearch Program, Bureau forEconomic Growth, Agriculture, and Trade,U.S. Agency for International Development.REFERENCESBarazani, O., Atayev A., Yakubov B.,Kostiukovsky, V, Popov, K., Golan-Goldhirsh,A. 2003. Genetic variability inTurkmen populations of Pistacia vera L.Genet. Resour. Crop Evol. 50: 383-389.Golan-Goldhirsh, A., Kostiukovsky, V.1998. Mediterranean Pistacia genusgermplasm collection at Sede Boker Israel.Acta Hort. 470: 131-137.Golan-Goldhirsh, A., Barazani, O.,Wang, Z.S, Khadka, D.K., Saunders,J.A., Kostiukovsky, V., Rowland, L.J.2004. Genetic relationships among MediterraneanPistacia species evaluatedBy RAPD and AFLP markers. PlantSyst. Evol. (in press).IPGRI. 1997. Descriptors for Pistachio(Pistacia vera L.). International PlantGenetic Resources Institute, Rome,Italy.Zohary D. 1995. The genus Pistacia L.In : Padulosi S., Caruso T., Barone E.(eds.) Taxonomy, distribution, conservationand uses of Pistacia genetic resources.IPGRI, Italy, pp. 1-11.Zohary M. 1952. A monographical studyof the genus Pistacia. Palestine J. Bot.(Jerusalem Series) 5: 187-228.O. Barazani and A. Golan-GoldhirshBen-Gurion University of The Negev, TheJacob Blaustein Institute for Desert Research,Albert Katz Department of Dryland Biotechnologies,Desert Plant Biotechnology Laboratory,Sede Boqer Campus 84990, IsraelE-mail: avigolan@bgumail.bgu.ac.il32 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

WHERE IN A PISTACHIO TREEIS Xanthomonas AND HOW DIDIT GET THERE?BACKGROUNDXanthomonas translucens has been identifiedas the probable causal agent of diebackof pistachio (Pistacia vera) in Australia.The bacteria have been regularlyisolated from twigs with internal stainingfrom diseased trees. The location of Xanthomonasin other parts of the tree andthe identification of possible entry pointsare the focus of research in progress atthe Sunraysia Horticultural Centre (Vic)and the University of Adelaide (SA).INVESTIGATIONIn spring 2001 we performed various surveysto determine the tissues in whichbacteria are most likely to be found. Hundredsof inflorescences and individualmale and female flowers, leaves, vegetativebuds and current season growth wereexamined for the presence of the bacteria.Bacteria were present in male inflorescences,leaves and current seasongrowth from highly diseased trees.We also felled diseased male and femaletrees and healthy male trees at Kyalite(NSW) to locate the bacteria, through laboratoryisolations, in different parts ofthe wood. Samples of wood from therootstocks up to one-year old twigs, andtissues from young and old sapwood andheartwood were also examined.Figure 1. Section of main trunk of a diseasedtree. Heartwood and young sapwood indicatedwith arrowsFigure 2. Lesion from a branch of a diseasedtree. Unstained and stained sapwood indicatedwith arrowsFigure 3. Pruned branch of a young potted treebeing inoculated with a drop of bacterialsuspensionFigure 4. Leaf scar of a young potted tree beinginoculated with bacterial suspensionX. translucens was located mainly in theyoung, unstained or stained sapwood ofthe main trunk (Figure 1), primary branchesand younger branches. The bacteriawere rarely found in discoloured heartwood(dark heartwood is normal in maturetrees) (Figure 1), and incidence in leaveswas very low. Root and soil samplesdid not yield Xanthomonas. Bacteria orpathogenic fungi were not found in thebark and cortex associated with lesionswhereas the stained young xylem beneaththe lesions yielded Xanthomonas (Figure2). Scanning electron microscopy isbeing used to confirm these results.Experiments were initiated in the glasshouseor shade house to determine if pruningwounds and/or scars are possibleentry points for the bacteria. We inoculatedpruned branches (Figure 3) and leafscars (Figure 4) on young trees with suspensionsof bacteria. Also, attempts aremade to introduce the bacteria into thetrees by injection (Figure 5) to simulateentry via an insect vector or wound, andto produce symptoms similar to those observedin the field. Results from these experimentswill also indicate how fast thebacteria move in the trees.In the coming months, roots and flowerswill be inoculated with the Xanthomonasbacteria to determine if those are alsopossible entry points.SUMMARYWe have determined where Xanthomonastranslucens is located in the trees.Results from the ongoing experiments willclarify how the bacteria got into the trees.Figure 5. Young potted tree being inoculated withan injection of bacterial suspensionMore research is needed to determine theways of dispersion of the bacteria in theorchard and to elucidate the cycle of thedisease.Other experiments will be set up to studypossible control measures.E. Facelli 1 , C. Taylor 2 , E. Scott 1 , R. Emmett 2and M. Sedgley 11School of Agriculture and Wine, Universityof Adelaide, Waite Campus, South Australia,Australia. 2 Department of Primary Industries,Mildura, Victoria Australia.E-mail: evellina.facelli@adelaide.edu.auCHESTNUT PRODUCTIONIN ANDALUSIAINTRODUCTIONLast data about Spanish chestnut productionestimates 18.259 t in Spain, half ofthe amount achieved in the seventies.That production corresponds to approximately50.000 ha out of the 128.537 haexisting in Spain. Though chestnut productionis located mainly in North Spain,an interesting area for chestnut productionis localised in Andalusia, South Spain(Pereira et al., 2001). Production in Andalusiais considered to be only 2.000 t butit could be underestimated according tomarketing data and it could reach 6.000 tin 6.000 ha. There are two main areas: i)Sierra de Huelva in Huelva province (Figure1), and ii) Serranía de Ronda (Figure2) and Sierra de Las Nieves, Málagaprovince. Other chestnut areas are locatedin Granada, where new orchards areFAO-CIHEAM - Nucis-Newsletter, Number 12 September 200433

Table 1. Climatic characteristics of chestnut areas in Asturias (www.inm.es), El Bierzo (www.inm.es), Extremadura (www.inm.es),Tenerife (www.inm.es), Andalusia (www.inm.es) and Galicia (Carballeira, 1983).Region Province Location Annual average Annual rainfall Average temperature Annual temperaturetemperature (°C) (mm) in January (°C) range (°C)Asturias Asturias Gijón 13,8 971 4,7 —Oviedo-El Cristo 12,9 973 4,2 —Aeropuerto Ranón 13,2 1140 5,4 —Bierzo León Ponferrada 13,0 675 1,1 17,5Extremadura Cáceres Guadalupe 14,7 755 3,3 17,6Canarias Tenerife Aeropuerto TenerifeNorte-Los Rodeos 16,5 557 12,8 —Andalucía Málaga Gaucín 15,0 1161 6,2 35,0Huelva Almonaster La Real 16,0 1137 4,9 34,9Aracena 14,0 1107 3,1 35,8Galicia Lugo Outeiro de Rei 12,0 1004 2,0 23,3Vilar do Caurel 7,9 1679 -1,3 24,5Ferreira 3,2 1086 2,4 25,7Fonsagrada 8,8 1754 -1,0 23,3Ourense Mesón de Pentes 10,6 1460 0,8 26,0Poboa de Trives 8,9 874 0,1 26,6A Rúa-Petín 15,6 763 5,1 26,7Table 2. Main characteristics of the cultivars of Andalusia (adapted from Pereira and Ramos, 2003).Cultivar Origin Harvesting time Use Male Nuts/kg Lateral Poly-embrionic Splitcatkin z nut (g) nuts (%) nuts (%)Capilla Málaga 20 - 26 Oct Industry, fresh, pollinizer L 66 17,00 11,00 16,00Comisaria Huelva 20 Oct - 2 Nov Industry, fresh, pollinizer L 80 13,37 6,00 0,50Corriente Málaga 18 Sep - 4 Oct Fresh, pollinizer L 65 18,58 15,50 4,50Dieguina Huelva 20 Oct - 1 Nov Marrón al natural, fresh L and M 87 13,61 8,00 2,50Helechal Huelva 22 - 29 Oct Industry, fresh B and L 80 14,14 7,00 1,50Pelón Huelva 19 Oct - 1 Nov Marmalades and purees, L 113 10,22 3,54 7,81pollinizerPilonga Málaga 17 Sep - 4 Oct Industry, fresh, pollinizer L 63 18,32 7,61 8,67Planta Alájar Huelva 20 - 29 Oct Fresh M 71 16,06 10,50 2,25Rubia Tardía Málaga 25 Oct Industry, fresh, pollinizer L 49 20,76 10,00 2,50Temprana Málaga 15 Sep - 4 Oct Fresh, pollinizer L 61 18,76 11,67 8,53Tomasa Málaga 20 - 29 Oct Fresh, pollinizer L 44 24,86 14,38 23,13Vázquez Huelva 20 Oct - 5 Nov Fresh, industry, pollinizer L 73 16,05 4,00 1,50zL = longuistaminate; M = mesostaminate; B = braquistaminateFigure 1. High density spacing in new orchardsof Serranía de Ronda, MálagaFigure 2. New orchard with irrigation in Güimar-Sierra, GranadaFigure 3. Old chestnut orchard in Sierra deHuelva, Huelvabeing established (Figure 3) though traditionallychestnut production came fromseedlings. Chestnut is an interesting alternativein Andalusia, mainly in Málaga,because of the precocity of two cultivars,‘Temprana’ and ‘Pilonga’, harvested atthe end of September. Most of the productionis commercialised by cooperativeswhere big quantities of chestnuts arecleaned and classified. Chestnuts aresold in Spain and in the international market.Recently, an important industry fortransforming chestnuts has been establishedin Huelva. Both in Huelva and Málaga,production is based on local cultivars.Growers graft them over seedlings producedin the same area. In Málaga, due tothe high prices of the nuts, trees are plantedso close as 5 m x 5 m, though it is necessaryto prune severely or remove partof the trees later. In Granada, no localcultivars have been found and new orchardsare based on the excellent cultivarsof Málaga (Figure 3). Weather conditionsfor chestnut areas in Spain are quitesimilar (Table 1), with a rainfall around1.000 mm per year and mild temperatu-34 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

Figure 4.- Capilla Figure 5.- Comisaria Figure 6.- CorrienteFigure 7.- Dieguina Figure 8.- Helechal Figure 9.- PelónFigure 10.- Pilonga Figure 11.- Planta Alájar Figure 12.- Rubia TardíaFigure 13.- Temprana Figure 14.- Tomasa Figure 15.- Vázquezres. Chestnut orchards in Tenerife showedlowest rainfall with 700 mm, followedby Castilla-León and Extremadura, around700 mm. In Andalusia, orchards are locatedin the most humid areas of SouthSpain.In 1995 a study was started to localiseand characterise cultivars of South Spain.An inventory was published in 2001 (Pereiraet al., 2001) and the results foundwere published in a guideline book (Pereiraand Ramos, 2003) and in a workwhere cultivars from Andalusia were comparedwith previous studies (Ramos andPereira, 2004). The paper is a summary ofthe main traits of the Andalusian cultivars.CULTIVARSUp to now 12 cultivars have been studiedin Andalusia which are used locally fornut production and propagated by grafting(Figures 4 to 15): ‘Capilla’, ‘Comisaria’,‘Corriente’, ‘Dieguina’, ‘Helechal’, ‘Pelón’,‘Pilonga’, ‘Planta Alájar’ ‘Rubia Tardía’,‘Temprana’, ‘Tomasa’ and ‘Vázquez’.Cultivars from Andalusia are differentfrom cultivars from other areas of Spain(Ramos and Pereira, 2004). In comparisonwith those from Galicia (Pereira andFernández, 1997) with better characteristicsin nut size, but with higher percentageof poly-embryonic and split nuts, andmore intrusions of the inner coat in cotyledonswhat makes it more difficult topeel them. Nuts coming from Andalusiancultivars showed less worms and diseaseswhat could be related with a drier environment.The best cultivar in Andalusia is ‘Pilonga’(Figure 10) due to its early ripening (endof September), big nut size and less percentageof poly-embryonic nuts (Table 2).The worst feature of ‘Pilonga’ is the highpercentage of split nuts, over 8 %. ‘Temprana’(Figure 13), cultivated in the samearea of ‘Pilonga’ is a little worse due to ahigher percentage of poly-embryonic nutsand intrusions of the inner coat in cotyledons.‘Capilla’, ‘Corriente’, ‘Rubia Tardía’FAO-CIHEAM - Nucis-Newsletter, Number 12 September 200435

and ‘Tomasa’, (Figures 4, 6, 12 and 14)are three excellent cultivars of Málaga,but they are harvested later than ‘Pilonga’and ‘Temprana’, at the end of October,when most of the Spanish chestnut productionis concentrated and prices areusually lower. Moreover, ‘Capilla’ and‘Tomasa’ showed the highest percentagesof split nuts and ‘Corriente’ the highestpercentage of poly-embryonic nuts. InHuelva four main cultivars should be promoted,‘Comisaria’, ‘Helechal’, ‘PlantaAlájar’ and ‘Vázquez’ (Figures 5, 8, 11and 15) due to their big nut size and lowpercentage of split nuts. ‘Planta Alájar’showed a higher percentage of poly-embryonicnuts. In comparison with cultivarsof Málaga, all cultivars in Huelva are harvestedat the end of October, main periodfor chestnut harvesting in Spain, whenprices are lower. Most of the cultivarsfrom Andalusia produce pollen, since onlyone accession studied of ‘Helechal’ wassterile. The most frequent type of malecatkin found was longuistaminate andonly ‘Planta Alájar’ and one accession of‘Dieguina’ showed mesostaminate catkins.‘Pelón’ (Figure 9) is a very spreadcultivar in Huelva that produces the smallestnuts in Andalusia what makes it lessinteresting for present market. These cultivarsare being collected and introducedin the Universidad de Santiago de Compostelato establish a core collection.REFERENCESCarballeira, A., 1983. Bioclimatologíade Galicia. Fundación Pedro Barrié dela Maza. ISBN: 84-85728-27.Pereira, S.; Fernandez, J. 1997. Loscultivares autóctonos de castaño (Castaneasativa Mill.) en Galicia. MonografíasI.N.I.A., 99: 1-533. ISBN: 84-7498-461-0Pereira, S.; B. Diaz, B.; Ciordia, M.; Ascasibar,J; Ramos, A.M.; Sau, F. 2001.Spanish chestnut cultivars. HortScience,36 (2): 344-347.Pereira, S.; Ramos, A.M. 2003. Característicasmorfológicas e isoenzimáticasde los cultivares de castaño (Castaneasativa Mill.) de Andalucía. Monografíasdel INIA, 13, 160 pages.Ramos, A.M.; Pereira, S. 2004. Geneticrelationship between Castanea sativaMill. cultivars from North-western toSouth Spain based on morphologicaltraits and isoenzymes. Genetic Resourcesand Crop Evolution (in press).S. Pereira, A.M. RamosUniversidad de Santiago de Compostela,Escola Politécnica Superior, Departament ofProducción Vegetal, Campus de Lugo, 27002Lugo, SpainE-mail: spereira@lugo.usc.esTHE WORLD MACADAMIAINDUSTRY AND RESEARCHACTIVITIESINTRODUCTIONMacadamia is one of the world’s finestnuts. The kernel is consumed as salted orunsalted snack food, as chocolate enrobedconfectionary, and as an ingredientin products such as biscuits or ice-cream.The kernel has a high oil content (70-80%) and most of the oil is mono-unsaturatedlipids with 65.0 and 18.5% oleic andpalmitoleic acid respectively (Cavaletto etal., 1966). Oil content has been shown tobe an important component of quality;kernels with a specific gravity of 1.0 (oilcontent of more than 72%) or less wereidentified as having superior eating androasting qualities and are classed as firstgrade kernels or the commercial standard.Total sugar content of raw M. integrifoliakernel ranges from between 3 to5% (Riperton et al. 1938), the majority ofwhich is sucrose.The Macadamia genus is a member ofthe Proteaceace, which is a characteristicGondwana family (Johnson and Briggs,1983). Of the nine species, seven are endemicto Australia, with the other two occurringon the islands of Indonesia. Onlytwo of these species, M. integrifolia andM. tetraphylla produce edible kernel. Thesespecies occur as a medium sizedevergreen tree in the lowland subtropicalrainforests of eastern Australia. The fruitconsists of a fleshy husk (pericarp) surroundingthe hard shell (testa) and theedible kernel (embryo and cotyledons)(Gross 1995).WORLD PRODUCTIONMacadamias represent less than 2% ofthe world tree nut market. The market ischaracterised by undersupply and pricefluctuations (Hargreaves 2003).AustraliaAustralia is the main macadamia producerwith approximately 40% of the worldproduction or 10.000 t of kernel (Table 1).Despite recent surveys indicating thatabout 1/3 of the industry is new plantings,industry growth that was forecast at 15%yearly has not been reached (Hargreaves2003) due to poor seasons in 2002 and2003. The major growing regions arearound Alstonville (28 o 51’S, 153 o 21’E) inNorthern NSW and Bundaberg (24 o 33’ S,152 o 00’ E) in South Eastern Queenslandwith approximately 45 and 35 % of theAustralian crop produced in these areasrespectively. There is renewed interest toidentify new growing areas in Australia formacadamia production. The major marketsfor Australian production are USA,Japan, Germany and China.USAMacadamias were first commercialized inHawaii and most of the USA productioncontinues to originate from these islands.The Hawaiian industry is characterised bymature orchards with new plantings atmuch smaller levels than for the otherproducing countries (Vidgen 2003). Domesticsales, including duty-free, are amajor market of USA production. Exportsare shipped predominately to Japan.South AfricaOver 45% of the trees in South Africa areless than 5 years of age and there is thepotential for significant growth of this industry(Lee 2003). Production of kernelhas grown by about 30% between 2000and 2003 to approximately 3000 t despitepoor production seasons in 2002 and2003. Half its production is exported toUSA, with Europe being the next majormarket (25%).OthersMacadamias are also produced in Kenya(1500 t), Malawi (1000 t), Brazil (500 t),New Zealand (35 t), Thailand and CostaRica.HORTICULTUREPropagationMacadamias are generally clonally propagated.Seedling rootstocks are commonin Australia using open pollinated seedfrom the cultivar H2. Seedlings from othercultivars have also been used to a smallerextent. Practical experience suggeststhat the roots system is not well developedin cuttings of some varieties in Australia.There is little information to guideTable 1. Macadamia worldwide kernel production (t) (www.inc.treenuts.org)Region 2002 Import Production Domestic Export 2003Carry-overCarry-overAustralia 800 0 9.900 2.000 7.900 800Africa 43 0 6.440 190 6.250 30USA 2.000 7.466 5.500 12.216 770 2.000Latin America 170 0 3.370 300 3.070 170Total 3.013 7.466 25.210 14.706 17.990 3.00036 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

ootstock choice, although recently an extensiverootstock trial of own rooted cutting,seedling and clonal rootstocks using12 cultivars was established in Australia(Hardner and McConchie, 2003). Suitablescion material is selected from the desiredcultivar and girdled to increase itscarbohydrate content. Scions are then eithergrafted or budded onto the rootstock.Initial results show that budding successrate is affected by the rootstock and scioncombination, which is consistent withanecdotal evidence from macadamia propagators.Orchard designOrchards are established at varying densitiesfrom 10mx10m (100 trees per ha) to7mx4m (357 trees/ha). Tree size variesamong cultivars (Hardner et al. 2002) andmay influence planting density. There hasbeen trend in recent years to establish orchardsat higher densities to offset highinitial capital investments at an earlierage. In addition, there has been somepractical experimentation with high densityplanting (5mx2m) (Bell and Bell, 2001)that produced high yields of between 8and 10 t/ha before canopy management,but there is little knowledge of the laterage performance of trees to evaluate thissystem.IrrigationOrchards located in Queensland are generallyirrigated, especially in the Bundabergregion where there is a significantlylower annual rainfall than in NorthernNSW where the majority of orchards arenot irrigated. Average annual rainfall forAlstonville in Northern NSW and Bundabergis 1600 mm and 1140mm respectively.Stephenson et al. (2003) reportedthat water stress at different phenologicalstages including floral development, immaturenut drop and nut maturation hadadverse effect on yield through decreasein nut number and nut size. Adverseeffects of irrigation in Northern NSW havealso been observed. Trochoulias andJohns (1992) found that NIS yield per unittrunk area decreased with irrigation by upto 10% at excessive rates of irrigation.Variable yield, water use and quality responsesbetween cultivars to irrigationhave been reported for macadamiasgrown in Bundaberg (Searle and Lu,2003).NutritionAn extensive account of macadamia nutritionwas undertaken as part of a reviewof macadamia by Nagao and Hirae(1992). Orchards in Australia have beenestablished on a range of soil types thathave differing chemical and physical properties,Stephenson et al. (1986) observedthe soil nutrient levels and fertiliserapplication on three common soil typesused to grow macadamias; red podsolic,krasnozem and sandy red earth, andfound varied nutrient levels and fertiliserapplications. Macronutrients are generallyapplied as granular fertilizers althoughin regions where irrigation is used fertigationmay also be used. Micronutrients areoften recommended as foliar sprays forrapid amendments of nutrient deficiencies.The methods of determining nutrientrequirements throughout the industryvary; however they generally involve oneor a combination of soil nutrient analysis,leaf nutrient analysis, crop removal replacementor crop logging. Foliar nutrientcontent varies with leaf age, leaf position,time of year and recent work by Huett etal. (2001) showed that content of severalleaf parameters including nitrogen andphosphorus increased with decreasedshading and concluded that the currentsampling protocols for determination ofleaf nitrogen and phosphorus levels werenot reliable.Flowering and fruit developmentInitial flowering generally occurs betweenthe 3 rd and 5 th year after planting. Flowersare hermaphroditic and borne on racemesthat contain between 100 to 300 flowerseach (Figure 1), between 6 to 35%of these flowers develop into immaturefruit (Urata 1954) and only 0.3% will remainat maturity (Ito 1980). It is traditionallyaccepted that flowers are located onwood that is at least 2 to 3 years of ageand there appears to be variation betweencultivars. The major pollinator appearsto be the European honeybee, althoughthe Australian native species of TrigonaFigure 1. Flowering raceme in M. integrifoliahas been observed. Flowering in Australianorchards occurs in September, thereare significant differences in timing of floweringbetween cultivars although thereis still high potential for cross-pollinationbetween cultivars (Boyton and Hardner,2002).Abscission of immature fruit occursthroughout fruit development, howeverthe maximum rates are observed from 3to 8 weeks after anthesis (Sakai and Nagao1984). Increases in fruit diameter andfruit fresh weight follow a single sigmoidalpattern in which growth continues until 14to 15 weeks after anthesis and 15 to 16weeks after anthesis for fruit diameterand fresh weight respectively. Truemanand Turnbull (1994) found that shell hardeningoccurred between 12 and 15weeks after anthesis and effectively ceasedincreases in fruit diameter althoughtotal fruit dry weight per branch increaseduntil 20 weeks post anthesis where monitoringceased. Increases in dry weight aftershell hardening are due to increasesin kernel mass, the majority of which is oilaccumulation (McConchie, et al. 1996). InAustralia, mature fruit drop begins fromMarch to April and can extend until Octoberfor some cultivars.HarvestingIn most Australian orchards, nut-in-huskis mechanically harvested from theground (Figure 2), although hand harvestingmay be used in excessively wet conditionsand in very steep areas of the orchard.Nuts are harvested every 4-6weeks to prevent deterioration of kernelon the ground (Mason and Wells, 1993).Tree shaking and Ethephon have beenused both experimentally and commerciallyto promote nut drop and so reducethe number of harvests, however theyhave not been widely adopted to date.Ethephon has been shown to be veryeffective but the response of leaf and nutdrop can vary with cultivar, time and methodof application and growing region.There are also concerns about the effectof Ethephon application on return andlong-term yields as variable responseshave been reported (Trueman, 2003,Penter et al., 2003).Figure 2. Mechanical harvesting of fallenmacadamia nuts in AustraliaFAO-CIHEAM - Nucis-Newsletter, Number 12 September 200437

Pests and DiseasesThe two major insect pests in Australiaare the larval macadamia nut-borer(Cryptophlebia viridula) and the fruit-spottingbug (Amblypelta spp.) (Huwer andMaddox, 2003). Nut-borer can cause prematurefruit drop or damage to the kernel,damage can be caused throughout nutdevelopment (Figure 3). Fruit spottingbug damages the kernel through surfacepits and also causes premature nut drop,the majority of damage being done beforeshell hardening occurs. Research on IPMstrategies includes the use of a nut-borerparasite, improved monitoring, trap cropping,and the use of IPM compatible pesticides(Huwer and Maddox, 2003). Furtherresearch is also needed to quantifythe impact of pest damage on the industry.Husk spot (pseudocercospera) is the majorpathogen in Australia. Considerableanecdotal evidence suggests that this diseaseinfects the husk of the developingfruit and leads to premature fruit drop.Currently the major defence againsthusk-spot is prescription application ofchemical as infection occurs long beforethe symptoms are apparent. However,considerably more work is required to documentthe lifecycle of the fungus, developrobust chemical and genetic defencesand estimate the economic impact of thedisease.Figure 3. Fruit damage due to nut-borer (courtesyof Gus Cambell NSW Agriculture)Canopy managementMacadamia is a vigorous tree and thereare concerns about balance between vegetativeand reproductive growth andshading on yield. Canopies are very dense,LAI measured in 11 year old macadamiacultivar ‘Haes’ 344 was between 14and 16, which are among the highest reportedfor fruit and nut crops (Meyers etal., 1999). Generally trees are trained intoa single vertical leader with laterals to carrythe crop in the first 3 years after plantingto maintain an open canopy (O’Hare etal., 1996). Experience suggests that narrowcrotch angles are susceptible tobreakage. New trials are being developedfor high-density plantings with intensivetree training and external tree supportsuch as trellises (Bell and Bell, 2001),trellis systems that promote irradiance tofruit are used for crops such as apples,however in macadamias yield has beenshown to increase with canopy area andlight interception, so open systems suchas this will need to be tested and comparedwith the economics of the system.Long term assessment of mature orchardshave found that, although macadamiaspossess one of the most densetree-crop canopies, there seems smalldecline in yield due to crowding (McFadyenet al., 2003) but there was largebetween year variation. However, hedgingof later age trees to maintain may reducethe risk of crop losses due to pestand disease.Post harvestThe husk is removed on farm as soon aspossible after harvesting, as heat of respirationfrom the husk can be large. Themoisture content of wet nut-in-shell harvestedfrom the orchard may range from10 to 25% depending on environmentalconditions before harvesting (Cavaletto1996) but is generally dried to below 10%moisture content on farm where kernel isconsidered safe from mould development.However, kernel remains susceptibleto both hydrolytic and oxidative rancidityat this level of moisture content.The nut-in-shell is transported to processingcompanies where it is slowly dried toapproximately 3.5% (1-1.5% kernel moisturecontent), cracked and then sorted toremove reject kernel (immature, discoloured,germinating, insect damaged ormould) and separate kernel into differentstyles defined by percentage of wholekernel and kernel size. Kernel is thenpackaged into foil vacuum-sealed bagsand stored in cool stores to reduce thedevelopment of oxidative rancidity. Currentlypost-harvest research in Australiafocuses on development of in-line sortingtechnology for reject kernel, control of afterroast-darkening and identification oftraits linked to consumer preference.Genetic improvementMost cultivars grown for production weredeveloped in Hawaii. Breeding and selectionprogrammes have also been undertakenin Australia, South Africa, California,Brazil and New Zealand. To date thereare approximately 700 names representingabout 500 genotypes (Hardnerand McConchie, 2003). However, there issome evidence that cultivar performanceis not stable across countries.Australian industry is currently investingheavily in macadamia genetic improvement.The major focus of this programmeis the development of improved cultivarsfor the Australian industry however theprogramme also encompasses conservationof wild macadamia germplasm, selectionof elite rootstocks and scion bree-ding. The native origin of macadamias givesthe Australian industry a significantadvantage over other countries. Threeex-situ conservations plantations havebeen established in eastern Australia withcuttings collected from over 300 wildtrees across the natural range of M. integrifoliaand M. tetraphylla. The industryhas also supported in-situ recovery of severaldegraded wild populations.The Australian macadamia breeding programmeaims to efficiently produce newcultivars for the Australian macadamia industryusing a structured quantitative geneticapproach. Breeding objectives aredeveloped with the industry and throughwhole of chain economic modelling toquantitatively assess the impact on profitabilityof changes in different traits. Thecurrent traits used in selection are treesize, cumulative yield, commercial kernelrecovery, proportion of whole kernel andkernel size. A breeding population of5.000 individuals has been establishedon 16 sites over 3 major growing regions(Bundaberg, Gympie and NNSW). Thefirst selection from these trials of candidatescultivars will be made in 2004 with releaseto the industry expected from 2012.ACKNOWLEDGEMENTSWe wish to thank the macadamia industriesof the different countries discussed,for their continued support of the researchprogrammes that have developed toinformation included in this review.REFERENCESBell D.J.D. and H.F.D. Bell. 2001. Experienceswith intensively managed highdensityorchards in macadamia. AustralianMacadamia Society News Bulletin28 (5):61-66.Boyton S. J. and C. M. Hardner. 2002.Phenology of flowering and nut productionin macadamia. Acta Horticulturae575: 381-387.Cavaletto C. G., Dela Cruz A., Ross E.and H. Y. Yamamoto. 1966. Factorsaffecting macadamia nut stability. I.Raw kernels. Food Technology 20(8):108-111.Gross C.L. 1995. Macadamia. Flora ofAustralia 16:419-425.Hardner C. M., and C. A. McConchie.2003. The future of macadamia geneticimprovement in Australia. In: Proceedingsof the 2 nd International MacadamiaSymposium 2003. Tweed Heads.Australian Macadamia Society, LismoreAustralia, pp. 85-89.Hardner C., Winks C., Stephenson R.,Gallagher E. and C. McConchie. 2002.Genetic parameters for yield traits inmacadamia. Euphytica 225: 255-264.38 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

Hargreaves G. 2003. World round-up –Australia. In: Proceedings of the 2 nd InternationalMacadamia Symposium2003. Tweed Heads. Australian MacadamiaSociety, Lismore Australia, pp. 6.Huett D. O., Gogel B. J., Meyers N. M.,McConchie C. A., McFadyen L. M. andS. C. Morris. 2001. Leaf nitrogen andphosphorus levels in macadamia inresponse to canopy position and lightexposure, their potential as leaf-basedshading indicators, and implications fordiagnostic leaf sampling protocols.Australian Journal of Agricultural Research52: 513-522.Huwer R. and C. Maddox. 2003. Integratedpest management in macadamias– opportunities and challenges.In: Proceedings of the 2 nd InternationalMacadamia Symposium 2003. TweedHeads. Australian Macadamia Society,Lismore Australia, pp. 106-110.Ito P J. 1980. Effect of style removal onfruit set in macadamia. Hort Science15: 520-521.Johnson L. A. S. and B. G. Briggs.1983. Proteales. IN: B. D. Morley, B. D.and H. R. Toelken (Eds.). FloweringPlants of Australia. Rigby, Adelaide.Pp. 238-244.Lee, P. 2003. World round-up – SouthAfrica (and Africa). IN: Proceedings ofthe 2 nd International Macadamia Symposium2003. Tweed Heads. AustralianMacadamia Society, Lismore Australia,pp. 7-11.Mason R. L. and I. A. Wells. 1983. Theeffect of harvest interval on the qualityof ground harvested macadamia nuts.Food technology in Australia 36: 373-378.McConchie C. A., Meyers N. M., AndersonK., Vivian-Smith A., O’Brien S. andS. Richards. 1996. Development andmaturation of macadamia nuts in Australia.IN: R. A. Stephenson and C. W.Winks (Eds). Challenges for Horticulturein the Tropics, Proceedings of theThird Australian Society of HorticulturalScience and the First Australian MacadamiaSociety Research Conference.18-22 August 1996 Pan Pacific HotelBroadbeach, Gold Coast, Australia, pp.234-238.McFadyen L., Morris S., Oldman M.,McConchie C., Huett D., Meyers N. andJ. Wood. 2003. Canopy management inmacadamia. In: Proceedings of the 2 ndInternational Macadamia Symposium2003. Tweed Heads. Australian MacadamiaSociety, Lismore Australia, pp.139-142.Meyers N. M., McFadyen L. M., HuettD. and C. A. McConchie. 1999. A comparisonof direct and indirect estimatesof leaf area index of Macadamia integrifolia(Maiden and Betcshe). IN: C. A.McConchie, L. M. McFadyen, N.M. Meyersand D. Huett (Eds.). Canopy managementfor consistent yield in macadamia.Final Report MC 95008, HorticulturalResearch and DevelopmentCorporation, Gordon, Australia.Nagao M. A. and H. H. Hirae. 1992. Macadamia:cultivation and physiology.Critical Reviews in Plant Science. 10:441-470.O’Hare P., Loebel R. and I. Skinner.1996. Growing macadamias. QueenslandHorticultural Institute, Departmentof Primary Industries, Queensland,Brisbane.Penter M., Snijder B. and F. Kruger.2003. The use of ethephon to facilitateharvesting in the South African macadamiaindustry. In: Proceedings of the2 nd International Macadamia Symposium2003. Tweed Heads. AustralianMacadamia Society, Lismore Australia,pp. 133-139.Ripperton J. C., Moltzau R. H. and D.W. Edwards. 1938. Methods of evaluatingthe macadamia nut for commercialuse and the variation occurring amongseedling plantings in Hawaii. HawaiiAgricultural Experimental Station bulletin79.Sakai W. S. and M. A. Nagao. 1985.Fruit growth and abscission in Macadamiaintegrifolia. Physiologia Plantarum64: 455-460.Searle C. and P. Lu. 2003. Whole-treewater use and irrigation scheduling inmacadamia. In: Proceedings of the 2 ndInternational Macadamia Symposium2003. Tweed Heads. Australian MacadamiaSociety, Lismore Australia, pp.125-130.Stephenson R A., Cull B.W., Price G.and J. Stock. 1986. Scientia Horticulturae.30: 83-95.Stephenson R. A., Gallagher E. C. andV. J. Doogan. 2003. Macadamia responsesto mild water stress at differentphenological stages. Australian Journalof Experimental Agriculture 54: 67-75.Trochoulias T. and G. G. Johns. 1992.Poor response of macadamia (Macadamiaintegrifolia Maiden and Betche)to irrigation in a high rainfall area ofsubtropical Australia. Australian Journalof Experimental Agriculture 32:507-512.Trueman S. J. 2003. Yield responses toethephon for unshaken and mechanicallyshaken macadamia. Australian Journalof Experimental Agriculture 43:1143-1150.Trueman S. J. and C. G. N. Turnbull.1994. Fruit set, abscission and dry matteraccumulation on girdled branches ofmacadamia. Annals of Botany 74: 667-674.Urata U. 1954. Pollination requirementsof macadamias. Hawaii agricultural ExperimentalStation Technical Bulletin22: 5-40.Vidgen, R. 2003. World round-up –Hawaii. In: Proceedings of the 2 nd InternationalMacadamia Symposium 2003.Tweed Heads. Australian MacadamiaSociety, Lismore Australia, pp. 11-12.C. Hardner, J. Wilkie and C. McConchieCSIRO Plant IndustryQueensland Bioscience Precinct306 Carmody RoadSt Lucia, 4067, AustraliaE-mail: craig.hardner@csiro.auCAROB TREE DEVELOPMENTIN CHINAINTRODUCTIONChina is the world’s largest producer ofrice, wheat, cotton, peanuts and rapeseed.It ranks second in corn and fourthin soybean production. Subtropical fruitssuch as citrus, litchi and loquat grow togetherwith apples, peaches, apricotsand pears of cool temperate climates.The recent interest for the carob tree isframed in the possibility of growing asagroforestry species and for the restorationof degraded areas, mainly, of threeSouthern Chinese provinces with subtropicalclimates: Sichuan (Yibasan andTongAn), Yunnan (Lufeng and YuanJiang) and Guangxi (Shangsi and PingGu)(Fig.1).Figure 1. -Areas where the carob tree has beenintroduced in China •The project “Introducing of nursery andcultivation techniques of Ceratonia siliquain China” is funded by the Chinese government.One of the aims of this project isto find a tree species suitable to dry areasof southwest of China that will contributeto the ecological rehabilitation. The projectwas started in 2001 and will be completedby 2005. During the year 2002 theChinese Academy of Forestry (CAF) contactedIRTA-Mas Bové (Catalonia, Spain)to start a possible collaboration on thisspecies between both institutions. Duringtwo years 2002 and 2004, IRTA sent toCAF more than 100.000 seeds, from importantcarob cultivars: five of Spain (‘Negra’,‘Duraió’, ‘Rojal’, ‘Banya de Cabra’and ‘Matalafera’) and two of Portugal(‘Mulata’ and ‘Galhosa’), with the purposeof introducing them in the nursery and, laterbe planted in the field (Fig 2). In thenew plantations two methods of plantingFAO-CIHEAM - Nucis-Newsletter, Number 12 September 200439

Figure 2.- New carob tree planting in Yibasancounty (Sichuan province)Figure 3.- Research group that visits the carobseed plantations in several areas of the Xichangcounty (Sichuan province)Figure 4.- Carob tree reforestation in degradedareas of Sichuan provincewere used: direct sowing in the field andsmall plants with containers. The last methodwould be the most suitable and to beused in the future.The necessity to know the possibilities ofdevelopment of the carob tree in China,as an answer to the new expectations generatedby this species, fostered the realizationof this technical collaborationbetween China and Spain (Fig. 3).ECOLOGYThe edaphoclimatic conditions of the Chineseareas where the carob has been introduced,Sichuan, Yunnan and Guangxiprovinces placed in the south, are differentfrom those of the Mediterranean Basinto which carob is well adapted. It highlightsthe great climatic variability, with altitudesof 800-1000 m above sea level,minimum temperatures of 0 or -1ºC andmaximum of 40ºC, although there is a biggap of temperature between day and night(about 10 ºC), and rainfall of 700-800 mmin the county of Xichang (Sichuan) and,higher in the other two provinces (Yunnanand Guangxi), about 1000 mm, distributedbetween June and September. Thecarob was planted on hilly lands and thesoils in overall are not very deep, clayloam,grey-brown colours, with neutral orslightly acid pH (6-7) and low levels ofphosphorus and calcium (Fig. 4).In the Mediterranean countries (Spain,Italy, Portugal, Greece, Morocco, Tunisia,etc.), the carob grows well in warm temperatureand subtropical areas, with altitudesbelow 600 m above sea level, lowrainfall in summer (300-500 mm/year),and tolerates hot and humid coastal zones.It can only withstand light frost; temperaturesbelow –4ºC can damage youngtrees and flowers of mature trees. Carobcan adapt to a wide range of soil typesfrom poor sandy soils and rocky hillsidesto deep soils, but they cannot withstandwaterlogging. In the Mediterranean Basincarob generally grows in marginal, calcareousand basic soils.CROP DEVELOPMENT RESTRICTIONSThe areas where the carob tree has beenintroduced in China, in the case of the Sichuanprovince, the following remarkscan be highlighted:The areas where the carob seeds havebeen planted are hilly, long away from thecoast, with high altitudes above sea level,high rainfall (800-1000 mm and concentratedon three summer months) and withvery poor soils, neutral or slightly acidpH, and these conditions are quite differentfrom the Mediterranean countries.Temperatures of the Yibasan and TongAnareas are warm and more similar to caroboriginal areas of Mediterranean Basin.Pests: Severe damage is caused inyoung trees by rabbits and some nativeants that gnaw trunks and new shoots.Diseases: High rainfall, concentrated onsummer months, can cause waterloggingproblems in young carobs and later promotefungus attacks of Phytophtora, inthe neck of the tree, and Oidium in theleaves of the most sensitive carob seedtrees.Pollination: the carob is a trioecious specieswith male, female and hermaphroditeflowers borne on different trees. It bloomsin summer-autumn and it seems to bemainly pollinated by insects, but also bywind. High rainfall and relative humidity inthe rainy summer season, could increasepoor fruit set during the pollination time(mainly the period that overlaps withblooming time).Mechanical harvesting: currently this factoris not very important because there islarge manpower and its cost is low.RESEARCH AND DEVELOPMENTNEEDSResearch and development carob worksin the Chinese Academy of Forestry isvery limited, outstanding some nurseryworks (seed germination, planting methods,etc.). Future work would have toassess the performance of the seedlingsplanted in the three Chinese provinces(about 100.000 seedlings) that adapt betterto the new environmental conditions,and select seedlings from more vigorouscultivars, female and hermaphrodite sexes,tolerant to diseases (Oidium), earlybearing and fruits with good quality. Laterit would be necessary to bud the besttrees on seedlings and to introduce themin collections and comparative trials to finishtheir agronomic and commercialevaluation. These works should be carriedout within the new agroforestry systemsframework in which the carob couldsolve reforestation problems in degradedareas and its bean production used bothfor human and animal feeding (sheep,goats, pigs, cows, horses, etc.). A climaticand soil characterization of the possiblelocations for this specie would also beinteresting, with the purpose of beingable to define its potential expansionarea in China, avoiding future problemsof lack of crop adaptation.CONCLUDING REMARKSThere are some favourable aspects onthe development of the carob cultivationin China, large surfaces, large manpowerfor harvesting, potential agroforestryuses, being able to use their productionboth for human and animal feeding. Onthe other hand, it is necessary to choosewell the growing areas, which should bewindy, with little risk of frost, and the cultivarsto be recommended should adaptwell to Chinese growing conditions becausethey are different to those of theiroriginal areas (Spain, Italy, Portugal,etc.). Based on the previously described,the carob tree in China it can be consideras an alternative crop in dry land areaswith warm or subtropical climates for diversificationon the agroforestry farms.ACKNOWLEDGEMENTSThese results emerged from a technicalvisit to China by the first author in December2003, which was supported by theChinese Academy of Forestry, Beijing(China).J. Tous 1 , and J. Chunqian 21Institut de Recerca i Tecnologia Agroalimentàries(IRTA), Dept. d’Arboricultura Mediterrània,Centre Mas Bové. Apartat 415. 43280Reus, Spain.2International Farm Forestry Training Center.Chinese Academy of Forestry, Beijing 100091,China.E-mail: joan.tous@irta.es40 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

NOTES AND NEWSIN MEMORIAM:MY PROFESSOR DALE KESTERA few weeks before Dale’s death the 21 stof November 2003 in Davis, California, Iwas talking to a group of technicians ofseveral growers’ associations and I startedmy talk recognising that I had beenfortunate in my professional life, and oneof the reasons was that I had been a studentwith Dale Kester. His influence hasbeen extremely positive for my work,even if we did not always agree on somesubjects, but his personal view of almondswas in any case a clear exampleof dedication and interest. As most almondresearchers, he had a passionatefor this species, as well as for his students,and was, as a consequence, ableto convey his passion for almond growingto us.It is now quite difficult to find somebodywith this ability to give such a generalperspective of almond. Dale, as well asthe other great figures of almond researchin the past century, Charles Grassellyand Antonio Felipe, was an expert bothon rootstocks and varieties, and this combinedview of the tree complexity wasalso the way to obtain a complex overviewof all problems related to almondgrowing. Until recently, when we probablyfind more articles on almond molecularmarkers than on the almond itself, it washardly impossible to find any article on almondswithout his name cited several timesin the bibliography list. As a matterof fact, his most cited article (Kester andGriggs, 1959. Fruit setting in almond: theeffect of cross-pollinating various percentagesof flowers. Proc. Amer. Soc. Hort.Sci. 74: 214-219) was one of the pointsabout which we sometimes had disagreed.Just after my arrival in Davis in December1972, he suggested four points upon whichI could start my research work. I only rememberone on leafing time and frost resistance,later taken by MustafaBüyükyilmaz, and the one I selected, onalmond self-compatibility. Once I hadmade my choice, he asked me if I couldread Portuguese, because he had an articlein that language he was unable toread and that could be important on almondself-compatibility. This was the pioneerwork of Almeida (1945), so often referredlater after my review. This hasbeen definitive for my future work in almond,because I continue in the samefield of work.Professor Dale Kester at California, USA,in August 1997Although after my stay in California weonly met on a few occasions, we continuedto talk on the same subject. I remembera trip to Davis in 1985 where heshowed me some pollination results difficultto explain according to the S allelerelationships of the different cultivars. Wecommented several possibilities and finallyI suggested a mutation from an Sallele to a nul allele. This hypothesis waslater confirmed (Kester et al., 1994. Amutation of ‘Nonpareil’ almond conferringunilateral incompatibility. J. Amer. Soc.Hort. Sci. 119: 1289-1292).During my stay there were other studentsworking under his guidance. They werethe mentioned Mustafa Büyükyilmaz fromTurkey, Juan Negueroles from Spain andMustafa Lasram from Tunisia. Also RachidHellali from Tunisia was partly workingwith him. They are all now recognisedprofessionals in fruit growing, research,education and administration, and Iguess that I am the only one who continuesto work in almond, but I do not regretit. On the contrary, it makes me feelthat probably I could boast of being morehis heir than the others.I cannot talk about Dale without mentioninghis wife Daphne. They were alwaystogether and she was a real support forhim. They opened their home to me bothwhen I was a student, as on Thanksgivingday, and when I visited Davis later. Bothwere friendly and made other people feelat ease with them. They visited Spain severaltimes, and very much enjoyed Spanishcuisine. I remember that on their secondvisit to Spain, they mentioned thatthey had attended meetings of the WeightWatchers in order to reduce weight asthey knew that in Spain they would eatvery well. They were delighted with the croquettesof Pili Royo, Antonio Felipe’s wife.His work always had the support of anothergood man that I would also like tobring to mind now, his technician DickAsay. They formed a real team and Dalerecognised it when Dick appeared so oftenin the scientific publications. This isanother example of the traits defining areal researcher: humble, adaptable andopen. He was humble, conscious of howmuch he did not know. Adaptable, in hisplace both in a scientific meeting and in agrowers’ talk. Open, willing to share hisknowledge.I saw him for the last time during the InternationalHorticultural Congress in Torontoin August 2002, and now I wouldhave preferred not to have seen him becausehe was not the same Dale. He hadsuffered Daphne’s loss and, although interestedas always in his work and all theproblems related to almond bud failure,he seemed a little lost to me. However,now I find him more clearly than ever inour daily work.R. Socias i CompanyCONGRESSESAND MEETINGSXIII GREMPA MEETINGON PISTACHIOSAND ALMONDSThe XIII GREMPA Meeting on Pistachiosand Almonds took place in Portugal inMirandela, the Garden-City of Terra-Quente (Hot land), from 1 to 5 June2003. Mirandela, is located in the Northeastof Portugal (Trás-os-Montes region).It has a Roman origin, but it was King D.Dinis who, in the XIV century, named thecity as Mirandela and ordered the constructionof a fortress wall surrounding it,part of which is still visible. The city stillhas other historical-architectonic structures,like the Roman bridge (“Ponte Velha”)with 20 asymmetrical arches, the ancientpalace of the Távoras (the noble familyaccused of attempting regicide in1759), the church of “Misericórdia” andseveral pillories of medieval origin,among other interesting features.The meeting was organized by the DirecçãoRegional de Agricultura de Trás-os-Montes (DRATM), the Instituto de BiologiaExperimental e Tecnológica (IBET)FAO-CIHEAM - Nucis-Newsletter, Number 12 September 200441

ging and dancing were also included andwhere the natural abilities of the participantscould be demonstrated.Because of their contribution to this meetingspecial thanks are due to FAO-CI-HEAM, the Director of the Regional AgriculturalServices of Trás-os-Montes andIBET, for their encouragement and supportduring all the organization. Also thePresidents of Câmara Municipal de Mirandelaand Alfândega da Fé, as well asAmendouro Company, Mr. António Martinsand Mss. Maria de Lurdes Lopes aremuch acknowledged for their help andgranted availability.Participants of the XIII Grempa Meeting on Pistachios and Almonds held at Mirandela, Portugaland the Universidade de Trás-os-Montese Alto Douro (UTAD), with the collaborationof the Inter-Regional FAO-CIHEAMCooperative Research Network on Nuts,and having Vitor Cordeiro and M. MargaridaOliveira as the conveners.The meeting was attended by more than80 participants from 14 countries: Algeria(1), Argentina (1), Australia (10), Belgium(1), France (3), Greece (1), Iran (5), Italy(8), Morocco (1), Portugal (22), Spain(17), Tunisia (3), Turkey (3) and USA (6).A Chinese delegation with 7 participantscould not attend the meeting because ofthe severe acute respiratory syndrome(SARS), although China was representedwith two abstracts. Also one participantfrom Macedonia and two other participantsfrom Algeria did not succeed to bepresent but managed to have their posterspresented.The participants represented several sectorsof activity linked to almond and pistachio,including farmers, industrials andresearchers. Important scientific advanceshave been reported and the meetingwas a good opportunity to exchangeknowledge and strategies among teamsfrom all around the world where almondand pistachio are grown. Besides thescientific sessions, numerous opportunitiesfor discussion and establishment ofnew collaborations were possible duringcoffee-breaks, meals or social events.In total, 75 communications were presented(39 oral communications and 36 posters),unequally distributed by the differentsessions, which included:“Cultivars, Rootstocks and Breeding” (10oral communications and 12 posters),“Flowering, Pollination and Fruit Set” (5orals, 5 posters), “Pests and Diseases” (3orals, 3 posters), “Harvesting, Marketingand Industry” (2 orals, 2 posters), “OrchardManagement” (4 orals, 4 posters)and finally “Physiology, Biology and Biotechnology”(15 orals, 10 posters). All theabstracts, including those of the participantsthat finally did not attend the meeting(in total 50 orals and 40 posters weresubmitted) were included in the Book ofAbstracts and distributed by the participants.Before closing the ceremony, aRound table was organized for discussionof common problems and strategies aimingto solve them. The proceedings ofthis meeting will be edited in a volume ofCahiers Options Méditerranéenes.In the first working day, the participantswere transported to the city Hall Auditoriumby the touristic train of Mirandela. Inthe afternoon of the second working day,a visit was organized to an almond privatecompany “Amendouro”, and to the villageof Moncorvo where the secular churchwas visited, as well as a traditional storewhere almonds are processed in a secretway!!! The day ended with a prosperousmeal offered by the Town-hall Presidentof Alfândega da Fé.On Wednesday, a full day was dedicatedto a long tour. The day started with a visitto almond orchards (at the ExperimentalCentre of Terra Quente), followed by a visitto old wine presses in a Douro farm(Quinta da Pacheca) and a travel by boat,up the Douro river, until Pinhão.On the last day, after the sessions andclosing ceremony, a farewell eveningbanquet organized at “Estalagem da Senhoradas Neves” gathered most of theparticipants in a happy event where sin-Finally, we would like to express ourthanks to the Scientific and OrganizingCommittee of the Meeting and to all participantsand contributors of the large almondand pistachio family. We hope thismeeting may have contributed for the exchangeof scientific knowledge and tostrengthen the collaboration of the participantswithin the network or in connectionwith it.M. Margarida OliveiraVitor CordeiroVI INTERNATIONALCONGRESS ON HAZELNUTThe 6 th International Congress on Hazelnutwas held in Tarragona, North-East ofSpain, on June 14-18, 2004. The Congressoffered an international forum tomeet each other and to exchange experiencesrelated to the latest results concerningagronomic, industrial, health benefitsand commercial aspects of hazelnutgrowing. The Congress was organizedby the Institut de Recerca i TecnologiaAgroalimentàries (IRTA) – CentreMas Bové in collaboration with the InternationalSociety for Horticultural Science(ISHS) and the FAO-CIHEAM Nut Network.This event was financially supportedby several institutions: Diputació deTarragona, Ministerio de Ciencia y Tecnología,INIA, European Union - projectPORTA, Generalitat de Catalunya –Agència de Gestió d’Ajuts Universitaris ide Recerca (AGAUR), MediterraneanAgronomic Institute of Zaragoza (IAMZ -CIHEAM), FAO-CIHEAM Nut Network,Caixa Tarragona, Town councils of Tarragona,Reus and Constantí; and the privatesponsors: Borges, Morella Nuts, IndústriesGarriga, Denomination of Origin“Avellana de Reus”, Unió Agrària Cooperativaand Coselva.42 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

Participants of the VI International Congress on Hazelnut held at the Palace of the Diputació of Tarragona, SpainSeveral authorities were represented inthe opening session: the Councillor ofAgriculture (Mr. Antoni Siurana) of theGeneralitat of Catalonia, the President ofthe Diputació of Tarragona (Mr. Joan Aregio),the Director of IRTA Dr. Josep Tarragó,the Chairman of the Nuts Sectionand Mediterranean Climate Fruits of TheInternational Society of HorticulturalScience (ISHS) Prof. Carlo Fideghelli, therepresentative of The Centre Internationalde Hautes Etudes Agronomiques Méditerranéennes(CIHEAM), Dr. Dunixi Gabiña,and other local authorities.The meeting was opened with a welcomingof the convener, Dr. Joan Tous, whoreferred to the main economic characteristicsof the Spanish hazelnut sector,with an average production of 9.000 t ofkernel per year and 23.000 ha, 95 % ofthis surface was concentrated in Cataloniaand, mainly in Tarragona province(18.000 ha), where the hazelnut is, in diversedistricts, an important source of income.In Catalonia region, there are7.000 small hazelnut farms with an averageorchard size of about 1 to 5 ha. Inaddition, most of these small farms needto be modernized. In this scenario, one ofthe most important aims is the reductionof the management cost and improve nutquality, to ensure greater returns forgrowers. The hazelnut sector is groupedin six OPA’s (Nut Growers Associations),all of them located in Catalonia. In Spain,hazelnut commercialization is mainly focusedon the chocolate and “snacks” industries.Opening session (sitting from left to right): Dr. J. Tous (Convener), Dr. J. Tarragó (Director of IRTA),Mr. Antoni Siurana (Conseller of Agriculture of the Generalitat of Catalonia), Mr. J. Aregio (Presidentof Diputació of Tarragona), Prof. Carlo Fideghelli (Chairman, ISHS-WG Nuts and Mediterranean Fruits),and Dr. D. Gabiña (CIHEAM)Dr. J. Tous also explained the history ofthe hazelnut Congresses in the world andnoted that the first Colloquium on Hazelnutin Spain was held in Reus in 1862, organisedby the Institut Agrícola of SantIsidre, and that there was discussed tointroduce the hazelnut tree as possiblealternative to grapevine in the coastalareas of Tarragona, due to ‘powdery mildew’fungus problems. Later, in the XXthcentury, in the year 1976, the 1st InternationalHazelnut Congress was held inReus (in this case together with the almond).Since then, different cities all overthe world have hosted the following:Avellino, Italy (1983); Alba, Italy (1992);Ordu, Turkey (1996), Corvallis, USAFAO-CIHEAM - Nucis-Newsletter, Number 12 September 200443

Table 1. Main Scientific conclusions of the VI International Congress on Hazelnut.Tarragona (Spain), 14-18 June, 2004.Session Main aspects Remarks and conclusions(Nr. of contributions)1. Germplasm and Genetic • Behaviour and adaptations of hazelnut cultivars • Current collections should be reviewed withImprovement (30) to new zones molecular markers.• Behaviour of several breeding obtentions• International data base of molecular markers(limited advances until today) is useful.• Molecular markers• Many presentations focussed on table varieties2.- Biology and Physiology (12) • Particular problems vary per country and zone • Phenological data of cultivars differ according(examples). sites and climatic conditions.• Nut fall in Australia• The knowledge of incompatibility (S)• Selection of pollinizers in Chile alleles in hazelnut cultivars is required.• Studies on root distribution in Portugal• Shedding effects on flowering in Italy3.- Propagation • Achievement of hardwood cuttings’ method • Rootstocks could be of great interest to solveand Rootstocks (6) specific problems, such as:• Improvement of agronomic characteristicsof ‘Negret’ cultivar with rootstocks (IRTA, Spain).4.- Orchard Management (24) • Nutrition and pruning • High N fertilizer doses could have a yield• Improvement of harvest facility depressing effect.(machinery, pruning, green covers)• Reference of P in leaves should be revised• Organic production in some areas.• The irrigation should be adaptedto the physiologic phases of the crop.• The Spanish strategy is to develop OrganisedTechnicians Groups (ADV) to help farmerswith new EU policy regulations (IPM)*.5.- Pest and Diseases (16) • Solution for specific problems in specific zones • New technologies of Teledetection (GPS or GIS)• General pest and diseases studies to study wide growing areas and their problems.for widespread areas• “Eastern Filbert Blight” fungus problem,• Increasing problems with traditional pests increases in Oregon hazelnut orchards.(bug species) which affect kernel quality• Possibilities for biological control will increase,• Biological control such as control of “Zeuzera” in Spain.6.- Post Harvest • Chemical composition • More research is needed on food securityand Quality (7) (old as well as new cultivars) (aflatoxin).• Drying to avoid aflatoxin problems, in Turkey7.- Health and nuts (2) • Nut composition and healthy effects • Most marketing efforts use the healthy strategy.• Scientific evidences of disease prevention • The nut kernels improve health and do notby the consumption of nuts seem to increase body weight.8.- Industry, Marketing • Crop and international price prediction • The Turkey policy could reduce surface by 25%.and Economics (10) • Consequences of national policies, • Market for organic production seemsmainly for the main producing country (Turkey) on the increase.• Organic production• High alternate bearing in the cropsof the main producing countries.• Consumer studies can give relevant informationto enhance strategies of producers, industries,distribution and research.• Other alternative nut species (Gevuina avellana,in Chile).* IPM (Integrated Pest Management)44 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

(2000) and finally Tarragona, Spain(2004), twenty eight years after the firstinternational event.Then, Prof. Carlo Fideghelli (Chair of theISHS Section for Nuts and MediterraneanClimate Fruits) gave the “ISHS MedalAward” to the convener to acknowledge hiseffort for the organisation of the Congress.The Congress was attended by 125 participantsfrom 20 countries: Turkey, Italy,USA (Oregón), Spain (Catalonia), France,Portugal, Australia, China, New Zealand,Poland, South America (Chile and Argentina),Ukraine, India, Albania, Austria,Georgia, Switzerland, The Netherlandsand Slovenia. A total amount of 107scientific papers were presented in thisCongress, 43 as oral communicationsand 64 as posters. The topics of this Congresswere related to the following issues:Plant material (30 papers), Biology andPhysiology (12), Propagation and Rootstocks(6), Orchard management (24),Pest and Diseases (16), Post harvest andQuality (7), Health benefits (2) and Industry,Marketing and Economics (10). In Table1 the main scientific conclusions ofthis Congress are summarized.Researchers had the opportunity to visitthe experimental fields of IRTA-Mas BovéResearch Center (collection of hazelnutgermplasm, rootstock and cultivar trialsand seedlings collection of Gevuina avellanaspecie), commercial hazelnut orchards,an exhibition of several harvestmachines and nut industries (Borges andCoselva). An Open day for the sector wasorganised the last journey with two workshops:“World hazelnut situation andperspectives in France, USA, Italy, Turkeyand Spain” and “Production and commercialaspects on hazelnut”. Also, mainscientific conclusions of the Congresswere presented to the sector.After a ‘proposal’ of Italian researchers,the next meeting on the 7 th ISHS InternationalCongress on Hazelnut, was decidedto be held in Viterbo (Italy), probablyin summer 2008.The five days of the congress were afriendly course of excellent scientific presentations,relaxed discussions and allowedto learn new aspects about our regionand local habits. Papers will be publishedin a volume of Acta Horticulturaein the near future.J. Tous (convener) and M. Rovira (secretariat)IRTA-Centre Mas Bové. Apartat 415. 43280Reus (Spain)E-mails: joan.tous@irta.es;merce.rovira@irta.esTO BE HELD:Almond and PistachioIV ISHS International Symposium on Pistachios and AlmondsDate: May 22-26, 2005Place: Tehran, IranConvener: Dr. A. JavanshahAddress: Iranian Pistachio Research InstitutePO Box 77175 / 435Rafsanjan, Islamic Republic of IranTel: 98 391 422 52 02Fax: 98 391 422 52 08E-mail: javanshah@pri.irhttp://www.pri.irChestnutIII ISHS International Symposium on ChestnutDate: October 20-23, 2004Place: Chaves (Vila Real), PortugalConvener: C. AbreuAddress: University of Tras os Montes e Alto DouroP.O. Box 2025000-911 Vila Real – PortugalTel: 351 259 350 508Fax: 351 259 350 480Email: cgabreu@utad.ptwww.utad.pt/eventos/chestnutcongressWalnutV ISHS International Symposium on WalnutDate: November, 7-14, 2004Place: Sorrento, Naples, ItalyConveners: D. Avanzato (MiPAF) and M.E. Malvolti (CNR)Addresses: Istituto Sperimentale per la Frutticoltura di Roma (MiPAF)Via di Fioranello, 52, 00134 Roma (Italy)Tel: 39 06 79348186Fax: 39 06 79340158E-mail: davanzato@mclink.itIstituto di Biología Agro-ambientale e Forestale (CNR)Viale Marconi 2, 05010 Porano, ItalyTel: 390763374688Fax: 390763374330E-mail: mimi@ias.tr.cnr.itAlmond orchard at Almería, SpainBunch of pistachio nutsFAO-CIHEAM - Nucis-Newsletter, Number 12 September 200445

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A Development Based Classificationfor Branch Architecture in Almond.Journal American Pomological Society,56 (2): 106-112.Gradziel, T.M.; Martínez, P., 2002. ShellSeal Breakdown in Almond is Associatedwith the Site of Secondary Ovule Abortion.Journal of the American Society forHorticultural Science, 127 (1): 69-74.Ikeda, T.; Ishikawa, H.; Sasaki, T., 2003.Regulation of Wolbachia density in theMediterranean flour moth, Ephestia kuehniella,and the almond moth, Cadra cautella.Zoological Science, 20 (2): 153-157.Jiang, Y.Q.;Ma, R.C., 2003. Generationand analysis of expressed sequence tagsfrom almond (Prunus dulcis Mill.) pistils.Sexual Plant Reproduction, 16 (4): 197-207.Kazantzis, I.; Nanos, G.D.; Stavroulakis,G.G., 2003. Effect of harvest time andstorage conditions on almond kernel oiland sugar composition. Journal of theScience of Food and Agriculture, 83 (4):354-359.Koukourikou Petridou, M.A., 2003. 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Plant Systematics and Evolution, 238 (1-4): 87-95.Ma, R.C.; Oliveira, M.M. 2002. Evolutionaryanalysis of S-RNase genes from Roseaceaespecies. Mol. Genet. Genomics,267: 71-78.Martinez, P.; López, M.; Alonso, J.M.; Ortega,E.; Batlle, I.; Socias i Company, R.;Dicenta, F.; Dandekar, A.M.; Gradziel,T.M., 2003. Identification of self-incompatibilityalleles in almond and related Prunusspecies using PCR. Genetics andBreeding of Tree Fruits and Nuts. ActaHorticulturae, 622: 397-401.Martínez, P.; Arulsekar, S.; Potter, D.;Gradziel, T.M., 2003. An extended interspecificgene pool available to peach andalmond breeding as characterized usingsimple sequence repeat (SSR) markers.Euphytica, 131 (3): 313-322.Martinez, P.; Arulsekar, S.; Potter, D.; Gradziel,T.M., 2003. Relationships amongpeach, almond, and related species asdetected by simple sequence repeat markers.Journal of the American Society forHorticultural Science, 128 (5): 667-671.Martinez, P.; Gradziel, T.M., 2003.Sexual polyembryony in almond. SexualPlant Reproduction, 16 (3): 135-139.Martinez, P.; Vaknin, Y.; Gradziel, T.M.;Dicenta, F., 2003. Karyotype analysis inalmond. Genetics and Breeding of TreeFruits and Nuts. Acta Horticulturae, 622:457-460.Martins, M.; Sarmento, D.; Batlle, I.; Vargas,F.J.; Oliveira, M.M. 2002. Molecularmarkers linked to tolerance/sensitivity toFusicoccum on almond. XIII CongresoNacional de Bioquímica, Lisboa, 2002.(Poster).Martins, M.; Tenreiro, R.; Oliveira, M.M.,2003. Genetic relatedness of Portuguesealmond cultivars assessed by RAPD andISSR markers. Plant Cell Reports, 22 (1):71-78.MirAli, N.; Nabulsi, I., 2003. Genetic diversityof almonds (Prunus dulcis) usingRAPD technique. Scientia Horticulturae,98 (4): 461-471.Misaki, R.; Fujiyama, K.; Yokoyama, H.;Ido, Y.; Miyauchi, K.; Yoshida, T.; Seki,T., 2003. Characterization of almond alpha-mannosidaseand its application forstructure analysis of sugar chain. Journalof Bioscience and Bioengineering, 96 (2):187-192.Ortega, E. ; Dicenta, F., 2003. Inheritanceof self-compatibility in almond breedingstrategies to assure self-compatibility inthe progeny. Theoretical and Applied Genetics,106 (5) : 904-911.Oukabli, A.; Lansari, A.; Wallali L.D.;Abousalim, A., 2001. Conséquences endogamiquessur la germination des graines,la croissance et le développementdes semis de la variété d’amandier autocompatibleTuono. Fruits 56 (3): 1-18.Oukabli A., Lansari, A.; Wallali, L.D.;Abousalim A.; Egea, J.; et Michaux Ferriere,N., 2000. Self and cross pollinationon pollen tube growth and fertilization inself-compatible almond Prunus dulcis‘Tuono’. 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Growth, osmotic adjustment,and nutrient acquisition of bitter almondunder induced sodium chloride salinity invitro. Communications in Soil Scienceand Plant Analysis, 34 (13-14): 1969-1979.Silva, C.; Tereso, S.; Nolasco, G.; Oliveira,M.M., 2003. Cellular location of Prunedwarf virus in almond sections by in situreverse transcription-polymerase chainreaction. Phytopathology, 93 (3): 278-285.Socias i Company, R.; Alonso, J.M.,2004. Cross-incompatibility of ‘Ferragnes’and ‘Ferralise’ and pollination efficiencyfor self-compatibility transmission in almond.Euphytica, 135 (3): 333-338.Socias i Company, R.; Felipe, A.J.; Aparisi,J.G., 2003. Almond bloom in a changingclimate. Journal American PomologicalSociety, 57 (2): 89-92.Testolin, R; Messina, R; Lain, O; Marrazzo,MT; Huang, WG; Cipriani, G., 2004.Microsatellites isolated in almond from anAC-repeat enriched library. MolecularEcology Notes, 4 (3): 459-461.Takeoka, G.R.; Dao, L.T., 2003. Antioxidantconstituents of almond [Prunus dulis(Mill.) D.A. Webb] hulls. Journal of Agriculturaland Food Chemistry, 51 (2): 496-501.Traynor, J., 2003. Almond pollinationcontractor uses microchips to discouragehive thefts. American Bee Journal, 143(12): 921-922.Tsipouridis, C; Thomidis, T, 2004.Rooting of ‘GF677’ (almond x peach hybrid)hardwood cuttings in relation tohydrogen hyperoxide, moisture content,oxygen concentration, temperature andpH of substrate. Australian Journal OfExperimental Agriculture, 44 (8): 801-805.Vezvaei, A., 2003. Isozyme diversity inIranian almond. Genetics and Breeding ofTree Fruits and Nuts (Acta Horticulturae),622: 451-456.Yi, W.G.; Law, S.E.; Wetzstein, H.Y.,2003. Fungicide sprays can injure thestigmatic surface during receptivity in almondflowers. Annals of Botany, 91 (3):335-341.Yi, W.G.; Law, S.E.; Wetzstein, H.Y.,2003. An In Vitro Study of FungicideEffects on Pollen Germination and TubeGrowth in Almond. HortScience, 38 (6):1086-1088.Young, C.T.; Schadel, W.E.; Pattee, H.E.;Sanders, T.H., 2004. The microstructureof almond (Prunus dulcis (Mill.) D.A.Webb cv. ‘Nonpareil’) cotyledon. Lebensmittel- Wissenschaft und - Technologie -Food Science and Technology, 37 (3):317-322.Zanetto, A.; Maggioni, L.; Tobutt, K.R.;Dosba, F., 2002. Prunus genetic resourcesin Europe: Achievement and perspectivesof a networking activity. GeneticResources and Crop Evolution, 49: 331-337.CAROBBurés, P.; Pavlicek, T.; Horova, L.; Nevo,E., 2004. Microgeographic genome sizedifferentiation of the carob tree, Ceratoniasiliqua, at ‘Evolution canyon’, Israel.Annals of Botany, 93 (5): 529-535.FAO-CIHEAM - Nucis-Newsletter, Number 12 September 200447

Correia, P.J.; Pestana, M.; Martins, M.A.,2003. Nutrient deficiencies in carob (Ceratoniasiliqua L.) grown in solution culture.Journal of Horticultural Science & Biotechnology,78 (6): 847-852.Correia, P.J.; Martins-Louçâo, M.A.,2004. Effect of nitrogen and potassiumfertilization on vegetative growth and floweringof mature carob trees (Ceratoniasiliqua): variations in leaf area index andwater use indices. Australian Journal ofExperimental Agriculture, 44: 83-89.Custodio, L; Martins-Loucao, MA; Romano,A., 2004. Influence of sugars on in vitrorooting and acclimatization of carobtree. Biologia Plantarum, 48 (3): 469-472.Jahns, S., 2003. A late Holocene pollendiagram from the Megaris, Greece, givingpossible evidence for cultivation of Ceratoniasiliqua L. during the last 2000 years.Veget. Hist. Archaeobot, 12: 127-130.Lo Gullo, M.A.; Nardini, A.; Trifilo, P.; Salleo,S., 2003. Changes in leaf hydraulicsand stomatal conductance following droughtstress and irrigation in Ceratonia siliqua(Carob tree). Physiologia Plantarum,117 (2): 186-194.Mediouni, J; Fukova, I; Frydrychova, R; etal., 2004. Karyotype, sex chromatin andsex chromosome differentiation in the carobmoth, Ectomyelois ceratoniae (Lepidoptera:Pyralidae). Caryologia, 57 (2):184-194.Ramon, L.; Mabberley, D.J., 2004. Theecological status of the carob-tree (Ceratoniasiliqua, Leguminosae) in the Mediterranean.Botanical Journal of the LinneanSociety, 144 (4): 431-436.Rhizopoulou, S, 2004. Aspects of cellwall extensibility in Ceratonia siliqua L.Flora, 199 (4): 327-333.Von Haselberg, C; Ludders, P; Stosser,R, 2004. Pollen tube growth, fertilizationand ovule longevity in the carob tree (Ceratoniasiliqua L.). J Appl Bot Food Qual,78 (1): 32-40.Wang, F.; Wang, Y.J.; Sun, Z., 2002.Conformational Role of Xanthan in its Interactionwith Locust Bean Gum. Journalof Food Science, 67 (7): 2609-2614.Zohary, D., 2002. Domestication of the carob(Ceratonia siliqua L.). Israel Journal ofPlant Sciences, 50, Suppl. S: S141-S145.Zunft, H.J.F.; Luder, W.; Harde, A.; Haber,B.; Graubaum, H.J.; Koebnick, C.;Grunwald, J., 2003. Carob pulp preparationrich in insoluble fibre lowers total andLDL cholesterol in hypercholesterolemicpatients. European Journal of Nutrition,42 (5): 235-242.CHESTNUTBolvansky, M., 2002. Variability and groupingof chestnuts (Castanea sativa mill.)grown in old orchards at the locality ModryKamen. Folia oecologica, 29: 95-106.Bolvansky, M.; Uzik, M., 2003. Phenotypicvariation of nut characteristics in halfsibfamilies of European chestnut (Castaneasativa). Biologia, Bratislava, 58/Supl.12: 27-34.Bounous, G.; Botta, R.; Beccaro, G., 2001.Valore nutritivo e pregi alimentary dellecastagne. Frutticoltura, LXIII (10): 37-44.Dane, F.; Lang, P.; Huang, H.; Fu, Y.,2003. Intercontinental genetic divergenceof Castanea species in eastern Asia andeastern North America. Heredity, 91 (3):314-321.Fonti, P.; Sell, J., 2003. Radial split resistanceof chestnut earlywood and its relationto ring with. Wood and Fiber Science,35 (2): 201-208.Lopez-Matas, MA; Nunez, P; Soto, A; etal., 2004. Protein cryoprotective activityof a cytosolic small heat shock proteinthat accumulates constitutively in chestnutstems and is up-regulated by low andhigh temperatures. Plant Physiol, 134 (4):1708-1717.Pereira, S.; Ramos, A.M., 2003. Característicasmorfológicas e isoenzimáticas delos cultivares de castaño. (Castanea sativaMill.) de Andalucía. Monografías INIA,Agrícola, 13: 160 pp. ISBN: 84-7498-486-6.Portela, E.; Roboredo, M.; Louzada, J.,2003. Assessment and description ofmagnesium deficiences in chestnut groves.Journal of Plant Nutrition, 26 (3):503-523.Rhoades, C.C.; Brosi, S.L.; Dattilo, A.J.;Vincelli, P., 2003. Effect of soil compactionand moisture on incidence of phytophthoraroot rot on American chestnut(Castanea dentata) seedlings. Forest Ecologyand Management, 184 (1-3): 47-54.HAZELNUTDe Salvador, F. R., 2001. La situazionevarietale della corilicoltura italiana. Frutticoltura,LXIII (10): 23-28.Del Castillo, M.L.R.; Flores, G.; Herraiz,M.; Blanch, G.P., 2003. Solid-phase microextractionfor studies on the enantiomericcomposition of filbertone in hazelnutOils. Journal of Agricultural andFood Chemistry, 51 (9): 2496-2500.Moure, A.; Franco, D.; Sineiro, J.; Dominguez,H.; Núñez, M.J., 2003. Simulationof multistage extraction of antioxidantsfrom Chilean hazelnut (Gevuina avellana)hulls. Journal of the American Oil ChemistsSociety, 80 (4): 389-396.Nas, M.N.; Read, P.E., 2004. A hypothesisfor the development of a defined tissueculture medium of higher plants andmicropropagation of hazelnuts. ScientiaHorticulturae, 101 (1-2): 189-200.Okay, Y.; Köksal, A.I.; Artik, N., 2003.Seasonal changes in fructose, glucoseand sucrose content of bark tissues of hazelnutgrown in the black sea region. 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static pollen applicator development andtests for almond, kiwi, date, and pistachio- An overview. Applied Engineering inAgriculture, 19 (2): 119-124.Mehlenbacher, S.A., 2003. Progress andprospects in nut breeding. Genetics andBreeding of Tree Fruits and Nuts. ActaHorticulturae, 622: 57-79.Trullols, E; Ruisanchez, I; Rius, FX; et al.,2004. Qualitative method for determinationof aflatoxin B-1 in nuts. J AOAC Int,87 (2): 417-423.Wang, SJ; Tang, JM, 2004. Radio frequencypost-harvest quarantine andphytosanitary treatments to control insectpest in fruits and nuts. Prod Pract QualAsse, 4: 17-53.Wiersma, P.A., 2003. Reproductive barriersin tree fruit crops and nuts. Geneticsand Breeding of Tree Fruits and Nuts.Acta Horticulturae, 622: 369-377.PECANAcuña Maldonado, L.E.; Smith, M.W.;Maness, N.O.; Cheary, B.S.; Carroll, B.L.;Johnson, G.V., 2003. 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Firstreport of Botrytis cinerea causing shootblight of pistachio in Greece. Plant Pathology,52 (6): 810.Epstein, L.; Beede, R.; Kaur, S.; Ferguson,L., 2004. Rootstock effects on pistachiotrees grown in Verticillium dahliae-infestedsoil. Phytopathology, 94 (4): 388-395.Fooladi, M.H.; Farahnaky, A., 2003. Aflatoxinremoval from pistachio nuts by naturalnatrolite. Journal of Food Science, 68(4): 1225-1228.Golan, A.; Barazani, O.; Wang, Z.S.;Khadka, D.K.; Saunders, J.A.; Kostiukovsky,V.; Rowland, L.J., 2004. Genetic relationshipsamong Mediterranean Pistaciaspecies evaluated by RAPD andAFLP markers. Plant Systematics andEvolution, 246 (1-2): 9-18.Haddad, CR; Louw, SV; Dippenaar-Schoeman, AS., 2004. An assessment ofthe biological control potential of Heliophanuspistaciae (Araneae: Salticidae) onNysius natalensis (Hemiptera: Lygaeidae),a pest of pistachio nuts. Biol Control,31 (1): 83-90.Katsiotis, A.; Hagidimitriou, M.; Drossou,A.; Pontikis, C.; Loukas, M., 2003. Geneticrelationships among species and cultivarsof Pistacia using RAPDs and AFLPs.Euphytica, 132 (3): 279-286.Kucukoner, E.; Yurt, B., 2003. Some chemicalcharacteristics of Pistacia vera varietiesproduced in Turkey. EuropeanFood Research and Technology, 217 (4):308-310.Ila, H.B.; Kafkas, S.; Topaktas, M., 2003.Chromosome numbers of four Pistacia(Anacardiaceae) species. Journal of HorticulturalScience & Biotechnology, 78 (1)35-38.FAO-CIHEAM - Nucis-Newsletter, Number 12 September 200449

Lee, N.A.; Wang, S.; Allan, R.D.; Kennedy,I.R., 2004. A rapid aflatoxin B-1 ELI-SA: Development and validation with reducedmatrix effects for peanuts, corn,pistachio, and soybeans. Journal of Agriculturaland Food Chemistry, 52 (10):2746-2755.Ma, Z.H.; Felts, D.; Michailides, T.J.,2003. Resistance to azoxystrobin in Alternariaisolates from pistachio in California.Pesticide Biochemistry and Physiology,77 (2): 66-74.Ma, Z.; Luo, Y.; Michailides, T.J., 2003.Nested PCR assays for detection of Moniliniafructicola in stone fruit orchardsand Botryosphaeria dothidea from pistachiosin California. Journal of Phytopathology- Phytopathologische Zeitschrift,151 (6): 312-322.Ma, Z.H.; Luo, Y.; Michailides, T.J., 2004.Spatiotemporal changes in the populationstructure of Botryosphaeria dothidea fromCalifornia pistachio orchards. Phytopathology,94 (4): 326-332.Martinez, J.J.Y.; Wool, D., 2003. Differentialresponse of trees and shrubs tobrowsing and pruning: the effects on Pistaciagrowth and gall-inducing aphids.Plant Ecology, 169 (2): 285-294.MirAli, N.; Nabulsi, I., 2003. Genetic diversityof Syrian grown pistachio cultivars(Pistacia vera L.) using RAPD technique.Adv. Hort. Sci., 17 (4): 215-222.MunneBosch, S.; Peñuelas, J., 2003.Photo- and antioxidative protection duringsummer leaf senescence in Pistacialentiscus L. grown under Mediterraneanfield conditions. Annals of Botany, 92 (3):385-391.Onay, A., 2003. Micropropagation of Pistachio.Micropropagation of Woody Treesand Fruits. Forestry Sciences, 75: 565-588.Onay, A.; Pirinc, V.; Yildirim, H.; Basaran,D., 2004. In vitro micrografting of maturepistachio (Pistacia vera var. Siirt). PlantCell Tissue and Organ Culture, 77 (2):215-219.Ozcan, M., 2004. Characteristics of fruitand oil of terebinth (Pistacia terebinthusL) growing wild in Turkey. Journal of theScience of Food and Agriculture, 84 (6):517-520.Reig-Arminana, J; Calatayud, V; Cervero,J; Garcia-Breijo, FJ; Ibars, A; Sanz, MJ,2004. Effects of ozone on the foliar histologyof the mastic plant (Pistacia lentiscusL.). Environmental Pollution, 132 (2):321-331.Parfitt, D.E., 2003. ‘Bonsai’ OrnamentalPistachio. HortScience, 38 (6): 1260-1261.Parfitt, D.E.; Arjmand, N.; Michailides,T.J., 2003. Resistance to Botryosphaeriadothidea in pistachio. Hortscience, 38 (4):529-531.Sepaskhah, A.R.; Karimi Goghary, S.,2003. Growth and chemical compositionof pistachio affected by salinities anddepths of water table. Communications inSoil Science and Plant Analysis, 34 (3-4):343-355.Vidrich, V.; Fusi, P.; Graziano, A.; Silvestrini,E.; Michelozzi, M.; Marco, F., 2004.Chemical composition of the essential oilof Pistacia lentiscus L. Journal of EssentialOil Research, 16 (3): 223-226.STONEPINEAlvarez, JB; Toledo, MJ; Abellanas, B, etal., 2004. Use of megagametophyte storageproteins as markers of the geneticdiversity in stone pine (Pinus pinea L.) inAndalucia, Spain. Genet Resour Crop Ev,51 (6): 621-627.Mutke, S.M.; Sada, B.; Iglesias, S.; Gil,L., 2003. Evaluación de la producción individualde piña en un banco clonal depino piñonero (Pinus pinea L.) en Madrid(in Spanish). Invest. Agrar.: Sist. Recur.For., 12 (1): 149-157.Mutke, S.; Gordo, J.; Climent, J.; Gil, L.,2003. Shoot growth and phenology modellingof grafted stone pine (Pinus pineaL.) in Inner Spain. Annals of ForestScience, 60 (6): 527-537.Ranaldi, F.; Giachetti, E.; Guerin, E.;Bacci, S.; Paoletti, E.; Boddi, V.; Vanni,P., 2003. Gravitational stress on germinatingPinus pinea seeds. Comptes RendusBiologies (France). 326 (6): 553-564.WALNUTAletà, N.; Ninot, A.; Voltas, J., 2003. Caracterizacióndel comportamiento agroforestalde doce genotipos de nogal (Juglanssp.) en dos localidades de Cataluña(in Spanish). Invest. Agrar.: Sist. Recur.For., 12 (1): 39-50.Amaral, J.S.; Casal, S.; Pereira, J.A.; Seabra,R.M.; Oliveira, B.P.P., 2003. Determinationof sterol and fatty acid compositions,oxidative stability, and nutritional valueof six walnut (Juglans regia L.) cultivarsgrown in Portugal. Journal of Agriculturaland Food Chemistry, 51 (26): 7698-7702.Breton, C; Cornu, D; Chriqui, D; et al.,2004. Somatic embryogenesis, micropropagationand plant regeneration of “EarlyMature” walnut trees (Juglans regia) thatflower in vitro. Tree Physiol, 24 (4): 425-435.Fady, B.; Ducci, F.; Aletà, N.; Becquey,J.; Díaz, R.; Fernández, F.; Jay-Allemand,C.; Lefèvre, F.; Ninot, A.; Panetsos,K.; Paris, P.; Pisanelli, A.; Rumpf, H.,2003. New Forests, 25: 211-225.Fukuda, T.; Ito, H.; Yoshida, T., 2003. Antioxidativepolyphenols from walnuts (Juglansregia L.). Phytochemistry, 63 (7):795-801.Jensen, P.N.; Sorensen, G.; Brockhoff,P.; Bertelsen, G., 2003. Investigation ofpackaging systems for shelled walnutsbased on oxygen absorbers. Journal ofAgricultural and Food Chemistry, 51 (17):4941-4947.50 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004

Kevers, C; Bisbis, B; Crevecoeur, M; etal., 2004. Wood formation in in vitro propagatedwalnut shoots in relation withroot formation and development. ActaBot Gallica, 151 (1): 45-53.Koyuncu, M.A.; Ekinci, K.; Sabrán, E.,2004. Cracking characteristics of walnut.Biosystems Engineering, 87 (3): 305-311.Mahoney, N.; Molyneux, R.J., 2004. Phytochemicalinhibition of aflatoxigenicity inAspergillus flavus by constituents of walnut(Juglans regia). Journal of Agriculturaland Food Chemistry, 52 (7): 1882-1889.McKenna, J.R.; Epstein, L., 2003. Susceptibilityof Juglans Species and InterspecificHybrids to Agrobacterium tumefaciens.HortScience, 38 (3): 435-439.Menard, M.; Baudry, A.; LeSaux, M.,2004. First report of bacterial canker ofwalnut caused by Brenneria nigrifluens inFrance. Plant Disease, 88 (2): 220.Ninot, A.; Aletà, N., 2003. Identificationand genetic relationship of Persian walnutgenotypes using isozyme markers.Journal American Pomological Society,57(3): 106-114.Orel, G.; Marchant, A.D.; McLeod, J.A.;Richards, G.D., 2003. Characterization of11 Juglandaceae genotypes based onmorphology, cpDNA, and RAPD. Hortscience,38 (6): 1178-1183.Sabatier, S.; Barthelemy, D.; Ducousso,I., 2003. Periods of organogenesis inmono- and bicyclic annual shoots of Juglansregia L. (Juglandaceae). Annals ofBotany, 92 (2): 231-238.Sakr, S.; Alves, G.; Morillon, R.L.; Maurel,K.; Decourteix, M.; Guilliot, A. FleuratLessard, P.; Julien, J.L.; Chrispeels, M.J.,2003. Plasma membrane aquaporins areinvolved in winter embolism recovery inwalnut tree. Plant Physiology, 133 (2):630-641.Solar, A.; Ivancic, A.; Stampar, F.; Hudina,M., 2002. Genetic resources for walnut(Juglans regia L.) improvement in Slovenia.Evaluation of the largest collectionof local genotypes. Genetic Resourcesand Crop Evolution, 45: 491-501.Solar, A.; Stampar, F., 2003. Genotypicdifferences in branching pattern and fruitinghabit in common walnut (Juglans regiaL.). Annals of Botany, 92 (2): 317-325.Vahdati, K; Leslie, C; Zamani, Z; et al.,2004. Rooting and acclimatization of in vitro-grownshoots from mature trees ofthree Persian walnut cultivars. Hortscience,39 (2): 324-327.Vettraino, A.M.; Belisario, A.; Maccaroni,M.; Vannini, A., 2003. Evaluation of rootdamage to English walnut caused by fivePhytophthora species. Plant Pathology,52 (4): 491-495.Vyas, D.; Sharma, S.K.; Sharma, D.R.,2003. Genetic structure of walnut genotypeusing leaf isozymes as variability measure.Scientia Horticulturae, 97 (2): 141-152.Yildiz, K.; Yilmaz, H., 2003. Effect ofTransplanting Rootstocks Before Graftingon Xylem Exudation and Graft Success inWalnut. Journal American PomologicalSociety, 57 (4): 146-148.THESISAlonso, J.M., 2004. Efecto de la consanguinidadsobre la expresión y la transmisiónde la autocompatibilidad en el almendro(Prunus amygdalus Batsch). PhDthesis. (in Spanish with summary in English).Lleida University. Escuela TécnicaSuperior de Ingeniería Agraria, 227 pages.Georges, A., 2003. Vessel-associated cellsin walnut trees (Juglans regia): Role ofplasma membrane H+-ATPase (in Frenchwith summary in English). Blaize PascalUniversity, France: 279 pages.Gharnit, N., 2003. Characterization of thecarob tree (Ceratonia siliqua L.) from theProvince of Chefchaouen (NW of Morocco).PhD thesis. (in French with summaryin English). Faculty of Sciences and Techniquesof Tangier, Université AbdelmalekEssaâdi (Morocco).López, M., 2004. Mejora del almendropara autocompatibilidad utilizando técnicasbiológicas y moleculares. PhD thesis.(in Spanish with summary in English).Lleida University. Escuela Técnica Superiorde Ingeniería Agraria, 270 pages.Martins, M. 2003. Análise molecular emamendoeira: da diversidade e estabilidadegenómicas à identificaçao de resistências.PhD thesis. (in Portuguese withsummary in English). Universidad de Lisboa,241 pages.MASTERSTürker, S. 2003., The effect of somerootstock x cultivar combinations of pistachioto the phonological and pomologicaltraits. Master Thesis. University of Harran,Graduate School of Natural andApplied Sciences. Department of Horticulture,86 pages.FAO-CIHEAM - Nucis-Newsletter, Number 12 September 200451

THE FAO-CIHEAM INTER-REGIONAL COOPERATIVE RESEARCH NETWORK ON NUTSNetwork Coordination Centre Network CoordinatorNut tree cropsAlmondStone PineInstitut de Recerca i Tecnologia Agroalimentàries IRTA.Centre de Mas BovéDepartament d’Arboricultura MediterràniaApartat 415. E 43280 - Reus (Spain)Tel: 34- 977 328424. Fax: 34- 977 344055E-mail: francisco.vargas@irta.esF. J. VargasSubnetworks Liaison Centre Liaison OfficerHazelnutAnkara University. Faculty of Agriculture.Department of Horticulture.06110 - Ankara (Turkey).Tel: 90 - 312 3170550. Fax: 90 - 312 3179119E-mail: ikoksal@agri.ankara.edu.trA.I.KöksalWalnut and PecanInstitut National de la Recherche Agronomique INRA.Unité de Recherches sur les Espèces Fruitières et la Vigne.B.P. 81 - 33883 - Villenave d’Ornon (France)Tel: 33 - 556 843277. Fax: 33 - 556 843274E-mail: mlafargue@bordeaux.inra.frM. LafarguePistachioChestnutUniversity of Harran, Faculty of AgricultureDepartement of Horticulture63200 - Sanliurfa (Turkey)Tel: 90 - 414 2472697. Fax: 90 - 414 2474480E-mail: beak@harran.edu.trUniversità degli Studi di Torino.Dipartamento di Colture Arboree. Cattedra di ArboriculturaVia Leonardo Da Vinci, 44. 10095 - Grugliasco (TO) (Italy)Tel. 39 - 011 6708653. Fax: 39 - 011 6708658.E-mail: bounous@agraria.unito.itB. E. AkG. BounousGenetic ResourcesInstitut de Recerca i Tecnologia Agroalimentàries IRTA.Centre de Mas Bové.Departament d’Arboricultura MediterràniaApartat 415. E 43280 Reus (Spain)Tel: 34 - 977 328424. Fax: 34 - 977 344055E-mail: ignasi.batlle@irta.esI. BatlleEconomicsServicio de Investigación Agraria.Diputación General de Aragón. Apartado 72750080 - Zaragoza (Spain)Tel. 34 - 976 576361. Fax: 34- 976 575501E-mail: albisu@mizar.csic.esL.M. AlbisuFAORegional Office for Europe REU:Viale delle Terme di Caracalla.00100 Roma (Italy)Tel: 39 - 06 57054405 Fax: 39 - 06 57055634E-mail: jutta.krause@fao.orgJutta KrauseCIHEAMInstituto Agronómico Mediterráneo de Zaragoza IAMZ.Apartado 202, 50080 Zaragoza (Spain)Tel: 34 - 976 71 60 00 Fax: 34- 976 71 60 01E-mail: gabina@iamz.ciheam.orgDunixi GabiñaIRTA- Mas BovéDepartament d'Arboricultura Mediterrània.Apartat, 415.E- 43280 REUS (Spain)Tel.: +34-977 32 84 24Fax: +34-977 34 40 55E-mail: ignasi.batlle@irta.esNetwork Coordinator: F.J. VargasEditor: I. BatlleEditorial staff: M. LannoyeTypeset by: Carácter Gráfico, S.L.E-mail: cg@ediho.esISSN 1020-079752 FAO-CIHEAM - Nucis-Newsletter, Number 12 September 2004