Scientific Papers Series B Horticulture

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Scientific Papers. Series B, Horticulture. Vol. LVII, 2013Print ISSN 2285-5653, CD-ROM ISSN 2285-5661, Online ISSN 2286-1580, ISSN-L 2285-5653SSR ANALYSIS CONCERNING THE RESISTANCE TO PPV(Plum pox virus) IN SEVERAL ROMANIAN APRICOT PROGENIESFrgua CIOMAGA 1 , Ligia ION 1 , Cristina MOALE 21 University of Agronomic Sciences and Veterinary Medicine of Bucharest, 59, Marasti Blvd,011464, Bucharest, Romania2 Research Station for Fruit Growing Constanta, Constanta, RomaniaAbstractCorresponding author email: ionnagyligia@yahoo.frPlum pox virus (PPV) is a devastating stone fruit disease of major importance, and better understanding of the geneticcontrol of resistance to this trait would be useful for more efficient development of resistant cultivars. Previous studieshave reported a locus major effect from PPV resistance on linkage group 1. The hybrids were grafted simultaneouslyand subsequently inoculated with the PPV-M and D strains. The symptom scoring on leaves was performed three timesover two vegetative cycles. The PPV resistant loci were mapped using composite interval mapping (CIM).This paperpresents data from PhD thesis part of the project POSDRU/107/1.5/S/76888, funded by European Social Fund throughthe Sectorial Operational Programme Human Resources Development 2007-2013.Key words: disease, markers, Prunus armeniaca, PPV, SSR, Sharka.INTRODUCTIONPlum pox virus (PPV) infection causes theSharka disease of Prunus, has spread from theBalkan countries throughout most of theEuropean subcontinent and around theMediterranean basin.Considerable economic loss and significantreduction in productive areas stimulatedbreeding programs aimed at enhancingresistance to the pathogen in such countries asGreece (Karayiannis et all., 2002), France(Audergon et all., 1994), Italy (Bassi et all.,1995), Spain (Egea et all., 1999) and the CzechRepublic (Polak, 1994).As a result of the intensive search for a sourceof resistance within available apricotgermplasm, some North American cultivars‘Stark Early Orange’ (SEO), ‘Goldrich’,‘Harlayne’, ‘NJA 45’, and others were found tohave natural resistance to PPV (Martinez-Gomez eta ll., 2000). These cultivars were usedas donors for a resistance trait in conventionalbreeding programs based on crosses betweenresistant and the best local cultivars susceptibleto virus.Recently, resistant selections have beenreleased for regional trials (Badenes and Llacer,2006; Dosba et all., 1992; Karayiannis et all,1999; Polak et all., 1997)177Several studies suggest that the resistance isconferred by a limited number (1–3) of genes(Dicenta et all., 2000; Dosba et all., 1992;Krska B et all., 2002; Karayiannis et all., 2007).Development of molecular marker maps forsegregating crosses is a significantaccomplishment toward understanding thegenetics of PPV resistance and developingmarkers that could potentially be useful inbreeding programs. Four molecular geneticmaps based on intraspecific crosses introducingPPV resistance from North American cultivars‘Stark Early Orange’ and ‘Goldrich’ have beenestablished to map a PPV resistance inapricot (Lambert et all., 2007; Sicard et all.,2007). On these maps, a major genomic regionassociated with PPV resistance was located onthe Prunus G1 at a distance of 20–40 cm. Intotal, five SSR markers linked to the targetedresistance locus were identified in this region.Three of them have been already successfullytested for marker assisted selection (MAS) in aset of susceptible/resistant cultivars.MATERIALS AND METHODSPlant materialWe are worked with 5 apricot population:
Pop 3 = Population 3 obtained by crossesbetween (‘Mari de Cenad, x,NJA 21’) x ‘KesthPshor’Pop 2 = Population 2 obtained by crosses‘Sirena’ x ‘NJA 42’Pop 4 = Population 4 obtained by crosses‘Cristal’ x ‚NJA 21’Pop 1 = Population 1 obtained by crosses ‘Maride Cenad’ x ‚SEO’Pop 5 = population 5 obtained by crossesbetween (‘Viceroy’, x,NJA 2’) x ‘NJA17’The apricot selection R9 P 53 (‘NJA 2’×‘Viceroy’) was crossed as a female parent to‘NJA 17’ a PPV resistant apricot cultivar.Theselection R13 VT 8/57 resistant to PPV (issuedfrom ‘Mari de Cenad x NJA 21) was crossed asa female parent to ‘Kesth Pshor’ (susceptible toPPV) in the frame of the Faculty ofHorticulture of University of AgronomicSciences and Veterinary Medicine of BucharestRomania in 2008. Crosses were performed byhand pollination with isolation of flowers afterthe petals and anthers removal from the flowerbuds. The F1 seeds were stratified at 5 o C for 3months and subsequent seedlings were grownin an insect-proof greenhouse.The young apricot populations sticks weregrafted onto inoculated GF305 (used likesusceptible rootstock) ready for testing to PPVresistance.DNA extractionGenomic DNA was extracted using themodified CTAB procedure [23]. DNAconcentrations were measured and workingsolutions of genomic DNA at 10 ng/μl and at100 ng/μl in 0.1× TE (0.01M Tris pH 8.0, 0,001M EDTA) buffer was prepared for the targetedSSR and AFLP analyses, respectively.SSR analysis‘Stark Early Orange’, ‘R13 VT 8/57’ NJA andR9 P 53 were screened with 3 SSR primercombinations from [2], associated withPPVresistance.PGS 1.21, PGS 1-24 and ppb22-195-F:CTCTTCTCGCCTCCCAATTT andR:GCTTAGCCCTGGGTACAAG andF:ATCTGCTCTTTCCCTCACCT withR:GATTATCCCTCAACCCATCC.PCR reactions, electrophoresis, and detectionof PCR products for the ‘Kesth Pshor’ × R13VT 8/57’ population were carried out accordingto conditions specified in (Zhebentyayeva etall., 2002). SSR primer combinations revealingpolymorphism were screened all apricotpopulations. The mix PCR consit in 10Xbuffer-2ul, MgCl2 (25 mM)-1.2 ul, dNTP 10mM – 0.16 ul, PGS 1.21-F – 0.6ul, PGS 1.21-R– 0.6 ul, PGS 1.24-F – 0.4 ul, PGS1.24-R – 0.4ul, ppb22-195-F-0.28 ul,ppb22-195-R – 0.28uland Taq 0.1 ul. A 2-μl aliquot of the PCRreaction was separated by electrophoresis on anagarose gel 2% in order to confirm theamplification of fragments of the expected sizeand DNA concentration. (Figure 1.).The PCR-products were diluted (45 ul H2O and5 ul DNA) and used to prepare the plate forsequencing.RESULTS AND DISCUSSIONSAll four SSR primer combinations amplifiedexpected size fragments in germoplasm understudy. SSR scores for genotypes are included insupporting documentation, Table 1. For all SSRloci, we recorded genotypes in order of theirelectrophoretic mobility from fastest to slowestband.The infection process was different for variousplant individuals; in some plants the presenceof PPV was detected after the first dormancyperiod, in some in the three vegetative cycles,while some plants were not infected at all.Selections ‘Kesth Pshor’ × ’R13 VT 8/57’ and’R9 P 53’ were screened with 3 SSR primercombinations. The products of PCR wereseparated by electrophoresis on an agarose gel2% in order to confirm the amplification offragments of the expected size and DNAconcentration. (Figure 1)Figure 1. Electrophoresis for PCR with SSR markers inapricot progeniesThese SSRs were polymorphic. This presenceof polymorphism is slightly higher than thatpreviously reported by Hurtado et al. (2002)178
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SCIENTIFIC PAPERSSERIES B. HORTICUL
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SCIENTIFIC COMMITTEE Bekir Erol AK
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Fructification - Florin Constantin
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Phenological Studies on Some Variet
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Scientific Papers. Series B, Hortic
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Table 2. The organoleptic appreciat
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processes become even slower and th
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- usingselectedbacteriafromm Leucon
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To organoleptic analysis of the win
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works were pickling the cucumbers i
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days after removal of weeds by burn
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it appears that the culture has gro
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Maintainance of the genetic structu
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Table 3. Interaction variety of dwa
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Experimental scheme is situated in
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improvement especially laboratory s
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Table 1 shows the type 2x3x2 trifac
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Neamtu G., Gheorghe Campeanu, Carme
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Experience consisted of the followi
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Table 7. Total production on plantV
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vineyard, several methods were used
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Table 4. Structure and number and r
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gets serious role through the ferme
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Table 3. Element content of the ber
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Figure 4. The length of internodes
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The process of fruit forming was de
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Compomist F1 hybrid reaches value o
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Figure 2. Experimental results repr
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and total number of tubers per plan
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and quality of organic potatoes for
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Petri dishes, the pathogenic fungi
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Variant 4R 2 11.0 4.0 15.0 7.0 22.0
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Analyzing the dynamics of the total
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Table 2. Indicative data on volumes
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Table 3. The Influence of the level
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Table 8. The Influence of irrigatio
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Figure 1. The aspect of apples from
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Conclusions on changes in chemicalc
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RESULTS AND DISCUSSIONSRaspberry is
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CONCLUSIONSAs a result of scientifi
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observations were made on phenologi
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Figure 1. Flowering period and blac
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content, A-acidity content) to the
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sugar accumulated in grapes and 1,0
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Figure 2. DryerFigure 3. Programmer
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Appearance of the apricots after 5
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Page 130 and 131: where: P F represent the coordinate
Page 134 and 135: Table 2. The production obtained at
Page 136 and 137: As shown in figures 5 and 6 the vit
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Page 142 and 143: If we take into account the both fa
Page 146 and 147: F1 - + infectedF2 - + infectedF3 +
Page 148: HORTICULTURALBIODIVERSITY ANDGENETI
Page 151 and 152: RESULTS AND DISCUSSIONSFruit charac
Page 156 and 157: The chorology maps of Artemisia alb
Page 158 and 159: Figure 5. Chorology of Artemisia le
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Page 165 and 166: The characterization of the grapevi
Page 167 and 168: Figure 4. Amino acid profile of the
Page 169 and 170: Table 4. The significance of differ
Page 171 and 172: The studied phenophases (table 1),
Page 173 and 174: REFERENCESCapusan Janina, 2013. Rez
Page 175 and 176: Neoaliturus fenestratus Herrich-Sch
Page 177: occurrence in western part of Roman
Page 181 and 182: well adapted in our country, and th
Page 186 and 187: Variations in acidity of the variet
Page 188: grape. Analele Universitatii din Cr
Page 191 and 192: Flow cytometry has proved to be an
Page 193 and 194: cell nuclei, using chromosome count
Page 195 and 196: Figure 1. Prekos F1Figure 4. Prekos
Page 197 and 198: Interpretation of the results conce
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Page 201 and 202: Dry matter (determinate refractomet
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Page 212 and 213: Likewise, vitamin C content of ‘G
Page 216 and 217: Figure 2. Air temperature (°C) in
Page 218: REFERENCESAnconelli S., Antolini G.
Page 221 and 222: Taking into the account these and t
Page 223 and 224: postharvest decays in fruits and ve
Page 225 and 226: manure semifermentated. In vegetati
Page 227 and 228: Figure 1. Prima Cl. 1022 varietyThe
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temperature required for a differen
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MATERIALS AND METHODSTo accomplish
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climate (IH4), becomes for this yea
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quantities of sugar accumulated in
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ibosomal to investigate 13 species
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Biogeography of Genus MangiferaTwo
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productionand uses. Center for trop
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analysis of variance to express the
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highlighted by a correlation coeffi
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without heat and in open field. Cro
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ut the fruit had a reduced number o
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Genetic autochthonous heritage wasi
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Table 1. The main characteristics o
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Data were subjected to statistical
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Also, the dry biomass and root leng
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“Clapp’s Favorite” in 2011 wh
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2012 the order was “Conference”
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microspore to mature pollen, determ
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microspores at certain date. Micros
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viin utilizate ca genitori potentia
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The objective of this paper, is to
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Table 1. Relationship between germi
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Figure 2. Tassels lengthIn case of
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ORNAMENTAL PLANTS,DESIGN ANDLANDSCA
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of Chisinau, also the old parks. Fo
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y cambium on the very center - meth
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flower is fully colored. Postharves
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Results from table 8 shows that Mar
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Pinus sylvestris ‘Gold Coin’, R
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Figure 4. Excelsa (H 18/1)Descripti
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Figure 1. Plants in cellular trays
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espectively 69%, followed by hybrid
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observations made on the plants of
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The values of pH diminished in all
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Figure 5. Content of total soluble
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Figure 1. Plan of Bran domain (Ion
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Carol the second, her brother, on h
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Figure 14. Tea house in the natural
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Cults, and in conformity with the c
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consult the scientific novelties an
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,,The restoration is the methodolog
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MATERIALS AND METHODSThe chorology
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Merce, 2011; Zamfirescu, 2010) (IAS
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experiment because it shows and ill
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among all the administrations, exce
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solution known as AKN. Content: pot
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As shown in the figure, Chrysal is
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MotivationsOfferprospectingCustomer
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conclude the presented case, with k
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Also, comparative studies, concerni
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done before. Also the speed of the
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as Marcellus theatre or Trajan foru
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ian regime, by huge contradictions
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Pyongyang, the hidden cityThe capit
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“ordinary” buildings as the gen
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- The Writer’s Rotunda“The cons
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Figure 4. Vegetation planRegarding
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Figure 9. BenchesVasesThere are 4 k
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REFERENCESPanoiu A., 2011, Evolutia
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RESULTS AND DISCUSSIONSResults of t
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Figure 5. Average growth rate of
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In 2012 there were higher onset tem
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fact that root injuries is the majo
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We recommend that the species and c
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and parameters of quality. The phys
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AESTHETICALLEVEL OFAPPROACHLEVEL OF
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REFERENCESArchibugi F., 1997. The E
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Figure 1. Optimized version of comp
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CONCLUSIONSThe optimized version (F
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etween 182-212 g/l, the lowest valu
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CONCLUSIONSThis study showed that 2
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RESULTS AND DISCUSSIONSFor this pur
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according to quality class.The calc
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third party countries who wish to b
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Stakeholder comments concerned nota
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Figure 2. The „Eco-leaf” logo o
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identified to facilitate its tracea
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- Analysis reports;This document ma
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Table 1. Sampling procedures applic
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Table 2. Example of sampling proced
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should be 5 kg.” According to thi
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One of the first and simplest condi
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of RIN transcription factor in ethy
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oriented towards the centre of the
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Figure 5. Macroscopic view of the c
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Related to the process of the evolu
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efore it is printed using the main
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Imposition of the pot is done every
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Figure 8. Test of Strength Vetiver
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Table 3 defines LSD test for genoty
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