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 2285-5653IDENTIFICATION OF PLOIDY LEVEL ON VARIETIESAND HYBRIDS OF KIWIFRUIT (ACTINIDIA SP.)Ramona COTRU 1 , Florin STNIC 1 , Giuseppe SCAPIGLIATI 21 University of Agronomic Sciences and Veterinary Medicine of Bucharest, 59 Marasti Blvd,District 1, 011464, Bucharest, Romania2 Department for Innovation in Biological systems, Food, Agriculture and Forestry (DIBAF),University of Tuscia, Via S. Camillo de Lellis snc, 01100, Viterbo, ItaliaAbstractCorresponding author email: ramona_cotrut@yahoo.comThe genus Actinidia includes over 66 species and over 118 taxon, that range in ploidy level from diploid to octoploid.Although there is basic information on ploidy levels of various species, sampling has been limited and little informationon specific cultivars and hybrids is available. The objective of this research was to determine ploidy levels among adiverse collection of species, hybrids, and cultivars using flow cytometry. Nuclei were extracted, stained with propidiumiodide (PI), and analyzed using a flow cytometer. Among the selections tested we found Actinidia arguta as diploid (2n= 2x) and tetraploid (2n = 4x), Actinidia deliciosa 'Hayward' as hexaploid (2n = 6x), Actinidia chinensis'Soreli' tetraploid (2n = 4x) and the interspecific hybrid A. arguta x A. deliciosa as diploid (2n = 2x). This informationprovides further insight into reproductive biology, substantiation of numerous hybrids and induced polyploids that willhelp facilitate the development of improved hybrids in the future.Key words: chromosomes, flow cytometry, interspecific hybrid, polyploidy, plant breeding.INTRODUCTIONvariation in ploidy make hybridization betweenActinidia species difficult. Better knowledgeCommercial kiwifruit production is based onand understanding of polyploidy of cultivarstwo Actinidia species, A. deliciosa C.F. Ling etand hybrids could facilitate kiwifruitA.R. Ferguson and A. chinensis Planch.improvement.Nevertheless, at present, A. arguta isPolyploidy has been an important process in thecommercially cultivated in Oregon in US, Chileevolution of plants and is an important factor inand New Zealand, and small-scale productionplant breeding because it can influencefor local consumption is conducted in manyreproductive compatibility, fertility, andregions under a relatively cool climatephenotypic traits (Chen and Ni, 2006; Jones(Ferguson and Huang, 2001). The genusand Ranney, 2009; Ranney, 2006; Soltis et al.,Actinidia Lindl. comprises 66 species and about2004). Hence, accurate and specific knowledge118 taxa with remarkably wide natural rangeof ploidy levels of species and cultivars isextending from the tropics (latitude 0º) to coldimportant information for kiwi breeders.temperate regions (50º N). Since the first seedThe base chromosome number for Actinidia isintroduction of A. deliciosa was made from 1n = 1x = 29. However, different subgeneraChina to New Zeeland in 1904, the economic contain species with a variety of ploidy levelspotential of kiwifruit has been extensively ranging from 2n = 2x = 58 to 2n = 6x =174;exploited (Ferguson and Bollard 1990).respectively 2n = 8x =232. The range in ploidyThere is considerable genetic diversity amongst levels within this genus also provides anthe species within the genus Actinidia, opportunity to indirectly substantiate hybridityparticularly in fruit skin type, skin colour, flesh when parents differ in ploidy levels.colour and flavour (Ferguson and Huang, Because many Actinidia species are polyploids2007).with high chromosome numbers, traditionalThis diversity provides many opportunities for cytology based on light microscopicdeveloping new types of kiwifruit. Dioecy, examination is a difficult and time-consuminglong generation cycles, high heterozygosity and process.189
Flow cytometry has proved to be an efficientmeans of estimating genome size andassociated ploidy level (Dolezˇel et al., 2007;Jones et al., 2007). Therefore, the objective ofthis study is to identify the ploidy levels of adiverse collection of species, hybrids, andcultivars of Actinidia developing a database foruse by kiwi breeders.MATERIALS AND METHODSThe plant material was constituted by acollection of plants from Actinidia arguta,Actinidia deliciosa, Actinidia chinensis and oneinterspecific crosses of Actinidia, total of130 plants were kept in a green house.Flow cytometric analysis was performed foreach of the Actinidia accessions by collectingleaf samples (3-5 per each), young fullyexpanded leaf collected from the shoot tips; tomaintain an adequate moister and ensure acorrect conservation tissues, the leaves wereplaced in plastic bags in a refrigerator at 4ºC.Using the method of Galbraith et al. (1983), 50mg Actinidia leaf tissue without midribs wasplaced in plastic Petri dishes chopped with arazor blade adding 0.5 ml of ice-cooledhomogenization (Otto I buffer) to homogenizethe tissues and release the nuclei, and kept onice for 5 min.After filtration through 42-μm nylon mesh thenuclear suspension were treated with 2.5 ml ofa staining solution containing 10 ml Tris, 50mM sodium citrate, 2 mM MgCl2, 1% (w/v)PVP 30, 0.1% (v/v) Triton X-100 and 1 mg/mlpropidium iodide (PI), pH 7.5, letting thesamples stand 5 min prior to flow cytometricanalysis.Samples were run on a Coulter Epics XL-MCLflow cytometer EXPO 32 ADC; for eachaccession was gated between 5000-10000events of a sample. Relative fluorescenceintensity was assessed using FL2, green lightsource detector (excitation 488 nm).Data were interpreted using WinMDI 2.9software. As an internal standard was usedPisum sativum cv. Ctirad. The measurementswere tripricated by using three leaf samplescollected from each plant.RESULTS AND DISCUSSIONSWe have evaluate for ploidy level six seletionsof Actinidia arguta, two of A. deliciosa‘Hayward’, one of A. chinensis ‘Soreli’ and oneinterspecific hybrid, A. arguta x A. deliciosa,using flow cytometry (Table 1).Table 1. Selection of species, hybrids of kiwi (Actinidiasp.)SpeciesGenotype No. plantsActinidia arguta R8P23 10Actinidia arguta R10P10 10Actinidia arguta R9P18 10Actinidia arguta R8P1 10Actinidia arguta R9P16 10Actinidia arguta x deliciosa P1 10Actinidia deliciosa’Hayward’ Hk 10Actinidia deliciosa’Hayward’ H8 30Actinidia chinensis’Soreli’ S 30Total 130The ploidy levels of Actinidia plants wereclearly distinguished by flow cytometry.Among Actinidiasp. tested, we found: Actinidiaarguta (2n=2x) R9P18, R8P23 as diploidplants, R10P10, R8P1, R9P16 astetraploid Actinidia arguta (2n=4x), plants; forActinidia deliciosa ‘Hayward’ (2n=6x) HK, H8as hexaploid; for Actinidia chinensis ‘Soreli’(2n=4x) tetraploid; for interspecific crosses A.arguta x A. deliciosa (2n=2x) P1 as diploid(Table 2).Table 2. Estimation of ploidy level of kiwi genotypes(Actinidia sp.)SpeciesGenotype Ploidy No.level chromosomesActinidia arguta R8P23 2x 58Actinidia arguta R10P10 4x 116Actinidia arguta R9P18 2x 58Actinidia arguta R8P1 4x 116Actinidia arguta R9P16 4x 116Actinidia arguta xdeliciosaP1 2x 58Actinidiadeliciosa’Hayward’Hk 6x 174Actinidiadeliciosa’Hayward’H8 6x 174Actinidiachinensis’Soreli’S 4x 116The results of an analysis are described as amono-parametric (fluorescence intensity/object) histograms peaks, each of them190
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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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where: P F represent the coordinate
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Table 2. The production obtained at
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As shown in figures 5 and 6 the vit
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Page 140 and 141: At two weeks from planting it was a
Page 142 and 143: If we take into account the both fa
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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
Page 162: Molecular differentiation showed a
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 and 178: occurrence in western part of Roman
Page 179 and 180: Pop 3 = Population 3 obtained by cr
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 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
Page 199 and 200: Photo 1- Redhaven (Control)The Roma
Page 201 and 202: Dry matter (determinate refractomet
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Page 208 and 209: During the growing season have been
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
Page 229 and 230: temperature required for a differen
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Page 235 and 236: quantities of sugar accumulated in
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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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