Grape Accessions Research Articles

Genetic diversity of wild and cultivated grapevine accessions from southeast Turkey.

Wild grapevine genetic diversity in southeast Turkey has not been documented to date. In the present work, in order to clarify the relationships between wild and cultivated grape accessions from southeastern Turkey, 22 nuclear and three chloroplast microsatellite loci were used on 21 wild grapevine Vitis vinifera L. ssp. sylvestris (Gmelin) and 13 cultivated grapevine Vitis vinifera ssp. sativa accessions. The number of alleles per SSR locus ranged from 4 (VVIn16) to 20 (VVIv67) and the mean allele number per locus was 10.09. Expected locus heterozygosity ranged from 0.586 (locus VVIb01) to 0.898 (locus (VVIv67)). The three cpSSR molecular markers presented variation in size both in cultivars and in wild Turkish accessions. Two size variants were detected for cpSSR3 (106 and 107 bp) for cpSSR5 (104 and 105 bp), and for cpSSR10 (115 and 116 bp). The six alleles in wild grapevines fell into three haplotypes B, C and D. A genetic structure according to accessions taxonomic status (wild or cultivated) was revealed by UPGMA analysis. This highlighted a clear separation between domesticated and wild accessions in Turkish germplasm. The results pointed out the need to further collect and characterize this wild and cultivated grapevine germplasm.

ANALYSIS OF GRAPE ROOTSTOCKS BY MICROSATELLITE MARKERS

The aim of this work was to determine the SSR profiles of 96 Vitis accessions (mainly rootstocks) for 19 loci, and to find genetic relatedness among them. Most molecular markers used in the last 30 years are not affected by the environment, giving a good tool for determination of genetic distances. Based on our earlier experience, SSR analysis was carried out on 96 grape accessions, mainly rootstocks. Analyses were carried out with 19 microsatellite loci. The loci were selected to cover all of the linkage groups to get a more precise impression of the genome of grape rootstocks. Results of SSR analyses and a dendrogram showing genetic relatedness are presented. Based on the results it can be established that most of the loci showed appropriate polymorphism. High similarity was detected between the clones of T5C and TK5BB. The Vitis sylvestris accessions showed close relatedness with the Vitis vinifera L. cultivars. Our results indicate a high polymorphism between grape rootstock accessions. Strong differences were detected between the so called ?clones? of T5C and TK5BB, suggesting, that they are not real clones, but different genotypes with highly similar morphological features.

BRAZILIAN GRAPE GERMPLASM BANK: PHENOLOGY AND INCIDENCE OF MAIN FUNGAL DISEASES

In the last years, viticulture has reached a very important role in Brazilian fruit production, not only in temperate zones, but also as an alternative for tropical regions. These different climates require cultivars with wide-ranging production cycles. In Brazil, phytosanitary treatment can reach 30% of production costs. Genetic breeding can play a role in the development of new cultivars with different production cycles and more tolerance to the main fungal diseases. The purpose of this work is to evaluate the phenology and disease incidence of grape accessions of the Brazilian germplasm bank in order to give support to the breeding program. For ten years, 700 accessions were evaluated and classified regarding growth cycle length as very early (0,6%), early (13,9%), medium (43,6%), late (41,8%) or very late (0,3%). In the same period, 1,100 accessions were evaluated for disease resistance. The Brazilian grape germplasm bank maintains resistance sources to the main fungal grape diseases that occur in the country. However, resistance to downy mildew and anthracnose were less widespread in the studied sample. More information about the Brazilian grape germplasm bank can be found on the web (http://www.cnpuv.embrapa.br/prodserv/ germoplasma/). These results are assisting the development of new grape cultivars, in order to give support to the evolution and expansion of Brazilian viticulture.

Characterization of Grape (Vitis vinifera L.) Berry Sunburn Symptoms by Reflectance

Sunburn can affect grape quality both for chemical modifications and by visual impact of the browning. Optical properties of 17 white grape accessions were investigated in the visible region using a noninvasive instrument. Reflectance spectra were obtained using a Jaz System spectrometer. Browning was induced by exposing grape bunches to direct sunlight at 12:30 p.m. for 5 h. During the experiment, the global solar radiation ranged from 2.6 to 2.7 MJ m-2 h-1 and the air temperature from 24.3 to 29.2 °C; the exposed berries reached a temperature of 34.2 °C in comparison to the 30.4 °C of the shaded ones. Differences between the spectra of controlled and exposed berries mainly emphasized the loss in chlorophyll and the formation of brown compounds. A positive correlation between the chlorophyll concentration and berry browning was proposed. Developing rapid, noninvasive, and low-cost methods based on reflectance spectroscopy could support grapevine variety characterization with respect to sunburn susceptibility as well as study of the physiological processes involved in the symptoms' appearance.

Genetic Characterization of Grape Cultivars from Apulia (Southern Italy) and Synonymies in Other Mediterranean Regions

Forty-five grape accessions, traditional and historically mentioned in Apulia (southeastern Italy), were genotyped at 13 microsatellite (SSR) markers and observed for their morphological features with the aim of characterizing and identifying the local grape diversity relevant for economic or historical significance and for endangered germplasm conservation. Twelve of the 45 accessions examined were found to be synonyms or somatic mutants, leaving 33 distinct genotypes. Attempts were then made to verify the true identities of the accessions investigated and to determine their appropriate denominations. This entailed comparing them with published allelic profiles and morphological features of cultivars from Apulia and from surrounding areas linked historically to the region. While confirming the identity of the major Apulian cultivars, further matches with varieties from other Mediterranean regions were revealed. Approximately half of the Apulian cultivars investigated were found to have a foreign counterpart mainly along the Adriatic Sea (Croatia), in Greece, or in other southern Italian regions. The new synonymies found with cultivars traditional to other areas shed light on the migration of cultivars following the settlement of colonies and the historical establishment of Mediterranean trade routes.

A Modern Ampelography: A Genetic Basis for Leaf Shape and Venation Patterning in Grape

Terroir, the unique interaction between genotype, environment, and culture, is highly refined in domesticated grape (Vitis vinifera). Toward cultivating terroir, the science of ampelography tried to distinguish thousands of grape cultivars without the aid of genetics. This led to sophisticated phenotypic analyses of natural variation in grape leaves, which within a palmate-lobed framework exhibit diverse patterns of blade outgrowth, hirsuteness, and venation patterning. Here, we provide a morphometric analysis of more than 1,200 grape accessions. Elliptical Fourier descriptors provide a global analysis of leaf outlines and lobe positioning, while a Procrustes analysis quantitatively describes venation patterning. Correlation with previous ampelography suggests an important genetic component, which we confirm with estimates of heritability. We further use RNA-Seq of mutant varieties and perform a genome-wide association study to explore the genetic basis of leaf shape. Meta-analysis reveals a relationship between leaf morphology and hirsuteness, traits known to correlate with climate in the fossil record and extant species. Together, our data demonstrate a genetic basis for the intricate diversity present in grape leaves. We discuss the possibility of using grape leaves as a breeding target to preserve terroir in the face of anticipated climate change, a major problem facing viticulture.

Polymorphisms inVvPelassociate with variation in berry texture and bunch size in the grapevine

Background and Aims The identification of the genes involved in traits of interest is one of the main aims of current plant genetic studies. Although berry texture is a quality trait of great importance in table grapes, no quantitative trait loci or related gene has been described for this trait so far. Methods and Results In this work, the gene VvPel, which codes for a pectate lyase in the grapevine, was selected as a candidate gene for an association study. This gene was sequenced in a core collection of 96 table grape accessions, which was also characterised for several bunch and berry morphological traits, including diverse texture parameters measured by means of a texture meter. The population structure, the nucleotide and haplotype diversity, the protein structure, the existence of selection, and the linkage disequilibrium (LD) were evaluated. Thirty-two single nucleotide polymorphisms (SNPs) and 15 haplotypes were identified in the VvPel sequence, and LD was low enough to provide a high-resolution power. Some of the polymorphisms associated significantly with texture parameters and with bunch size, explaining part of the variation found for those traits. Conclusions Among the polymorphisms found in the gene VvPel, S1027 and S405/S441 in relation to berry texture, and S48/S1030 in relation to bunch size, are the most promising and would need to be investigated further as putative causal polymorphisms or markers for the respective traits. Significance of the Study The present work is the first genetic study on berry texture in the grapevine, and constitutes a starting point for a deeper understanding of the genetic basis of the natural variation for berry texture and bunch size.

Microsatellite analysis to rationalize grape germplasm in India and development of a molecular database

Three hundred and seventeen grape accessions from the National Active Grape Germplasm Site in India were analysed with 25 microsatellite markers. A total of 411 alleles were detected, of which 42% were rare alleles. Unique alleles for 56 genotypes were also identified. The analysis of microsatellite data identified 63 duplicate accessions and only 254 accessions were unique genotypes. Several cases of misnomers, synonymy and homonymy were identified. Parental genotype for a few clonal selections was ascertained. Population structure analysis grouped 254 unique genotypes into four major clusters. The analysis also revealed the presence of admixtures with only 79% of pure ancestry. A core collection comprising 80 genotypes was identified, which represented all the alleles and genetic diversity. A user-friendly and interactive computer application software was developed for storage and the retrieval of molecular data. A molecular database for the 254 genotypes was created. This analysis will help in the rationalization and better management of germplasm. Information on genetic diversity and population structure will form the basis for varietal improvement programmes.

VvGAI1 polymorphisms associate with variation for berry traits in grapevine

Gibberellins have important effects over different developmental processes in plants and the exogenous application of GA3 has been extensively used in grapevine to improve certain quality traits (to control berry size, set and weight, to decrease cluster compactness and to induce the absence of seeds in grapes). In this study, we performed association analyses using a core collection of 127 table grape accessions in order to identify causal SNPs in the gene VvGAI1, a negative regulator of gibberellins response. Seventeen quantitative descriptors, related with fertility, bunch and berry traits were used for the association analysis with VvGAI1. Fifteen polymorphisms and 15 haplotypes were identified, and some of them associated significantly with important traits for table grape: berry texture, juice yield, and bunch weight. The genetic structure found in the collection of varieties was consistent with their origins, and was included in the association test. In addition, nucleotide and haplotypic diversity, linkage disequilibrium and the possible existence of selection over the gene were also evaluated to support the association test.

Genetic diversity and differentiation within and between cultivated ("Vitis vinifera" L. ssp "sativa") and wild ( "Vitis vinifera" L. ssp "sylvestris") grapes

Genetic characterization of 502 diverse grape accessions including 342 cultivated ( V. vinifera ssp. sativa ) and 160 wild ( V. vinifera ssp. sylvestris ) grapes showed considerable genetic diversity among accessions. A total of 117 alleles were detected across eight SSR loci with the average of 14 alleles per locus. The genetic diversity of wild grapes was slightly lower than that observed in the cultivated grapes probably due to small populations and severe natural selection leading to drift and loss of alleles and heterozygosity in wild grapes. The distance cluster analysis (CA) supported the classical ecogeographic groups with moderate genetic differentiation among them. There was a greater affinity of Occidentalis grape to wild grape from the Caucasus than other groups. However, a number of low to moderate frequency alleles that are present in the cultivated grape are not represented in the wild grape.

Genetic relationship between Chinese wild Vitis species and American and European Cultivars based on ISSR markers

abstract Inter-simple sequence repeat (ISSR) markers were employed to detect the genetic diversityamong 70 grape accessions including 52 clones of 17 Chinese wild grape species, seveninterspecific hybrids, 10 Vitis vinifera L. cultivars, and one strain of Vitis riparia L. A total of119 polymorphic bands with an average of 11.9 per primer were observed. The unweightedpair-group method (UPGMA) analysis indicated that the 70 clones or accessions hada similarity range from 0.08 to 0.93, indicating that abundant diversities exist among theseaccessions. Based on cluster analysis and principal coordinate analysis, all accessions couldbe divided into two major groups, the Chinese wild grape group, and the American andEuropean cultivar group. The largest distance was found among V. riparia MichX, Vitispiasezkii, V. vinifera L. interspecific hybrid (Vitis binifera V. labrusca) and the wild grapesnative to China. 2012 Elsevier Ltd. All rights reserved. 1. IntroductionGrape(Vitis vinifera L.)is one of theoldest and mostimportant fruit crops in theworld.The Vitisgenus contains more than70 species, with centers of origin in south Europe, Asia Minor, East Asia, and North and Central America (Wan et al., 2008).China is one of the major germplasm centers of Vitis species and more than 35 Vitis species originate in China (Wan et al.,2008). Chinese wild Vitis has many useful characteristics, including (i) high quality wine making attributes, such as highsugar content and moderate acidity (Li and He, 2000); (ii) high level of resistance to fungal diseases, including PowderyMildew [Uncinula necator (Schw.) Burr.], Anthracnose [Elsinoe ampelina (de Barry) Shear], Ripe Rot [Glomerella cingulata(Ston.) Spauld et Schrenk], and Crown Gall [Agrobacterium tumefaciens Smith and Townsend] (He et al., 1991; Wang et al.,1995, 2007; Wang, 1979); (iii) highly efficient photosynthesis in V. quinquangularis (Zhu et al., 1994). In addition, Chinesewild Vitis can be easily crossed with American Vitis species, and the undesirable “foxy” flavor compounds in the berries ofAmerican Vitis species is nonexistent in Chinese wild Vitis species (He, 1999b; Wan et al., 2008). The selection for thesedesirable characteristics has enlarged the scope of applications for grape breeding, which results in many intermediate andtransitional types of Vitis and causes difficulties in germplasm research of the wild Chinese Vitis.

Genetic diversity and relationships of indigenous and newly bred Bulgarian grape cultivars assessed by nuclear and chloroplast markers

<p style="text-align: justify;"><strong>Aim</strong>: Assessment of the genetic diversity and relationships in a group of 31 Bulgarian grape accessions through microsatellite markers.</p><p style="text-align: justify;"><strong>Methods and results</strong>: Thirty-one accessions, including 20 old rare local and 11 newly bred varieties were characterized through 13 nuclear and 5 chloroplast microsatellite loci. The genetic diversity (0.81±0.01) obtained for the investigated group of cultivars was comparable to those reported for other grapevine germplasms. The low PI value (1.0x10<sup>-16</sup>) allowed proper genetic identification and determination of synonyms. Microsatellite analysis of the 31 accessions resulted in 26 unique genotypes and 2 groups of synonyms. Four cases of supposed synonymy with local Bulgarian and foreign cultivars were rejected. Three chlorotypes, B, C and D, were defined among the studied cultivars, with a prevalence of chlorotype C (62%).</p><p style="text-align: justify;"><strong>Conclusion</strong>: The high genetic diversity found in the set of old rare grapevines demonstrated their importance as a rich source of alleles for breeding. The pattern of chlorotype distribution observed among local varieties confirmed the previous results and supports the hypothesis of an Eastern origin of local Bulgarian cultivars.</p><p style="text-align: justify;"><strong>Significance and impact of the study</strong>: The obtained results provide an important support for the preservation of grape biodiversity in Bulgaria as well as for the clarification of genetic relationships between local and foreign cultivars.</p>

Characterization of Polyphenolic Metabolites in the Seeds of Vitis Germplasm

The composition and content of polyphenols in the seeds of 91 grape accessions from 17 Vitis species were characterized. Eleven compounds, including 2 gallic derivatives, 3 monomeric flavan-3-ols, 3 flavonols, resveratrol, and procyanidin B1 and B2, were identified via HPLC-MS and quantified by HPLC-DAD. In addition, seventeen dimeric and trimeric flavan-3-ols were also quantified. Tremendous variation was observed both among and within species for these compounds. Monomeric flavan-3-ols were the most abundant polyphenols in seeds, followed by dimeric and trimeric flavan-3-ols, which collectively accounted for more than 96% of the total polyphenols. V. palmata, V. vinifera, and V. vulpina had significantly higher content of total polyphenols than other species. A number of Vitis accessions with high content of various types of seed polyphenols were identified, and they can serve as potential germplasm for improving the composition and content of seed polyphenols in cultivated grapes.

Polyphenolic composition and content in the ripe berries of wild Vitis species

To explore wild genetic resources for improving fruit and processing quality of cultivated grape cultivars, the polyphenolic composition and content in the ripe berries of 147 grape accessions from 16 Vitis species for two consecutive years were characterised. A total of 48 polyphenolic compounds, including 28 anthocyanins, 6 flavonols, 6 flavanols, 6 hydroxycinnamic derivatives, and 2 hydroxybenzoic acids, were identified via HPLC-MS and quantified by HPLC-DAD. These wild grape species had unique presence of abundant di-glucoside derivatives of anthocyanins. In addition, anthocyanins in most wild species were predominantly nonacylated. The mean contents for anthocyanins, flavanols, flavonols, hydroxycinnamic derivatives, and hydroxybenzoic acids were 9.610, 0.769, 0.093, 0.441 and 0.027mgg−1 FW, respectively. They were about 2 to 10 folds higher than their respective counterparts in the most widely cultivated grape species Vitis vinifera. As expected, most of these groups of compounds were correlated negatively with berry weight, but positively with the content of total soluble solids.

Genetic diversity in table grapes based on RAPD and microsatellite markers

The objective of this work was to analyze the genetic diversity of 47 table grape accessions, from the grapevine germplasm bank of Embrapa Semiárido, using 20 RAPD and seven microsatellite markers. Genetic distances between pairs of accessions were obtained based on Jaccard's similarity index for RAPD data and on the arithmetic complement of the weighted index for microsatellite data. The groups were formed according to the Tocher's cluster analysis and to the unweighted pair‑group method with arithmetic mean (UPGMA). The microsatellite markers were more efficient than the RAPD ones in the identification of genetic relationships. Information on the genetic distance, based on molecular characteristics and coupled with the cultivar agronomic performance, allowed for the recommendation of parents for crossings, in order to obtain superior hybrids in segregating populations for the table grape breeding program of Embrapa Semiárido.

Genetic diversity of Moroccan grape accessions conserved ex situ compared to Maghreb and European gene pools

Grape diversity present in Morocco and the part of this diversity used nowadays are poorly documented. In order to choose diversified genotypes, to select them so that their agronomic interest will be tested, a group of 21 autochthonous cultivars preserved in the germplasm collections of SODEA and 18 Moroccan cultivars from “Domaine de Vassal” INRA grape collection was compared to a group of cultivars from neighbouring countries (Algeria and Tunisia), and from a core collection optimizing simple sequence repeat (SSR) allelic diversity of grape. Data from 20 nuclear and 3 chloroplastic SSR markers were obtained for this set of 211 cultivars. A total of 156 alleles (mean of 7.8 alleles per locus) were detected for the nSSRs and 7 alleles for the cpSSR in the Moroccan group. Chlorotype diversity in Moroccan and Algerian group were similar, but slightly lower than in the Tunisian group and the core collection. Similarly, the nSSR diversity was high in the core collection and low in the Moroccan and the Algerian groups compared to the two other groups. Clustering of cultivars based on nSSR data reflected their geographical origin and, to a certain extent, the use of the cultivars. The specificity of the Moroccan plant material was attested by the Bayesian analysis using Structure, while differences of the core collection were clearly revealed both by the Bayesian and a multivariate analysis. These results confirm the differentiation of the material from Maghreb and more specifically of Moroccan material, having evolved independently from Europe.

Genetic diversity of table grape based on morphoagronomic traits

The conservation and characterization of grape (Vitis spp) genetic resources in germplasm banks have been the basis of its use in breeding programs that result in development of new cultivars. There are at least 10,000 grape cultivars kept in germplasm collection. The genetic diversity in 136 table grape accessions from the state of Bahia, Brazil, was evaluated. Continuous and discrete morphoagronomic traits were assessed. The clustering analysis by the Tocher otimization method resulted in 30 clusters (considering continuous morphoagronomic traits), and 9 clusters (taking into consideration multicategorical traits). There was no agreement between clusters obtained by both, continuous or discrete phenotypic descriptors, independent of the cluster method analysis used. A satisfactory genetic variability among the table grape accessions was observed.

Genetic diversity in Anatolian wild grapes (Vitis viniferasubsp.sylvestris) estimated by SSR markers

Anatolia (Asia Minor) is considered to be the cradle of viticulture, but wild grape accessions from this region have not been subjected to any genetic analysis. We present the first genetic characterization of wild grapes (Vitis viniferasubsp.sylvestris) from this region. Using 15 nuclear microsatellites, we genotyped 84 wild grape accessions collected from three Anatolian locations. The unweighted pair group method with arithmetic mean analysis revealed four genetic clusters that partially separated Anatolian wild grape populations. In general, accessions from geographically closer locations showed higher genetic similarities than those from more distant locations. In some cases, accessions from one population showed close genetic relationships to accessions from a different population. The genetic diversity between natural populations from both ends of the Mediterranean basin has shown higher genetic diversity in the Anatolian Peninsula than peripheral populations, suggesting that this area could be one centre of diversity of the species. The genetic relationship between wild and cultivated grapevine from Anatolian Peninsula indicated a clear separation between them. However, we cannot discarded a local genetic contribution. Overall, this study reveals useful information for management and potential utilization of Anatolian wild grape germplasm.

Comparison of genetic diversity between cultivated and wild grape varieties originating from the Near-Caspian zone of Azerbaijan

<p style="text-align: justify;"><strong>Aim</strong>: The Caucasus is considered as a possible primary centre of origin of grapevine. The aim of our research work was to study genetic diversity among cultivated Caucasian grape varieties and wild relatives originating from Near-Caspian areas of Azerbaijan on the basis of simple sequence repeat (SSR) markers.</p><p style="text-align: justify;"><strong>Methods and Results</strong>: Microsatellite allele data obtained in 31 local table grape varieties of <em>Vitis vinifera</em> L. ssp. <em>sativa</em> (Absheron region) and 34 wild varieties of <em>Vitis vinifera</em> L. ssp. <em>sylvestris</em> (Nabran, Davachi and Guba regions) were used to estimate population genetic parameters such as the polymorphism information content (PIC), the probability of identity (PI) and the frequency of null alleles (r). The genetic differences among the wild populations and between the wild and the cultivated gene pools were assessed using POPGENE, Arlequin and GENETIX programs. The selected fifteen microsatellite (SSR) markers revealed a high degree of polymorphism within and among the grape populations analyzed. The genetic similarity index ranged from 0.02 to 0.933.</p><p style="text-align: justify;"><strong>Conclusions</strong>: Among the selected markers, VVMD28 and VVMD36 displayed the highest diversity level regarding the expected heterozygosity and PIC (highest values) and PI (lowest values). In consequence, we postulate that these two markers are the most appropriate ones for the identification of grape accession and the determination of genetic diversity among cultivated and wild grape genotypes. Clustering analysis based on SSR markers data led to a good separation between cultivated and wild accessions and between wild accessions originating from different regions.</p><p style="text-align: justify;"><strong>Significance and impact of the study</strong>: The fifteen microsatellite markers used in this study were highly informative for the identification and analysis of genetic structure of Azerbaijan grapevine populations and clarified the relationships among grape accessions.</p>

Genetic diversity of a brazilian wine grape germplasm collection based on morphoagronomic traits

The objectives of this study were to evaluate the performance of cultivars, to quantify the variability and to estimate the genetic distances of 66 wine grape accessions in the Grape Germplasm Bank of the EMBRAPA Semi-Arid, in Juazeiro, BA, Brazil, through the characterization of discrete and continuous phenotypic variables. Multivariate statistics, such as, principal components, Tocher's optimization procedure, and the graphic of the distance, were efficient in grouping more similar genotypes, according to their phenotypic characteristics. There was no agreement in the formation of groups between continuous and discrete morpho-agronomic traits, when Tocher's optimization procedure was used. Discrete variables allowed the separation of Vitis vinifera and hybrids in different groups. Significant positive correlations were observed between weight, length and width of bunches, and a negative correlation between titratable acidity and TSS/TTA. The major part (84.12%) of the total variation present in the original data was explained by the four principal components. The results revealed little variability between wine grape accessions in the Grape Germplasm Bank of Embrapa Semi-Arid.