Analysis of genetic diversity among some Persian walnut genotypes (Juglans regia L.) using morphological traits and SSRs markers

There is a high diversity among cultivated walnut trees in Iran due to its long time of seed propagation and vast area of cultivation. In this study some morphological characters as well as Simple Sequence Repeat (SSRs) markers were used to analyze the genetic diversity and relationships among 31 Iranian walnut genotypes along with four foreign cultivars. The nut weight ranged from 7.52 to 17.73 g, kernel weight from 4.00 to 9.83 g, and kernel percentage ratio from 38.78 to 67.05% among studied genotypes. In SSRs analysis, nine primer pairs were tested that produced 39 alleles ranging from 2 to 8, with a mean value of 5.10 allele per primer. The Iranian genotypes showed relatively high diversity both for their SSRs loci and morphological traits. Although the foreign cultivars (‘Serr’, ‘Vina’, ‘Franquette’ and ‘Lara’) clustered with each other, they also laid close and within the Iranian genotype. The results of the study provided us with valuable diversity among our genotypes which could be used for breeding studies and also showed the power of genetic markers for analysis and evaluation of this diversity.     

DNA fingerprinting of olive varieties in Istria (Croatia) by microsatellite markers

The Istria region, where olives have been cultivated for many centuries, is characterized by a considerable variety of microclimates. The study of varieties traditionally cultivated in Croatian Istria and their relationships with varieties in historically and geographically connected regions is very important in order to identify native olive germplasm, well adapted to local conditions, and to characterize the oil of regional origin. Twelve olive microsatellite markers were used for identification and differentiation of a set of 27 olive accessions grown in Istria (Croatia). Among the 27 accessions, 18 different SSR profiles were discriminated. All 12 microsatellite markers analysed were polymorphic, revealing a total of 81 alleles. The number of alleles per locus ranged from four to nine. This is the first molecular characterization of olive germplasm in Croatian Istria. The analysis clarified the genetic relationships of varieties native to Croatian Istria with introduced olive varieties, as well as with varieties in the neighbouring Slovene Istria region. Numerous varieties in neighbouring regions showed high similarity and a few cases of synonymy (‘Bilica’-‘Bjankera’; ‘Buga’-‘?rna’) and one Croatian-Slovenian homonymy (‘Bu?a’-‘Buga’) were observed. The results provide useful information for a native germplasm survey and can be used for the construction of a unique database comprising all olive varieties in the Istrian region of Croatia and Slovenia.     

Molecular characterization and genetic relationship among almond cultivars assessed by RAPD and SSR markers

RAPDs and SSRs were used to study the genetic diversity of Iranian almond cultivars and their relationship to important foreign cultivars and three related species. Eight unidentified almond Shahrodi cultivars and three wild almonds (Prunus communis, Prunus orientalis and Prunus scoparia) were also included. Of the primers tested, 42 (out of 80) RAPD and 18 (out of 26) SSR primers were selected for their reproducibility and high polymorphism. A total of 664 polymorphic RAPD bands were detected out of 729 bands. The number of presumed alleles revealed by the SSR analysis ranged from 3 to 10 alleles per locus with a mean value of 6.64 alleles per locus. Both techniques discriminated the genotypes very effectively, but only RAPDs were able to discriminate the cultivars Monagha and Sefied. Results demonstrated an extensive genetic variability within the tested cultivars as well as the value of SSR markers developed in peach for characterization of almond and related species of Prunus. Dice similarity coefficient was calculated for all pair wise comparisons and was used to construct a UPGMA dendrogram. For both markers a high similarity in dendrogram topologies was obtained although some differences were observed. All dendrograms, including that obtained by the combined use of both the marker data, depicted the phenetic relationships among the cultivars and species, depending upon their geographic region and/or pedigree information. Almond cultivars clustered with accession of P. communis showing their close relationship. P. orientalis and P. scoparia were clustered out of the rest of P. dulcis.     

Evaluation of the genetic diversity of Asian peach accessions using a selected set of SSR markers

A total of 94 peach accessions from the Zhejiang province of China were analyzed using 34 polymorphic single-locus simple sequence repeat (SSR) markers. Genetic distance analysis divided the accessions into two major clusters, one with mainly local accessions from the Fenghua region. Preselected lines from some crosses were exclusively clustered with the local ones, possibly related to maintaining the taste quality of the fruit. The number of alleles per locus at most loci was two or three, with an average of 2.85. The value of observed heterozygosity varied from 0.05 to 0.84 (average of 0.48). Diversity within the introduced accessions was higher than that of the Fenghua local accessions. Of the accessions analyzed in this study, 94% were individually identified. Those that could not be differentiated were all derived from the ‘Yulu’ cultivar, being either mutations or of identical origin.     

Effectiveness of SSR molecular markers in evaluating the phylogenetic relationships among eight Actinidia species

The aim of the present study is to evaluate a set of microsatellites and their effectiveness in determining the phylogenetic relationships among Actinidia species. A set of 14 genotypes belonging to eight Actinidia species were subjected to PCR using SSRs as molecular markers. The PCR products were analyzed on denaturing polyacrylamide gels. The discriminating ability of SSRs was based on the number of alleles and the indices DI, I and PIC. All SSR primer pairs were polymorphic and identified a total number of 61 alleles corresponding to an average of 7.8 alleles per locus. Data showed that the di-nucleotide microsatellites were more polymorphic, than the tri-nucleotide and penta-nucleotide, and were more efficient in establishing genetic similarities. The genetic similarity of the genotypes calculated with Jaccard and/or Dice similarity coefficients varied from 0.100 to 0.579, indicating a broad genetic base for the genetic material tested. Both similarity matrices clustered either with UPGMA or NJ, produced similar topology with minor clustering differences among genotypes. Thus, the eight species were clustered in two main groups and each main group in two subgroups. Data showed a close genetic relationship among Actinidia chinensis and Actinidia deliciosa species. The same conclusions were, also, drawn using the principal coordinate analysis.     

Characterization of variability and genetic similarity of European pear using microsatellite loci developed in apple

Seven microsatellite loci (SSR) developed in apple were used for the identification of 63 European pear cultivars. A total of 46 fragments were amplified, with an average of 6.6 alleles per SSR. Only one microsatellite amplified more than one locus. The mean expected and observed heterozygosities over the six single-locus SSRs averaged 0.68 and 0.44, respectively, and the number of effective alleles per loci was 3.43. The amplified fragments produced 61 different fingerprinting patterns that allowed to unequivocally distinguish all the varieties analyzed. Only two varieties, which derive from mutations, could not be distinguished from the original variety. Cluster analysis of the estimated genetic similarity, grouped the varieties according to their pedigree and their geographic origin. The variability detected with the SSRs in European pear varieties was low when compared with the variability detected in other fruit crops in the Rosaceae.     

Isolation and characteristics of eight novel polymorphic microsatellite loci from the genome of garlic (Allium sativum L.)

Here we characterized eight novel polymorphic SSR markers, developed from an enriched genomic library of garlic (Allium sativum L.). These SSRs produced a total of 64 alleles across 90 garlic accessions, with an average of 8 alleles per locus. Values for observed (H_O) and expected (H_E) heterozygosity ranged from 0.16 to 0.77 (mean = 0.44) and from 0.22 to 0.86 (mean = 0.65), respectively. Six loci deviated significantly (P < 0.05) from Hardy–Weinberg equilibrium (HWE). The averages of gene diversity and PIC values were 0.65 and 0.62, respectively. The mean genetic similarity coefficient was 0.4380, indicating that among garlic accessions existed wide genetic variation. Based on 64 alleles obtained by 8 SSRs, a phenogram was constructed to understand the relationships among the 90 accessions. These newly developed SSRs should prove very useful tools for genotypes identification, assessment of genetic diversity and population structure in garlic.     

Rose (Rosa hybrida L.) EST-derived microsatellite markers and their transferability to strawberry (Fragaria spp.)

The ‘Genome database for Rosaceae (GDR)’ provides a large collection of expressed sequence tags (ESTs) harboring simple sequence repeats (SSRs) from several Rosaceae genera, including Rosa (rose). Primer pairs flanking SSR were designed for 312 unique Rosa ESTs based on GDR database. Eight rose (Rosa hybrida L.) genotypes were tested for PCR amplification, and 287 (92%) of the primer pairs generated allele-specific PCR bands that were readily scored. From 183 (63.7%) primer pairs that evidenced polymorphic alleles among the eight rose cultivars, 20 pairs evidencing EST sequence homology to known gene functions and high levels of polymorphism were selected and utilized for DNA fingerprinting and genetic diversity assessments of 47 rose hybrids. A total of 202 polymorphic bands were scored and generated unique fingerprints for each rose hybrid. The Nei–Li genetic similarity coefficients among 1081 pair-wise comparisons of 47 cultivars exhibited a broad range of genetic variations from 0.30 (‘Grand King’ and ‘Carnival’) to 0.99 (‘First Red’ and ‘Red Champ’). UPGMA cluster analysis divided 47 hybrids into five major groups and two sub-groups. The cross-species transferability of 273 EST-SSR primer pairs was evaluated using four genotypes of the strawberry, a genus member of the Rosaceae family. PCRs on the DNA samples of strawberry were successful for 165 primer pairs; among these, 123 pairs amplified 243 polymorphic bands. As surrogates of the marker transfer, the phenetic relationship among the four strawberry genotypes was evaluated. Genetic similarity coefficients varied from 0.78 (‘Maehyang’ and ‘Janghyee’) to 0.64 (‘Janghyee’ and ‘Pragana’). The results of cluster analysis showed that the three octaploid strawberry cultivars were quite similar, whereas the diploid ‘Pragana’ was related distantly at the genomic DNA level. The EST-SSR markers developed in the present study can be efficiently utilized for genetic diversity studies in Rosaceae.     

Intra-cultivar variability of three major olive cultivars grown in different areas of central-southern Italy and studied using microsatellite markers

A collection of 70 olive samples, originating from diverse areas in central-southern Italy (Abruzzo, Apulia, Calabria, and Umbria) and corresponding to 3 major cultivars denominations (‘Carolea’, ‘Coratina’ and ‘Frantoio’), was genotyped at 10 microsatellite loci. In total, 44 alleles with a mean number of 4.4 alleles per locus were detected. The molecular analysis, allowed the study to show a clear genetic diversity between the three cultivars ‘Carolea’, ‘Coratina’ and ‘Frantoio’ and to state that ‘Carolea’ is a polyclonal cultivar, while ‘Coratina’ and ‘Frantoio’, are probably monoclonal ones. The analysis of intra-varietal polymorphism, through the SSR analysis, proved to be very useful both for varietal identification and for intra-varietal ones. Our work shows that the current designations of olive cultivars fall short of describing the genetic variability among economically important plant material. A thorough investigation of the existing variability will prove of major importance for both management and economic production of olive trees.     

Genetic diversity in local Tunisian pears (Pyrus communis L.) studied with SSR markers

Few records are available about local Tunisian pear cultivars characterized by low chilling requirements and adaptation to dry conditions. In this work, seven SSRs derived from apple were successfully transferred to 25 local Tunisian pear genotypes and 6 common varieties of Pyrus communis cultivated in Europe. The 7 SSRs used amplified a total of 36 fragments. All the microsatellites except one seem to amplify more than one locus in some of the genotypes studied. Only 12 different fingerprinting patterns could be distinguished among the 25 Tunisian cultivars studied indicating a high number of synonymies. The mean expected and observed heterozygosities in the 25 Tunisian cultivars analyzed averaged 0.71 indicating a high level of genetic diversity among the local Tunisian pear germplasm. These markers will be useful to optimize the conservation of this highly threatened germplasm.     

Genome composition and genetic diversity of Musa germplasm from China revealed by PCR-RFLP and SSR markers

Banana (Musa spp.) is one of the most important crops in the world. In this study, 216 banana accessions, 184 from the National Banana Germplasm Collection of China (NBGCC) and 32 from the International Network for the Improvement of Banana and Plantain (INIBAP), were used to determine the genome composition of banana plants in these collections and to estimate their genetic diversity. The genome composition was examined using PCR-RFLP markers. The molecular data for all but one accession (ITC 1231) from INIBAP were in agreement with the initial records based on phenotypic characteristics. Microsatellite (SSR) markers were used to investigate the genetic variability and relationships among these banana accessions. Ten of the 47 primer pairs tested consistently produced reproducible and discrete fragments. We identified a total of 92 alleles, ranging from 5 to 15 per locus. The genetic similarity between the accessions ranged between 0.1 and 1, when estimated using Jaccard's coefficient. The UPGMA method based on genetic similarities, grouped the NBGCC accessions according to those containing the ‘A’ and ‘B’ genomes. However, this analysis could not separate all the accessions, especially the somatic mutations, using the primers in this study. These data indicated that limited genetic variation exists within these accessions and the collections of NBGCC should include a much wider range of banana plant material.     

桃同株花果茎叶杂色材料的获得和特征特性初探

在观赏桃育种过程中获得了1 份特殊材料‘PCM-1’,花为粉-白杂色,叶片为紫红-绿杂色,嫩茎表面有紫红色条带或细点,幼果表面有不规则红色斑块或细点。从杂色性状的形态学特性、无性繁殖表现和有性遗传规律对其进行了初步探索,并进行了其亲本和无性系的SSR 鉴定。连续6 年观察发现‘PCM-1’的杂色性状稳定;徒手切片观察到杂色均出现在表层的第一层细胞。嫁接繁殖表明杂色性状稳定（97.4%）,但无性系中也能分离出较低比例的纯色株系（2.6%）;花、果、叶、嫩茎4 个部位的杂色具有同步性。以‘PCM-1’为母本,与花色纯白、叶色纯绿的父本杂交,后代中能够出现12.2% ~ 24.7%的杂色单株。通过SSR 鉴定发现‘PCM-1’的母本为‘红粉佳人’;经324 个SSR 标记鉴定,‘PCM-1’（杂色）、‘PCM-1R’（纯红）和‘PCM-1G’（纯绿）3 个无性系间未发现等位基因差异。     

利用苹果SSR引物分析山楂属植物遗传关系

SSR引物在不同物种间具有通用性,从141对苹果属(Malus spp.)SSR引物中筛选出10对适合于山楂属(Crataegus spp.)植物的SSR引物,并对8个种37份山楂种质资源的遗传关系进行了分析。10对SSR引物共检测到91个多态性谱带,每个位点的等位基因数为3～13个,平均为9.1个。位点杂合度为0.432～0.790,平均为0.688。10对SSR引物可以将20份山楂资源区分开,17份不能区分的资源分为3组,第1组为3个伏山楂品种,第2组和第3组分别包括大果山楂的2个和12个品种。基于SSR标记构建的聚类树状图将供试37份山楂资源分成2个类群,第1类群包括6个山楂野生种,第2类群包括供试的所有伏山楂、山楂和大果山楂资源。该聚类结果与传统形态学分类一致。     

桃野生种和地方品种种质资源亲缘关系的AFLP分析

利用AFLP技术对94份野生桃种质资源进行了DNA多态性分析,从64对E+2/M+3引物组合中筛选出9对引物用于扩增基因组DNA,共获得清晰可辨的236个标记,其中多态性标记为141个,多态性检出率为59.9%。UPGMA聚类结果显示,在遗传距离约为0.3267时,普通桃、陕甘山桃、光核桃3个种被聚为一类,与这3个种相近的为新疆桃,其次为甘肃桃,最远为山桃(白花山桃、红花山桃)。在遗传距离0.2178处,五月鲜扁干、白黏胡桃和珲春桃3份材料各自聚为一类。在遗传距离约0.1997处,普通桃又可明显分为4个组:组I包括原产于南方和西北地区的地方品种,且大部分南方蟠桃主要聚在此类;组II仅有油桃(甘肃酒泉)一个野生材料;组III包括野生毛桃和大部分原产于北方的地方品种,北方硬肉桃主要聚在这一类;组IV由白碧桃和白花山碧桃2个观赏桃品种组成。     

SSR fingerprinting Chinese peach cultivars and landraces (Prunus persica) and analysis of their genetic relationships

Thirty-two Chinese peach landraces/cultivars, a major subset of the core Chinese peach collection, were fingerprinted using seven pairs of SSR primers to assess their genetic diversity and relatedness. The seven primer pairs detected eight loci and revealed an allele richness of 3.125 (average alleles per locus), an expected heterozygosity (He) of 0.450, and a Shannon index of 0.728 among the landraces/cultivars. This level of genetic diversity is lower compared to other fruit trees and Prunus congenus species (cherry and apricot), but it is comparable to previous reports in peaches. A greater level of genetic diversity was observed in landraces than in cultivars, indicating that peach landraces are valuable for germplasm collection. All cultivars and landraces, except two, were unambiguously identified based on multi-locus genotypes. Eight unique alleles were detected among this group of Chinese peaches. UPGMA clustering analysis separated the 32 cultivars/landraces into two distinct groups, which is generally in accordance with the known pedigree information. The results provide accurate genetic information for defined acquisition policy in the repositories, improving the integrity and efficiency of germplasm management and giving evidences for protection of breeder's intellectual rights.     

Assessment of genetic diversity among mango (Mangifera indica L.) genotypes using RAPD markers

Knowledge about the extent of genetic diversity/relatedness in mango germplasm is vital for developing coherent strategies for future gains in productivity. The genetic diversity/relatedness among mango cultivars/genotypes developed in Pakistan has not been investigated previously. We have assessed the genetic diversity among 25 mango genotypes/cultivars using randomly amplified polymorphic DNA (RAPD). Sixty random ten-mer primers were surveyed, out of which 45 yielded amplicons in all the genotypes. Genetic similarity between genotypes/cultivars was in the range of 64–89% with an average of 74%. Similarly, the genetic relatedness among all variants derived from a mango cultivar Chaunsa was in the range of 81.18–88.63%. These coefficients were utilized to construct a dendrogram using the unweighted pair group of arithmetic means (UPGMA). The genotypes were grouped into three (A, B, C) clusters. Generally, genotypes originating from Pakistan were grouped in cluster ‘A’ while cluster ‘B’ primarily composed of southern India as well as Florida cultivars. Kensington Pride was the most distantly related genotype which grouped with Maya and Yakta, forming a distinct cluster ‘C’.     

桃种质资源对桃蚜的抗性评价

通过对桃属(Prunus L．)植物5个种419份资源进行抗桃蚜（Myzus persicae）性的田间自然鉴定和初选出的34份抗性资源的人工接种鉴定，结果表明：山桃(P．davidiana)的抗性最强，甘肃桃(P．kansuensis)的抗性最弱 不同品种的抗蚜性差异很大，按照品种群划分，其蚜害指数寿星桃20．00、白桃45．70、油桃77．00、黄桃60．16、蟠桃77．86 筛选出15份抗性资源，这些资源可分为三类 山桃、寿星桃、碧桃。     

早熟桃新品种‘锦玉’

 ‘Jinyu’is a early-ripening peach cultivar derived from the cross of‘Zhaohui’בOkayama Wase’. Its average fruit weight is 167 g,and the biggest one is 210 g. The fruit is oblong with red blush. The flesh is white,melting,sweet and semi-freestone. The soluble solids content is 13.2%. The average fruit development period is 72 days.     

应用SSR标记进行部分黄肉桃种质鉴定和亲缘关系分析

以116份黄肉桃种质和19份其它桃或桃近缘种种质为试材,利用定位于李属参考图谱上的SSR标记,进行遗传多样性评价及亲缘关系分析。SSR扩增结果表明,筛选出的28对SSR引物共扩增出206个等位基因,其中多态性等位基因159个（占76.56%）。聚类结果表明,在相似系数为0.82时,黄肉桃资源在聚类图上呈两组：美洲、欧洲和亚洲育成黄桃品种聚为一组（Ⅰ）,中国地方黄桃品种中唯‘西安杏瓤桃’和‘天津黄肉’出现在这一组;来源于云南、华北、西北地区的中国地方黄桃品种聚为另一组（Ⅱ）。在相似系数为0.87时,育成黄桃品种（Ⅰ）分为8个亚组,美洲育成品种分布在所有8个亚组中,欧洲育成品种分布在Ⅰ-2、Ⅰ-3和Ⅰ-4亚组,而亚洲育成品种仅分布在Ⅰ-3和Ⅰ-4亚组;亲本相同的育成品种在聚类图中临接或出现在较近的位置,育成品种的聚类结果与品种的系谱关系基本吻合。在相似系数为0.87时,中国地方黄桃品种分为7个亚组,西南黄桃和西北黄桃存在一定的组群界限,来自西南的黄桃地方资源主要分布在Ⅱ-1和Ⅱ-5亚组,而来自西北的黄桃地方品种主要出现在Ⅱ-2、Ⅱ-4和Ⅱ-7亚组。品种群的聚类结果表明,亚洲育成品种与美洲育成品种间亲缘关系最近,其次为欧洲育成品种;中国地方黄桃品种与育成品种间亲缘关系较远。