Genetic variability in Brazilian wheat cultivars assessed by microsatellite markers

Wheat (Triticum aestivum) is one of the most important food staples in the south of Brazil. Understanding genetic variability among the assortment of Brazilian wheat is important for breeding. The aim of this work was to molecularly characterize the thirty-six wheat cultivars recommended for various regions of Brazil, and to assess mutual genetic distances, through the use of microsatellite markers. Twenty three polymorphic microsatellite markers (PMM) delineated all 36 of the samples, revealing a total of 74 simple sequence repeat (SSR) alleles, i.e. an average of 3.2 alleles per locus. Polymorphic information content (PIC value) calculated to assess the informativeness of each marker ranged from 0.20 to 0.79, with a mean of 0.49. Genetic distances among the 36 cultivars ranged from 0.10 (between cultivars Ocepar 18 and BRS 207) to 0.88 (between cultivars CD 101 and Fudancep 46), the mean distance being 0.48. Twelve groups were obtained by using the unweighted pair-group method with arithmetic means analysis (UPGMA), and thirteen through the Tocher method. Both methods produced similar clusters, with one to thirteen cultivars per group. The results indicate that these tools may be used to protect intellectual property and for breeding and selection programs.     

Genetic diversity of different apricot geographical groups determined by SSR markers.

Forty apricot cultivars with different geographic origins belonging to the germplasm collections of St. Istvan University (Budapest, Hungary) and the Instituto Valenciano de Investigaciones Agrarias (IVIA) (Valencia, Spain) were studied by means of SSR markers. The aim of the study was to determine the genetic relationships among genotypes from different eco-geographical groups. Sixteen primer pairs flanking microsatellite sequences in the peach genome were assayed. Eleven of them were polymorphic in the set of cultivars studied and allowed every genotype to be unambiguously distinguished. Genetic diversity in the population studied was analyzed using several variability parameters. A total of 34 alleles were detected with a mean value of 3.1 alleles/locus. The expected heterozygosity mean was 0.46 and the observed heterozygosity was 32% on an average leading to a high value of the Wright's fixation index (0.32). Additionally, UPGMA cluster analysis based on Nei's genetic distance grouped genotypes according to their geographic origins and pedigrees. SSR markers have proved to be an efficient tool for fingerprinting cultivars and conducting genetic-diversity studies in apricot.     

Genetic identification of 91 poplar cultivars based on SSR markers

Accurate genetic identification and relationship analysis of poplar cultivars is necessary to establish commercial poplar plantations and select suitable breeding strategies. In this study, 91 poplar cultivars belonging to four sections (Aigeiros, Tacamahaca, Populus and Turanga) and inter/intra-sectional hybrids were genotyped using 18 polymorphic simple sequence repeat (SSR) markers. In total, 222 alleles were amplified with an average of 12.3 alleles per marker. The mean polymorphic information content and power of discrimination were 0.706 and 0.813, respectively. Five SSR markers (ORPM_103, ORPM_247, GCPM_1048, GCPM_1255 and LG_X_19) constituted a core fingerprint and were sufficient to identify all the tested cultivars. With some notable exceptions, cultivars of the same species generally clustered together in cluster (UPGMA) and ordination (PCO) analyses. Flow cytometry indicated that 11 poplar cultivars were triploid. Among these, seven had three alleles at some loci, suggesting that SSR markers could indicate the ploidy level to some extent. This study provides useful genetic information for the identification and protection of poplar cultivars in China and offers a guideline for the selection of poplar crossing parents based on ploidy level and genetic relationships.     

Genetic variability of Old Portuguese bread wheat cultivars assayed by IRAP and REMAP markers

Retrotransposons (RTNs) constitute informative molecular markers for plant species as a result of their ability of integrating into a multitude of loci throughout the genome and thereby generating insertional polymorphisms between individuals. Inter-retrotransposon amplified polymorphisms (IRAPs) and the retrotransposon-microsatellite amplified polymorphisms (REMAPs) are marker systems based on long terminal repeats (LTRs) RTNs, developed for plants, that have been widely used for evolution, genetic diversity, DNA fingerprinting of cultivars and varieties, genetic mapping linkage and for detection of genetic rearrangements induced by polyploidisation. In the present study, we aimed to analyse the genetic variability among 48 Old Portuguese bread wheat cultivars using both IRAP and REMAP markers. Five IRAP and six REMAP primer combinations were used. IRAP produced 103 polymorphic fragments in a total of 113 bands. On average, 22.6 bands were amplified per IRAP primer combination. The bands ranged in size from 250 to 5000 bp. The REMAP primer combinations allowed the amplification of 53 bands, 51 of them polymorphic. An average of 8.8 REMAP bands was scored per primer combination. The REMAP bands ranged from 250 to 3000 bp. Both marker systems presented high percentages of polymorphism. However, IRAP markers were suitable for detecting genetic variability at the individual level and did not differentiate higher taxa. The REMAP maker system allowed the clustering by botanical variety and identified most of the homonym bread wheat cultivars.     

中国香菇主栽品种遗传多样性的SSR分析及指纹图谱构建

【目的】香菇(Lentinulaedodes)是世界第二大食用菌,研究我国现有栽培种群体的遗传多样性和遗传构成以及准确鉴定品种是新品种开发和产业健康发展的基础。【方法】采用多态性的SSR(Simple sequence repeat)分子标记对中国历年来使用主栽品种进行遗传多样性及群体结构分析,比较其谱系来源,解析中国主栽品种的群体多样性构成,并构建指纹图谱用于品种鉴定。【结果】24对多态性SSR引物对51份香菇菌株都具有多态性。聚类分析在相似系数0.69处可将栽培种群体分为4个类群,野生种驯化或参与杂交获得的菌株位于类群Ⅲ和Ⅳ,其他菌株位于另外两个类群Ⅰ和Ⅱ。群体结构分析可将栽培群体分为6个遗传构成,显示L808、L135等代表性菌株在各自的构成中参与了其他菌株的选育过程,解释了以其为亲本的部分品种的谱系来源。依据筛选出的9对条带清晰的SSR引物组合构建了多位点SSR指纹图谱,可对45个香菇商业菌种进行辨识。【结论】我国香菇主栽品种亲缘关系较近,育种多围绕L808、L135、9015等核心代表性品种进行,本研究可为选育具有自主知识产权、适应不同栽培模式的新品种提供依据;指纹图谱的构建也能为香菇品种的准确鉴定提供保证。     

Genetic diversity and structure of local apple cultivars from Northeastern Spain assessed by microsatellite markers

A set of 493 old and local Spanish accessions of apple (Malus x domestica Borkh) maintained at three collections in Northeastern Spain was studied using 16 simple sequence repeats in order to estimate their genetic diversity and to identify the genetic structure and relationships among their accessions. An additional diverse set of 45 apple cultivars, including old Spanish and international cultivars, was added as reference. Genetic analyses performed by Bayesian model-based clustering revealed a very strong differentiation of two major groups. The first one clustered 159 individuals (52?% of unique genotypes) including local accessions and six old Spanish cultivars. The second major group was formed by 145 individuals, including 38 international reference cultivars and one old Spanish cultivar. Nested Bayesian clustering was applied to those two groups and two and four sub-groups were found at each one, respectively. The identification of private and unique alleles, and the remarkable differences in allelic richness among groups and sub-groups constitute further evidence of a clear genetic structure. The results obtained through the factorial correspondence and analyses of molecular variance confirmed those obtained by Bayesian analyses, revealing moderate but significant differentiation among the two major groups (F _ST?=?0.076) and the six sub-groups (F _ST?=?0.111). Our results highlight that the genetic diversity encompassed by currently cultivated apple accounts only for a small fraction of that existing within the species, and that an important part (??60?%) of the local material analyzed constitutes a good example of genetic distinctness with respect to the main cultivars used in European orchards.     

Genetic variability in local Brazilian horse lines using microsatellite markers

Genetic variability at 11 microsatellite markers was analyzed in five naturalized/local Brazilian horse breeds or genetic groups. Blood samples were collected from 328 animals of the breeds Campeira (Santa Catarina State), Lavradeira (Roraima State), Pantaneira (Pantanal Mato-Grossense), Mangalarga Marchador (Minas Gerais State), as well as the genetic group Baixadeiro (Maranh?o State), and the exotic breeds English Thoroughbred and Arab. We found significant genetic variability within evaluated microsatellite loci, with observed heterozygosis varying between 0.426 and 0.768 and polymorphism information content values of 0.751 to 0.914. All breeds showed high inbreeding coefficients and were not in Hardy-Weinberg equilibrium. The smallest genetic distance was seen between the Pantaneira and Arab breeds. The principal component analyzes and Bayesian approach demonstrated that the exotic breeds have had a significant influence on the genetic formation of the local breeds, with introgression of English Throroughbred in Pantaneira and Lavradeira, as well as genetic proximity between the Arab, Pantaneira and Mangalarga Marchador populations. This study shows the need to conserve traits acquired by naturalized horse breeds over centuries of natural selection in Brazil due to the genetic uniqueness of each group, suggesting a reduced gene flow between them. These results reinforce the need to include these herds in animal genetic resource conservation programs to maximize the genetic variability and conserve useful allele combinations.     

Genetic variability and relationship between MT-1 elephant grass and closely related cultivars assessed by SRAP markers

Genetic variability and relationships among elephant grass cultivars were estimated by the SRAP (sequence-related amplified polymorphism) assay. A total of 60 individuals collected from five cultivars in China were analysed. Sixty-two selected primer combinations generated 1395 bands, with an average of 22.5 per primer combination. The average value of percentage of polymorphic bands (PPB) was 72.8% at species level. The PPB was from 15.2% to 75%, with an average of 39.6% at cultivar level. H _POP , within-cultivar Shannon’s index was 1.738 at cultivar level; at species level, the Shannon’s index (H _SP ) was 3.880. An assessment of diversity between cultivars [(H _SP − H _POP )/H _SP ] indicated that most of the diversity (55.2%) was detected among cultivars, and only 44.8% was within cultivars in total genetic variation. According to UPGMA dendrogram, the five cultivars were clustered into three main groups. One group included MT-1 and Mott with a bootstrap support of 100%, another consisted of Huanan and N51 with a bootstrap support of 81%, and last one was only Guimu-1. The results indicate that the MT-1 and Mott have a closest genetic relationship; Huanan and N51 possess a relatively close relationship, and Guimu-1 is the most distinct from the other four cultivars.     

Genetic relationships and population structure of local olive tree accessions from Northeastern Spain revealed by SSR markers

The olive (Olea europaea L. subsp. europaea) is an old traditional crop which was domesticated from the wild at various locations around the Mediterranean basin, resulting in a vast number of accessions worldwide. This is the case of the northeast part of Spain (Aragon), where 163 genotypes were prospected and characterised. The majority of these genotypes (90 %) seem to be autochthonous accessions of this area and the rest are synonyms of accessions from other areas of Spain. In this study, 11 nuclear SSR markers were used to discriminate between the accessions and to understand the genetic relationship among them. All primers produced a successful amplification giving a total of 176 fragments in the genotypes studied, with an average of 16 alleles per SSR, ranging from 5 (DCA13) to 24 (DCA11). Allele size ranged from 120 bp at locus UDO99 to 284 bp at locus DCA15. The dendrogram generated using the variability observed classified most of the genotypes according to their geographical origin (Huesca, Teruel and Zaragoza provinces), confirming the particular evolution of different olive ecotypes. Structure software was used to investigate the genetic population structure among olive accessions, showing that the maximum rate of change in the log probability of the data occurred at K = 2. In addition, the SSR markers have consequently shown their usefulness for cultivar identification in olive, for establishing the genetic closeness among its accessions, and for establishing genealogical relationships.     

Chloroplast DNA variability in old and recently introduced potato cultivars

Chloroplast DNA (cpDNA) variability has been examined in a range of tetraploid European potato cultivars. The potato genotypes studied included primitive cultivars such as Lumpers (1806), Yam (1836), Myatts Ashleaf (1847) and more recently bred cultivars such as Brodick (1990). Three cpDNA pheno-types were detected and these probably represent original introductions from South America into Europe. The most common cp phenotype was the T type cpDNA (Hosaka & Hanneman, 1988) characteristic of cultivars descended from cv. Rough Purple Chili. The presence of the T type cpDNA in the cultivar Yam indicates that this genotype which is of Andigena origin shares a common cytoplasm with other 5. tuberosum ssp. tuberosum clones which have a Chilean type cytoplasm. The implications of these results are discussed in relation to the origin of the T type cpDNA. Methods for increasing the cytoplasmic diversity of the cultivated potato gene pool are proposed.     

基于SSR标记的六倍体小黑麦遗传多样性分析

采用已筛选出的多态性较好的分布于六倍体小黑麦21对染色体上的100对SSR引物对从国际玉米小麦改良中心(CIMMYT)引进的339份六倍体小黑麦进行了遗传多样性分析。结果显示,100对引物共检测出323个等位变异,变幅是1~6个,平均等位变异丰富度为3.23;多态信息含量(PIC)的变幅为0~0.748,平均多态信息含量为0.465;平均遗传多样性指数(H')为0.1439,说明供试的339份六倍体小黑麦品种的SSR遗传多样性较为丰富。同时基于Nei's遗传距离对339份材料进行聚类分析,可以将供试材料分为6个类群。其中,第Ⅵ类群与其他类群的遗传距离较大、遗传差异较明显。研究结果为将六倍体小黑麦的优良基因应用于普通小麦的遗传改良提供了理论依据。     

Genetic variation and comparison of orchardgrass (Dactylis glomerata L.) cultivars and wild accessions as revealed by SSR markers

Orchardgrass is a highly variable, perennial forage grass that is cultivated throughout temperate and subtropical regions of the world. Despite its economic importance, the genetic relationship and distance among and within cultivars are largely unknown but would be of great interest for breeding programs. We investigated the molecular variation and structure of cultivar populations, compared the level of genetic diversity among cultivars (Baoxing, Anba, Bote, and Kaimo), subspecies (Dactylis glomerata ssp Woronowii) and advanced breeding line (YA02-116) to determine whether there is still sufficient genetic diversity within presently used cultivars for future breeding progress in China. Twenty individuals were analyzed from each of six accessions using SSR markers; 114 easily scored bands were generated from 15 SSR primer pairs, with an average of 7.6 alleles per locus. The polymorphic rate was 100% among the 120 individuals, reflecting a high degree of genetic diversity. Among the six accessions, the highest genetic diversity was observed in Kaimo (H = 0.2518; I = 0.3916; P = 87.3%) and 02-116 had a lower level of genetic diversity (H = 0.1806; I = 0.2788; P = 58.73%) compared with other cultivars tested. An of molecular variance revealed a much larger genetic variation within accessions (65%) than between them (35%). This observation suggests that these cultivars have potential for providing rich genetic resource for further breeding program. Furthermore, the study also indicated that Chinese orchardgrass breeding has involved strong selection for adaptation to forage production, which may result in restricted genetic base of orchardgrass cultivar.     

Genetic variability and diversification process in local pear cultivars from northwestern Spain using microsatellites

This is the first known large-scale molecular study of simple sequence repeats loci based on pear cultivars from the northwestern Iberian Peninsula. Most of the Spanish pear crop (one of the largest in Europe) is based on the Spanish cultivar Blanquilla and various other foreign cultivars. However, local cultivars can still be found in old orchards in northwestern Spain. Between 1978 and 1981, the Centro de Investigaciones Agrarias de Mabegondo (Xunta de Galicia) established a Germplasm Bank of local pear cultivars containing 221 accessions. In the current study, these were analysed and compared with 20 commercial cultivars of Pyrus spp. using 19 microsatellites. We identified 127 genotypes out of 221 accessions with an average of 43% clonality in this collection. Genotypes were analysed using a model-based Bayesian procedure (Structure), factorial correspondence analysis and molecular variance analysis, and Jaccard coefficients were estimated. Four reconstructed populations were identified by Structure, one related to Asian cultivars, two to French and English cultivars and one to Galician cultivars. The four identified groups of pears had evolved independently. This study explains the diversification process in pear cultivars from northwestern cultivars based on hybridization (16%), selection of triploids (38%) and the introgression of commercial cultivars in the collection (4%).     

Genetic diversity of elite sweet sorghum genotypes assessed by SSR markers

To determine genetic diversity among 47 elite sweet sorghum (Sorghum bicolor ssp. bicolor L.) genotypes, 46 simple sequence repeat (SSR) markers evenly distributed on all 10 chromosomes were selected. All SSR markers used were polymorphic among the genotypes studied. A total of 228 alleles were identified with an average of 4.96 alleles per marker. Furthermore, the genotypes studied showed medium genetic diversity. Clustering analysis grouped the 47 genotypes into 5 distinct clusters.     

Genetic diversity of African maize inbred lines revealed by SSR markers

Knowledge of genetic diversity (GD) and relationships among maize inbred lines is indispensable in a breeding program. Our objectives were to (1) investigate the level of genetic diversity among maize inbred lines and (2) assess their genetic structures by applying simple sequence repeat (SSR) markers. Fifty-six highland and mid-altitude maize inbred lines obtained from CIMMYT programs in Ethiopia and Zimbabwe were genotyped using 27 SSR loci. All of the genotypes studied could unequivocally be distinguished with the combination of the SSRs used. In total, 104 SSR alleles were identified, with a mean of 3.85 alleles per locus. The average polymorphism information content (PIC) was 0.58. GD expressed as Euclidean distance, varied from 0.28 to 0.73 with an average of 0.59. Cluster analysis using unweighted pair group method with arithmetic average (UPGMA) suggested five groups among the inbred lines. Most of the inbred lines adapted to the highlands and the mid-altitudes were positioned in different clusters with a few discrepancies. The pattern of groupings of the inbred lines was mostly consistent with available pedigree information. The variability detected using SSR markers could potentially contribute towards effective utilization of the inbred lines for the exploitation of heterosis and formation of genetically diverse source populations in Ethiopian maize improvement programs.     

利用RAPD技术对葱属品种遗传关系的分析

利用随机扩增多态性DNA(RAPD)技术分析了我国栽培的几个主要葱属品种。所用的20个引物,有11个能扩增出重现性好且稳定的谱带,10个品种共扩增出102条带,其中68条具多态性。根据DNA谱带计算的10个品种相似系数范围为0.571～0.947。对10个品种的UPGMA聚类分析表明,章丘大葱(Allium fistulosum var. gigantum cv. Zhangqiu Welsh onion)、天津五叶齐（A. fistulosum var.gigantum cv. Tianjin Wuyeqi Welsh onion）、上海分葱（A. fistulosum var. caepitosum cv.Shanghai Spring onion）、南京冬葱（A. fistulosum var. caepitosum cv. Nanjing Winter onion）和韩国大葱（A. fistulosum var .gigantum cv. Korea Welsh onion）关系较近。根据聚类结果,将章丘大葱、天津五叶齐、上海分葱、南京冬葱、韩国大葱、细香葱（A. schoenoprasum）以及楼葱（A. fistulosum var. viviparum）归为葱组;而将胡葱（A. ascalonicum）、洋葱（A. cepa）及洋葱和大葱的杂交种归为洋葱组。最后根据RAPD标记,讨论了各品种之间的亲缘关系。     

Genetic relatedness of Portuguese almond cultivars assessed by RAPD and ISSR markers

Randomly amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to analyse the genetic diversity of Portuguese Prunus dulcis cultivars and their relationship to important foreign cultivars. Of the primers tested, 6 (out of 60) RAPD and 5 (out of 18) ISSR primers were selected for their reproducibility and high polymorphism. Out of 124 polymerase chain reaction fragments that were scored, 120 (96.8%) were polymorphic. All the plants could be discriminated and constitute a very heterogeneous group. Five unidentified almond plants found in the region of Foz Côa (north Portugal) and wild almond (P. webbii) from Italy and Spain were also included. Four main groups of plants could be distinguished: P. dulcis cultivars; one Foz Côa plant; P. webbii; and P. persica (outgroup). The segregating Foz Côa plant may represent a feral individual or a hybrid between P. dulcis and P. webbii.Abbreviations dNTP Deoxynucleotide triphosphate - CTAB Cetyltrimethylammonium bromide - ISSR Inter-simple sequence repeats - PCR Polymerase chain reaction - RAPD Randomly amplified polymorphic DNA - RASTM Regional Agricultural Services of Trás-os-Montes - TE Tris-EDTA buffer - UPGMA Unweighted pair group method with arithmetical averagesCommunicated by P. Puigdoménech     

半野生大豆种质资源SSR位点遗传多样性分析

利用12对SSR引物对67份半野生大豆种质进行了遗传多样性的检测分析,结果表明,12个位点共检测到184个等位基因变异,平均每个位点等位基因数目为15．41个,平均多态性信息量,平均遗传多样性指数,平均遗传距离分别为0．849,0．706,0．118,根据SSR分析结果,按欧式距离将67份半野生大豆种质聚类并划分为5个组群。     

Genetic diversity among alfalfa (Medicago sativa) cultivars coming from a breeding program, using SSR markers

Alfalfa (Medicago sativa) is an autotetraploid, allogamous and heterozygous species whose cultivars are synthetic populations. The breeders apply selection pressure for some agronomic traits within a breeding pool to increase the frequency of favorable individuals. The objective of this study was to investigate the differentiation level among seven cultivars originating from one breeding program, and between these cultivars and the breeding pool, with eight SSR markers. These highly polymorphic and codominant markers, together with recent population genetic statistics extended to autotetraploids, offer tools to analyse genetic diversity in alfalfa. The number of alleles per locus varied between 3 and 24. All loci were at a panmictic equilibrium in the cultivars, except one, probably because of null alleles. With seven SSR loci, each cultivar was at panmictic equilibrium. The mean gene diversity was high, ranging from 0.665 to 0.717 in the cultivars. The parameter F _ST indicated a low but significant diversity among cultivars. Among 21 pairs of cultivars, 15 were significantly different. The breeding pool also had a high diversity, and was significantly different from each cultivar except the most recent one. Considering the characteristics of the breeding program and the mode of cultivar elaboration, we found that they were unable to generate a large variety differentiation. Estimation of population genetics parameters at SSR loci can be applied for assessing the differences between cultivars or populations, either for variety distinction or the management of genetic resources.     

Genetic diversity and structure of landrace accessions,elite lineages and cultivars of common bean estimated with SSR and SNP markers

Molecular Biology Reports - Common bean (Phaseolus vulgaris L.) is an important source of proteins, fibers and minerals for humans, being grown mainly in developing countries and representing a...