Genetic diversity of the azuki bean (Vigna angularis (Willd.) Ohwi & Ohashi) gene pool as assessed by SSR markers

To facilitate the wider use of genetic resources including newly collected cultivated and wild azuki bean germplasm, the genetic diversity of the azuki bean complex, based on 13 simple sequence repeat (SSR) primers, was evaluated and a core collection was developed using 616 accessions originating from 8 Asian countries. Wild germplasm from Japan was highly diverse and represented much of the allelic variation found in cultivated germplasm. The SSR results together with recent archaeobotanical evidence support the view that Japan is one center of domestication of azuki bean, at least for the northeast Asian azuki bean. Cultivated azuki beans from China, Korea, and Japan were the most diverse and were genetically distinct from each other, suggesting a long and relatively isolated history of cultivation in each country. Cultivated azuki beans from eastern Nepal and Bhutan were similar to each other and quite distinct from others. For two primers, most eastern Nepalese and Bhutanese cultivated accessions had null alleles. In addition, wild accessions from the Yangtze River region of China and the Himalayan region had a null allele for one or the other of these primers. Whether the distinct diversity of azuki bean in the Himalayan region is due to introgression or separate domestication events requires further study. In contrast, western Nepalese azuki beans showed an SSR profile similar to that of Chinese azuki beans. The genetic distinctness of cultivated azuki beans from Vietnam has been revealed for the first time. The specific alleles indicate that Vietnamese azuki beans have been cultivated in isolation from Chinese azuki beans for a long time. Wild germplasm from the Himalayan region showed the highest level of variation. Based on the results, Himalayan germplasm could be considered a novel gene source for azuki bean breeding. A comparison with mungbean SSR analysis revealed that the mean gene diversity of cultivated azuki bean (0.74) was much higher than that of cultivated mungbean (0.41). The reduction in gene diversity due to domestication, the domestication bottleneck, in azuki bean is not strong compared with that in mungbean.     

小豆种质资源农艺性状综合鉴定与评价

通过田间观测和室内分析,对来自全国各生态区的262份小豆优异种质资源连续2年进行了农艺性状鉴定及综合评价。结果表明:262份小豆微核心种质资源在考察的16个形态学性状中均具有丰富的遗传变异类型;不同年份间生态环境尤其是光照条件对小豆农艺性状影响较大;主成分分析确定了3类影响因子,表明小豆资源的选育要集中在生长势良好(生育日数较短、株高较矮),单株荚数和单荚粒数多的地方品种;聚类分析结果显示在相似系数为0.40时可将参试材料分为5大类群,各类群间性状差异明显;群体性状表现与地理来源之间没有直接的关系。     

Morphological variation and evolutionary significance of <Emphasis Type="Italic">Coccinia grandis</Emphasis> (L.) Voigt: an under-exploited cucurbitaceous vegetable crop

Morphological variation was analyzed in wild accessions and cultivars of the vegetatively propagated dioecious Coccinia grandis. Variations of 43 morphological characters, 19 qualitative and 23 quantitative traits, were analyzed among 40 female accessions, including 25 cultivars and 15 wild accessions. Multivariate statistical analyses were used to group accessions according to their morphological similarity. Principal component (PC) analysis revealed that the first three PCs accounted for 50% of the total variance, and differences among the accessions were evidenced principally in relation to fruit characteristics such as fruit weight, fruit length and the number of seeds in each fruit. Analysis of variance carried out in the entire germplasm revealed significant differences within the germplasm, whereas ANOVA carried out between the wild accessions and the cultivars proved the null hypothesis that there are no significant differences between the two groups, and differences were observed only in fruit characters that are targets of human selection. Principal component analysis, UPGMA cluster analysis and discriminant factor analysis revealed strong overlaps between the two groups indicating the ongoing process of evolution and selection in the species.     

Evaluating Agronomic Performance and Investigating Molecular Structure of Drought and Heat Tolerant Wild Alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis> L.) Collection from the Southeastern Turkey

Drought is a major stress factor for agricultural production including alfalfa production. One way to counterbalance the yield losses is the introgression of drought tolerant germplasm into breeding programs. As an effort to exploit such germplasm, 16 individual plants were selected from the Southeastern Turkey from their natural habitat and clonally propagated in field trials with an ultimate goal to use the germplasm as parents for releasing a synthetic cultivar. Forage yield and forage quality traits were evaluated and molecular genetic diversity among genotypes were determined using inter simple sequence repeat markers. Genotypes showed a variation from growth habit to yield and quality traits indicating sufficient phenotypic variation for diverse breeding efforts (for grazing or harvesting) and long term selection schemes. A large amount of genetic variation was observed even with a limited number of marker and genotypes. However, no pattern of spatial genetic structure was observed for the scale of the study when genetic variation is linked to the geographic origin. We conclude that ex situ natural variation provides a wealth of germplasm that could be incorporated into breeding programs aiming to improve drought tolerance. We also suggest an extensive collection of seeds/plant tissue from unique plants with desirable traits rather than putting more efforts to create a spatial germplasm sampling efforts in narrow regions.     

Patterns of genetic diversity in the Andean gene pool of common bean reveal a candidate domestication gene

Most studies on the genetic diversity of common bean (Phaseolus vulgaris L.) have focussed on accessions from the Mesoamerican gene pool compared to the Andean gene pool. A deeper knowledge of the genetic structure of Argentinian germplasm would enable researchers to determine how the Andean domestication event affected patterns of genetic diversity in domesticated beans and to identify candidates for genes targeted by selection during the evolution of the cultivated common bean. A collection of 116 wild and domesticated accessions representing the diversity of the Andean bean in Argentina was genotyped by means of 114 simple sequence repeat (SSR) markers. Forty-seven Mesoamerican bean accessions and 16 Andean bean accessions representing the diversity of Andean landraces and wild accessions were also included. Using the Bayesian algorithm implemented in the software STRUCTURE we identified five major groups that correspond to Mesoamerican and Argentinian wild accessions and landraces and a group that corresponds to accessions from different Andean and Mesoamerican countries. The neighbour-joining algorithm and principal coordinate clustering analysis confirmed the genetic relationships among accessions observed with the STRUCTURE analysis. Argentinian accessions showed a substantial genetic variation with a considerable number of unique haplotypes and private alleles, suggesting that they may have played an important role in the evolution of the species. The results of statistical analyses aimed at identifying genomic regions with consistent patterns of variation were significant for 35 loci (~20 % of the SSRs used in the Argentinian accessions). One of these loci mapped in or near the genomic region of the glutamate decarboxylase gene. Our data characterize the population structure of the Argentinian germplasm. This information on its diversity will be very valuable for use in introgressing Argentinian genes into commercial varieties because the majority of present-day common bean varieties are of Andean origin.     

食用豆类种质资源粗蛋白及粗淀粉含量的评价

对"十五"期间收集、整理并存入国家种质库的1696份食用豆类种质资源的主要品质性状进行了鉴定评价.结果表明,9种食用豆类种质资源的粗蛋白、粗淀粉平均含量分别为24.93%、45.11%,不同食用豆类种质资源中各品质性状变异程度不同,其中菜豆和鹰嘴豆的粗蛋白和豇豆的粗淀粉含量变异最大.不同来源的食用豆类种质资源品质性状比较结果表明,国内绿豆种质资源粗蛋白含量明显优于国外种质资源.通过鉴定筛选出一批品质性状优良的食用豆类种质,并对使用的评价标准进行了讨论.     

Genetic characterization of an elite coffee germplasm assessed by gSSR and EST-SSR markers

Coffee is one of the main agrifood commodities traded worldwide. In 2009, coffee accounted for 6.1% of the value of Brazilian agricultural production, generating a revenue of US$6 billion. Despite the importance of coffee production in Brazil, it is supported by a narrow genetic base, with few accessions. Molecular differentiation and diversity of a coffee breeding program were assessed with gSSR and EST-SSR markers. The study comprised 24 coffee accessions according to their genetic origin: arabica accessions (six traditional genotypes of C. arabica), resistant arabica (six leaf rust-resistant C. arabica genotypes with introgression of Híbrido de Timor), robusta (five C. canephora genotypes), Híbrido de Timor (three C. arabica x C. canephora), triploids (three C. arabica x C. racemosa), and racemosa (one C. racemosa). Allele and polymorphism analysis, AMOVA, the Student t-test, Jaccard's dissimilarity coefficient, cluster analysis, correlation of genetic distances, and discriminant analysis, were performed. EST-SSR markers gave 25 exclusive alleles per genetic group, while gSSR showed 47, which will be useful for differentiating accessions and for fingerprinting varieties. The gSSR markers detected a higher percentage of polymorphism among (35% higher on average) and within (42.9% higher on average) the genetic groups, compared to EST-SSR markers. The highest percentage of polymorphism within the genetic groups was found with gSSR markers for robusta (89.2%) and for resistant arabica (39.5%). It was possible to differentiate all genotypes including the arabica-related accessions. Nevertheless, combined use of gSSR and EST-SSR markers is recommended for coffee molecular characterization, because EST-SSRs can provide complementary information.     

Genetic diversity and relationship of global faba bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.) germplasm revealed by ISSR markers

Genetic diversity and relationships of 802 faba bean (Vicia faba L.) landraces and varieties from different geographical locations of China and abroad were examined using ISSR markers. A total of 212 repeatable amplified bands were generated with 11 ISSR primers, of which 209 were polymorphic. Accessions from North China showed highest genetic diversity, while accessions from central China showed low level of diversity. Chinese spring faba bean germplasm was clearly separated from Chinese winter faba bean, based on principal component analysis and UPGMA clustering analysis. Winter accessions from Zhejiang (East China), Jiangxi (East China), Sichuan (Southwest China) and Guizhou (Southwest China) were quite distinct to that from other provinces in China. Great differentiation between Chinese accessions and those from rest of the world was shown with a UPGMA dendrogram. AMOVA analyses demonstrated large variation and differentiation within and among groups of accessions from China. As a continental geographic group, accessions from Europe were genetically closer to those from North Africa. Based on ISSR data, grouping results of accessions from Asia, Europe and Africa were obviously associated with their geographical origin. The overall results indicated that the genetic relationship of faba bean germplasm was closely associated with their geographical origin and their ecological habit.     

Characterization and utilization of microsatellites in the <Emphasis Type="Italic">Coffea canephora</Emphasis> genome to assess genetic association between wild species in Kenya and cultivated coffee

Coffee is an important beverage crop in the world and has a significant contribution to Kenya’s economy. Here, we analyzed the genome-wide distribution of microsatellites in the Coffea canephora genome. A total of 159,041 SSRs were identified, with an overall density of 308 SSRs per Mb. Tetra-nucleotide repeats are the most abundant, accounting for 32 % of the total SSRs. AT-rich motifs are dominant across all SSR repeat units, while GC-rich motifs were generally rare. A set of 100 SSRs was selected to amplify 96 coffee accessions, including 10 wild accessions collected from Mt. Marsabit (Kenya). Of these SSRs, 33 % generated clear polymorphic bands among all tested accessions, with an average of 3.9 alleles per SSR locus. Wild coffee species from Mt. Marsabit showed a close genetic similarity with cultivated accessions in Kenya, suggesting that the wild species in Mt. Marsabit played an important role in the domestication of cultivated coffee in Kenya. Significantly low pairwise genetic divergence was observed between cultivated and wild accessions in Kenya, suggesting a relatively narrow level of genetic basis among coffee germplasm in Kenya. In addition, cultivated and wild coffee accessions in Kenya show a great divergence from those in other countries. Our results not only provide molecular tools for genetic studies in coffee but are also helpful for conservation and coffee breeding programs in Kenya.     

Variability and divergence in Pongamia pinnata (L.) Pierre germplasm – a candidate tree for biodiesel

Three explorations were undertaken in South East Coastal zone of India covering parts of Andhra Pradesh (AP) and Orissa states to collect Pongamia pinnata (L.) Pierre germplasm during March–June 2007. A total of 123 accessions were collected and seed data recorded were analyzed for morphometric traits viz., seed length, seed width, seed thickness, 100‐seed weight and oil content. Variation in the collected germplasm was analyzed using anova , simple measures of variation and D² statistics. Significant genetic variability between seed traits and oil content and association among the seed traits was recorded. Phenotypic variance was higher than genotypic variance for all the characters indicating dominant role of environment. High heritability (broad sense) for 100‐seed weight (97.6%) and oil content (86.7%) indicated the reliability of these characters as selection criteria for plus trees. Genetic gain was maximum for 100‐seed weight (62.6%) followed by oil content (30.5%). D² analysis grouped the accessions into 12 clusters. Cluster XII and cluster IX were the most diverse based on the intercluster distance. Based on the observed diversity, Chittoor, Srikakulam and Adilabad districts of AP are most suitable for collecting diverse germplasm lines and also for in situ conservation.     

Genetic diversity in Indian cucumber based on microsatellite and morphological markers

Genetic variation among 44 cucumber accessions was assessed using morphological and SSR markers. High genetic variability was observed for days to 50% female flowering (37–46 days from sowing), number of fruits per plant (1.4–6.0), individual fruit weight (0.04–0.552 kg) and root length (14.25–32.8 cm). The pair-wise Jaccard similarity coefficient ranged between 0.25 and 0.85 indicating that the accessions represent genetically diverse populations. The allelic diversity of polymorphic markers ranged from 0.001 to 0.9396 with an average of 0.31 based on polymorphic information content. The clustering pattern of SSR markers was not in consonance with the groupings based on quantitative traits. The accession of Indian state i.e.; Madhya Pradesh and Uttar Pradesh were diverged from the accessions of other parts of India. The study provides information for future exploration and collection of cucumber germplasm in India and utilization of diverse germplasm for developing cultivars/hybrids for specific traits.     

Simple sequence repeat diversity in diploid and tetraploid Coffea species.

Thirty-four fluorescently labeled microsatellite markers were used to assess genetic diversity in a set of 30 Coffea accessions from the CENICAFE germplasm bank in Colombia. The plant material included one sample per accession of seven East African accessions representing five diploid species and 23 wild and cultivated tetraploid accessions of Coffea arabica from Africa, Indonesia, and South America. More allelic diversity was detected among the five diploid species than among the 23 tetraploid genotypes. The diploid species averaged 3.6 alleles/locus and had an average polymorphism information content (PIC) value of 0.6, whereas the wild tetraploids averaged 2.5 alleles/locus and had an average PIC value of 0.3 and the cultivated tetraploids (C. arabica cultivars) averaged 1.9 alleles/locus and had an average PIC value of 0.22. Fifty-five percent of the alleles found in the wild tetraploids were not shared with cultivated C. arabica genotypes, supporting the idea that the wild tetraploid ancestors from Ethiopia could be used productively as a source of novel genetic variation to expand the gene pool of elite C. arabica germplasm.     

Validation of a PCR test to predict the presence of flavor volatiles mesifurane and γ-decalactone in fruits of cultivated strawberry (<Emphasis Type="Italic">Fragaria</Emphasis> × <Emphasis Type="Italic">ananassa</Emphasis>)

Flavor improvement is currently one of the most important goals for strawberry breeders. At the same time, it is one of the most complex traits to improve, involving the balanced combination of several desired characteristics such as high sweetness, moderate acidity, and the appropriate combination of aroma compounds that are beginning to be delineated in consumer tests. DNA-informed breeding will expedite the selection of complex traits, such as flavor, over traditional phenotypic evaluation, particularly when markers linked to several traits of interests are combined during the breeding process. Natural variation in mesifurane and γ-decalactone, two key volatile compounds providing sweet Sherry and fresh peach-like notes to strawberry fruits, is controlled by the FaOMT and FaFAD1 genes, respectively. In this study, we have optimized a simple PCR test for combined analysis of these genes and determined a prediction accuracy above 91% using a set of 71 diverse strawberry accessions. This high accuracy in predicting the presence of these important volatiles combined with the simplicity of the analytical methodology makes this DNA test an efficient tool for its implementation in current strawberry-breeding programs for the selection of new strawberry cultivars with superior flavor.     

Developing a common bean core collection suitable for association mapping studies

Because of the continuous introduction of germplasm from abroad, some collections have a high number of accessions, making it difficult to explore the genetic variability present in a germplasm bank for conservation and breeding purposes. Therefore, the aim of this study was to quantify and analyze the structure of genetic variability among 500 common bean accessions to construct a core collection. A total of 58 SSRs were used for this purpose. The polymorphism information content (PIC) in the 180 common bean accessions selected to compose the core collection ranged from 0.17 to 0.86, and the discriminatory power (DP) ranged from 0.21 to 0.90. The 500 accessions were clustered into 15 distinct groups and the 180 accessions into four distinct groups in the Structure analysis. According to analysis of molecular variance, the most divergent accessions comprised 97.2% of the observed genetic variability present within the base collection, confirming the efficiency of the selection criterion. The 180 selected accessions will be used for association mapping in future studies and could be potentially used by breeders to direct new crosses and generate elite cultivars that meet current and future global market needs.     

Divergent Selection for Secondary Traits in Upland Tetraploid Switchgrass and Effects on Sward Biomass Yield

Switchgrass (Panicum virgatum L.) is currently undergoing intensive breeding efforts to improve biomass yield. Consideration must be made regarding the relative importance of spaced plantings to sward plots for evaluation and selection for increased biomass yield. It has previously been suggested that selection schemes using secondary plant morphological traits as selection criteria within spaced plantings may be an efficient method of making genetic gain. The objective of this study was to empirically test the effects of direct selection for plant height, tiller count, flowering date, and visual selection for biomass yield within spaced plantings on biomass yield and morphology traits within sward plots. Divergently selected populations for each trait were developed from the WS4U upland tetraploid germplasm and evaluated for biomass yield at five locations in Wisconsin during two growing seasons. Significant variation was observed between maternal parents of the selected populations for both selected and nonselected traits. Despite substantial differences between parent plant populations for plant morphology, significant differences were not observed for sward-plot biomass yield or sward-plot morphology relative to the base population. Late flowering selections yielded 2.0 Mg/ha greater biomass than early flowering selections (29 % increase). Plant height within sward plots was observed to have a strong positive correlation with biomass yield. Tiller count was observed to have a weak correlation with biomass yield. Based on the observed results, it is recommended that greater emphasis be placed on evaluation of biomass yield using sward plots.     

AFLP analysis of genetic diversity within and among Coffea arabica cultivars

Genetic diversity of Coffea arabica cultivars was estimated using amplified fragment length polymorphism (AFLP) markers. Sixty one Coffea accessions composed of six arabica cultivars, including Typica, Bourbon, Catimor, Catuai, Caturra and Mokka Hybrid, plus two diploid Coffea species, were analyzed with six EcoRI- MseI primer combinations. A total of 274 informative AFLP markers were generated and scored as binary data. These data were analyzed using cluster methods in the software package NTSYSpc. The differences among cultivars at the DNA level were small, with an average genetic similarity of 0.933. Most accessions within a cultivar formed a cluster, although deviant samples occurred in five of the six cultivars examined due to residual heterozygosity from ancestral materials. Among the six cultivars fingerprinted, the highest level of genetic diversity was found within the cultivar Catimor, with an average genetic similarity of 0.880. The lowest level was found within Caturra accessions, with an average genetic similarity of 0.993. Diversity between C. arabica and two other Coffea species, Coffea canephora and Coffea liberica, was also estimated with average genetic similarities of 0.540 and 0.413, respectively, suggesting that C. canephora is more closely related to C. arabica than is C. liberica. The genetic variation among arabica cultivars was similar to the variation within cultivars, and no cultivar-specific DNA marker was detected. Although arabica cultivars appear to have a narrow genetic base, our results show that sufficient polymorphism can be found among some arabica cultivars with a genetic similarity as low as 0.767 for genetic/QTL mapping and breeding. The assessment of genetic diversity among arabica cultivars provided the necessary information to estimate the potential for using marker-assisted breeding for coffee improvement.     

Heritable variation in the inflorescence replacement program of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Owing to their sessile habits and trophic position within global ecosystems, higher plants display a sundry assortment of adaptations to the threat of predation. Unlike animals, nearly all higher plants can replace reproductive structures lost to predators by activating reserved growing points called axillary meristems. As the first step in a program aimed at defining the genetic architecture of the inflorescence replacement program (IRP) of Arabidopsis thaliana, we describe the results of a quantitative germplasm survey of developmental responses to loss of the primary reproductive axis. Eighty-five diverse accessions were grown in a replicated common garden and assessed for six life history traits and four IRP traits, including the number and lengths of axillary inflorescences present on the day that the first among them re-flowered after basal clipping of the primary inflorescence. Significant natural variation and high heritabilities were observed for all measured characters. Pairwise correlations among the 10 focal traits revealed a multi-dimensional phenotypic space sculpted by ontogenic and plastic allometries as well as apparent constraints and outliers of genetic interest. Cluster analysis of the IRP traits sorted the 85 accessions into 5 associations, a topology that establishes the boundaries within which the evolving Arabidopsis genome extends and restricts the species’ IRP repertoire to that observable worldwide.     

Morphological diversity of Ethiopian barley (Hordeum vulgare L.) in relation to geographic regions and altitudes

A total of 199 germplasm accessions collected from 10 administrative regions of Ethiopia and four released cultivars, which were used for estimating of error variance, of barley in Ethiopia were field evaluated for nine agronomic traits at Holetta and Bekoji Agricultural Research Centers of Ethiopia during the 2006 main cropping season using non-replicated augmented design plots consisting of four incomplete blocks. The objectives were to assess the extent and pattern of morphological variation in the barley accessions with respect to regions and altitude of collection, to classify the genotypes tested into relatively homogenous groups and to identify the major traits contributing to the overall observed diversity in the germplasm. Genotype variance estimate of regions and altitudes indicated wide variation among accessions depending on the traits involved. The presence of high morphological variation within regions and altitudes particularly above 2000 m a.s.l. indicated the potential of each region and high altitude zones for barley improvement and conservation in the country. The clustering of accessions did not show grouping on the basis of regions of origin. Traits like thousand kernel weight, plant height, days to head and days to maturity accounted for most of the gross variance among the barley accessions and played role in differentiating accessions collected from different regions and altitude classes into principal components. In general because of environmental factors on the observed morphological variation future germplasm collection should consider to explore wide geographical and climatic differences within the country.     

中俄茄子种质资源遗传多样性研究

利用25对SSR分子标记和24个表型性状对105份中俄茄子材料进行遗传多样性分析。表型变异分析结果表明:24个表型性状在中俄材料间均表现出了不同程度的多样性,但是同一性状在中俄材料中多样性不同;主成分分析可将24个表型性状概括为果形因子、颜色因子、果实外观因子、叶片形态因子、果萼刺和花药条纹6个指标,其中果实特征占主要成分;利用UPGMA法进行聚类,遗传相似系数在0.4~0.8之间,平均值0.6。25对多态性SSR标记,扩增出122个条带,含有等位基因82个,其中有效等位数24.8个,PIC值为0.3~0.7。分子聚类的遗传相似系数在0.5~1之间,平均值是0.7。表型聚类和分子标记聚类的结果相似,105份茄子种质资源间的类群划分与地理来源之间没有直接关系,但与茄子的果实性状有一定的相关性。