Genetic relationship in Vigna mungo (L.) Hepper and V. radiata (L.) R. Wilczek based on morphological traits and SDS-PAGE

Thirty seven diverse genotypes of Vigna mungo and three of V. radiataresembling to V. mungo for seed characters were studied to determine the extent of genetic variation based on morphological characters. In addition, 4black seeded genotypes in each species and8 green seeded V. radiata genotypes were included for SDS-PAGE analysis. Seed proteins were analyzed through slab type SDS-PAGE. High variance was observed for plant height, days to flowering, days to maturity, number of branches/plant, number of pods/plant, pod length, seeds per pod,biomass yield/plant, grain yield/plant and harvest index (%) during 2 consecutive years. First four components of PCA with eigenvalues >1 contributed 78.7 and79.1% of the total variance amongst 40genotypes during 1998 and 1999. The populations with high PC₁ values were expected to be high yielding characterised by earlyness, high seed weight and harvest index, all traits strictly related to reproductive phase. Four clusters of genotypes were observed during both the years and genetic diversity was in association for both the years. Based on SDS-PAGE, specific bands were suggested to be used for identifying Vigna radiatafrom mixed germplasm with Vigna mungo. The SDS-PAGE proved to be a powerful tool for differentiating Vigna radiata and Vigna mungo, whereas a low level inter-specific genetic diversity was observed and no clear differentiation was observed both for agronomic characteristics and for geographical origin. This revised version was published online in July 2006 with corrections to the Cover Date.     

Species relationships in the subgenus Ceratotropis (genus Vigna) as revealed by RAPD analysis

Summary The genetic variation among 23 accessions of 5 species in the subgenus Ceratotropis, genus Vigna, were investigated by random amplified polymorphic DNA (RAPD) analysis. A total of 404 fragments amplified with 24 primers were scored and analyzed by cluster analysis. The accessions used were separated into two main groups with an average of 70% differences. Within the main groups, five subgroups were recognized, which are in complete agreement with taxonomic species. Wild forms were always grouped with their most closely related cultivated forms and they showed variation in each species. The largest intraspecific variation was found in V. radiata (mungbean), in which wild forms (V. radiata var. sublobata) were highly different from each other and from cultivated forms. V. angularis (adzuki bean) showed the least variation and thus, was probably differentiated in relatively recent times.     

The effectiveness of evaluating wild species: searching for sources of resistance to bruchid beetles in the genus Vigna subgenus Ceratotropis

A species level germplasm collection representing 76% of known taxa in the genus Vigna subgenus Ceratotropis was evaluated for resistance to two species of bruchid beetles, Callosobruchus chinensis and C. maculatus. Seven taxa consisting of 29 accessions were found to be resistant to C. chinensis and 4 taxa consisting of 24 accessions were found to be resistant to C. maculatus. This compared with no resistant accessions being found in several hundred landrace accessions of mungbean, V. radiata var. radiata, in the same subgenus. Sometimes resistance was found in all accessions of a particular taxon, such as complete resistance to both C. chinensis and C. macualtus in V. umbellata. Other taxa showed intra taxon variation for resistance such as V. reflexo-pilosa andV. minima. The levels and patterns of resistance among taxa were diverse. The results suggest that various factors cause resistance to bruchid in the subgenus Ceratotropis. While the number of eggs laid on seeds generally reflected seed size, one small seeded cultivar of V. mungo var. mungo, black gram, had an unusually high number of eggs laid per seed. No correlation was found between seed size and levels of resistance. The species level germplasm collection, which reflects the core collection concept in trying to maximize genetic diversity in a limited number of accessions, has enabled a large number of potentially useful sources of resistance to bruchid beetles to be found efficiently. This revised version was published online in July 2006 with corrections to the Cover Date.     

Partly fertile interspecific hybrid between a black gram × green gram derivative and an adzuki bean

A true‐breeding black gram (Vigna mungo) × green gram (Vigna radiata) derivative was reciprocally crossed with adzuki bean, Vigna angularis. Pollinated pistils were treated with gibberellic acid (GA₃) (70 p.p.m.) 24 and 48 h after pollination. One fertile pod containing two hybrid seeds was obtained, when V. angularis was used as the male parent. One of these seeds germinated and a partly fertile interspecific hybrid was obtained. Its hybridity was confirmed at the seedling stage based on the presence of the green epicotyl colour of the adzuki bean. Pubescence of epicotyl, stem, leaf margins and pod, and the colour of the mature pod, characteristics of the black gram × green gram derivative, were also expressed in the hybrid. In addition, the hybrid exhibited two novel traits — the presence of racemose inflorescence and plant regeneration ability — not present in the parents. The study of this hybrid and four offspring revealed that green epicotyl colour of adzuki bean, and the pubescence of stem and leaf margins of the black gram × green gram derivatives were dominantly inherited traits. Colours of epicotyl and stem bases were found to be linked.     

利用SSR标记分析野生小豆及其近缘野生植物的遗传多样性

利用28对SSR引物对96份野生小豆资源、小豆近缘野生植物及栽培小豆品种进行遗传多样性分析,共检测到255个等位变异,平均每对SSR引物9.10个,多态信息含量的变异范围从0.374到0.865,平均为0.722。野生小豆材料和近缘植物Vigna minima遗传变异丰富。来自不同地域的野生小豆材料具有大量特异等位变异,基于非加权成组配对算术平均法的聚类分析可将不同地理来源的野生小豆单独分群,且与主坐标分析的结果相一致。4份栽培小豆材料与日本野生小豆遗传距离较近,表明目前国内小豆育种中较多使用了含有日本血缘的小豆材料,以及国内野生小豆资源的搜集和利用工作落后于日本。本研究对国内野生小豆资源的搜集和保存具有指导意义,并可以为这些资源的评价、利用和优异基因的发掘提供参考。     

中国绿豆种质资源初选核心种质构建

以国家作物种质资源数据库中5 072份国内绿豆资源为材料, 根据14个农艺性状, 利用地理来源(省)和性状群进行分组, 分别采用比例法、平方根法和多样性指数法确定取样数及聚类选择和随机选择2种个体选择法构建了13个不同的绿豆初选核心样本, 对不同的取样方法及总资源间进行了品种间平均相似性系数、性状符合度、遗传多样性指数和数量性状变异系数的比较。结果表明, 聚类选择取样优于随机取样, 按照性状群分组优于按省分组; 在聚类选择条件下采用多样性指数法确定取样数优于平方根法和比例法。最终确定按性状群分组, 利用多样性指数确定取样数, 聚类选择个体为绿豆核心种质构建的最佳方案。用此方案, 构建了包含719份绿豆种质的初选核心种质, 取样比例为14.2%, 性状符合度达100%。     

Comparative analysis of diversity based on morpho-agronomic traits and microsatellite markers in common bean

Morpho-agronomic traits and microsatellite markers were used to survey genetic diversity in 115 common bean genotypes that included 70 Indian landraces, 24 released varieties and 21 exotic accessions. Twelve morpho-agronomic traits, namely, days to 50% flowering, leaflet length, leaflet width, pod length, pod width, number of pods per plant, days to maturity, seed length, seed width, number of seeds per pod, 100 seed weight and seed yield per plant were studied. Field data of two consecutive years were subjected to multivariate analysis as proposed by Mahalanobis’s D²-statistics, Tochers method of clustering and combined analysis of variance. Seventeen microsatellite markers were also used to examine genetic diversity at molecular level that showed polymorphic information content (PIC) in the range of 0.00–0.684. Dendrograms based on Euclidean distances and UPGMA analysis showed the presence of majority of released varieties into single cluster, which pointed toward their low genetic base in comparison to indigenous landraces and exotic germplasm. Significant correlation existed between morphological genetic distance and microsatellite genetic distance tested by Mantel test (r = 0.876).     

Patterns of RFLP-based genetic diversity in germplasm pools of common wheat with different geographical or breeding program origins

A set of 292 accessions of common wheat (Triticum aestivum L.) representing 21 germplasm pools based on geographical or breeding program origins was assayed for RFLP diversity. Thirty cDNA and genomic DNA probes and the HindIII restriction enzyme were employed for RFLP analysis. About 61% of all 233 scored bands were present in 75% or more of the accessions. All but one of the 30 probes revealed polymorphism, and the average number of distinct patterns per probe over all accessions was 9.5.Polymorphic Information Content (PIC) values within a pool varied from 0 to 0.9 and depended on the identities of both the germplasm pool and the probe. Rare banding patterns with a relative frequency of ≤0.2 within a pool were detected. These rare patterns were more likely to occur in pools exhibiting high levels of heterogeneity. The highest level of polymorphism was observed in the Turkish landraces from Southwest Asia. The Eastern U.S. soft red winter wheat germplasm pool was more genetically diverse than the other advanced germplasm pools, and nearly as diverse as the Turkish landrace pool. RFLP-based genetic relationships between germplasm pools generally tracked expectations based on common geographical origin, breeding history and/or shared parentages. The Chinese wheat landraces from Sichuan, Tibet, and Yunnan provinces were distinct from other pools. Similarity matrices for among-pool genetic distance estimates based on either band frequencies or banding pattern frequencies showed good correlation with matrices derived from Nei and Li's mean genetic similarity estimates (r=−0.82^** and r=−0.73^**, respectively. This revised version was published online in July 2006 with corrections to the Cover Date.     

Random amplified polymorphic DNA (RAPD) markers variability among cultivars and landraces of common beans (Phaseolus vulgaris L.) of south-Brazil

To evaluate the variability among cultivars and landraces of common bean(Phaseolus vulgaris L.), 15 cultivars and 18 landraces of common bean (Phaseolus vulgarisL.), a undefined species of Phaseolus,two landraces of Vigna angularis L., and a landrace of soybean (Glycine maxL.), were screened with fifteen oligonucleotide primers in PCR reactions. An average of 20.3 RAPD bands were scored per primer. A total of 304 amplification products were scored of which 88.8% were polymorphic among Phaseolus genotypes. Based on the RAPD markers, four major clusters were formed. Three clusters corresponded to the soybean, to the two Vigna angularis landraces, and to the Phaseolus sp. landrace, respectively. The fourth cluster include all the landraces and cultivars of Phaseolus vulgaris. This large group could be separated into three subgroups that were correlated with the phaseolin patterns and the average seed weight of the genotypes. The analysis shows that most of the landraces collected in South Brazil (17 out of 18) belong to the Andean gene pool, and most of the cultivars (13 out of 15) belong to the Middle American gene pool. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity of the black gram [Vigna mungo (L.) Hepper] gene pool as revealed by SSR markers

In this study, 520 cultivated and 14 wild accessions of black gram (Vigna mungo (L.) Hepper) were assessed for diversity using 22 SSR markers. Totally, 199 alleles were detected with a mean of 9.05 alleles per locus. Wild black gram showed higher gene diversity than cultivated black gram. Gene diversity of cultivated accessions among regions was comparable, while allelic richness of South Asia was higher than that of other regions. 78.67% of the wild gene diversity presented in cultivated accessions, indicating that the domestication bottleneck effect in black gram is relatively low. Genetic distance analysis revealed that cultivated black gram was more closely related to wild black gram from South Asia than that from Southeast Asia. STRUCTURE, principal coordinate and neighbor-joining analyses consistently revealed that 534 black gram accessions were grouped into three major subpopulations. The analyses also revealed that cultivated black gram from South Asia was genetically distinct from that from West Asia. Comparison by SSR analysis with other closely related Vigna species, including mungbean, azuki bean, and rice bean, revealed that level of gene diversity of black gram is comparable to that of mungbean and rice bean but lower than that of azuki bean.     

Cross‐genera amplification of informative microsatellite markers from common bean and scarlet runner bean for assessment of genetic diversity in mungbean (Vigna radiata)

Mungbean (V. radiata) is an important Asiatic legume supplying inexpensive protein to a vast majority of vegetarian masses. To increase markers repertoire in mungbean, a study was conducted to analyse 384 microsatellite markers derived from common bean, scarlet runner bean and adzuki bean for their transferability and polymorphism. The results showed that 87 (24.71%) primer pairs could amplify DNA loci of 20 mungbean genotypes including one accession of V. trilobata, while 52 showed reliable banding and polymorphism. These showed different degrees of variability at each locus producing 250 alleles with the number of alleles varying from 2 to 9. The major allele frequency varied from 0.17 to 0.95, while the polymorphic information content of SSRs ranged between 0.09 and 0.86 with an average of 0.60 ± 0.16. UPGMA revealed three major clusters accommodating ~95% of the accessions while one accession of V. trilobata (‘NSB‐007’) did not group with any other genotype describing the discriminating power of informative microsatellites. This study identified a set of useful microsatellite markers to accelerate the genetic studies and breeding programme of mungbean.     

Inheritance of seed resistance to bruchids in cultivated mungbean (Vigna radiata, L. Wilczek)

Bruchid beetles or seed weevils are the most devastating stored pests of grain legumes causing considerable loss to mungbean (Vigna radiata (L.) Wilczek). Breeding for bruchid resistance is a major goal in mungbean improvement. Few sources of resistance in cultivated genepool were identified and characterized, however, there has been no study on the genetic control of the resistance. In this study, we investigated the inheritance of seed resistance to Callosobruchus chinensis (L.) and C. maculatus (F.) in two landrace mungbean accessions, V2709BG and V2802BG. The F₁, F₂ and BC generations were developed from crosses between the resistant and susceptible accessions and evaluated for resistance to the insects. It was found that resistance to bruchids in seeds is controlled by maternal plant genotype. All F₁ plants derived from both direct and reciprocal crosses exhibited resistance to the bruchids. Segregation pattern of reaction to the beetles in the F₂ and backcross populations showed that the resistance is controlled by a major gene, with resistance is dominant at varying degrees of expressivity. Although the presence of modifiers was also observed. The gene is likely the same locus in both V2709BG and V2802BG. The resistant gene is considered very useful in breeding for seed resistance to bruchids in mungbean.     

Patterns of genetic diversity in landraces of common bean (Phaseolus vulgaris L.) from Galicia

Knowledge of patterns of genetic diversity among existing cultivars helps to broaden the genetic base of new cultivars and maximizes the use of available germplasm resources. This study examined the organization of diversity for morphological traits in 66 landraces of cultivated common bean (Phaseolus vulgaris) from Galicia and its relationship with phaseolin seed protein diversity. Data on growth habit, seed and pod traits obtained from field evaluations at two locations during the 1989—91 cropping seasons were subjected to multivariate statistical analysis. Cluster analysis based on 14 quantitative variables and five qualitative variables identified 11 groups. The landraces were also characterized by phaseolin electrophoresis. The results allowed separation of these landraces into Middle and Andean American groups, which could be further divided into at least eight groups within the Andean American cultivars and three within the Mesoamerican cultivars. These groups in turn corresponded to the previously described races Nueva Granada and Peru of South American origin, and races Durango, Jalisco and Mesoamerica from the Middle American domestication centre. These results confirm the existence of two major groups of germplasm in the cultivated common bean landraces from Galicia, Mesoamerican vs. Andean American.     

绿豆SSR标记的开发及遗传多样性分析

SSR标记以其数量丰富、多态性好、共显性遗传等优点在基础研究和育种工作中发挥了重要作用,但目前绿豆基因组中的SSR标记依然较少。本研究将磁珠富集法和测序技术相结合高通量检测绿豆基因组SSR位点,鉴定出3,275,355个SSR位点,开发了2742个SSR标记。选取其中157个SSR进行PCR验证,发现有90个(57.33%)标记在10份材料中表现出多态性。挑选40个条带清晰、多态性高、染色体上均匀分布的标记对90份绿豆资源进行遗传多样性分析,单个位点检测到的等位变异数为2～8个,平均为3.0个,有效等位基因数为1.31～4.21个,平均为2.16。Nei’s基因多样性指数在0.23～0.76之间,平均为0.51。多态性信息含量为0.22～0.72,平均为0.43。聚类分析将90份材料分为2个类群,包含4个组。第I组主要由北方资源组成,第Ⅱ组种质来源较为分散,第Ⅲ组主要由山东的资源构成,第Ⅳ组包含多数河北的种质资源。本研究开发的多态性SSR标记不仅可以用于绿豆种质资源的遗传多样性分析,也将在高密度遗传图谱构建、基因定位和分子标记辅助育种中发挥重要作用。     

Potential loss of unique genetic diversity in tomato landraces by genetic colonization of modern cultivars at a non-center of origin

We report the genetic diversity of tomato landraces from Myanmar, with reference to worldwide tomato accessions. Ten simple sequence repeat markers were screened and used to create diversity profiles of tomato germplasm. A total of 133 alleles were amplified from the germplasm investigated; alleles specific to landraces from Myanmar were also observed. Higher genetic diversity for the Myanmar landraces highlighted the broad genetic base of tomato germplasm. Principal component analysis showed that most of the Myanmar landraces were divergent from other accessions. The different approaches used to analyse landrace diversity documented the broader genetic base of germplasm from Myanmar, which is a non‐center of origin for tomato. Genetic diversity in the landraces may be attributable to diverse production systems used by different ethnic groups in Myanmar, ranging from natural hydroponics in eastern areas, to slash‐and‐burn fields in mountain areas and shifting cultivation in many parts of the country. The availability of uniform and high‐yielding cultivars/hybrids threatens the valuable landraces in Myanmar. We emphasize the need to evaluate tomato germplasm from Myanmar for different traits and sustainable use with simultaneous conservation.     

用于中国小豆种质资源遗传多样性分析SSR分子标记筛选及应用

以375份小豆核心种质为试验材料, 利用从小豆及其近缘种SSR引物中筛选出的13对引物进行遗传多样性分析。检测结果显示, 小豆种质资源具有丰富的遗传多样性, 共检测到133个等位变异, 每对SSR引物检测到等位变异4~19个, 平均10.23个, 国内各省多态信息含量(PIC)平均为0.561, 多态位点比例(P)平均为93.523%。聚类结果表明, 小豆资源遗传关系与生态分区间有明显的联系, 且东北地区资源与中南部资源遗传关系较近。湖北、安徽、陕西3省资源的PIC较高, 且基本位于主坐标三维图的中心区域, 推断湖北、安徽、陕西是中国栽培小豆的起源地或多样性中心。该结果有助于更好地对小豆种质资源进行收集、保护和利用。     

黑龙江省大豆遗传结构及遗传多样性分析

利用22个表型性状和60个微卫星(simple sequence repeat, SSR)位点对黑龙江省140份代表性种质(78份地方品种和62份育成品种)进行分析, 根据UPGMA (unweighted pair group method with arithmetic mean)和Model-base对SSR数据进行遗传结构划分。结果表明, 参试品种可分为2大类群, 第II类群的各项多样性指标均高于第I类群, 2个类群遗传距离为0.2427;PCO结果显示这2个类群分布在不同区域, 这与地理来源和育成年代密切有关。依据品种类型分为育成品种和地方品种两组, 后者的各项多样性指标均高于前者, 两组间的遗传距离为0.1131。依据表型数据的PCO分析表明, 分布区域与品种类型有关, 与SSR结构分类的结果吻合度低, 两组品种主要在3个主成分的6个表型性状上有所不同。它们不是2个相对独立的遗传群体, 根据分子标记和表型分类各有特点;建议在种质遗传多样性研究中将分子数据和表型数据结合起来。     

Phenotypic variation in a core collection of common bean (Phaseolus vulgaris L.) in the Netherlands

Forty accessions, forming a core collection of mainly bush type of the common bean (Phaseolus vulgaris L.) germplasm in the Netherlands, were evaluated for 14 qualitative and quantitative traits at the Agricultural University, Wageningen (WAU), the Netherlands in 1992. These and an additional 117 Dutch accessions, mainly collected in private home gardens, were also evaluated for phaseolin seed protein pattern, and morphological and agronomic traits at the International Center for Tropical Agriculture (CIAT, Spanish acronym), Cali, Columbia between 1987 and 1997. Multivariate and principal component analyses at both WAU and CIAT indicated existence of one large group with no discernable patterns among Dutch common bean collections of landraces, garden forms and cultivars. However, when phaseolin, an evolutionary, biochemical marker, was used as an initial classification criterion followed by use of morphological markers, the two major gene pools; Andean and Middle American with two races in each (Chile and Nueva Granada in Andean, and Durango and Mesoamerica in Middle American) were identified. The Andean gene pool was predominant (136 of 157 accessions), especially the race Nueva Granada (126 accessions) characterized by the bush determinate growth habit type I and T phaseolin. The new core collection comprised 31 accessions. Bean races Chile, Durango, and Mesoamerica were represented by 10, 7, and 14 accessions, respectively. Of the 9 French or snap bean accessions six possessed characteristics of race Mesoamerica and three belonged to Durango race. Occurrence of these and a large number of other recombinants strongly suggested considerable hybridization and gene exchange between Andean and Middle American gene pools, thus blurring the natural boundaries and forming a large single group of common bean germplasm in the Netherlands. The inter-gene-pool recombinants of both dry and French beans should be of special interest to breeders for use as bridging-parents for development of broad-based populations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Agronomical and quality traits of runner bean germplasm and implications for breeding

White runner bean landraces are greatly appreciated in the North highlands of Spain due to their excellent culinary seed quality. Runner bean cultivars are grown like pole beans. Diversity within a runner bean collection of 31 accessions from the Iberian Peninsula (Spain and Portugal) was examined using morphological, agronomical and seed quality traits. Landraces showed significant differences for most of the agronomical and seed quality traits studied except for seeds per pod, water absorption, seed coat tenderness and floury texture. Runner bean landraces showed sufficient variability to select inbred lines for future breeding. Genotype × environment interaction was significant for days to first flowering, days to first dry pod, seeds per pod and seed length. The majority of physical and nutritional seed quality traits studied which are important to determine the commercial value of a variety were not subject to environmental influences. Different selection pressures affecting to the runner bean genetic material could have occurred in several regions of the Iberian Peninsula. Extra-large and high yielding runner bean germplasm was identified and represents a valuable source of genetic diversity that has potential for development of improved cultivars to be chosen for commercialisation. This revised version was published online in July 2006 with corrections to the Cover Date.     

Resistance to cowpea mottle carmovirus in Vigna vexillata

Cowpea mottle carmovirus (CPMoV) causes grain yield losses of up to 75% in cowpea (Vigna unguiculata [L.] Walp.). There is no resistance to this virus among cultivated cowpea lines, but a high level of resistance exists in Vigna vexillata, a wild Vigna species. Fifty‐four accessions of V. vexillata germplasm collection at IITA were tested for resistance to CPMoV. Seedlings were mechanically inoculated with the virus and susceptibility or resistance was assessed by visual scoring of disease symptoms and serological analysis using antigen‐coated plate enzyme‐linked immunosorbent assay (ACP‐ELISA). All but three V. vexillata lines belonging to the variety angustifolia were resistant to CPMoV. Crosses were made between two resistant V. vexillata lines and the three susceptible lines. Segregation patterns observed in the F₂ and the backcross populations of all the crosses showed that resistance to CPMoV in V. vexillata is controlled by a single dominant gene, and the level of resistance conferred by this gene in V. vexillata is very high.