Genetics of disease resistance in urdbean (Vigna mungo (L.) Hepper) to the leaf spots caused by Colletotrichum truncatum (Schw.) Andrus & Moore

Summary The resistance sources among various test cultivars of urdbean to Colletotrichum truncatum, a leaf spotting pathogen, were identified and genetics of resistance was worked out by studying F₁, F₂ and F₃ generations of crosses between resistant cultivars and the susceptible cv. Kulu 4 and of those among the resistant parents. The resistance was found to be controlled by single dominant genes and the resistance genes were non-allelic.     

Interspecific hybridization between rice bean (Vigna umbellata) and its wild relative (V. minima): Fertility-sterility relationships

Summary The pre-and post-fertilization barriers in the interspecific crosses between Vigna umbellata and V. minima were investigated. In the reciprocal crosses (V. minima as the parent) the entry of pollen tubes into the ovary was delayed by about 4 h, and no seed set was observed. However, no pre-fertilization barriers were encountered in crosses involving V. minima as the parent and V. umbellata as the parent (normal cross). The delay/absence of divisions in the endosperm and the failure of embryo to divide were the post-fertilization barriers responsible for somatoplastic sterility in normal crosses which yielded a few hybrid seeds. The hybrid seeds showed poor germinability. The F₁ hybrids were intermediate between the parents in most morphological characters, and are completely sterile for pollen. Backcrossing of F₁ hybrid with both the parents did not restore fertility in the progenies. V. minima is considered as the tertiary gene pool of the rice bean.     

小豆种子活力的初步研究

早熟、丰产、抗逆性强的小豆新品种的选育以及大田生产中选用优质小豆种子均需要了解种子的活力水平。本研究选用3个小豆品种的新、陈种子,应用电导率法和发芽法进行种子活力测定,结果表明:小豆种子试样间在电导率、发芽势、发芽率、幼苗生长量和活力指数等方面均存在差异。不同品种间种子活力水平表现为红小豆LSH1>绿小豆>红小豆8937;同一品种的活力水平表现为新种子>陈种子;电导率值与种子活力指数呈显著负相关(r=-0.8667)。本研究认为,电导率法是测定小豆种子活力的一种简便、快速、有效的方法。     

Interspecific hybridization between Vicia faba (L.) and Vicia narbonensis (L.): Early pod growth and embryo-sac development

Summary Fertilized embryo-sac development and pod growth was studied in one Vicia faba cultivar, one Vicia narbonensis population and their reciprocal crosses. The initial development of endosperm and embryo was at least four days faster in V. narbonensis than in V. faba. Pods and ovules developed also faster in V. narbonensis than in V. faba. The growth rate of the hybrid pods followed the growth rate of the mother species, but was slower than that of the pods from selfed flowers. In the cross V. narbonensis × V. faba the ovules stopped growing 9 days after pollination, while in the reciprocal cross they stopped growing 15 days after pollination. Hybrid embryo-sacs from V. faba × V. narbonensis were aborted before they reached the stage of 256 endosperm nuclei or 200 embryo cells. Selfed V. faba embryo-sacs reached this stage in less than 9 days after pollination. In the reciprocal cross the embryo-sacs were aborted before they reached the stage of 128 endosperm nuclei or 80 embryo cells. Selfed V. narbonensis embryo-sacs reached this stage at the 4th day after pollination. Given that at these stages the embryo has less than 200 cells it was concluded that an in-ovule embryo culture technique should be developed to obtain hybrid plants.     

Interspecific hybridization between Vigna unguiculata (L.) Walp. and V. vexillata (L.) A. Rich. through in vitro embryo culture

Immature embryos resulting from the cross V. vexillata × V. unguiculata were cultured on MS medium supplemented with 2,4-D (2 mg/l) and resulted in embryogenic calli. Thirteen hybrid regenerants were obtained via organogenesis by subculturing the calli on MS medium supplemented with BAP (2 mg/l) + adenine sulphate (40 mg/l) + CH (500 mg/l) + cowpea tender pod extract (10%). The interspecific regenerants showed intermediate morphological traits between the parents for leaf shape, pod colour and seed coat colour. The hybrid plants inherited stem, leaf and pod hairiness of the wild species which could serve as a mechanical barrier against viral vectors. Electrophoretic studies of two isozyme systems, peroxidase and esterase, also confirms the hybrid nature of the regenerants as they expressed unique bands of both parents. Cytological study of the meiotic chromosomes revealed high frequency of univalent formation in the hybrids suggesting that the genomes of the parental species are structurally differentiated. The hybrid regenerants exhibited high enzyme activity for three enzymes viz., peroxidase, polyphenol oxidase and phenyl alanine ammonia lyase over the cultivated parent which may be useful in conferring resistance against viral pathogens. This revised version was published online in July 2006 with corrections to the Cover Date.     

Development of an interspecific Vigna linkage map between Vigna umbellata (Thunb.) Ohwi & Ohashi and V. nakashimae (Ohwi) Ohwi & Ohashi and its use in analysis of bruchid resistance and comparative genomics

To facilitate transfer of bruchid resistance to azuki bean (Vigna angularis) from its relatives an interspecific mapping population was made between rice bean, V. umbellata, and the related wild species V. nakashimae. The V. umbellata parent is completely resistant and V. nakashimae is completely susceptible to the bruchid beetle pests, azuki bean weevil (Callosobruchus chinensis) and cowpea weevil (C. maculatus). There is very low cross compatibility between V. umbellata and azuki bean. Therefore, V. nakashimae, that crosses with both V. umbellata and V. angularis without the need for embryo rescue, is used as a bridging species. A genetic linkage map was constructed based on an interspecific F₂ mapping population between V. umbellata and V. nakashimae consisting of 74 plants. A total of 175 DNA marker loci (74 RFLPs and 101 SSRs) were mapped on to 11 linkage groups spanning a total length of 652 cM. Segregation distortion was observed but only three markers were not linked to any linkage group due to severe segregation distortion. This interspecific genome map was compared with the genome map of azuki bean. Of 121 common markers on the two maps, 114 (94.2%) were located on the same linkage groups in both maps. The marker order was highly conserved between the two genome maps. Fifty F₂ plants that produced sufficient seeds were used for quantitative trait locus (QTL) analysis and locating gene(s) for C. chinensis and C. maculatus resistance in V. umbellata. The resistance reaction of these F₂ plants differed between C. chinensis and C. maculatus. Both resistances were quantitatively inherited with no F₂ plants completely susceptible to C. chinensis or C. maculatus. One putative QTL for resistance to each of these bruchid species was located on different linkage groups. Other putative QTLs associated with resistance to both C. chinensis and C. maculatus were localized on the same linkage group 1. Linked markers associated with the bruchid‐resistant QTL will facilitate their transfer to azuki bean breeding lines.     

Application of SSR markers and real-time PCR for variety identification in azuki-bean [Vigna angularis (Willd.) Ohwi and Ohashi]

The azuki bean in Korea consists of seven domestic varieties which have been developed and registered for the public during last 25 years. Here, we present a simple but reliable method to screen and identify Korean azuki bean varieties. A method based on simple sequence repeat (SSR) markers is widely used for prominent gene identification and variety discrimination. In molecular biology, real-time polymerase chain reaction (PCR) is a laboratory technique based on the polymerase chain reaction that is used to amplify and simultaneously quantify a targeted DNA molecule. It enables easy detection of a specific sequence in a DNA sample without performing electrophoresis and further processes. For separation of seven Korean azuki bean varieties, 110 unique azuki bean SSR markers from an (AG)n-enriched library were selected, synthesized and used for polymerase chain reaction (PCR). Data were taken through acrylamide gel electrophoresis and automated multi-capillary electrophoresis system for selection of specific markers and then changed into proper formats for data mining analysis. Ten primer pairs that showed high polymorphism were chosen for the indepth study. These ten primers were re-amplified with real-time PCR and checked the cycle threshold (Ct) and temperature (Tm) for comparison of amplification sequence in seven varieties. Consequently, a total of 20 alleles and 6 SSR primers were detected from the standard PCR amplification. Within these 6 primers, 7 alleles of 3 SSR primers were isolated for variety identification. From real-time PCR results, 3 SSR primers were selected as efficient markers for discrimination of seven Korean azuki bean varieties. The approach described here could be applied in monitoring our varieties and can be adapted in the azuki bean breeding program.     

我国小豆种质资源形态多样性鉴定与分类研究

对224份小豆种质资源的形态多样性进行了研究鉴定, 结果表明, 我国小豆种质资源具有丰富的形态多样性, 平均多样性指数为1.035, 高于国外材料(0.827)20.1%. 通过多变量的主成分分析, 第一主成分和第二主成分一共代表了小豆形态多样性的56%. 基于形态性状, 把224份小豆种质聚类并划分为三大组群, 第一组群, 生育期较长, 植株较     

Cross-Compatibility of Cultivated Adzuki Bean (Vigna angularis) and Rice Bean (V. umbellata) with Their Wild Relatives

The wild Ceratotropis species which occur in Japan consist of Vigna angularis var. nipponensis, V. nakashimae, V. riukiuensis and V. reflexo-pilosa. The main aim of the present study is to detect cross-compatibility of adzuki bean and rice bean with these wild relatives. The cross of V. nakasbimae with adzuki bean was successful in both directions and viable seeds were produced, while the cross with rice bean was not successful. V. riukiuensis is cross-compatible with adzuki bean and rice bean only when V. riukiuensis is used as male parent and the hybrid seeds showed normal germination. The cross between V. reflexopilosa and the two cultigens adzuki bean and rice bean did not succeed in either direction. The present results suggest that V. nakashimae and V. riukiuensis could be used as gene sources for the adzuki bean breeding programme through interspecific hybridization. Successful hybrids between adzuki bean and rice bean cannot easily be produced. However, since V. riukiuensis is cross-compatible with both adzuki bean and rice bean, V. riukiuensis could be useful as a bridge species between adzuki bean and rice bean.     

以地理来源分组和利用表型数据构建中国小豆核心种质

中国国家种质库里保存着4 877份小豆(Vigna angularis)种质资源,但仅有部分资源在小豆的改良中被利用。构建小豆核心种质, 不但能简化管理, 而且可以提高遗传资源在小豆改良中的利用效率。根据我国小豆的地理来源进行分组, 在分组的基础上利用表型数据, 以类平均法聚类构建了中国现有小豆核心种质, 共435份资源, 占资源总量的8.92%, 涵盖98.3%的表型变异类型, 控制不同性状表型相关的相互适应的遗传复杂性在核心种质中也得到了适当的保持。不同性状的均值t测验、方差F测验、表型频率分布、χ²测验等分析表明, 所构建的小豆核心种质可以作为整体资源的代表性样本, 在小豆种质资源的充分利用中将发挥重要作用。     

Adaptation and competition in mixtures of cowpea (Vigna unguiculata (L.) Walp.)

Summary Six pure lines and four mixtures were grown in six environments throughout lowland Papua New Guinea. There were no significant transgressive increases in the grain yield of mixtures above their pure line components, and mixture yields were adequately predicted by the mean of components. In yield stability it was found that the individual buffering of pure lines was of more importance than population buffering, and that the magnitude of population buffering varied with the particular combination of components. Competitive effects in all the mixtures were of the compensating type. Dramatic changes in mixture composition resulting from natural selection precluded their use in local agriculture. The outcome of competition in mixtures was strongly influenced by the growing environment, such that selective index was correlated to general fertility.     

ISSR and SCAR markers linked to the mungbean yellow mosaic virus (MYMV) resistance gene in blackgram [Vigna mungo (L.) Hepper]

A recombinant inbred line (RIL) mapping population (F₈) was generated by crossing Vigna mungo (cv. TU 94‐2) with Vigna mungo var. silvestris and screened for mungbean yellow mosaic virus (MYMV) resistance. The inter simple sequence repeat (ISSR) marker technique was employed to identify markers linked to the MYMV resistance gene. Of the 100 primers screened, 54 showed amplification of which 36 exhibited polymorphism between the parents TU 94‐2 (resistant) and V. mungo var. silvestris (susceptible). Individual plants from 53 RIL populations were analysed and one marker (ISSR811₁₃₅₇) was identified as tightly linked to the MYMV resistant gene at 6.8 cM. Both the phenotype as well as the ISSR811₁₃₅₇ marker segregated in a 1 : 1 ratio. The ISSR811₁₃₅₇ marker was sequenced and sequence characterized amplified region (SCAR) primers were designed (YMV1‐F and YMV1‐R) to amplify the marker. Screening for the SCAR marker in the RIL population distinguished the MYMV resistant and susceptible plants, agreeing well with the phenotypic data. The ISSR811₁₃₅₇ marker was validated using diverse blackgram genotypes differing in their MYMV reaction. The marker will be useful for the development of MYMV‐resistant genotypes in blackgram.     

The domestication and improvement of cowpeas (Vigna unguiculata (L.) Walp.)

Summary Physiological and morphological characteristics of the two wild and three domesticated subspecies of cowpeas are compared. The wild accessions are alike in having small, hard seeds borne in dehiscent pods, but differ in other characteristics. We suggest that the wild subsp. dekindtiana, from the seasonally-arid tropics, is more likely to have been the progenitor of modern cowpeas than the other wild subspecies (subsp. mensensis), but that subsp. dekindtiana was first cultivated in the humid tropics where its pods are slow to dehisce. Domestication has been associated with changes in the structure of pod valves and seed coats which reduce pod dehiscence and seed hardness. Pods and seeds have increased in size, mainly by increases in the rate of dry weight accumulation, and their increase has been only partly paralleled by increase in the area of subtending leaves. There has been no increase in the maximum photosynthetic rate of leaves, but the duration of their photosynthetic activity has increased. Domesticates are less sensitive than are wild plants to some environmental controls, such as in the response of germination to temperature, but in their flowering responses to daylength both wild and cultivated forms retain sensitivity under conditions where this is of adaptive value.     

Effects of photoperiod and temperature on flowering in mungbean (Vigna radiata (L.) Wilczek)

Summary Forty strains of mungbeans, including all of the entries in the 1st and 2nd International Mungbean Nurseries (IMN), were grown in plant growth chambers in photoperiods of 12, 13, 14, 15, and 16 hours and mean temperatures of 18, 23, and 28°C. Results show that (1) mungbean strains differ in their flowering response to photoperiod and to mean temperature; (2) increasing the photoperiod of reducing the mean temperature delayed flowering, the amount of delay varied with the strain: (3) variations in mean temperature may alter the effect of the photoperiod on flowering in particular strains. Twenty-five strains which flowered in all photoperiod-temperature treatments could be divided into four groups, each group being successively later in flowering. The remaining 15 strains could be divided into four groups, according to their failure to flower within 105 days in specific photoperiod-temperature treatments. Flowering response in the growth chamber is useful in explaining flowering response in the 2nd IMN at three locations varying from 14°N to 49°N latitude.Contribution No 7516 from the Missouri Agricultural Experiment Station.     

Interspecific hybridization in Cucumis (L.). I. Need for genetic variation,biosystematic relations and possibilities to overcome crossability barriers

Summary The genetic variation in pickling and slicing cucumbers (C. sativus L.) seems insufficient to enable the breeder to solve serious cultivation problems. Wild allies of C. sativus possess a wide range of interesting characters which could be incorporated by means of species crosses. In the past, attempts to achieve such crosses have failed. Important characters of Cucumis species are mentioned and biosystematic problems discussed. Results of previous research on species crosses prompted an examination of the prospects of further research.     

Inheritance of seed size in cowpea (Vigna unguiculata (L.) Walp.)

Summary The inheritance of seed weight in cowpea was examined in a field planting of the parents, reciprocal F₁s, F₂s and backcrosses to both parents of a cross between TVu 1977-OD (small seeded) and ACC 70002 (large).Seed weight was inherited quantitatively and small seed was partially domiminant to large seed size. Gene action was predominantly additive but dominance and additive × additive epistatic effects were also significant.Broad and narrow sense heritabilities were 85.1±5.3% and 75.4±18.6% respectively. The minimum number of loci involved in the inheritance of seed size was eight, and each gene pair contributed up to 1.02 g increase to seed weight. The estimate of genetic advance from F₂ to F₃ generations with 5% selection intensity was 3.58 g.International Institute of Tropical Agriculture, Semi-Arid Food Grain Research and Development Project/National Cowpea Improvement Program, B.P. 1783, Ouagadougou, Upper Volta.Department of Primary Industries, Hermitage Research Station, Warwick 4370, Queensland, Australia.International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), ICRISAT Center, Patancheru, P.O. 502 324, A.P., India.     

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.     

Interspecific hybridization in the genus Gossypium through embryo culture

Summary With embryo rescue and culture techniques interspecific hybrids have been obtained amongst diploid cultivated (Gossypium arboreum, G. herbaceum) and wild species (G. stocksii, G. anomalum) of cotton. The early abscission of the young bolls was prevented by repeated application of growth regulators followed by the culture of immature hybrid embryos (15 days after pollination). The best growth and development were obtained on modified Murashige and Skoog's medium with supplements. The plants were stouter when the cultures were first incubated in the dark for 15–25 days, followed by exposure to light. The hybrid plants transferred to soil developed further and matured, and were more or less intermediate between their respective parents.     

Linkage relationships among loci controlling morphological traits in cowpea (Vigna unguiculata (L.) Walp.)

Summary Linkage among loci controlling various morphological traits in cowpea were determined using F₂ progenies. Data were collected on individual plants of four crosses segregating for several loci. Recombination estimates between the following pairs of loci were as follows: Sw (swollen vs normal stem base)-Fbc (cream vs green flower buds) (41±4.8%), Pu^s(purple vs green stems)-Cbr (cocoa-brown vs straw-yellow dry pods) (31±5.7%), Pu^p(purple vs green immature pods)-Cbr (30±5.7%), Pu^s-Pu^p (4±1.5%), Ndt (non-determinate vs determinate)-Pd (peduncle colour) (26±2.8%), Ndt-Hg (semi-erect vs erect plant type) (26±2.8%), P^t(purple vs green pod tips)-Bk (greyish-black vs straw-yellow dry pods) (19±2.4%) and Hg-Bpd (normal vs branching peduncle (24±9.5%). Four linkage groups (LG) were identified in these studies. The proposed LG I contained loci Sw and Fbc; LG II loci Pu^s, Pu^p, and Cbr; LG III loci Pd, Ndt, Gh, and Bpd; and LG IV loci P^tand Bk.     

Genetic characterization of Malawian cowpea (Vigna unguiculata (L.) Walp) landraces: diversity and gene flow among accessions

Cowpea has been cultured from ancient timesin Africa and Asia and is now widespreadthroughout the tropics and subtropics.Cowpea accessions can be distinguishedphenotypically from one another by theirgrowth habit, time to maturity, yield, andseed size and colour. Data on geneticdiversity of farmer-developed accessionsare lacking. The main objective of thisstudy was to determine the pattern andextend of RAPD marker variation within andamong cowpea populations from differentagroecological zones and to determine thedegree of genetic relationships and geneflow among different landraces used bylocal farmers. Twenty of the 30 RAPDprimers tested allowed amplifications ofrandom polymorphic (RAPD) loci. Overall,80% of the scored loci were polymorphic. The degree of band sharing was used toevaluate genetic distance betweenaccessions and to construct a phylogenetictree. The genetic distance values amongaccessions varied between 0.09 to 0.57.Although some small clusters groupedaccessions of the same growth habits, ageneral lack of agreement betweenclustering and morphological features wasobserved. The analysis of molecularvariance revealed that within-region ortypes (among accessions) variationaccounted for 96% of the total molecularvariance. This high within-accessionvariability is being sustained by anuncontrolled gene flow among populations.