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Genetic diversity of Cambodian melon was evaluated by the analysis of 12 RAPD and seven SSR markers using 62 accessions of melon landraces, and compared with 231 accessions from other areas for genetic characterization of Cambodian melon. Among 62 accessions, 56 accessions were morphologically classified as small-seed type whose seed length was shorter than 9mm as like groups Conomon and Makuwa. Gene diversity of Cambodian melon was 0.228 which was equivalent to those of groups Conomon and Makuwa and smaller than those of Vietnamese and Central Asian landraces. Phylogenetic tree constructed from genetic distance matrix classified 293 accessions into three major clusters. Small-seed type accessions from east and southeast Asia formed clusters I and II, which were distantly related with cluster III consisted of large-seed type melon from other areas. All of Cambodian melon belonged to cluster I except three accessions, along with those from Thailand, Myanmar, Yunnan (China) and “Dua thom” from northwestern part of Vietnam, indicating genetic similarity in these areas. In addition, Cambodian melon was not differentiated among geographical populations. Conomon and Makuwa were classified into cluster II, together with melon groups from plain areas of Vietnam. The presence of two groups of melon in southeast Asia was also indicated by population structure and PCO analysis. These results indicated close genetic relationship between Cambodia and the neighboring countries, and suggested that Cambodian melon is not directly related with the establishment of Conomon and Makuwa.

A total of 401,293 germination test results accumulated over 30 years for 110,819 seed lots of 50 species and 4 sub-species stored at –1°C and 30% relative humidity was analysed to assess seed longevity. The Kaplan- Meier estimator was used to perform comprehensive analysis, including censored observations due to seed lot regeneration or substitution. The Weibull distribution was used to model the observed and censored death times for each survival curve and its parameters were used as the common indicator. Interspecific differences in seed longevity, the time until the germination percentage falls below 85% of its initial value, were observed. The shortest and longest seed longevity periods were 8.4 and 127.1 years for Panicum maximum and Cucumis sativus, respectively. Moreover, intra-specific differences in seed longevity were observed at different levels of domestication and geographical distribution of origin. For example, the average seed longevity of six-row Hordeum vulgare landraces that originated from southern Europe showed 2.5 times that of the average of this species. Here, reference materials for setting appropriate intervals of germination testing are presented.

Vigna riukiuensis plant – a rare type of vigna, found only in Taiwan and the islands of Okinawa prefecture, Japan – possesses intrinsic property of high level of salt and heat tolerance. To underst...

The genus Vigna contains important crops such as cowpea and mungbean. Wild Vigna showing higher salt tolerance than Vigna crops were screened and their tolerance mechanisms are discussed. Primary screening using 7 Vigna crops and 23 wild Vigna under 300 mM NaCl selected V. luteola, V. marina and V. vexillata. A study under different salt concentrations revealed the highest survival ability of V. marina. Diversity of salt tolerance in each species was revealed using a total of 230 accessions. Growth and physiological responses under 150 mM NaCl were then compared using two selected accessions from each species. The pattern of Na+ accumulation in roots, stems and leaves suggested that V. vexillata (V1) and V. luteola (L8, L9) are ‘Na+ excluder’ type, while V. marina (M1, M4) is ‘Na+ includer’ type. V. luteola (L8, L9) showed the highest dry matter production under control condition and well-maintained shoot dry weight under salt stress. Interestingly, V. luteola (L9) accumulated the highest Na+ in roots (3000 μM g−1) and increased root dry weight under salt stress, which might work as Na+ reservoir restricting Na+ transition to the leaves, leading to the increased photosynthetic rate. V. luteola has great potential in areas where moderate salt damage occurs. V. marina (M1, M4) accumulated Na+ at high level in roots, stem, and leaves. Under salt stress, they increased stomatal conductance, transpiration rate, and photosynthetic rate, which suggested the adaptational regulation of aquaporin gene expression. V. marina will be useful as food, pasture and phytoremediation legumes in highly salt-damaged areas.

Black gram (Vigna mungo var. mungo) is an important pulse crop in Asia. The cowpea weevil (Callosobruchus maculatus) is a stored-seed insect pest (seed weevil/bruchid) that causes serious postharvest losses in pulse crops, including black gram. In this study, we constructed a high-density linkage map for black gram and identified quantitative trait loci (QTLs) for C. maculatus resistance. A recombinant inbred line (RIL) population of 150 lines from a cross between BC48 [cultivated black gram (var. mungo); bruchid-susceptible] and TC2210 [wild black gram (var. silvestris); bruchid-resistant] were used to construct a linkage map of 3,675 SNP markers from specific-locus amplified fragment sequencing. The map comprised 11 linkage groups spanning 1,588.7 cM with an average distance between adjacent markers of 0.57 cM. Seeds of the RIL population grown in 2016 and 2017 were evaluated for C. maculatus resistance through two traits; the percentage of damaged seeds (PDS) and infestation severity progress (AUDPS). Inclusive composite interval mapping identified three QTLs each for PDS and AUDPS. Two QTLs, qVmunBr6.1 and qVmunBr6.2, mapped about 10 cM apart on linkage group 6 were common between PDS and AUDPS. Comparative genome analysis revealed that qVmunBr6.1 and qVmunBr6.2 are new loci for C. maculatus resistance in Vigna species and that genes encoding a lectin receptor kinase and chitinase are candidates for qVmunBr6.2. The high-density linkage map constructed and QTLs for bruchid resistance identified in this study will be useful for molecular breeding of black gram.

The moth bean (Vigna aconitifolia), possibly the most primitive crop of the genus Vigna, is a highly drought- and heat-resistant legume grown in arid areas. Moth bean domestication involved phenotypic changes, including reduction of seed dormancy and pod shattering, increased organ size, and earlier flowering and maturity. However, the genetics of the domestication process in moth bean is not known. In this study, we constructed a genetic linkage map for moth bean and used the map to identify quantitative trait loci (QTL) for domestication-related traits of an F2 population of 188 individuals produced from a cross of wild moth bean (TN67) and cultivated moth bean (ICPMO056). The genetic linkage map comprised 11 linkage groups (LG) of 172 simple sequence repeat markers and spanned a total length of 1016.8 centiMorgan (cM), with an average marker distance of 7.34 cM. A comparative genome analysis showed high genome synteny between moth bean and mungbean (Vigna radiata), adzuki bean (Vigna angularis), rice bean (Vigna umbellata), and yardlong bean (Vigna unguiculata). In total, 50 QTLs and 3 genes associated with 20 domestication-related traits were identified. Most of the QTLs belonged to five LGs (1, 2, 4, 7, and 10). Key traits related to domestication such as seed dormancy and pod shattering were controlled by large-effect QTLs (PVE > 20%) with one or two minor QTLs, whereas all other traits were controlled by one–seven minor QTLs, apart from seed weight, which was controlled by one major and seven minor QTLs. These results suggest that a small number of mutations with large phenotypic effects have contributed to the domestication of the moth bean. Comparative analysis of QTLs with related Vigna crops revealed that there are several domestication-related large-effect QTLs that had not been used in moth bean domestication. This study provides a basic genetic map and identified genome regions associated with domestication-related traits, which will be useful for the genetic improvement of the moth bean and related Vigna species.

Vigna nakashimae is one of the closely related species of Vigna angularis (Adzuki bean). Two strain of ‘Ukushima’ and ‘G418’ were identified as salt tolerance strains in Vigna nakashimae from gene bank collection. F2 populations from an inter- or intra-specific cross between the sensitive and tolerant strains are useful for the detection of salt tolerance QTL in Vigna nakashimae. Although Vigna angularis reference genome is available and useful for genetic analysis by genotyping-by-sequencing/RADseq in closely related species, it is not enough for isolation of responsible genes. To reveal sequence variation in Vigna nakashimae “Ukushima” and “G418”, the whole genome sequencing was performed using Illumina HiSeq X Ten system (411,174,986 and 478,116,282 read). NGS data was deposited in the DNA Data Bank of Japan (DDBJ) under accession number DRA009307. Keywords: Vigna nakashimae, Closely wild relatives of Adzuki bean, Next-generation sequencing, HiSeq, Whole genome sequencing

This chapter reviews the taxonomic treatment of mungbean (Vigna radiata) and its relatives and presents photographs of key characters and a phylogenetic tree. After Tomooka et al. (2002a) listed 21 species in the subgenus Ceratotropis, six new species have been described. Phylogenetic analysis revealed the positions of two new species, Vigna indica and Vigna sahyadriana, but the remaining four new species, Vigna konkanensis, Vigna pandeyana, Vigna sathishiana, and Vigna yadavii still need to be analyzed. We transferred Vigna subramaniana from the section Ceratotropis to section Aconitifoliae. Two unidentified gene bank accessions, NI1135 and JP245506, have been referred for assessment with regard to the relationships with the new species. The gene pool for mungbean has been expanded from Tomooka et al. (2005), with special emphasis on Vigna vexillata and NI1135. Drought and salt stress studies are cited to show the importance of systematic screening for stress tolerances, which is the first step toward exploiting novel genes expected in wild relatives.

Mungbean (Vigna radiata (L.) Wilcz.) is an important leguminous crop cultivated mainly in Asia. Its wild relatives are considered useful genetic resources for mungbean breeding. However, the taxonomic history of mungbean and its wild relatives is complicated and some confusion is still present in recent publications. In this study, we examined the rDNA-ITS sequences and morphological characteristics of 83 gene bank accessions closely related to mungbean. As a result, we classified the 83 accessions into five species and one unclassified accession. The proper taxon name for each species was determined (Vigna grandiflora (Prain) Tateishi et Maxted, Vigna mungo (L.) Hepper, V. radiata, Vigna subramaniana (Babu ex Raizada) Raizada, and Vigna trinervia (Heyne ex Wight et Arn.) Tateishi et Maxted) based on a review of their taxonomic history and morphological comparisons between gene bank accessions and type specimens. A new taxonomic treatment is proposed and a morphological key has been prepared. In this treatment, Vigna radiata (L.) Wilcz. var. setulosa (Dalz.) Ohwi et Ohashi is treated as a synonym of Vigna radiata (L.) Wilcz. var. sublobata (Roxb.) Verdc., and Vigna hainiana Babu, Gopinathan et Sharma is a synonym of V. subramaniana. Accession ‘NI1135’ was revealed to be most closely related to, but is considerably differentiated from, mungbean (V. radiata) based on its rDNA-ITS sequences. It also has distinguishing morphological characters. Plants with similar morphologies and DNA profiles might be distributed in the Indian Himalaya. However, since only one accession is available at present, the taxonomic classification of ‘NI1135’ needs to be reconsidered in the future.

Seed size is one of the most important traits in leguminous crops. We obtained a recessive mutant of blackgram that had greatly enlarged leaves, stems and seeds. The mutant produced 100% bigger leaves, 50% more biomass and 70% larger seeds though it produced 40% less number of seeds. We designated the mutant as multiple-organ-gigantism (mog) and found the mog phenotype was due to increase in cell numbers but not in cell size. We also found the mog mutant showed a rippled leaf (rl) phenotype, which was probably caused by a pleiotropic effect of the mutation. We performed a map-based cloning and successfully identified an 8 bp deletion in the coding sequence of VmPPD gene, an orthologue of Arabidopsis PEAPOD (PPD) that regulates arrest of cell divisions in meristematic cells. We found no other mutations in the neighboring genes between the mutant and the wild type. We also knocked down GmPPD genes and reproduced both the mog and rl phenotypes in soybean. Controlling PPD genes to produce the mog phenotype is highly valuable for breeding since larger seed size could directly increase the commercial values of grain legumes.

A rapid quantification method for leaf sodium (Na+) and potassium (K+) concentrations was developed using a compact ion meter. Leaf ion concentrations were evaluated for species of Vigna vexillata (L.) A. Rich. after two weeks of treatment with 0–200 mM of sodium chloride. To compare the ion extraction efficiency, extraction solutions of distilled water and ammonium acetate were tested. The ion concentrations of extracts obtained by both solutions were measured using an ion meter, and the values were validated using ion chromatography. For both extraction solutions, the ion meter values were highly correlated with those of the ion chromatograph. However, correlations between ion meter and chromatograph values were largely different for Na+ and K+. The rapid quantification of ion concentrations using an ion meter developed in this study was successfully utilized for evaluating differences in leaf Na+ concentrations, K+ concentrations, and K+/Na+ ratio in species of V. vexillata.

Vigna nakashimae is one of the closely related species of Vigna angularis (Adzuki bean). Two strain of ‘Ukushima’ and ‘G418’ were identified as salt tolerance strains in Vigna nakashimae from gene bank collection. F2 populations from an inter- or intra-specific cross between the sensitive and tolerant strains are useful for the detection of salt tolerance QTL in Vigna nakashimae. Although Vigna angularis reference genome is available and useful for genetic analysis by genotyping-by-sequencing/RADseq in closely related species, it is not enough for isolation of responsible genes. To reveal sequence variation in Vigna nakashimae “Ukushima” and “G418”, the whole genome sequencing was performed using Illumina HiSeq X Ten system (411,174,986 and 478,116,282 read). NGS data was deposited in the DNA Data Bank of Japan (DDBJ) under accession number DRA009307.

SUMMARYLegumes have evolved a unique manner of seed dispersal in that the seed pods explosively split open with helical tension generated by sclerenchyma on the endocarp. During domestication, azuki bean (Vigna angularis) and yard-long bean (Vigna unguiculata cv-gr. Sesquipedalis) have reduced or lost the sclerenchyma and lost the shattering behavior of seed pods. Here we performed fine-mapping with back-crossed populations and narrowed the candidate genomic region down to 4 kbp in azuki bean and 13 kbp in yard-long bean. Among genes located in these regions, we found MYB26 genes encoded truncated proteins in both the domesticated species. We also found in azuki bean and other legumes that MYB26 is duplicated and only the duplicated copy is expressed in seed pods. Interestingly, in Arabidopsis MYB26 is single copy and is specifically expressed in anther to initiate secondary wall thickening that is required for anther dehiscence. These facts indicated that, in legumes, MYB26 has been duplicated and acquired a new role in development of pod sclerenchyma. However, pod shattering is unfavorable phenotype for harvesting and thus has been selected against by human.

Adzuki bean brown stem rot (BSR), caused by Cadophora gregata f. sp. adzukicola, and adzuki bean Fusarium wilt (AFW), caused by Fusarium oxysporum f. sp. adzukicola, are serious problems in Hokkaido, Japan, and have been managed using cultivars with multiple resistance. However, a limited number of adzuki bean varieties are resistant to these pathogens because of the frequent appearance of new races; thus, new sources of resistance have been sought in related Vigna spp., particularly in the section Angulares to which adzuki bean belongs. An analysis of selected Vigna accessions (JP81231 to JP235420) conserved in the Genebank of the National Institute of Agrobiological Sciences, Tsukuba, Japan, revealed wide variation in resistance spectra and resistance combinations, and eight disease response groups (A to H) were identified. Four of eight were newly detected response groups, suggesting the existence of novel resistance genes. Of 252 accessions from 26 species, 28 accessions in Vigna angularis var. nipponensis, V. hirtella, V. minima, and V. tenuicaulis (section Angulares) from group D, which were cross-compatible with adzuki bean, are expected to be potential sources of multiple resistance genes. They were resistant to all races of BSR and AFW pathogens examined.

The screening of mini-core collection of azuki bean accessions (Vigna angularis (Willd.) Ohwi & Ohashi) for comparative uptake of (137)Cs in their edible portions was done in field trials on land contaminated by the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident. Ninety seven azuki bean accessions including their wild relatives from a Japanese gene bank, were grown in a field in the Fukushima prefecture, which is located approximately 51 km north of FDNPP. The contamination level of the soil was 3665 ± 480 Bq kg(-1) dry weight ((137)Cs, average ± SD). The soil type comprised clay loam, where the sand: silt: clay proportion was 42:21:37. There was a significant varietal difference in the biomass production, radiocaesium accumulation and transfer factor (TF) of radiocaesium from the soil to edible portion. Under identical agricultural practice, the extent of (137)Cs accumulation by seeds differed between the accessions by as much as 10-fold. Inter-varietal variation was expressed at the ratio of the maximum to minimum observed (137)Cs transfer factor for seeds ranged from 0.092 to 0.009. The total biomass, time to flowering and maturity, and seed yield had negative relationship to (137)Cs activity concentration in seeds. The results suggest that certain variety/varieties of azuki bean which accumulated less (137)Cs in edible portion with preferable agronomic traits are suitable to reduce the (137)Cs accumulation in food chain on contaminated land.

The azuki bean weevil (Callosobruchus chinensis L.) is an insect pest responsible for serious postharvest seed loss in leguminous crops. In this study, we performed quantitative trait locus (QTL) mapping of seed resistance to C. chinensis in moth bean (Vigna aconitifolia [Jaqc.] Maréchal). An F2 population of 188 plants developed by crossing resistant accession ‘TN67’ (wild type from India, male parent) and susceptible accession ‘IPCMO056’ (cultivated type from India, female parent) was used for mapping. Seeds of the F2 population from 2014 and F2:3 populations from 2016 and 2017 were bioassayed with C. chinensis, and the percentage of damaged seeds and progress of infestation severity were measured. Segregation analysis suggested that C. chinensis resistance in TN176 is controlled by a single dominant gene, designated as Rcc. QTL analysis revealed one principal and one modifying QTL for the resistance, named qVacBrc2.1 and qVacBrc5.1, respectively. qVacBrc2.1 was located on linkage group 2 between simple sequence repeat markers CEDG261 and DMB-SSR160 and accounted for 50.41% to 64.23% of resistance-related traits, depending on the trait and population. Comparative genomic analysis suggested that qVacBrc2.1 is the same as QTL Brc2.1 conferring C. chinensis resistance in wild azuki bean (V. nepalensis Tateishi and Maxted). Markers CEDG261 and DMB-SSR160 should be useful for marker-assisted selection for C. chinensis resistance in moth bean.

Wild relatives of crop plants are thought as reservoir of prominent genetic resources for abiotic stress tolerance. However, insufficient information on genetic variation and phenotypic traits restricts their use for crop breeding. This study focused on wild species of genus Vigna (family Fabaceae) originated from highly humid to arid regions. To clarify the diversity of drought tolerance during the vegetative stage, 69 accessions, including 15 domesticated, and 54 wild accessions, were evaluated under two drought conditions of non-terminal and terminal stresses. In the non-terminal drought condition, the plants were grown in pipes of different heights where surface soil water content decreased faster in pipes with greater height. Relative shoot biomass was used for tolerance evaluation and we identified 19 drought tolerant accessions. Almost of them were wild accessions showing higher relative shoot biomass than that in the domesticated accessions. Domesticated species were mostly classified as drought susceptible but could be improved using tolerant conspecific wild ancestors with cross-compatibility. The tolerance was related with higher plant water status presumably due to small water consumption. However, the variation of drought tolerance could not be explained by simple tolerance factor alone, and other tolerance mechanisms such as deep rooting and increasing in root biomass were found in the tolerant accessions. In the terminal drought condition, the plants were grown in small pots, and the watering was stopped to expose them extreme and rapid soil water scarcity. The tolerance was evaluated as the number of days until wilting. However, the accessions found to be tolerant in the pot experiment were not the same as those in the pipe experiment. In this condition, plant water status was not related with the length of days to wilting. This indicates that different mechanisms are necessary for adaptation to each of the non-terminal and terminal drought conditions. Many accessions were tolerant to one of the conditions, although we identified that some accessions showed tolerance in both experiments. The great diversity in drought tolerance in the genus Vigna might serve to both improve crop drought tolerance and understand the mechanisms of adaptation in drought-prone environments.

Wild relatives of crop plants are thought as reservoir of prominent genetic resources for abiotic stress tolerance. However, insufficient information on genetic variation and phenotypic traits restricts their use for crop breeding. This study focused on wild species of genus Vigna (family Fabaceae) originated from highly humid to arid regions. To clarify the diversity of drought tolerance during the vegetative stage, 69 accessions, including 15 domesticated, and 54 wild accessions, were evaluated under two drought conditions of non-terminal and terminal stresses. In the non-terminal drought condition, the plants were grown in pipes of different heights where surface soil water content decreased faster in pipes with greater height. Relative shoot biomass was used for tolerance evaluation and we identified 19 drought tolerant accessions. Almost of them were wild accessions showing higher relative shoot biomass than that in the domesticated accessions. Domesticated species were mostly classified as drought susceptible but could be improved using tolerant conspecific wild ancestors with cross-compatibility. The tolerance was related with higher plant water status presumably due to small water consumption. However, the variation of drought tolerance could not be explained by simple tolerance factor alone, and other tolerance mechanisms such as deep rooting and increasing in root biomass were found in the tolerant accessions. In the terminal drought condition, the plants were grown in small pots, and the watering was stopped to expose them extreme and rapid soil water scarcity. The tolerance was evaluated as the number of days until wilting. However, the accessions found to be tolerant in the pot experiment were not the same as those in the pipe experiment. In this condition, plant water status was not related with the length of days to wilting. This indicates that different mechanisms are necessary for adaptation to each of the non-terminal and terminal drought conditions. Many accessions were tolerant to one of the conditions, although we identified that some accessions showed tolerance in both experiments. The great diversity in drought tolerance in the genus Vigna might serve to both improve crop drought tolerance and understand the mechanisms of adaptation in drought-prone environments.

In this paper, we discuss stress-adapted wild Vigna plants and several important Vigna crops. Seeds, young shoots and/or tubers of many wild Vigna species are edible and are eaten by people in some locations where they grow. We propose the concept of ‘neo-domestication’ of stress-adapted wild edible Vigna species. As the causative change of the mutation resulting in a domestication gene is usually ‘loss-of-function’ type, ‘neo-domestication’ could be achieved by conventional mutation breeding together with screening by TILLING. The ‘neo-crops’ could play an important role in areas unsuitable for growing other crops to increase world food production. We also propose that wild Vigna species can be ‘new model plant species’ for the genetic study of natural adaptation to stresses (e.g. salt, acid, alkali, drought, flood, pests and diseases). To facilitate this, the Vigna Genome Project has been initiated. In addition, sustainable cropping systems may be enhanced by analysis of the nitrogen-fixing systems of stress-adapted Vigna species. Stress-adapted symbiotic bacteria produce nodules on stress-adapted Vigna species. Therefore, analyses of the genetic diversity of symbiotic bacteria and the process of symbiosis under stress environments should be conducted.

Zombi pea [Vigna vexillata (L.) A. Rich] is an underutilized legume crop, with highly diverse wild species that carry genes conferring resistance to several biotic and abiotic stresses. The objective of this study was to construct a zombi pea genetic linkage map and locate quantitative trait loci related to salt resistance. A zombi pea F2 population (159 individuals) developed from a cross between the salt-resistant wild zombi pea accession JP235908 (var. ovata; female parent) and the salt-susceptible cultivated zombi pea accession TVNu240 (var. macrosperma; male parent) was genotyped with simple sequence repeat (SSR) and restriction site-associated DNA sequencing (RAD-seq) markers. A genetic linkage map with 10 linkage groups (LGs) was constructed based on 379 markers (136 SSR and 243 RAD-seq markers). The map spanned 793.25 cM, with a mean distance between adjacent markers of 2.14 cM. A comparative genome analysis revealed high macro-synteny between zombi pea and mung bean/adzuki bean, although several chromosomal translocations likely occurred in zombi pea. The salt tolerance of the F2:3 population was evaluated at the seedling stage under hydroponic conditions. Inclusive composite interval mapping identified three quantitative trait loci (QTLs) (qSaltol1.1 on LG1, qSaltol2.1 on LG2, and qSaltol6.1 on LG6) related to the salt resistance of the F2 population. The qSaltol1.1, qSaltol2.1, and qSaltol6.1 QTLs explained 13.3%, 7.6%, and 8.1% of the salt-resistance phenotypic variance, respectively. A comparative genome analysis revealed that qSaltol1.1 may correspond to the Saltol1.1 QTL conferring salt tolerance in beach cowpea [Vigna marina (Burm.) Merr.], which is a halophytic species. Moreover, two genes encoding a plasma membrane H+-ATPase and a gene encoding a cation/proton exchanger may contribute to the salt resistance of zombi pea and beach cowpea. These findings may be relevant for the breeding of salt-resistant zombi pea plants.

QTL mapping in F 2 population [ V. luteola × V. marina subsp. oblonga ] revealed that the salt tolerance in V. marina subsp. oblonga is controlled by a single major QTL. The habitats of beach cowpea (Vigna marina) are sandy beaches in tropical and subtropical regions. As a species that grows closest to the sea, it has potential to be a gene source for breeding salt-tolerant crops. We reported here for the first time, quantitative trait loci (QTLs) mapping for salt tolerance in V. marina. A genetic linkage map was constructed from an F2 population of 120 plants derived from an interspecific cross between V. luteola and V. marina subsp. oblonga. The map comprised 150 SSR markers. The markers were clustered into 11 linkage groups spanning 777.6 cM in length with a mean distance between the adjacent markers of 5.59 cM. The F2:3 population was evaluated for salt tolerance under hydroponic conditions at the seedling and developmental stages. Segregation analysis indicated that salt tolerance in V. marina is controlled by a few genes. Multiple interval mapping consistently identified one major QTL which can explain about 50 % of phenotypic variance. The flanking markers may facilitate transfer of the salt tolerance allele from V. marina subsp. oblonga into related Vigna crops. The QTL for domestication-related traits from V. marina are also discussed.

A cDNA clone from amaranth perisperm that encodes an isoamylase (ISA)-type starch debranching enzyme 3 was isolated and analyzed for the first time. The cDNA consisted of 2,715 bp with a single open reading frame of 2,346 bp, encoding a protein of 781 amino acid residues. The deduced amino acid sequence of CrISA3 shared 63–71 % identity with those of other plant ISA3s. We also investigated the genetic diversity of ISA3 in three species of grain amaranth. A comparison of their ISA3 coding sequences revealed an extremely high level of conservation and only 11 single nucleotide polymorphisms were detected. The expression of the CrISA3 gene in amaranth developmental seeds and several tissues was investigated by qRT-PCR analysis. The results showed that CrISA3 was rapidly expressed at the early stage during seed maturation. It was also expressed in non-storage tissues (leaf, petiole, stem, and root) as well as in storage tissue. This observation demonstrates that CrISA3 may play an important role in perisperm starch accumulation at the early developmental stages. In addition, our results indicate that CrISA3 plays important roles in the synthesis of storage and transitory starches. The characterization of the CrISA3 gene will contribute to further studies on starch biosynthesis in Amaranthus.

The National Institute of Agrobiological Sciences (NIAS) Genebank coordinates the conservation of plant, microorganism, and animal genetic resources related to food and agriculture in Japan. It also coordinates the distribution of genetic resources in the public domain for research, breeding, and educational purposes. To operate the NIAS Genebank efficiently, we have developed a genetic resources database, data management software, and web-based data retrieval systems to make the data available worldwide. This article describes the NIAS Genebank’s Core Collections of global and Japanese soybean (Glycine max (L.) Merr.), Japanese azuki bean (Vigna angularis (Willd.) Ohwi et Ohashi), and Japanese wheat (Triticum aestivum L. s. l.), all of which are available through the Genebank website. This article also describes new features of the NIAS Genebank database, such as the ability to select single-seed-derived germplasm of soybean in the plant search system and to download photographic data on accessions. By using the downloaded plant image PDF files, users can obtain detailed passport and agronomic information by clicking on the image of an accession of interest.

The objective of this study was to identify quantitative trait loci (QTL) affecting fitness of hybrids between wild soybean (Glycine soja) and cultivated soybean (Glycine max). Seed dormancy and seed number, both of which are important for fitness, were evaluated by testing artificial hybrids of G. soja × G. max in a multiple-site field trial. Generally, the fitness of the F1 hybrids and hybrid derivatives from self-pollination was lower than that of G. soja due to loss of seed dormancy, whereas the fitness of hybrid derivatives with higher proportions of G. soja genetic background was comparable with that of G. soja. These differences were genetically dissected into QTL for each population. Three QTLs for seed dormancy and one QTL for total seed number were detected in the F2 progenies of two diverse cross combinations. At those four QTLs, the G. max alleles reduced seed number and severely reduced seed survival during the winter, suggesting that major genes acquired during soybean adaptation to cultivation have a selective disadvantage in natural habitats. In progenies with a higher proportion of G. soja genetic background, the genetic effects of the G. max alleles were not expressed as phenotypes because the G. soja alleles were dominant over the G. max alleles. Considering the highly inbreeding nature of these species, most hybrid derivatives would disappear quickly in early self-pollinating generations in natural habitats because of the low fitness of plants carrying G. max alleles.

Powdery mildew (PM) is a common and serious disease of mungbean [Vigna radiata (L.) Wilczek]. A few quantitative trait loci (QTL) for PM resistance in mungbean have been reported. The objective of this study was to locate QTL for PM resistance in two resistant accessions V4718 and RUM5. Simple sequence repeat markers were analyzed in an F2 population from a cross between Kamphaeng Saen 1 (KPS1; susceptible to PM) and V4718 (resistant to PM), and in F2 and BC1F1 populations from a cross between Chai Nat 60 (CN60; susceptible to PM) and RUM5 (resistant to PM). Progenies of 134 F2:3 and F2:4 lines derived from KPS1 × V4718, and 190 F2:3 and 74 BC1F1:2 lines derived from CN60 × RUM5 and CN60 × (CN60 × RUM5), respectively, were evaluated for response to PM under field conditions. Multiple interval mapping identified a major QTL on linkage group (LG) 9 and two minor QTL on LG4 for the resistance in V4718, and detected two major QTL on LG6 and LG9 and one minor QTL on LG4 for the resistance in RUM5. Comparative linkage analysis of the QTL for PM resistance in this study and in previous reports suggests that the resistance QTL on LG9 in V4718, RUM5, ATF3640 and VC6468-11-1A are the same locus or linked. One QTL on LG4 is the same in three sources (V4718, RUM5 and VC1210A). Another QTL on LG6 is the same in two sources (RUM5 and VC6468-11-1A). In addition, one QTL in V4718 on LG4 appears to be a new resistance locus. These different resistance loci will be useful for breeding durably PM-resistant mungbean cultivars.

The National Institute of Agrobiological Sciences' (NIAS) Genebank is the coordinating institute in Japan for the conservation of plant, microorganism and animal genetic resources related to food and agriculture. The NIAS Genebank manages this activity in collaboration with sub-banks located across Japan and acts as the central bank. It is important for genebanks to maintain the viability of genetic resources during conservation. Generally, seeds are stored at low temperature and low humidity. Germination percentage is used to monitor seed viability. We have developed data-management software to manage germination testing data in the NIAS Genebank. The software can monitor the pattern of the loss of seed viability of each accession and also support the selection of accessions that need regeneration of seeds. Web-based plant accessions search software displays the latest data on the availability and germination percentages of accessions.

Since chloroplasts and mitochondria are maternally inherited and have unique features in evolution, DNA sequences of those organelle genomes have been broadly used in phylogenetic studies. Thanks to recent progress in next-generation sequencer (NGS) technology, whole-genome sequencing can be easily performed. Here, using NGS data generated by Roche GS Titanium and Illumina Hiseq 2000, we performed a hybrid assembly of organelle genome sequences of Vigna angularis (azuki bean). Both the mitochondrial genome (mtDNA) and the chloroplast genome (cpDNA) of V. angularis have very similar size and gene content to those of V. radiata (mungbean). However, in structure, mtDNA sequences have undergone many recombination events after divergence from the common ancestor of V. angularis and V. radiata, whereas cpDNAs are almost identical between the two. The stability of cpDNAs and the variability of mtDNAs was further confirmed by comparative analysis of Vigna organelles with model plants Lotus japonicus and Arabidopsis thaliana.

Breeding salt tolerant plants is difficult without utilizing a diversity of wild crop relatives. Since the genus Vigna (family Fabaceae) is comprised of many wild relatives adapted to various environmental conditions, we evaluated the salt tolerance of 69 accessions of this genus, including that of wild and domesticated accessions originating from Asia, Africa, Oceania, and South America. We grew plants under 50 mM and 200 mM NaCl for two weeks and then measured the biomass, relative quantum yield of photosystem II, leaf Na+ concentrations, and leaf K+ concentrations. The accessions were clustered into four groups: the most tolerant, tolerant, moderately susceptible, and susceptible. From the most tolerant group, we selected six accessions, all of which were wild accessions adapted to coastal environments, as promising sources of salt tolerance because of their consistently high relative shoot biomass and relative quantum yield. Interestingly, variations in leaf Na+ concentration were observed between the accessions in the most tolerant group, suggesting different mechanisms were responsible for their salt tolerance. Phylogenetic analysis with nuclear DNA sequences revealed that salt tolerance had evolved independently at least four times in the genus Vigna, within a relatively short period. The findings suggested that simple genetic changes in a few genes might have greatly affected salt tolerances. The elucidation of genetic mechanisms of salt tolerances in the selected accessions may contribute to improving the poor salt tolerance in legume crops.

Yardlong bean [Vigna unguiculata (L.) Walp. ssp. unguiculata cv.-gr. sesquipedalis] is a vegetable legume crop evolved from cultivated grain cowpea (V. unguiculata ssp. unguiculata cv.-gr. unguiculata) which is domesticated from wild cowpea. It has a dramatic change in pod length and pod fiber content, and a complete loss of pod shattering, as compared to its wild progenitor. In this study, we identified quantitative trait loci (QTLs) controlling pod fiber content and pod shattering in two populations (BC1F1 and F2) derived from a cross between yardlong bean and wild cowpea. BC1F1:2 and F2:3 families were grown under field condition in which insoluble dietary fiber (cellulose, hemicelluloses and lignin) contents in mature pods, and pod shattering were evaluated. Correlation analysis showed positive relationship among the types of fiber, and between the fiber and shattering. Inclusive composite interval mapping revealed that a major QTL on linkage group 7 (LG7) controlled cellulose, hemicellulose and lignin contents in pod, and pod shattering. The other QTLs related to pod fibers on LG1 and LG4 also co-localized with the QTLs for pod shattering. Comparative genome analysis with azuki bean (Vigna angularis) suggested that the QTL region on LG7 for cellulose, hemiclellulose, lignin and pod shattering in yardlong bean contains genes encoding MYB transcription factor, MYB83, regulating biosynthesis of the three fibers, while the QTL region for cellulose and shattering of pod on LG1 harbors gene encoding cellulose synthase A7 (CESA7). These genes may be important targets for functional study to reveal major factors regulating pod fiber biosynthesis and pod shattering in yardlong bean.

The genus Vigna (Fabaceae) consists of five subgenera, and includes more than 100 wild species. In Vigna, 10 crops have been domesticated from three subgenera, Vigna, Plectrotropis, and Ceratotropis. The habitats of wild Vigna species are so diverse that their genomes could harbor various genes responsible for environmental stress adaptation, which could lead to innovations in agriculture. Since some of the gene bank Vigna accessions were unidentified and they seemed to be novel genetic resources, these accessions were identified based on morphological traits. The phylogenetic positions were estimated based on the DNA sequences of nuclear rDNA-ITS and chloroplast atpB-rbcL spacer regions. Based on the results, the potential usefulness of the recently described species V. indica and V. sahyadriana, and some wild Vigna species, i.e., V. aconitifolia, V. dalzelliana, V. khandalensis, V. marina var. oblonga, and V. vexillata, was discussed.