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.     

Genetic variability in flooding tolerance of mungbean (Vigna radiata L. Wilczek) genotypes

The study was an exploratory in nature conducted using a large number of mungbean (Vigna radiata L. Wilczek) genotypes of diverse growth habit and adaptive characters. Soil flooding induced changes in eleven morpho-physiological characters of one-week old seedlings of 530 mungbean genotypes was compared in the study. The first and second principal components (PC) of principal component analysis (PCA) results accounted for 58 and 14%, respectively of the total variations of mungbean genotypes. The variation for first PC was composed mainly of relative dry weight (DW) of shoot and leaf as well as total DW. The second PC distinguished the genotypes that produced larger root system. There were seven clusters distinguished in the cluster analysis. The genotypes in cluster 4 and 6 performed better in respect of relative total DW and relative root DW, respectively and hence having flooding tolerance. The genotypes in clusters 7 and 1 performed very poorly and those of under clusters 3, 2 and 5 were moderate to poor. D² analysis indicated that the clusters differed significantly from each other. Discriminant function analysis (DFA) reaffirmed that more than 90% of the genotypes were correctly assigned to clusters. Both PCA and DFA confirmed that the relative total DW followed by shoot and leaf DW as well as leaf area were the major discriminatory variables and the root : shoot ratio and root DW were the secondary important variables to distinguish genotypes into groups. In this study, multivariate analyses were used in identifying the mungbean genotypes of desirable traits for flooding tolerance.     

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.     

Detection of quantitative trait loci for mungbean yellow mosaic India virus (MYMIV) resistance in mungbean (Vigna radiata (L.) Wilczek) in India and Pakistan

Yellow mosaic disease (YMD) is one of the major diseases affecting mungbean (Vigna radiata (L.) Wilczek). In this study, we report the mapping of the quantitative trait locus (QTL) for mungbean yellow mosaic India virus (MYMIV) resistance in mungbean. An F₈ recombinant inbred line (RIL) mapping population was generated in Thailand from a cross between NM10-12-1 (MYMIV resistance) and KPS2 (MYMIV susceptible). One hundred and twenty-two RILs and their parents were evaluated for MYMIV resistance in infested fields in India and Pakistan. A genetic linkage map was developed for the RIL population using simple sequence repeat (SSR) markers. Composite interval mapping identified five QTLs for MYMIV resistance: three QTLs for India (qYMIV1, qYMIV2 and qYMIV3) and two QTLs for Pakistan (qYMIV4 and qYMIV5). qYMIV1, qYMIV2, qYMIV3, qYMIV4 and qYMIV5 explained 9.33%, 10.61%, 12.55%, 21.93% and 6.24% of variation in disease responses, respectively. qYMIV1 and qYMIV4 appeared to be the same locus and were common to a major QTL for MYMIV resistance in India identified previously using a different resistant mungbean.     

Inheritance pattern of mungbean yellow mosaic disease resistance and gene action for different traits in mungbean (Vigna radiata (L.) Wilczek) under protected and unprotected field conditions

Understanding of mungbean yellow mosaic disease resistance is crucial to develop resistant varieties to combat worldwide threat of the disease epidemics. This study was aimed to determine nature of disease resistance in terms of number of genes governing it and genetics of related traits. Experimental site was located on 31.43°N and 73.06°E with an elevation of 186 m, and evaluation trials were conducted during spring season due to high disease epidemics in this season. Two contrasting genotypes, that is, NM 6‐68‐2 (resistant) and NM 1‐32‐1 (highly susceptible), were crossed to raise six populations, that is, P₁, P₂, F₁, F₂, BC₁ and BC₂ for evaluation under protected and unprotected field conditions using chi‐square test and generation mean and variance analysis. It was discovered that disease resistance was governed by two major genes with additive effects. Disease resistance can easily be incorporated through backcrossing and direct selection following hybridization. Direct selection should be practised at earlier generation for plant height, chlorophyll contents and number of seeds per pod due to preponderance of additive effects whereas at later generations for seed yield per plant, 100 seed weight, harvest index, number of pods per plant and pod length due to involvement of duplicate epistasis.     

Genetical relationships between reactions to bacterial leaf spot,yellow mosaic and Cercospora leaf spot diseases in mungbean (Vigna radiata)

Summary Studies on the inheritance pattern of bacterial leaf spot (BLS), yellow mosaic (YM) and Cercospora leaf spot (CLS) reactions in crosses of BLS and YM resistant/tolerant but CLS susceptible × CLS resistant but BLS and YM susceptible parents indicated that resistances to BLS and CLS were governed by single dominant genes, whereas YM tolerance was a monogenic recessive character. The studies also indicated that these three genes were inherited independently. The simple inheritance pattern and independent assortment of the genes governing resistance/tolerance to these diseases suggest that the usual breeding methods will be adequate to develop multi-disease resistant mungbean cultivars.Paper XII in the series Studies on resistance in crops to bacterial diseases in India.     

绿豆遗传连锁图谱的整合

利用绿豆及其近缘种的701对SSR引物,对现有绿豆遗传连锁图谱进行补充,结果在高感豆象绿豆栽培种Berken和高抗豆象绿豆野生种ACC41两亲本间筛选到多态性SSR引物104对。群体分析后,结合其他分子数据,使用作图软件Mapmaker/Exp 3.0b,获得一张含有179个遗传标记和12个连锁群,总长1831.8cM、平均图距10.2cM的新遗传连锁图谱,包括97个SSR标记,91个来自绿豆近缘种;RFLP标记76个;RAPD标记4个;STS标记2个。对32个绿豆、小豆共用SSR标记在遗传连锁图谱的分布分析发现,二个基因组间有一定程度的同源性,共用标记在连锁群上的排列顺序基本上一致,只有部分标记显示绿豆和小豆基因组在进化过程中发生了染色体重排;利用新图谱对ACC41的抗绿豆象主效基因重新定位,仍定位于I(9)连锁群,与其相邻分子标记的距离均小于8cM,其中与右翼SSR标记C220的距离约2.7cM。与原图谱比较,新定位的抗性基因与其相邻标记的连锁更加紧密。     

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.     

Inheritance of photoperiod response in mungbean (Vigna radiata [L.] Wilczek)

Summary A dominant or partially dominant gene for sensitivity to photoperiod in mungbean strain PI 180311 was identified and labeled Ps. The gene was expressed when strain P1180311, or crosses involving PI 180311 were grown in 16- or 14-hour photoperiods but was not expressed when the populations were grown in a 12-hour photoperiod or in the field at Columbia, Missouri. Dominance × dominance epistatic effects were indicated as governing days to flower in absence of the obvious effect of the Ps gene. A gene A for purple hypocotyl was independent of the gene Ps.Contribution No. 7894 from the Missouri Agriculture Experiment Station.     

The major quantitative trait locus for mungbean yellow mosaic Indian virus resistance is tightly linked in repulsion phase to the major bruchid resistance locus in a cross between mungbean [Vigna radiata (L.) Wilczek] and its wild relative Vigna radiata ssp. sublobata

Mungbean yellow mosaic Indian virus (MYMIV) and bruchid infestation are severe production constraints of mungbean in South Asia, a major global mungbean production area. Marker-assisted selection for resistance against these disorders while maintaining or even improving agronomic traits is an important step toward breeding elite mungbean varieties. This study employed recombinant inbred lines (F12) derived from a cross between MYMIV-tolerant Vigna radiata NM92 and bruchid-resistant V. radiata ssp. sublobata TC1966 to identify chromosomal locations associated with disease and insect pest resistance and seed traits. A linkage map comprising 11 linkage groups was constructed with random amplified polymorphic DNA (RAPD), sequence characterized amplified regions (SCAR), cleaved amplified polymorphic DNA (CAP), amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. Quantitative trait loci (QTLs) for MYMIV and bruchid resistance, 100 seed weight and seed germination rate were identified. Three major QTLs for MYMIV and one major bruchid resistance locus were mapped on LG 9. The resistance alleles were contributed by the MYMIV tolerant parent NM92 and the bruchid resistant parent TC1966 respectively. One of the MYMIV QTLs was tightly linked in repulsion phase to the bruchid resistance locus. In addition, three minor QTLs for MYMIV resistance were found, where the resistance alleles were contributed by TC1966. Lines combining MYMV resistance alleles from both parents have greater resistance to MYMIV than the tolerant parent. Two minor bruchid resistance QTLs were identified in TC1966. Furthermore, three QTLs each for 100 seed weight and germination rate were detected. The markers defining the QTLs identified in this study will be useful in marker-assisted breeding of improved mungbean varieties in the future.     

绿豆几个表型性状的遗传特性分析

种皮色泽和花青甙显色是绿豆(Vigna radiata)的重要性状,可在纯度鉴定、辅助选择育种等研究中发挥作用。本研究以绿豆核心种质种皮色泽、花青甙显色性状分析为基础,配制杂交组合,根据不同世代的表型调查,探讨绿豆种皮颜色、光泽和花青甙显色等性状的遗传特征。结果表明,绿种皮对黄种皮为单基因控制,绿色为显性;黑种皮对绿种皮为单基因控制,黑色为显性。绿豆种皮光泽不符合简单性状的遗传规律,且不同组合间后代的表型分布存在差异。绿豆种皮色与光泽间并无连锁关系。植株各部位的花青甙显色对不显色为显性,均符合简单单基因的分离规律。进一步分析表明,不同部位花青甙显色的相关性不一致,其中控制幼茎色基因与控制复叶基部花青甙显色基因紧密连锁在一起,或为一个基因,而控制龙骨瓣花青甙显色的基因与其他相关基因的遗传连锁程度最低。本文对绿豆种皮相关性状及植株花青甙显色性状的遗传分析将为这些性状的育种利用及深入研究奠定基础。     

Genetics of the resistance to powdery mildew disease in mungbean (Vigna radiata (L.) Wilczek)

Powdery mildew disease caused by the fungus Erysiphe polygoni D.C. is an important disease of mungbean. Loss can be more serious if the disease attacks at seedling stage. In this paper, we report genetics of the resistance to powdery mildew disease in mungbean using a recombinant inbred line (RIL) population derived from a cross between the susceptible parent “KPS1” and the resistant parent “VC6468-11-1A”. Five hundred and ninety-two RILs were developed by random descending from 200 F₂ plants. The population was evaluated against the fungus in field and greenhouse conditions. The data were analyzed following a nested design for selfpollinating plants to determine genetic heritability of powdery mildew resistance. The severity of the infestation was measured by using disease index (DI) and area under disease progress curve (AUDPC). Both values showed continuous distribution in the RILs. Under field conditions, the narrow-sense heritability of DI and AUDPC were 0.67 and 0.48, respectively. While under greenhouse conditions, the values were 0.68 and 0.62, respectively. The results suggested that the resistance is quantitatively inherited with high heritability and predominantly additive gene action. To develop powdery mildew-resistant mungbean varieties, the plant breeder can select for resistant lines by using standard selection procedures for self-pollinating crops, viz. pedigree selection, bulk selection, early generation testing, and single-seed descent.     

Identification and inheritance of a new gene for powdery mildew resistance in mungbean (Vigna radiata L. Wilczek)

Powdery mildew (PM) is one of the important foliar diseases of mungbean. Resistance sources have been identified in India and the inheritance studies showed that complete resistance (RO) was controlled by two dominant genes, Pm1 , Pm2 . The breakdown of complete resistance (RO) into moderate resistance (R2) by race-2 (Akola) has been reported. It is assumed that the change in resistance reaction is due to a mutation in the pathogen. The present investigation was carried out with a view to screen germplasm, cultivars and mutants for identification of complete resistance (RO) sources against race-2 and to study their inheritance. 'Mulmarada', a local mungbean cultivar from Maharashtra state of India was identified as a complete resistance (RO) source for race-2. The inheritance of Mulmarada's resistance (RO) was studied. The F₁ and the segregation in F₂ and F₃ showed that the complete resistance (RO) in 'Mulmarada' is controlled by a single dominant gene, which is different from the earlier identified Pm1 and Pm2 resistance genes. Mulmarada's resistance gene is designated as Pm3 for PM resistance.     

Development of a SCAR marker linked with a MYMV resistance gene in mungbean (Vigna radiata L. Wilczek)

Yellow mosaic disease (YMD) caused by mungbean yellow mosaic virus (MYMV) is the most important disease of mungbean, causing great yield loss. The present investigation was carried out to study the inheritance and identify molecular markers linked with MYMV resistance gene by using F₁, F₂ and 167 F_2 : 8 recombinant inbred lines (RILs) developed from the cross ‘TM‐99‐37’ (resistant) × Mulmarada (susceptible). The F₁ was susceptible, F₂ segregated in 3S:1R phenotypic ratio and RILs segregated in 1S:1R ratio in the field screening indicating that the MYMV resistance gene is governed by a single recessive gene. Of the 140 RAPD primers, 45 primers showing polymorphism in parents were screened using bulked segregant analysis. Three primers amplified specific polymorphic fragments viz. OPB‐07_600, OPC‐06₁₇₅₀ and OPB‐12₈₂₀. The marker OPB‐07₆₀₀ was more closely linked (6.8 cM) with a MYMV resistance gene as compared to OPC‐06₁₇₅₀ (22.8 cM) and OPB‐12₈₂₀ (25.2 cM). The resistance‐specific fragment OPB‐07₆₀₀ was cloned, sequenced and converted into a sequence‐characterized amplified region (SCAR) marker and validated in twenty genotypes with different genetic backgrounds.     

Genetic analysis of resistance in barley to barley yellow dwarf virus

The inheritance of resistance to barley yellow dwarf virus (BYDV) was studied in the selected 24 spring and winter barley cultivars that showed a high or intermediate resistance level in 1994‐97 field infection tests. The polymerase chain reaction diagnostic markers YLM and Ylp were used to identify the resistance gene Yd2. The presence of the Yd2 gene was detected with both markers in all the resistant spring barley cultivars and lines from the CIMMYT/ICARDA BYDV nurseries. The results of field tests and genetic analyses in winter barley corresponded with marker analyses only when the Ylp marker was used. Genes non‐allelic with Yd2 were detected by genetic analyses and the Ylp marker in moderately resistant spring barley cultivars ‘Malvaz’, ‘Atribut’ and ‘Madras’, and in the winter barley cultivars ‘Perry’ and ‘Sigra’. Significant levels of resistance to BYDV were obtained by combining the resistance gene Yd2 with genes detected in moderately resistant cultivars. The utilization of analysed resistance sources in barley breeding is discussed.     

Genetic control of resistance to soilborne wheat mosaic virus in Brazilian cultivars of Triticum aestivum L. Thell.

In Southern Brazil, under ideal conditions Soilborne Wheat Mosaic Virus (SBWMV) induces yield reductions to the wheat crop of over 50%. The only effective way of controlling the disease is through resistance. However, the inheritance of resistance is not been fully understood. The purpose of this work was to study the genetic control of resistance to SBWMV. Crosses were carried out between the resistant wheat cultivar Embrapa 16 and the susceptible cultivars BR 23 and IAC 5-Maringá, at the National Wheat Research Centre, Passo Fundo, RS, Brazil. The parents, F₁, F₂, and backcrosses were sown in plots in a field where soil was naturally infested with the vector of the virus, the fungus Polymyxa graminis, in order to promote natural infection. All plants were individually evaluated for severity and type of lesions, according to a scale of 0 to 5, where, 0 = absence of symptoms and 5 = plants severely affected by mosaic plus dwarfing and rosetting. The statistical analyses of the data showed broad sense heritability values between 43% and 55%. The data suggested the presence of two genes controlling resistance to SBWMV in the segregating population of both crosses. This revised version was published online in August 2006 with corrections to the Cover Date.     

Inheritance of seed phytate in mungbean (Vigna radiata (L.) Wilczek)

Phytic acid (PA) is the main storage organic form of phosphorus (P) in seeds of cereals and legumes. It is a strong inhibitor against the absorption of nutrients in monogastric animals. The objective of this study was to determine the variation of total P (TP) in seeds of mungbean germplasm and investigate the inheritance of seed P compound and phytate contents. TP content in seeds of 250 accessions were found to range from 2.34 to 5.75 mg/g. The inheritance was studied in the F₂ population derived from a cross between 2 accessions with the lowest and highest PA contents, viz. V1658BBR and V1141BG. Broad-sense heritability estimates of TP, inorganic P (IP), and phytate P (PhyP) contents were 80.8, 78.6 and 80.7%, respectively. The 9:7 segregation ratio of F₂ population in this study indicated that high TP and PhyP were controlled by dominant alleles at 2 independent loci of major genes showing duplicated recessive epistasis.     

Development of PCR-based markers for a major locus conferring powdery mildew resistance in mungbean (Vigna radiata)

Powdery mildew (PM) can cause significant yield loss in mungbean and several loci conferring resistance to this disease have been identified. A restriction fragment length polymorphism (RFLP) marker (VrCS65) linked closely to one of these loci was used to screen a mungbean bacterial artificial chromosome (BAC) library and positive BAC clones identified were used to develop simple sequence repeat (SSR or microsatellite) and sequence tagged site (STS) markers. Four of the new PCR markers (including two SSRs and two STSs) co-segregated with the original RFLP marker VrCS65, and another SSR marker (VrCS SSR2) was located 0.5 cM away from it. These PCR-based and locus-specific markers could be useful in breeding cultivars with enhanced resistance to PM and in the further characterization of the locus including the isolation of gene(s) responsible for the resistance.     

Physiological Analysis of Yield Variation in Mungbean (Vigna radiata [Roxb.] Wilczek)

Two mungbean varieties, Guj-1 and PIMS-1 differing in their productivity potential, were examined to investigate their photosynthetic system at various stages of growth in relation to yield with two different dates of sowing. Vertical leaves were found to be beneficial. On the other hand, excessive leaf area during the later stages of growth may be detrimental to yield. Lower LAD at seedling stage and overall high NAR values may be reflected in higher grain yield. NAR declined as season progressed and this, in turn, may be related with increase in LAI. The grain yield was considerably more at the first date of sowing as compared to second date of sowing, the effect was being more pronounced in var. PIMS-1 . Various environmental factors recorded at two different dates of sowing and at various stages of growth indicated that these factors may have influenced the yield potential at two different dates of sowing.