Fine mapping of QTL conferring Cercospora leaf spot disease resistance in mungbean revealed TAF5 as candidate gene for the resistance.

KEY MESSAGE: This paper reports fine mapping of qCLS for resistance to Cercospora leaf spot disease in mungbean and identified LOC106765332encoding TATA-binding-protein-associated factor 5 (TAF5) as the candidate gene for the resistance Cercospora leaf spot (CLS) caused by the fungus Cercospora canescens is an important disease of mungbean. A QTL mapping using mungbean F2 and BC1F1 populations developed from the "V4718" (resistant) and "Kamphaeng Saen 1" (KPS1; susceptible) has identified a major QTL controlling CLS resistance (qCLS). In this study, we finely mapped the qCLS and identified candidate genes at this locus. A BC8F2 [KPS1 × (KPS1 × V4718)] population developed in this study and the F2 (KPS1 × V4718) population used in a previous study were genotyped with 16 newly developed SSR markers. QTL analysis in the BC8F2 and F2 populations consistently showed that the qCLS was mapped to a genomic region of ~ 13 Kb on chromosome 6, which contains only one annotated gene, LOC106765332 (designated "VrTAF5"), encoding TATA-binding-protein-associated factor 5 (TAF5), a subunit of transcription initiation factor IID and Spt-Ada-Gcn5 acetyltransferase complexes. Sequence comparison of VrTAF5 between KPS1 and V4718 revealed many single nucleotide polymorphisms (SNPs) and inserts/deletions (InDels) in which eight SNPs presented in eight different exons, and an SNP (G4,932C) residing in exon 8 causes amino acid change (S250T) in V4718. An InDel marker was developed to detect a 24-bp InDel polymorphism in VrTAF5 between KPS1 and V4718. Analysis by RT-qPCR showed that expression levels of VrTAF5 in KPS1 and V4718 were not statistically different. These results indicated that mutation in VrTAF5 causing an amino acid change in the VrTAF5 protein is responsible for CLS resistance in V4718.

A single base substitution in BADH/AMADH is responsible for fragrance in cucumber (Cucumis sativus L.), and development of SNAP markers for the fragrance.

Sequence analysis revealed that an SNP (A1855G) in CsBADH of cucumber accession PK2011T202 causes amino acid change in a highly conserved motif, Y163C. Gene mapping showed association between the SNP and the fragrance. Pandan-like fragrance is a value-added trait in several food crops such as rice, vegetable soybean and sorghum. The fragrance is caused by the volatile chemical 2-acetyl-1-pyrroline (2AP). Mutation(s) in betaine aldehyde dehydrogenase 2 (BADH2; also known as aminoaldehyde dehydrogenase 2) gene causes defective BADH2 and results in biosynthesis of 2AP. Recently, cucumber cultivars possessing pandan-like fragrance were discovered in Thailand. In this study, we report an association between CsBADH and the fragrance in cucumber accession "PK2011T202". Gene expression analysis of CsBADH in leaves of PK2011T202 and "301176" (non-fragrant) at various growth stages revealed that CsBADH was expressed in both accessions. Sequence comparison of CsBADH showed that PK2011T202 possesses a single base substitution (A1855G) in exon 5 which causes an amino acid change in a highly conserved motif of BADH, Y163C. Single nucleotide-amplified polymorphism markers were developed to detect the SNP polymorphism between the wild-type and fragrance alleles. Since CsBADH is located on chromosome 1, quantitative trait locus (QTL) mapping was conducted for this chromosome using an F2 and a backcross populations developed from the cross between PK2011T202 and 301176. QTL analysis in both populations showed that the major QTL for fragrance, qFgr, was co-localized with the CsBADH. We concluded that the defect function of CsBADH is responsible for fragrance in cucumber PK2011T202.

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.

QTL mapping for salt tolerance and domestication-related traits in Vigna marina subsp. oblonga, a halophytic species.

QTL mapping in F2 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.

Dual-Purpose Vermicompost for the Growth Promotion and Suppression of Damping-Off Disease on Potted Vegetable Soybean.

Vermicompost is applied as a soil amendment to promote plant growth and yield. It also helps to significantly reduce the incidence of soil-borne diseases. However, its efficiency depends on the type of earthworm from which it is formed. The current study aims to compare the effects of two vermicompost types derived from African nightcrawler (AF) and Tiger worm (TG) as a soil amendment to evaluate its potential for suppressing damping-off disease both in vitro and in vivo. It also aims to determine the effects of both vermicompost types on the growth and yield-related traits of potted Thai vegetable soybean [Glycine max (L.) Merrill] variety "Chiang Mai 84-2" grown under greenhouse conditions when amended to the soil at various application rates (1%, 2%, 3% w/w). AF vermicompost exhibited better suppression of damping-off disease than TG vermicompost in vitro and under greenhouse conditions. AF vermicompost performed significantly greater suppressive efficacy on the mycelial growth of Athelia rolfsii in vitro than TG vermicompost, indicated by 50% and 16% inhibition, respectively. Damping-off incidence on vegetable soybean seedlings grown in soil amended with AF vermicompost was significantly lower (21%) than in soil amended with TG vermicompost (32%) under greenhouse conditions. With an increased application rate of 1% to 2% or 3% for each vermicompost type, plant yields significantly enhanced, with no significant variations among the 2% and 3% rates. Applying vermicompost at 2% w/w through soil amendment was the appropriate rate for promoting the growth and yield of potted vegetable soybeans. The results can be used to guide the application of vermicompost to control soil-borne plant diseases, promote plant growth, and enhance yields, especially in terms of organic crop production. Further research is needed to evaluate its potential for other potted crops and protect against soil-borne diseases.

Effects of winged bean tuber (WBT) modified starches substituted for cassava chip in concentrate diets on rumen fermentation, nutrient utilization, and blood metabolites in Thai native beef cattle.

Objective: This study examined the effects of substituting winged bean tuber steam (WBTS) modified starches for cassava chips (CSC) in the concentrate diet on rumen fermentation, nutrient utilization, and blood metabolites in Thai-native beef cattle. Methods: Four Thai-native bulls were assigned randomly as a 4 × 4 Latin square design, which represents the amount of CSC replaced with WBTS in the concentrate mixture diets at 0, 10, 20, and 30%. Results: Increasing levels of WBTS replacement for CSC in the concentrate diets had a quadratic effect on total dry matter (DM) intake (p<0.05). Replacement of WBTS at 20% and 30% for CSC did not alter total DM intake compared to 0% WBTS, whereas 10% WBTS replacement could significantly increase total DM intake by 0.41 kg DM/day compared to the control group. In addition, neutral detergent fiber (NDF) digestibility showed a quadratic increase (p < 0.05) when CSC was substituted at various levels of WBTS in the concentrate diet (p < 0.05). Replacement of CSC with WBTS at 10% and 20% showed higher NDF digestibility when compared to 0% replacement. There was a quadratic increase in blood glucose at 4 h post-feeding, and the average blood glucose value was significantly lower (p < 0.01) when substituting CSC with WBTS. Substituting WBTS for CSC at 10% in the concentrate diet showed the highest blood glucose concentration when compared to other treatments. Replacing CSC with WBTS at 10% and 20% shows a higher concentration of C3 than those of other treatments (0% or 30%). The nitrogen (N) intake (NI) increased linearly (p<0.05) when substituting WBTS for CSC at all levels in the diet. Additionally, N retention (NR) and the ratio of N retention to N intake increased (p<0.05) when substituting WBTS for CSC at 10%, 20%, and 30% compared to 0%. The gross energy intake (GEI), digestible energy intake (DEI), and energy efficiency (DEI/GEI) were quadratically increased when substituted with various levels of WBTS for CSC in the concentrate diet. Conclusion: The findings indicate that substituting 10% of CSC in the concentrate diet with WBTS may be sufficient as an alternative feed resource for improving nutrient utilization and metabolic efficiency in beef cattle diets.

Effect of Replacing Corn Meal with Winged Bean Tuber (Psophocarpus tetragonolobus) Pellet on Gas Production, Ruminal Fermentation, and Degradability Using In Vitro Gas Technique.

The objective of this study is to evaluate the effects of replacing corn meal in ruminant diets with winged bean (Psophocarpus tetragonolobus) tubers (WBT) on ruminal fermentation, gas production parameters, and in vitro degradability. The study employed a completely random design (CRD) in its execution. The experimental design employed was a completely randomized design (CRD), featuring eleven levels of corn meal substitution with winged bean tubers pellet (WBTP) at 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100%. The levels were grouped into four categories of replacement: control (0% in the diet), low levels (10%, 20%, and 30% in the diet), medium levels (40%, 50%, 60%, and 70% in the diet), and high levels (80%, 90%, and 100% in the diet). The experimental results indicated that substituting corn meal with WBTP at moderate and high levels in the diet could improve the performance of the fermentation process by increasing the gas production rate constant from the insoluble fraction (p < 0.01). The IVDMD exhibited a higher degree of in vitro degradation after 12 h (h), with the mean value being higher in the high group compared to the medium until the high group (p < 0.05). At the 4 h mark, the groups that substituted corn meal with WBTP exhibited a decrease in pH value (p < 0.05) in comparison to the control group. The substitution of corn meal with WBTP resulted in the lowest protozoal count after 8 h in the median group (p < 0.05). A significant difference in the effect of WBTP on total volatile fatty acid (TVFA) concentration was observed at 8 h after incubation (p < 0.05). The medium and high levels of WBTP replacement resulted in the lowest TVFA concentration at 8 h (p < 0.05). The mean proportion of acetic acid (C2) linearly declined and was lowest when a high level of WBTP replaced cornmeal (p < 0.05). The concentration of propionic acid (C3) at 8 h after incubation and average values were linearly significantly different when various levels of WBTP were utilized. Replacing corn meal with WBTP at a high level showed the highest concentration of C3. Moreover, substituting medium and high concentrations of WBTP for corn meal resulted in a significant reduction in both the C2:C3 ratio at 8 h and the mean value (p < 0.05). In conclusion, WBTP exhibits a nutritional composition that is advantageous and may be an energetic substitute for corn meal.

Combining ability of tropical × temperate maize inducers for haploid induction rate, R1-nj seed set, and agronomic traits.

In vivo maternal haploid induction in isolation fields is proposed to bypass the workload and resource constraints existing in haploid induction nurseries. A better understanding of combining ability and gene action conditioning traits related to hybrid inducers is necessary to set the breeding strategy including to what extent parent-based hybrid prediction is feasible. This study aimed to evaluate the following in tropical savanna in the rainy and dry seasons for haploid induction rate (HIR), R1-nj seed set, and agronomic traits: 1) combining ability, line per se, and hybrid performance of three genetic pools; 2) genetic parameters, the modes of gene action, and heterosis; and 3) the relationships of inbred-general combining ability (GCA) and inbred-hybrid performance. Fifty-six diallel crosses derived from eight maize genotypes were evaluated in the rainy season of 2021 and the dry season of 2021/2022. Reciprocal cross effects including the maternal effect barely contributed to the genotypic variance for each trait observed. HIR, R1-nj seed set, flowering dates, and ear position were highly heritable and additive inherited, while ear length showed dominant inheritance. The equal importance of additive and dominance effects was found for yield-related traits. Temperate inducer BHI306 was the best general combiner for the HIR and R1-nj seed set, followed by two tropical inducers, KHI47 and KHI54. The ranges of heterosis were trait-dependent and slightly influenced by the environment, where hybrids in the rainy season consistently had higher heterosis than those in the dry season for each trait observed. Both hybrid groups derived from tropical × tropical and tropical × temperate inducers showed taller plants, larger ear size, and higher seed sets than the corresponding parents. However, their HIRs were still below the standard check of BHI306. The implications of genetic information, combining ability, and inbred-GCA and inbred-hybrid relationships on breeding strategies are discussed.

Comparison of Cassava Chips and Winged Bean Tubers with Various Starch Modifications on Chemical Composition, the Kinetics of Gas, Ruminal Degradation, and Ruminal Fermentation Characteristics Using an In Situ Nylon Bag and an In Vitro Gas Production Technique.

This research assessed the impact of cassava chips (CSC) and winged bean tubers (WBT) with various starch modification methods on the chemical composition, ruminal degradation, gas production, in vitro degradability, and ruminal fermentation of feed using an in situ and in vitro gas production technique. Experimental treatments were arranged for a 2 × 5 factorial, a completely randomized design with two sources of starch and five levels of modification treatments. Two sources of starch were CSC and WBT, while five modification treatments of starch were: no modification treatment, steam treatment, sodium hydroxide (NaOH) treatment, calcium hydroxide (CaOH2) treatment, and lactic acid (LA) treatment. The starch modification methods with NaOH and CaOH2 increased the ash content (p <0.05), whereas the crude protein (CP) content was lower after treatment with NaOH (p < 0.05). Steam reduced the soluble fraction (a) and effective dry matter degradability of WBT in situ (p <0.05). In addition, the WBT steaming methods result in a lower degradation rate constant in situ (p <0.05). The degradation rate constants for the insoluble fraction (c) in the untreated CSC were higher than those of the other groups. Starch modification with LA reduced in vitro dry matter degradability at 12 and 24 h of incubation (p <0.05). The starch modification method of the raw material showed the lowest pH value at 4 h (p <0.05). The source of starch and starch modification methods did not influence the in vitro ammonia nitrogen concentrations, or in vitro volatile fatty acids. In conclusion, compared to the CSC group and untreated treatment, treating WBT with steam might be a more effective strategy for enhancing feed efficiency by decreasing or retarding ruminal starch degradability and maintaining ruminal pH.

In Vitro Evaluation of Winged Bean (Psophocarpus tetragonolobus) Tubers as an Alternative Feed for Ruminants.

The purpose of the current study is to determine the effects of the replacement of cassava chips with winged bean (Psophocarpus tetragonolobus) tubers (WBTs) on gas production parameters, in vitro degradability, and ruminal fermentation in ruminant diets. The study was performed using a 3 × 4 factorial arrangements and was designed using a completely random method. Factor A employed three various roughage sources that were frequently used by locals to feed ruminants: Oryza sativa L. (a1), Brachiaria ruziziensis (a2), and Pennisetum purpureum (a3). The levels of WBTs substitutions for cassava chips at 0%, 33%, 66%, and 100% in the diet were selected as factor B. The experiment's findings revealed that replacing the cassava chips in the diet with WBTs at levels of 66 and 100% enhanced the fermentation process by producing a high gas volume at 96 h when Ruzi grass (RZ) was used as the main source of roughage (p < 0.01). The interaction between the roughage source and WBTs showed that organic matter (OM) degradability improved markedly in the case of RZ grass in combination with WBTs at all levels. Both the total volatile fatty acids (TVFAs) at 8 h of incubation and the average value decreased when a complete substitution of casava chips with WBT (WBT 100%) was employed or when employing rice straw as the main source of roughage (p < 0.01). There was no interaction between WBTs and roughage source on the ammonia-nitrogen (NH3-N) concentration (ml/dL) and rumen microbial count (p > 0.05). In summary, WBTs can be used effectively when combined with grass (Ruzi and Napier). The implementation of WBTs as a novel alternative feed may effectively replace cassava chips without affecting rumen function.

Unique Salt-Tolerance-Related QTLs, Evolved in Vigna riukiuensis (Na+ Includer) and V. nakashimae (Na+ Excluder), Shed Light on the Development of Super-Salt-Tolerant Azuki Bean (V. angularis) Cultivars.

Wild relatives of crops have the potential to improve food crops, especially in terms of improving abiotic stress tolerance. Two closely related wild species of the traditional East Asian legume crops, Azuki bean (Vigna angularis), V. riukiuensis "Tojinbaka" and V. nakashimae "Ukushima" were shown to have much higher levels of salt tolerance than azuki beans. To identify the genomic regions responsible for salt tolerance in "Tojinbaka" and "Ukushima", three interspecific hybrids were developed: (A) azuki bean cultivar "Kyoto Dainagon" × "Tojinbaka", (B) "Kyoto Dainagon" × "Ukushima" and (C) "Ukushima" × "Tojinbaka". Linkage maps were developed using SSR or restriction-site-associated DNA markers. There were three QTLs for "percentage of wilt leaves" in populations A, B and C, while populations A and B had three QTLs and population C had two QTLs for "days to wilt". In population C, four QTLs were detected for Na+ concentration in the primary leaf. Among the F2 individuals in population C, 24% showed higher salt tolerance than both wild parents, suggesting that the salt tolerance of azuki beans can be further improved by combining the QTL alleles of the two wild relatives. The marker information would facilitate the transfer of salt tolerance alleles from "Tojinbaka" and "Ukushima" to azuki beans.

A Rhizobacterium, Streptomyces albulus Z1-04-02, Displays Antifungal Activity against Sclerotium Rot in Mungbean.

Sclerotium rot causes damping-off and stem rot in seedlings and mature mungbeans, which negatively impacts cultivation. The use of a rhizobacterium to control soil-borne diseases is an alternative method to the excess use of synthetic fungicides; therefore, this study aims to screen rhizosphere actinobacteria with fungicidal activities against Sclerotium rolfsii, the pathogen that causes sclerotium rot in mungbeans. Primary screening showed that the Streptomyces sp. isolate Z1-04-02 displayed the highest effectiveness against S. rolfsii in dual culture plates, with a percentage inhibition of 74.28%. An assay containing enzymes that degrade cell walls, of the cell-free culture filtrate (CF) of Z1-04-02, showed that the activities of chitinase and ß-1,3-glucanase were 0.0209 and 1.0210 U/mL, respectively, which was significantly higher than that of the control (media alone). The cell-free CF of Z1-04-02, incubated at 37 °C and 100 °C, using agar well diffusion, effectively inhibited the growth of S. rolfsii with inhibition percentages of 37.78% and 27.78%, respectively. Solid-phase microextraction (SPME) was applied to trap volatiles released from Z1-04-02 and gas chromatography-mass spectrometry (GC/MS); volatile antifungal compounds were tentatively identified as bicyclic monoterpene (1R)-(-)-myrtenal. The application of the cell-free CF, and the spore suspension of Z1-04-02, showed disease severity indexes (DSIs) of 12.5% and 8.25%, respectively, which were significantly lower than those showing inoculation by S. rolfsii alone. The identification of this strain by morphology, biochemistry tests, and 16s rDNA sequences revealed that Z1-04-02 was Streptomyces albulus. This finding revealed that S. albulus Z1-04-02 displayed diverse fungicidal activities against S. rolfsii, and it has the potential to act as a biological control agent in terms of inhibiting sclerotium rot in mungbeans.

Mapping QTLs Controlling Soybean Rust Disease Resistance in Chiang Mai 5, an Induced Mutant Cultivar.

Soybean rust (SBR) caused by the fungus Phakopsora pachyrhizi is an important folia disease of soybean (Glycine max). In this study, we identified QTLs controlling SBR in Chiang Mai 5 (CM5), an SBR-resistant cultivar developed by induced mutation breeding. A recombinant inbred line (RIL) population of 108 lines developed from a cross between Sukhothai 2 (SKT2, a susceptible cultivar) and CM5 was evaluated for SBR resistance under field conditions in Thailand. QTL analysis for the resistance in the RIL population identified a single QTL, qSBR18.1, for resistance. qSBR18.1 was mapped to a 212-kb region on chromosome 18 between simple sequence repeat markers Satt288 and sc21_3420 and accounted for 21.31-35.09% depending on the traits evaluated for resistance. The qSBR18.1 interval overlapped with genomic regions containing resistance to P. pachyrhizi 4 (Rpp4), a locus for SBR resistance. Three tightly linked genes, Glyma.18G226250, Glyma.18G226300, and Glyma.18G226500, each encoding leucine-rich repeat-containing protein, were identified as candidate genes for SBR resistance at the qSRB18.1. The qSBR18.1 would be useful for breeding of SBR resistance.

Seasonal Variation of Tropical Savanna Altered Agronomic Adaptation of Stock-6-Derived Inducer Lines.

Tropicalization is one of the major objectives in breeding haploid inducers to address the poor adaptation of temperate haploid inducers in doubled haploid production in tropical maize. Gaining a better understanding of weather profiles in targeted agroecology is important. This study aimed to investigate the seasonal variation of tropical savanna climate and its impact on agronomic traits and haploid induction rate (HIR) of Stock-6-derived haploid inducer lines. A total of 14 haploid inducers were evaluated across two typical growing seasons between 2020 and 2021. Weather data were collected on daily minimum and maximum temperatures, relative humidity, precipitation, and solar radiation whereas phenotypic data were recorded on plant phenology, tassel attributes, plant stature, ear components, inducer seed rate (ISR), and HIR. The effects of season, genotype, and genotype by season were significant for all traits except season factor on ISR. Seasonal variation existed where the dry season was more suitable for haploid induction and inducer maintenance, as haploid inducers revealed better agronomic performance and seed set, delayed flowering dates, and higher HIR. Since the crossover performance of haploid inducers over seasons was detected, further implications on genotype selection in each season are discussed.

The First Genetic Linkage Map of Winged Bean [Psophocarpus tetragonolobus (L.) DC.] and QTL Mapping for Flower-, Pod-, and Seed-Related Traits.

Winged bean [Psophocarpus tetragonolobus (L.) DC.] (2n = 2× = 18) is a tropical legume crop with multipurpose usages. Recently, the winged bean has regained attention from scientists as a food protein source. Currently, there is no breeding program for winged bean cultivars. All winged bean cultivars are landraces or selections from landraces. Molecular markers and genetic linkage maps are pre-requisites for molecular plant breeding. The aim of this study was to develop a high-density linkage map and identify quantitative trait loci (QTLs) for pod and seed-related traits of the winged bean. An F2 population of 86 plants was developed from a cross between winged bean accessions W054 and TPT9 showing contrasting pod length, and pod, flower and seed colors. A genetic linkage map of 1384 single nucleotide polymorphism (SNP) markers generated from restriction site-associated DNA sequencing was constructed. The map resolved nine haploid chromosomes of the winged bean and spanned the cumulative length of 4552.8 cM with the number of SNPs per linkage ranging from 36 to 218 with an average of 153.78. QTL analysis in the F2 population revealed 31 QTLs controlling pod length, pod color, pod anthocyanin content, flower color, and seed color. The number of QTLs per trait varied between 1 (seed length) to 7 (banner color). Interestingly, the major QTLs for pod color, anthocyanin content, and calyx color, and for seed color and flower wing color were located at the same position. The high-density linkage map QTLs reported in this study will be useful for molecular breeding of winged beans.

A SNP in GmBADH2 gene associates with fragrance in vegetable soybean variety "Kaori" and SNAP marker development for the fragrance.

Fragrance in soybean is due to the presence of 2-acetyl-1-pyrroline (2AP). BADH2 gene coding for betaine aldehyde dehydrogenase has been identified as the candidate gene responsible for fragrance in rice (Oryza sativa L.). In this study, using the RIL population derived from fragrant soybean cultivar "Kaori" and non-fragrant soybean cultivar "Chiang Mai 60" (CM60), STS markers designed from BADH2 homolog were found associating with 2AP production. Genetic mapping demonstrated that QTL position of fragrance and 2AP production coincides with the position of GmBADH2 (Glycine max betaine aldehyde dehydrogenase 2). Sequence comparison of GmBADH2 between Kaori and non-fragrant soybeans revealed non-synonymous single-nucleotide polymorphism (SNP) in exon 10. Nucleotide substitution of G to A in the exon results in an amino acid change of glycine (GGC; G) to aspartic acid (GAC; D) in Kaori. The amino acid substitution changes the conserved EGCRLGPIVS motif of GmBADH2, which is essential for functional activity of GmBADH2 protein, to EGCRLDPIVS motif, suggesting that the SNP in GmBADH2 is responsible for the fragrance in Kaori. Five single nucleotide-amplified polymorphism (SNAP) markers which are PCR-based allele specific SNP markers were developed for fragrance based on the SNP in GmBADH2. Two markers specific to A allele produced a band in only Kaori, while three markers specific to G alleles produced a band in only CM60. The simple PCR-based allele specific SNAP markers developed in the present study are useful in marker-assisted breeding of fragrant soybean.

Dual-Purpose of the Winged Bean (Psophocarpus tetragonolobus (L.) DC.), the Neglected Tropical Legume, Based on Pod and Tuber Yields.

Winged beans (Psophocarpus tetragonolobus (L.) DC.) are grown as a vegetable legume crop in Thailand. All parts of the plant, including pods, seeds, leaves, flowers, and tubers are edible and are rich in protein and nutrients. Although the major consumption of winged bean is based on pod and tuber yields, only the people of Myanmar and Indonesia utilize winged bean tubers as food materials. The usefulness of the winged bean as an alternative crop for staple food and feed can shed some light on the impact of winged bean. Therefore, the evaluation of the dual purpose of the winged bean based on pod tuber yields is the objective of this study. In this study, ten-winged bean accessions-six accessions obtained from introduced sources and four accessions obtained from local Thai varieties-were laid out in randomized complete block design (RCBD) with three replications at the Agronomy Field Crop Station, Faculty of Agriculture, Khon Kaen University, Khon Kaen, Thailand from September 2019 to April 2020 and from October 2020 to April 2021. Data, including total pod weight, number of pods, pod length, 10-pod weight, and tuber weight were recorded, and the proximate nutrient and mineral contents in the tubers were also determined. The results revealed that the principal effects of year (Y) and genotype (G) were significant for total pod weight and the number of pods. Moreover, the Y × G interactions were principal effects upon the total pod weights and tuber weights. The results indicated that superior genotype and appropriate environmental conditions are key elements in successful winged bean production for both pod and tuber yields. The winged bean accessions W099 and W018 were consistent in both experimental years for pod and tuber yields at 23.6 and 18.36 T/ha and 15.20 and 15.5 T/ha, respectively. Each accession also proved high in tuber protein content at 20.92% and 21.04%, respectively, as well as significant in fiber, energy, and minerals. The results suggest that the winged bean accessions W099 and W018 can be used for dual-purpose winged bean production in Thailand.

Selection Gain of Maize Haploid Inducers for the Tropical Savanna Environments.

Lacking elite haploid inducers performing high haploid induction rate (HIR) and agronomic performance is one of fundamental factors hindering the rapid adoption of doubled haploid technology in maize hybrid breeding, especially under tropical savanna climate. Breeding haploid inducers for specific agro-ecology, thus, is indispensable yet challenging. We used temperate inducer Stock6 as genetic source for haploid induction ability and eight tropical maize genotypes as principal donors for agronomic adaptation. Three cycles of modified ear-to-row with 5% intra-family selection were applied in a population set of 78 putative haploid inducer families emphasized on agronomic performance, R1-nj anthocyanin intensity, and inducer seed set. Genetic gains, variance components, and heritability on given traits were estimated. Hierarchical clustering based on five selection criteria was performed to investigate the phenotypic diversity of putative families. Cycle effect was predominant for all observed traits. Realized genetic gain was positive for HIR (0.40% per cycle) and inducer seed set (30.10% or 47.30 seeds per ear per cycle). In this study, we reported the first haploid inducers for regions under tropical savanna climate. Three inducer families, KHI-42, KHI-54, and KHI-64, were promising as they possessed HIR about 7.8% or 14 haploid seeds per tester ear and inducer seed rate about 95.0% or 208 inducer seeds per ear. The breeding method was effective for enhancing the seed set and the expression of R1-nj anthocyanin marker of inducers, yet it showed a low effectiveness to improve haploid induction rate. Introgression of temperate inducer Stock6 into tropical gene pool followed by phenotypic selections through modified ear-to-row selection on inducer seed set and R1-nj marker did not compromise the agronomic traits of tropical inducer families. Implications and further strategies for optimizing genetic gain on HIR are discussed.

Candidate gene mapping reveals VrMLO12 (MLO Clade II) is associated with powdery mildew resistance in mungbean (Vigna radiata [L.] Wilczek).

Loss/reduction of function of Mildew Locus O (MLO) genes clade V and MLO clade IV has been shown to be responsible for powdery mildew (PM) resistance in several plant species. Mungbean (Vigna radiata) genome possesses 18 MLO genes, VrMLO1 - VrMLO18. A previous study using mungbean F2 and BC1F1 populations derived from a cross between "CN60″ (susceptible) and "RUM5″ (resistance) demonstrated that QTL qPMRUM5-3 is a major QTL for PM resistance caused by Erysiphe polygoni and is the same with major QTL qPMV4718-3 that confers PM resistance in "V4718″ (resistance). In this study, bioinformatics analysis revealed VrMLO12 locates in the qPMRUM5-3 region. Fine mapping in the F2 and BC1F1 populations using newly developed DNA markers including gene-specific markers demonstrated association between VrMLO12 and the PM resistance. Sequence analyses of VrMLO12 revealed that compared to susceptible mungbeans, RUM5 and V4718 possess SNPs in exon 10 and exon 13. The SNPs caused amino acid changes of VrMLO12, A387S and A476 G, respectively. The change occurred in transmembrane 6 domain and calmodulin binding domain (CaMBD) of the VrMLO12 protein, respectively. qRT-PCR showed that transcript expression level of VrMLO12 in RUM5 challenged with and without by E. polygoni was significantly higher than that in CN60. Phylogenetic analysis revealed that in contrast to previous findings that MLO proteins associated with PM resistance belong to MLO clade V and MLO clade IV, VrMLO12 belongs to MLO clade II. The result suggested that VrMLO12 may function differently from the other MLOs that associated with PM susceptibility. Our findings provide insight into the PM resistance in mungbean and tools for mungbean breeding.

Whole genome sequencing data of Vigna nakashimae var. Ukushima and G418.

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.