Identification of genes for drought resistance and prediction of gene candidates in soybean seedlings based on linkage and association mapping

Drought is one of the primary abiotic stress factors affecting the yield,growth,and development of soybeans.In extreme cases,drought can reduce yield by more than 50%.The seedling stage is an important determinant of soybean growth:the number and vigor of seedlings will affect growth and yield at harvest.Therefore,it is important to study the drought resistance of soybean seedlings.In this study,a recombinant inbred line(RIL)population comprising 234 F_(6:10)lines(derived from Zhonghuang35×Jindou 21)and a panel of 259 soybean accessions was subjected to drought conditions to identify the effects on phenotypic traits under these conditions.Using a genetic map constructed by single nucleotide polymorphism(SNPs)markers,18 quantitative trait loci(QTL)on 7 soybean chromosomes were identified in two environments.This included 9 QTL clusters identified in the RIL population.Fifty-three QTL were identified in 19 soybean chromosomes by genome-wide association analysis(GWAS)in the panel of accessions,including 69 significant SNPs(-log_(10)(P)≥3.97).A combination of the two populations revealed that two SNPs(-log_(10)(P)≥3.0)fell within two of the QTL(qPH7-4 and qPH7-6)confidence intervals.We not only re-located several previously reported drought-resistance genes in soybean and other crops but also identified several non-synonymous stress-related mutation site differences between the two parents,involving Glyma.07 g093000,Glyma.07 g093200,Glyma.07 g094100 and Glyma.07 g094200.One previously unreported new gene related to drought stress,Glyma.07 g094200,was found by regional association analysis.The significant SNP CHR7-17619(G/T)was within an exon of the Glyma.07 g094200 gene.In the RIL population,the DSP value of the"T"allele of CHR7-17619 was significantly(P<0.05)larger than the"G"allele in different environments.The results of our study lay the groundwork for cloning and molecular marker-assisted selection of droughtresistance genes in soybeans at the seedling stage.

Differences between soybean genotypes in physiological response to sequential soil drying and rewetting

Soybean genotypes show diverse physiological responses to drought, but specific physiological traits that can be used to evaluate drought tolerance have not been identified. In the present study we investigated physiological traits of soybean genotypes under progressive soil drying and rewetting, using a treatment mimicking field conditions.After a preliminary study with eight soybean genotypes, two drought-tolerant genotypes and one susceptible genotype were grown in the greenhouse and subjected to water restriction. Leaf expansion rate, gas exchange, water relation parameters, total chlorophyll(Chl), proline contents of leaves, and root xylem p H were monitored in a time course, and plant growth and root traits were measured at the end of the stress cycle. Drought-tolerant genotypes maintained higher leaf expansion rate, net photosynthetic rate(Pn), Chl content,instantaneous water use efficiency(WUEi), % relative water content(RWC), water potential(ψw), and turgor potential(ψp) during progressive soil drying and subsequent rewetting than the susceptible genotypes. By contrast, stomatal conductance(gs) and transpiration rate(Tr)of tolerant genotypes declined faster owing to dehydration and recovered more sharply after rehydration than the same parameters in susceptible ones. Water stress caused a significant increase in leaf proline level and root xylem sap p H of both genotypes but tolerant genotypes recovered to pre-stress levels more quickly after rehydration. Tolerant genotypes also produced longer roots with higher dry mass than susceptible genotypes. We conclude that rapid perception and adjustment in response to soil drying and rewetting as well as the maintenance of relatively high Pn, %RWC, and root growth constitute the mechanisms by which drought-tolerant soybean genotypes cope with water stress.

QTL mapping of drought tolerance traits in soybean with SLAF sequencing

Drought stress is an important factor affecting soybean yield.Improving drought tolerance of soybean varieties can increase yield and yield stability when the stress occurs.Identifying QTL related to drought tolerance using molecular marker-assisted selection is able to facilitate the development of drought-tolerant soybean varieties.In this study,we used a high-yielding and drought-sensitive cultivar‘Zhonghuang 35’and a drought-tolerant cultivar‘Jindou 21’to establish F6:9 recombinant inbred lines.We constructed a highdensity genetic map using specific locus amplified fragment sequencing(SLAF-Seq)technology.The genetic map contained 8078 SLAF markers distributing across 20 soybean chromosomes with a total genetic distance of 3780.98 c M and an average genetic distance of0.59 c M between adjacent markers.Two treatments(irrigation and drought)were used in the field tests,the Additive-Inclusive Composite Interval Mapping(ICIM-ADD)was used to call QTL,and plant height and seed weight per plant were used as the indicators of drought tolerance.We identified a total of 23 QTL related to drought tolerance.Among them,seven QTL(q PH2,q PH6,q PH7,q PH17,q PH19-1,q PH19-2,and q PH19-3)on chromosomes 2,6,7,17,and 19 were related to plant height,and five QTL(q SWPP2,q SWPP6,q SWPP13,q SWPP17,and q SWPP19)on chromosomes 2,6,13,17,and 19 were related to seed weight and could be considered as the major QTL.In addition,three common QTL(q PH6/q SWPP6,q PH17/q SWPP17,and q PH19-3/q SWPP19)for both plant height and seed weight per plant were located in the same genomic regions on the same chromosomes.Three(q PH2,q PH17,and q PH19-2)and four novel QTL(q SWPP2,q SWPP13,q SWPP17,and q SWPP19)were identified for plant height and seed weight per plant,respectively.Two pairs of QTL(q PH2/q SWPP2 and q PH17/q SWPP17)were also common for both plant height and seed weight per plant.These QTL and closely linked SLAF markers could be used to accelerate breeding for drought tolerant cultivars via MAS.