Genetic analysis and identification of two soybean mosaic virus resistance genes in soybean [Glycine max (L.) Merr]

Soybean mosaic virus (SMV) commonly affects soybean production worldwide, and the SC18 strain has been widespread in China. This study aimed to characterize and map the SC18 resistance genes present in soybean cultivars ‘Kefeng No. 1’ and ‘Qihuang 22’. Inheritance analysis revealed that two independent single dominant genes in Kefeng No. 1 and Qihuang 22 confer resistance to SC18. Using simple sequence repeat (SSR) markers and bulked segregant analysis, the Kefeng No. 1 and Qihuang 22 resistance genes were located on soybean chromosomes 2 and 13, respectively. We further screened two populations of recombinant inbred lines with 32 SSR markers in the target region, where the resistance gene in Kefeng No. 1 was fine mapped to an 80‐kb region containing six putative genes. Sequence and expression analyses of these genes revealed that SMV resistance in Kefeng No. 1 was probably attributable to three of the candidate genes (i.e. Glyma.02G127800, Glyma.02G128200 and Glyma.02G128300). Collectively, the results of this study will greatly facilitate the cloning of SC18 resistance genes and marker‐assisted breeding of SMV‐resistant soybean cultivars.     

Mapping quantitative trait loci for yield-related traits in soybean (Glycine max L.)

Development of soybean cultivars with high seed yield is a major focus in soybean breeding programs. This study was conducted to identify genetic loci associated with seed yield-related traits in soybean and also to clarify consistency of the detected QTLs with QTLs found by previous researchers. A population of 135 F_2:3 lines was developed from a cross between a vegetable soybean line (MJ0004-6) and a landrace cultivar from Myanmar ({"type":"entrez-nucleotide","attrs":{"text":"R18500","term_id":"772110","term_text":"R18500"}}R18500). They were evaluated in the experimental field of Kasetsart University, Kamphaeng Saen, Nakhon Pathom, Thailand in a randomized complete block design with two replications each in 2011 and 2012 growing seasons. The two parents exhibited contrasting characteristics for most of the traits that were mapped. Analysis of variance showed that the main effects of genotype and environment (year) were significant for all studied traits. Genotype by environment interaction was also highly significant for all the traits. The population was genotyped by 149 polymorphic SSR markers and the genetic map consisted of 129 SSR loci which converged into 38 linkage groups covering 1156 cM of soybean genome. There were 10 QTLs significantly associated with seed yield-related traits across two seasons with single QTLs explaining between 5.0% to 21.9% of the phenotypic variation. Three of these QTLs were detected in both years for days to flowering, days to maturity and 100 seed weight. Most of the detected QTLs in our research were consistent with earlier QTLs reported by previous researchers. However, four novel QTLs including SF1, SF2 and SF3 on linkage groups L and N for seed filling period and PN1 on linkage group D1b for pod number were identified in the present study.     

Tagging the juvenile locus in soybean [Glycine max (L.) Merr.] with molecular markers

Soybean cultivars carrying the `long juvenile trait' show a delayed flowering response under short day conditions. The incorporation of this character into genotypes of agronomic interest may allow a broader range of sowing dates and latitudes for a single cultivar adaptation. The objective of this work was to identify molecular markers linked to the juvenile locus in soybean. Experiments were carried out using two pairs of near isogenic lines(NILs) differing in the presence of the long juvenile trait, and RAPD markers. Four hundred primers were first screened to find polymorphism associated with the trait. Additional differences between NILs were sought by digesting the genomic DNA with five restriction enzymes. Polymorphic fragments detected between NILs were tested for linkage to the juvenile locus in the corresponding F₂ segregating populations. Marker bc357-HaeIII was linked (χ²L = 46.316) to the juvenile locus with an estimated recombination frequency of 0.13 ± 0.03in one of the genetic backgrounds studied. The fragment was cloned, sequenced and converted into a SCAR marker. Moreover,bc357-HaeIII was used as RFLP probe. Both, SCAR and RFLP generated markers linked to the juvenile locus in the two genetic backgrounds analysed. Results presented in this work can be utilised for both, the localisation of the gene associated with the character and for tagging the juvenile trait in soybean breeding programs. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genomewide association analysis of salt tolerance in soybean [Glycine max (L.) Merr.]

Salinity is a common abiotic stress causing soybean [Glycine max (L.) Merr.] yield loss worldwide. The use of tolerant cultivars is an effective and economic approach to coping with this stress. Towards this, research is needed to identify salt‐tolerant germplasm and better understand the genetic and molecular basis of salt tolerance in soybean. The objectives of this study were to identify salt‐tolerant genotypes, to search for single‐nucleotide polymorphisms (SNPs) and QTLs associated with salt tolerance. A total of 192 diverse soybean lines and cultivars were screened for salt tolerance in the glasshouse based on visual leaf scorch scores after 15–18 days of 120 mM NaCl stress. These genotypes were further genotyped using the SoySNP50K iSelect BeadChip. Genomewide association mapping showed that 62 SNP markers representing six genomic regions on chromosomes (Chr.) 2, 3, 5, 6, 8 and 18, respectively, were significantly associated with salt tolerance (p < 0.001). A total of 52 SNP markers on Chr. 3 are mapped at or near the major salt tolerance QTL previously identified in S‐100 (Lee et al., 2014). Three SNPs on Chr. 18 map near the salt tolerance QTL previously identified in Nannong1138‐2 (Chen, Cui, Fu, Gai, & Yu, 2008). The other significant SNPs represent four putative minor QTLs for salt tolerance, newly identified in this study. The results above lay the foundation for fine mapping, cloning and molecular breeding for soybean salt tolerance.     

Genetic analysis for the leaf pubescence density and water status traits in soybean [Glycine max (L.) Merr.]

Leaf pubescence density (PD) is an important component for the adaptation of soybean [ Glycine max (L.) Merr.] to drought-prone environment. Quantitative trait loci (QTL) controlling PD on the upper surface of leaf blade (PDU), PD on the lower surface of leaf blade (PDL), leaf wilting coefficient (WC) and rate of excised leaf drying (ELD) were identified using recombinant inbred lines (RILs) population from the cross between soybean cultivars 'kefeng1' and 'nannong1138-2' at the field soil drought stress stage from the mid-end of stem elongation to onset of flowering. A total of 20 QTLs were detected on molecular linkage groups (MLGs) A2, D1b, E, H, G and I with individual QTL explained 4.49–23.56% of phenotypic variation by composite interval mapping. The QTLs for PD on MLG H were mapped to near Ps locus while the QTLs on MLG D1b were located near Rsc-7 . Three genome regions for PD and water status traits on MLGs A2, D1b and H were associated. This study revealed that leaf surface PD may play an important role in the soybean drought tolerance.     

QTL mapping of phosphorus deficiency tolerance in soybean (Glycine max L. Merr.)

Phosphorus (P) deficiency is a major abiotic stress that limits plant growth and crop productivity throughout the world. In the present study, 184 recombinant inbred line (RIL) families developed from soybean varieties Kefeng No. 1 and Nanong 1138-2 were used to identify quantitative trait loci (QTL) associated with P deficiency tolerance. Seven traits of plant height (HT), weight of fresh shoot (FSW), weight of fresh root (FRW), weight of dry root (DRW), length of main root (RL), phosphorus content in leaf (LP), phosphorus content in root (RP), were used as parameters to assess the phosphorus deficiency tolerance. The QTL mapping for the seven traits was performed using the program WinQTLCart. Seven QTLs were detected and mapped on two linkage groups for three traits of weight of fresh shoot, phosphorus contents in leaf and in root. The QTLs that had LOD scores more than three were detected for all of the three traits above. Most of the QTLs explained more than 10% of the total variation. The two QTLs for phosphorus content in leaf explained more than 20% of the total variation, respectively. Five QTLs were mapped on linkage group F2, and two on linkage F1. It was suggested that the genes related to phosphorus deficiency tolerance located on linkage group F in soybean.Contributed equally to this work.     

SSR marker tightly linked to the Ti locus in Soybean [Glycine max (L.) Merr.]

Soybean is a major source of protein meal in the world. Soybean kunitz trypsin inhibitor (SKTI) protein is a responsible for the inferior nutritional quality of unheated or incompletely heated soybean meal. The primary objective of this research was to identify DNA markers linked to the Ti locus controlling presence and absence of kunitz trypsin inhibitor protein. Two mapping populations were developed. Population 1 was derived from a cross between cultivar Jinpumkong2 (TiTi) and C242 (titi). Population 2 was made from a mating between cultivar Clark (TiTi) and C242. The F₁ plants were grown in the greenhouse to produce F₂ seeds. Each F₂ seed from F₁ plants was analyzed electrophoretically to determine the presence of the SKTI protein band. One-thousand RAPD primers, 342 AFLP primer sets, and 35 SSR primers were used to map Ti locus in population 1 and 2. The presence of SKTI protein was dominant to the lack of a SKTI protein and kunitz trypsin inhibit protein band was controlled by a single locus. Twelve DNA markers (4 RAPD, 4 AFLP, and 3 SSR) and Ti locus were found to be genetically linked in population 1 consisted with 94 F₂ individual plants. Three SSR markers (Satt409, Satt228, and Satt429) were linked with Ti locus within 10 cM. Satt228 marker was tightly linked with Ti locus. Satt228 marker was tightly linked within 0–3.7 cM of the Ti locus and may be useful in a marker assisted selection program.     

Major QTLs associated with green stem disorder insensitivity of soybean (Glycine max (L.) Merr.)

Green stem disorder (GSD) is one of the most serious syndromes affecting soybean (Glycine max) cultivation in Japan. In GSD, stems remain green even when pods mature. When soybean plants develop GSD, seed surfaces are soiled by tissue fluid and seed quality is deteriorated during machine harvesting. We performed quantitative trait locus (QTL) analyses for GSD insensitivity using recombinant inbred lines (RILs; n = 154) derived from a cross between an insensitive line (‘Touhoku 129’) and a sensitive leading cultivar (‘Tachinagaha’) during a 6-year evaluation. Three effective QTLs were detected. The influences of these QTLs were in the following order: qGSD1 (LG_H) > qGSD2 (LG_F) > qGSD3 (LG_L). At these three QTLs, ‘Touhoku 129’ genotypes exhibited more GSD insensitivity than ‘Tachinagaha’ genotypes. The lower incidence of GSD for ‘Touhoku129’ was attributable primarily to these three QTLs because RILs harboring a ‘Touhoku 129’ genotype at the three QTLs exhibited a GSD incidence similar to that of ‘Touhoku 129.’ Although a limitation of this study is that only one mapping population was evaluated, this QTL information and the flanking markers of these QTLs would be effective tools for resolving GSD in soybean breeding programs.     

Genetic analysis of antixenosis resistance to the common cutworm (Spodoptera litura Fabricius) and its relationship with pubescence characteristics in soybean (Glycine max (L.) Merr.)

The common cutworm (CCW, Spodoptera litura Fabricius) is one of the most serious pests of soybean (Glycine max (L.) Merr.). Previously, two quantitative trait loci (QTLs) for antibiosis resistance to CCW, CCW-1 and CCW-2, were detected in the resistant cultivar Himeshirazu. In this study, we conducted an anti-xenosis bioassay using a recombinant inbred population derived from a cross between a susceptible cultivar Fukuyutaka and Himeshirazu to perform QTL analysis. Two QTLs for antixenosis resistance, qRslx1 and qRslx2, were identified on Chrs 7 and 12, and the resistant alleles of qRslx1 and qRslx2 were derived from Himeshirazu and Fukuyutaka, respectively. The position of qRslx1 is similar to that of CCW-1. We also analyzed pubescence characteristics because they have been reported to be associated with soybean insect resistance. Two QTLs for pubescence length (on Chrs 7 and 12) and two QTLs for pubescence density (on Chrs 1 and 12) were identified. The pubescence QTLs on Chrs 7 and 12 were located near qRslx1 and qRslx2, respectively. These results suggest that the antixenosis resistance could be controlled genetically by the identified QTLs and that the pubescence characteristics might contribute to the soybean antixenosis resistance to CCW.     

Analysis of quantitative trait loci underlying the period of reproductive growth stages in soybean (<Emphasis Type="Italic">Glycine max</Emphasis> [L.] Merr.)

Quantitative trait loci (QTLs) underlying reproductive growth stages are important for molecular breeding of soybeans [Glycine max (L.) Merr.]. Most of these QTLs identified so far derived from a single environment, and thus may be influenced by specific environmental conditions. In this study (from 2004 to 2005), analysis of QTLs underlying the period to reach a given reproductive growth stage was performed in three different environments (Harbin, Heilongjiang Province, China). QTL analysis was achieved with a recombination inbred line (RIL) population consisting of 153 lines. The RIL population derived from a cross between an American semi-dwarf cultivar (cv. Charleston) and a Chinese line with a short growth stage (cv. Dongnong 594). The growth stage data of soybean was recorded for each day. QTLs for all eight reproductive growth stages of soybean (R1 to R8) were analyzed by a composite interval mapping method combined with a mixed genetic model. Fifty-four QTLs displayed main effects and 56 QTL pairs showed epistatic effects. Two marker intervals (Satt173–Satt581, Satt402–Satt267), located on the linkage group O and D1a respectively, strongly influenced plant developmental processes during reproductive growth stages. The findings of this study open the possibility to modulate the structure of soybean growth stages by marker-assisted selection and pyramiding QTL analysis. H.-M. Qiu and D.-W. Xin contributed equally to this work.     

Effect of different planting times on the quantitative variation of total seed isoflavone content and composition in Korean soybean cultivars (Glycine max (L.) Merr.)

Journal of Crop Science and Biotechnology - There is great interest in the enhancement of isoflavones as one of the functional ingredients in soybean. This study aimed to investigate the effects of...     

Soybean (Glycine max (L.) Merr.) growth and development response to CO2 enrichment under different temperature regimes

The carbon dioxide (CO₂) concentration of the global atmosphere has increased during the last decades. This increase is expected to impact the diurnal variation in temperature as well as the occurrence of extreme temperatures. This potentially could affect crop production through changes in growth and development that will ultimately impact yield. The objective of this study was to evaluate the effect of CO₂ and its interaction with temperature on growth and development of soybean (Glycine max (L.) Merr., cv. Stonewall). The experiment was conducted in controlled environment chambers at the Georgia Envirotron under three different temperatures and two CO₂ regimes. The day/night air temperatures were maintained at 20/15, 25/20 and 30/25 °C, while the CO₂ levels were maintained at 400 and 700 ppm, resulting in six different treatments. Plants were grown under a constant irradiance of 850 μmoles m⁻² s⁻¹ and a day length of 12 h; a non-limiting supply of water and mineral nutrients were provided. Five growth analyses were conducted at the critical development stages V4, R3, R5, R6 and R8. No differences in start of flowering were observed as a function of the CO₂ level, except for the temperature regime 25/20 °C, where flowering for the elevated CO₂ level occurred 2 days earlier than for the ambient CO₂ level. For aboveground biomass, an increase in the CO₂ level caused a more vigorous growth at lower temperatures. An increase in temperature also decreased seed weight, mainly due to a reduction in seed size. For all temperature combinations, final seed weight was higher for the elevated CO₂ level. This study showed that controlled environment chambers can be excellent facilities for conducting a detailed growth analysis to study the impact on the interactive effect of changes in temperature and CO₂ on soybean growth and final yield.     

An intersubgeneric hybrid between Glycine tomentella Hayata and the soybean,G. max (L.) Merr.

Summary The objective of the present paper is to provide information on the morphology and cytology of an intersub-generic hybrid (2n=59) between Glycine tomentella Hayata (2n=78) and G. max (L.) Merr. (2n=40) obtained through in vitro immature seed culture. The hybrid plant was slow in vegetative growth and twinning like the female parent but morphologically was intermediate between both parents for several traits. At metaphase I, the average chromosome associations and ranges for 25 cells were 44.0 I (37–51)+7.5 II (3–11). The plant was completely pollen and seed sterile. The present investigation suggests that wild perennial Glycine species can be exploited as either the male or female parent in wide hybridization programs with the soybean, G. max.     

大豆胁迫相关蛋白GmSAP3基因的克隆和表达分析

胁迫是影响植物生长发育和农作物产量的重要因素.胁迫相关蛋白(stress associated protein,SAP)是一类锌指蛋白,广泛参与植物发育和胁迫响应.为了研究大豆SAP基因的功能,本研究以'商豆1201'为材料克隆了GmSAP3基因,并进行了生物信息学分析,采用RT-PCR分析该基因在大豆不同组织和温度胁迫下的表达模式.结果 表明,GmSAP3基因CDS序列全长为513 bp,编码170个氨基酸,GmSAP3蛋白等电点pI为6.79,分子量为18.30568kD;序列分析发现,GmSAP3含有2个保守结构域(zf-A20和ZnF-AN1),序列比对和进化分析发现GmSAP3与GmSA P26亲缘关系最为接近.RT-PCR结果表明,GmSAP3在大豆不同组织中均有表达,其中根和叶片表达较高;GmSAP3受高温胁迫诱导表达,推测该基因在大豆胁迫响应中具有重要作用.本研究为GmSAP3基因功能研究提供了一定的理论支持.     

Genotype x Bradyrhizobium japonicum strain interactions in dinitrogen fixation and agronomic traits of soybean (Glycine max L. Merr.)

Summary Bradyrhizobium japonicum strain G49 has been the only inoculum used in French soils. Soybean (Glycine max L. Merr.) cultivars were selected and tested according to their performances with this rhizobial strain. The aim of the present study was to determine the consequences of strain substitution on N₂ fixation abilities of various genotypes. Three genotypes and cultivar Weber, in combination with B. japonicum strain G49 or SMGS1, were cultivated in pots and tested for nitrogenase activity under differing nitrogen nutrition conditions. The reliability of ARA (acetylene reduction activity) measurement for assessing symbiotic nitrogen fixation under the experimental conditions used was checked. Genotypic variability for symbiotic fixation activity was observed with each strain under soil culture conditions; important genotype x strain interactions were also involved. These results were corroborated for the protein yield and other yield component performances of the various genotype-strain associations. Thus, in France, the replacement of strain G49 with another one might result in the alteration of the relative agronomic performances of the soybean cultivars, since N₂ fixation is considered as a major factor of soybean productivity.     

Quantitative trait locus analysis for soybean (Glycine max) seed protein and oil concentrations using selected breeding populations

Soybean seed protein and oil concentrations are important traits that directly affect the quality of soyfoods. Many studies and breeding programmes have been conducted to find major quantitative trait loci (QTL) that regulate protein and oil concentrations and to develop soybean cultivars with high protein and/or oil content. The purpose of this study was to identify these QTL using a selected breeding population. The population was tested in field conditions over a period of 3 years. Seed protein and oil concentrations were measured each year. Single‐nucleotide polymorphisms (SNPs) were used to construct genetic map using a 180K SoyaSNP array, which identified 1,570 SNPs. We identified 12 QTL for seed protein, 11 for seed oil concentration and four for both traits. Among these, 17 QTL were closely mapped to previously reported QTL, whereas ten sites were novel. Several QTL were detected across at least two experimental years. These loci are good candidate QTL for optimal seed protein and oil concentrations. Our results demonstrate that favourable target QTL can be successfully identified using selected breeding populations.     

Spatial field variations in soybean (Glycine max [L.] Merr.) performance trials affect agronomic characters and seed composition

The objective of this study was to investigate field trends in agronomic characters of soybean (Glycine max [L.] Merr.) performance trials. Field experiments were arranged as lattice designs, and for each plot within an experiment, an environmental covariate representing the spatial trend was calculated using neighbour analysis. Correlations between environmental covariates of different characters were then used to analyse the degree of similarity in their spatial trends. Grain yield, seed protein content, and seed size were more influenced by spatial variations than time to flowering, time to maturity, and oil content. Significant correlations between environmental covariates were found, which demonstrate the similarity of trend patterns in different characters within particular experiments. In field trials exhibiting a high degree of spatial heterogeneity, correlations between environmental covariates of grain yield and protein content were highly positive. This indicates that field conditions, which promote grain yield, would also enhance protein content. Moreover, the observed trend patterns considerably affected the calculation of phenotypic coefficients of correlation between grain yield and seed protein content. The results suggest that spatial analysis should be applied to all characters of interest, when field conditions are not homogeneous.     

New genes for resistance to Xanthomonas campestris pv. glycines in soybean [Glycine max (L.) Merr.] and their inheritance

Bacterial leaf pustule (BLP) caused by Xanthomonas campestris pv. glycines is an important disease of soybean. A new resistant source, P-4-2, showing an immune reaction in controlled conditions to BLP was crossed with the susceptible cultivar Monetta to study the inheritance of resistance. All F1 plants were susceptible. The F2 population segregated 15 susceptible: 1 resistant plants indicating the presence of duplicate recessive genes controlling resistance. This was further confirmed in the F3 generation. Two recessive genes conferring high levels of resistance in soybean to Xanthomonas campestris pv. glycines under controlled conditions are being reported here for the first time. This revised version was published online in July 2006 with corrections to the Cover Date.     

Mineral Nitrogen Availability and Isoflavonoid Accumulation in the Root Systems of Soybean (Glycine max (L.) Merr.)

Isoflavonoids, as plant-to-bacteria signal molecules, play an important role in the establishment of the soybean ( Glycine max (L.) Merr.)– Bradyrhizobium japonicum nitrogen (N) fixing symbiosis. They are essential to the development of effective root nodules and responsible for inducing the nod genes of B. japonicum . Because N affects a broad range of infection events, especially the symbiotic events occurring within 18 h of inoculation, it is reasonable to hypothesize that mineral N disrupts the interorganismal signal exchange between soybean host plants and B. japonicum . High performance liquid chromatographic (HPLC) analysis of root extracts of soybean, inoculated with B. japonicum or not, grown with various levels of mineral N in the rooting medium were performed to test this hypothesis. The results of these studies indicated that: (1) at early plant growth stages (before the onset of N fixation), a strong negative relationship between N application and soybean root isoflavonoid (genistein and daidzein) concentrations existed; (2) although isoflavonoid (genistein and daidzein) concentrations in both inoculated and non-inoculated soybean root systems were generally decreased by N application, at very low N levels (10 mg N l⁻¹) genistein in the non-inoculated plant roots was not decreased relative to the 0 N plants; (3) averaged over all mineral N treatment levels and sampling times, inoculation of soybean with B. japonicum increased root daidzein concentrations (P > 0.05), but did not affect genistein. Overall, N application reduced the isoflavonoid concentration of soybean root systems, which probably plays a part in the regulation of soybean nodule formation by available N.