Identification and confirmation of quantitative trait loci for stachyose content in soybean seed

Stachyose is an unfavorable sugar in soybean meal that causes flatulence for non‐ruminant animals. Understanding the genetic control of stachyose in soybean will facilitate the modification of stachyose content at the molecular level. The objective of this study was to identify quantitative trait loci (QTL) associated with seed stachyose content using simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers. A normal stachyose cultivar, ‘Osage’, was crossed with a low stachyose line, V99‐5089, to develop a QTL mapping population. Two parents were screened with 33 SSR and 37 SNP markers randomly distributed on chromosome 10, and 20 SSR and 19 SNP markers surrounding a previously reported stachyose QTL region on chromosome 11. Of these, 5 SSR and 16 SNP markers were used to screen the F_3:4 lines derived from ‘Osage’ x V99‐5089. Seed samples from F_3:5 and F_3:6 lines were analyzed for stachyose content using high‐performance liquid chromatography (HPLC). Composite interval mapping analysis indicated that two stachyose QTL were mapped to chromosome 10 and 11, explaining 11% and 79% of phenotypic variation for stachyose content, respectively. The SSR/SNP markers linked to stachyose QTL could be used in breeding soybean lines with desired stachyose contents. Chi‐square tests further indicated that these two QTL probably represent two independent genes for stachyose content. Therefore, a major QTL was confirmed on chromosome 11 and a novel QTL was found on chromosome 10 for stachyose content.     

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.     

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.     

Association analysis for detecting significant single nucleotide polymorphisms for phosphorus-deficiency tolerance at the seedling stage in soybean [Glycine max (L) Merr.]

Tolerance to low-phosphorus soil is a desirable trait in soybean cultivars. Previous quantitative trait locus (QTL) studies for phosphorus-deficiency tolerance were mainly derived from bi-parental segregating populations and few reports from natural population. The objective of this study was to detect QTLs that regulate phosphorus-deficiency tolerance in soybean using association mapping approach. Phosphorus-deficiency tolerance was evaluated according to five traits (plant shoot height, shoot dry weight, phosphorus concentration, phosphorus acquisition efficiency and use efficiency) comprising a conditional phenotype at the seedling stage. Association mapping of the conditional phenotype detected 19 SNPs including 13 SNPs that were significantly associated with the five traits across two years. A novel cluster of SNPs, including three SNPs that consistently showed significant effects over two years, that associated with more than one trait was detected on chromosome 3. All favorable alleles, which were determined based on the mean of conditional phenotypic values of each trait over the two years, could be pyramided into one cultivar through parental cross combination. The best three cross combinations were predicted with the aim of simultaneously improving phosphorus acquisition efficiency and use efficiency. These results will provide a thorough understanding of the genetic basis of phosphorus deficiency tolerance in soybean.     

Identification of quantitative trait loci underlying soybean (Glycine max [L.] Merr.) seed weight including main,epistatic and QTL × environment effects in different regions of Northeast China

Seed weight (SW) is the important soybean (Glycine max [L.] Merr.), yield component and also affected the quality of soybean‐derived foods. The aim of this study was to identify the quantitative trait loci (QTL) underlying SW through 112 recombinant inbred lines (RILs) derived from the cross between “Zhongdou27” (G. max, designated by its bigger seed size, 21.9 g/100 seeds) and “Jiunong 20” (G. max, smaller seed size, 17.5 g/100 seeds). Phenotypic data were collected from this RIL population after it was grown in the sixteen tested environments. A total of eight QTL (QSW1‐1, QSW2‐1, QSW2‐2, QSW5‐1, QSW15‐1, QSW17‐1, QSW19‐1 and QSW20‐1) were identified, and they could explain 4.23%–14.65% of the phenotypic variation. Among these eight QTL, three QTL (QSW1‐1 located on the interval of Sat_159‐Satt603 of chromosome (Chr) 1 (LGD1a), QSW19‐1 located on the interval of Sat_340‐Satt523 of Chr 19 (LGL) and QSW20‐1 located on Sat_418‐Sat_105 of Chr 20 (LGI)) were newly identified and could explain 4.235%–10.08%, 8.45%–13.49% and 8.08%–10.18% of the phenotypic variation, respectively. Six of the eight identified QTL including QSW2‐2, QSW5‐1, QSW15‐1, QSW17‐1, QSW19‐1 and QSW20‐1 exhibited a significant additive (a) effect, while two QTL (QSW2‐1 and QSW19‐1) only displayed significant additive‐by‐environment (ae) effects. A total of four epistatic pairwise QTL for SW were identified in the different environments. These eight QTL and their genetic information obtained here were valuable for molecular marker‐assisted selection and the realization of a reasonable SW breeding programme in soybean.     

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.     

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.     

大豆蛋白质有关性状的QTL定位

以科丰1号×南农1138-2组合衍生的184个重组自交家系(简称RIKY)和(Essex×ZDD2315)×ZDD2315衍生的114个BC₁F₂家系(简称BIEX)为材料,对蛋白质含量、蛋油总量与油脂含量,11S、7S、11S/7S,11S-1~11S-4, 7S-1~7S-6等4组16个性状利用WinQTL Cartographer Ver.2.5的复合区间作图法(CIM)、多区间作图法(MIM)和IciMapping Ver.2.0的完备区间作图法(ICIM)进行QTL分析, 结果表明：(1) 在RIKY和BIEX群体分别定位到17⁺个和21⁺个QTL,合计38⁺个QTL;在RIKY有蛋白、油脂、蛋油总量QTL11个,在11S和7S亚基组上分别只有1⁺和3⁺个;在BIEX有前性状QTL2⁺个,有后性状QTL分别9⁺和6⁺个;(2) 两群体16个性状上均没有检测到共享的QTL,说明两群体的蛋白质有关性状具有完全不同的遗传基础;RIKY的两个亲本间蛋白、油脂和蛋油总量有明显遗传差异,但在亚基组上遗传差异不大,而BIEX则反之;(3) 4组总、分性状中,两群体一致表现出蛋白、油脂和蛋油总量和11S、7S和11S/7S比值两组在总、分性状间共享QTL(共同遗传基础), 而11S亚基组和7S亚基组两组性状在总、分性状间无共享的QTL;(4) 蛋白质有关性状QTL定位结果和分离分析结果共同表明这类性状主效基因和微效基因均占较大比重,要考虑两者兼用的育种方法。     

大豆不同生育时期叶绿素含量QTL的定位及其与产量的关联分析

利用来自波高×南农94-156的151个RI家系检测与4个不同生育时期叶绿素含量（累积量、净增量）有关的QTL,并分析其与籽粒产量、表观生物学产量和表观收获指数的关系。结果表明,与叶绿素累积量有关的QTL位于D1a+Q、F、G、H、L和M连锁群上,每个QTL可解释表型变异的6.9%~23.4%。V6和R2期没有检测到2个年份均表达的QTL,而在R4期检测到4个在2个年份均表达的QTL（qccF.1、qccG.2、qccH.1和qccM.1）,R6期仅检测到1个QTL（qccH.1）在2个年份均表达,该QTL在R4也表达。与叶绿素含量净增量有关的QTL位于B2和L连锁群上,在V6-R2时期没有检测到与叶绿素净增量有关的QTL,在B2和L连锁群上的两个QTL（qccB2-1.1和qccL.1）在R2-R4和R4-R6时期均表达,qccB2-1.1可解释表型变异的6.4%~9.8%,而qccL.1所解释表型变异达29.5%~31.3%。但这两个QTL在R2-R4和R4-R6时期表达的性质不同,且与2年均表达的籽粒产量QTL共位。这印证了生育后期叶绿素含量与籽粒产量间存在的极显著正相关。     

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.     

Detection of QTLs for salt tolerance in Asian barley (Hordeum vulgare L.) by association analysis with SNP markers

Two hundred ninety-six Asian barley (Hordeum vulgare L.) accessions were assessed to detect QTLs underlying salt tolerance by association analysis using a 384 single nucleotide polymorphism (SNP) marker system. The experiment was laid out at the seedling stage in a hydroponic solution under control and 250 mM NaCl solution with three replications of four plants each. Salt tolerance was assessed by leaf injury score (LIS) and salt tolerance indices (STIs) of the number of leaves (NL), shoot length (SL), root length (RL), shoot dry weight (SDW) and root dry weight (RDW). LIS was scored from 1 to 5 according to the severity of necrosis and chlorosis observed on leaves. There was a wide variation in salt tolerance among Asian barley accessions. LIS and STI (SDW) were the most suitable traits for screening salt tolerance. Association was estimated between markers and traits to detect QTLs for LIS and STI (SDW). Seven significant QTLs were located on chromosomes 1H (2 QTLs), 2H (2 QTLs), 3H (1 QTL), 4H (1 QTL) and 5H (1 QTL). Five QTLs were associated with LIS and 2 QTLs with STI (SDW). Two QTLs associated with LIS were newly identified on chromosomes 3H and 4H.     

A fine‐scale genetic linkage map reveals genomic regions associated with economic traits in walnut (Juglans regia)

A genetic linkage map of walnut containing 2,220 single nucleotide polymorphisms (SNPs) in 16 linkage groups (LGs) was constructed using an F₁ mapping population from a cross between “Chandler” and “Idaho,” two contrasting heterozygous parents. Five quantitative yield traits, lateral fruitfulness, harvest date and three nut traits (shell thickness, nut weight and kernel fill) were then mapped on to linkage groups. A significant quantitative trait locus (QTL) in LG 11 with negative additive effects suggested heterozygote superiority in the expression of lateral bearing. A set of three QTLs explaining ~10% of the variation in harvest date was located in LG 1. Shell thickness, nut weight and kernel fill were under the control of two to three linked pleiotropic QTLs in LG 1 segregating from “Idaho.” The marginal positive additive effects of QTLs for harvest date, shell thickness and nut weight and small negative additive effects for kernel fill suggested that the QTLs had a marginal effect on the expression of these traits.     

Genetic dissection of the relationship between plant architecture and yield component traits in soybean (Glycine max) by association analysis across multiple environments

Plant architecture and yield components are critical for the determination of seed yield in soybean. In this study, we performed genetic association analysis to dissect the relationships between plant architecture and yield component traits. Two hundred and nineteen accessions were employed, and eight agronomic traits were evaluated in six environments. Our results revealed strong positive correlations of plant architecture traits with yield components and the significant association of 4 SNPs with plant architecture traits and of 7 SNPs with yield component traits in two or more environments. Eight SNPs were co‐associated with two traits. Based on the phenotypic effects of the alleles of the detected SNPs, the best alleles were mined for twenty‐three distinct SNPs. Fifteen typical carrier materials harbouring the best allele effects were also mined. Twenty parental combinations were proposed by pyramiding possible alleles per SNP in one individual (excluding possible epistatic effects). These proposed combinations with the best alleles and carrier materials will aid in the improvement of targeted traits and marker‐assisted selection (MAS) efficiency in soybean breeding.     

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.     

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.     

大豆根区逆境耐性的种质鉴定及其与根系性状的关系

依根系类型从黄淮海和长江中下游地区301份代表性材料中选取62份，以株高、叶龄、地上部干物重、地下部干物重为指标，采用平均隶属函数值方法鉴定了苗期耐旱性、苗期耐铝毒性，加上主茎节数、分枝数、单株荚数、单株粒数、百粒重等性状鉴定了后期耐旱性，并通过钒钼黄比色法测定植株P含量鉴定了苗期耐低磷性。筛选出1级苗期     

大豆百粒重QTL定位

大豆百粒重是产量构成的重要因素之一,与产量呈正相关。本研究以溧水中子黄豆和南农493-1的504个F2正反交单株及其亲本间具有多态性的150个SSR标记信息构建连锁图谱,2008年分别在江苏南京和山东临沂两地种植其衍生的正反交F2:4家系,鉴定其百粒重,应用Win QTL CartographerV2.5复合区间作图法和两地正反交联合分析进行QTL定位。结果表明,复合区间作图法检测到16个主效QTL,联合分析检测到24个主效QTL、环境效应与细胞质效应、1个环境×QTL互作和12个细胞质×QTL互作。其中,两方法共同检测到10个主效QTL,正反交群体在两地中共同检测到3个主效QTL;Meta分析发现与其他研究一致的4个QTL。这些结果为大豆产量遗传与标记辅助育种实践提供理论基础。     

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.