Identification and fine mapping of qSW2 for leaf slow wilting in soybean

Drought is one of the abiotic stresses limiting the production of soybean(Glycine max).Elucidation of the genetic and molecular basis of the slow-wilting(SW)trait of this crop offers the prospect of its genetic improvement.A panel of 188 accessions and a set of recombinant inbred lines produced from a cross between cultivars Liaodou 14 and Liaodou 21 were used to identify quantitative-trait loci(QTL)associated with SW.Plants were genotyped by Specific-locus amplified fragment sequencing and seedling leaf wilting was assessed under three water-stress treatments.A genome-wide association study identified 26 SW-associated single-nucleotide polymorphisms(SNPs),including three located in a 248-kb linkage-disequilibrium(LD)block on chromosome 2.Linkage mapping revealed a major-effect QTL,qSW2,associated with all three treatments and adjacent to the LD block.Fine mapping in a BC_(2)F_(3) population derived from a backcross between Liaodou 21 and R26 confined qSW2 to a 60-kb interval.Gene expression and sequence variation analysis identified the gene Glyma.02 g218100,encoding an auxin transcription factor,as a candidate gene for qSW2.Our results will contribute significantly to improving drought-resistant soybean cultivars by providing genetic information and resources.

Genome-wide association with transcriptomics reveals a shade-tolerance gene network in soybean

Shade tolerance is essential for soybeans in inter/relay cropping systems.A genome-wide association study(GWAS)integrated with transcriptome sequencing was performed to identify genes and construct a genetic network governing the trait in a set of recombinant inbred lines derived from two soybean parents with contrasting shade tolerance.An improved GWAS procedure,restricted two-stage multi-locus genome-wide association study based on gene/allele sequence markers(GASM-RTM-GWAS),identified 140 genes and their alleles associated with shade-tolerance index(STI),146 with relative pith cell length(RCL),and nine with both.Annotation of these genes by biological categories allowed the construction of a protein–protein interaction network by 187 genes,of which half were differentially expressed under shading and non-shading conditions as well as at different growth stages.From the identified genes,three ones jointly identified for both traits by both GWAS and transcriptome and two genes with maximum links were chosen as beginners for entrance into the network.Altogether,both STI and RCL gene systems worked for shade-tolerance with genes interacted each other,this confirmed that shadetolerance is regulated by more than single group of interacted genes,involving multiple biological functions as a gene network.

A novel procedure for identifying a hybrid QTL-allele system for hybrid-vigor improvement, with a case study in soybean(Glycine max)yield

“Breeding by design” for pure lines may be achieved by construction of an additive QTL-allele matrix in a germplasm panel or breeding population, but this option is not available for hybrids, where both additive and dominance QTL-allele matrices must be constructed. In this study, a hybrid-QTL identification approach, designated PLSRGA, using partial least squares regression(PLSR) for model fitting integrated with a genetic algorithm(GA) for variable selection based on a multi-locus, multi-allele model is described for additive and dominance QTL-allele detection in a diallel hybrid population(DHP). The PLSRGA was shown by simulation experiments to be superior to single-marker analysis and was then used for QTL-allele identification in a soybean DPH yield experiment with eight parents. Twenty-eight main-effect QTL with 138 alleles and nine QTL × environment QTL with 46 alleles were identified, with respective contributions of 61.8% and 23.5% of phenotypic variation. Main-effect additive and dominance QTL-allele matrices were established as a compact form of the DHP genetic structure. The mechanism of heterosis superior-to-parents(or superior-to-parents heterosis, SPH) was explored and might be explained by a complementary locus-set composed of OD+(showing positive over-dominance, most often), PD+(showing positive partial-to-complete dominance, less often) and HA+(showing positive homozygous additivity, occasionally) loci, depending on the parental materials. Any locus-type, whether OD+, PD + and HA+, could be the best genotype of a locus. All hybrids showed various numbers of better or best genotypes at many but not necessarily all loci, indicating further SPH improvement. Based on the additive/dominance QTL-allele matrices, the best hybrid genotype was predicted, and a hybrid improvement approach is suggested. PLSRGA is powerful for hybrid QTL-allele detection and cross-SPH improvement.

Cd Distribution and Subcellular Localization in Leaf and Its Effects on Growth of Soybean(Glycine max) Seedlings

In order to investigate Cd accumulation, subcellular distribution, and local-ization in soybean seedlings leaves, soybean seedlings were cultivated in solution containing different concentrations of Cd. The results showed that most Cd associ-ated with the cellwal s and soluble fractions, and a minor part of Cd presented in mitochondria fractions, nuclear and chloroplast fractions, especial y exposure to high Cd concentrations. Under 20.00 mg/L Cd stress, Cd subcellular distribution fol owed a sequence as： soluble fractions （55.00%）＆gt;cellwal s （30.0%）＆gt;mitochondria fractions （8.21%）＆gt;nuclear and chloroplast fractions （6.79%）. Deposited Cd black particles were observed in cellwal s, chloroplasts, nuclei, and vacuoles through electrical microscope slice. This fact indicated that the cellwal s of soybean leaves were the first protecting organel es from Cd toxicity, and the cellwal s and soluble fractions were the main place for Cd storage. Due to Cd accumulated in the organel es, the intercellular space was enlarged and the subcellular structure was damaged, especial y for the chloroplasts.

Natural variations and geographical distributions of seed carotenoids and chlorophylls in 1167 Chinese soybean accessions

Understanding the composition and contents of carotenoids in various soybean seed accessions is important for their nutritional assessment.This study investigated the variability in the concentrations of carotenoids and chlorophylls and revealed their associations with other nutritional quality traits in a genetically diverse set of Chinese soybean accessions comprised of cultivars and landraces.Genotype,planting year,accession type,seed cotyledon color,and ecoregion of origin significantly influenced the accumulation of carotenoids and chlorophylls.The mean total carotenoid content was in the range of 8.15–14.72μg g–1 across the ecoregions.The total carotenoid content was 1.2-fold higher in the landraces than in the cultivars.Soybeans with green cotyledons had higher contents of carotenoids and chlorophylls than those with yellow cotyledons.Remarkably,lutein was the most abundant carotenoid in all the germplasms,ranging from 1.35–37.44μg g–1.Carotenoids and chlorophylls showed significant correlations with other quality traits,which will help to set breeding strategies for enhancing soybean carotenoids without affecting the other components.Collectively,our results demonstrate that carotenoids are adequately accumulated in soybean seeds,however,they are strongly influenced by genetic factors,accession type,and germplasm origin.We identified novel germplasms with the highest total carotenoid contents across the various ecoregions of China that could serve as the genetic materials for soybean carotenoid breeding programs,and thereby as the raw materials for food sectors,pharmaceuticals,and the cosmetic industry.

Mapping and validation of a dominant salt tolerance gene in the cultivated soybean(Glycine max) variety Tiefeng 8

Salt is an abiotic stress factor that strongly affects soybean growth and production. A single dominant gene has been shown to confer salt tolerance in the soybean cultivar Tiefeng 8.The objective of the present study was to genetically map the salt-tolerance gene in an F2:3population and a recombinant inbred line(RIL) population derived from a cross between two cultivated soybeans, Tiefeng 8(tolerant) and 85-140(sensitive). The F2:3families and RILs were treated with 200 mmol L-1Na Cl to evaluate salt tolerance. The F2:3population showed 1(42 tolerant): 2(132 segregating): 1(65 sensitive) segregation, indicating a single dominant gene for salt tolerance in Tiefeng 8. A sequence-characterized amplified region(SCAR) marker from a previously identified random amplified polymorphic DNA(RAPD)marker and four insertion/deletion polymorphism(In Del) markers were developed within the mapping region. Using these markers along with SSR markers, the salt-tolerance gene was mapped within 209 kb flanked by SCAR marker QS08064 and SSR marker Barcsoyssr_3_1301 on chromosome 3. Three markers that cosegregated with the salt tolerance gene and SCAR marker QS08064 were used to genotype 35 tolerant and 23 sensitive soybean accessions. These markers showed selection efficiencies of 76.2% to94.2%. The results indicate that these markers will be useful for marker-assisted breeding and facilitating map-based cloning of the salt tolerance gene in soybean.

Analysis of QTLs for the Trichome Density on the Upper and Downer Surface of Leaf Blade in Soybean [Glycine max (L.) Merr.]

Trichomes （plant hairs） are present on nearly all land plants and are known to play important roles in plant protection, specifically against insect herbivory, drought, and UV radiation. The identification of quantitative trait loci （QTL） associated with trichome density should help to interpret the molecular genetic mechanism of soybean trichome density. 184 recombinant inbred lines （RILs）, derived from a cross between soybean cultivars Kefeng 1 and Nannong 1138-2 were used as segregating population for evaluation of TDU （trichome density on the upper surface of leaf blade） and TDD （trichome density on the downer surface of leaf blade）. A total of 15 QTL were detected on molecular linkage groups （MLG） A2, Dla, Dlb, E and H by composite interval mapping （CIM） and among all the QTL, qtuA2-1, qtuD 1 a-1, qtuD lb-2, qtuH-2 qtuE-1, qtdDlb-2, and qtdH- 2 were affirmed by multiple interval mapping （MIM）. The contribution ofphenotypic variance of qtuH-2 was 31.81 and 29.4% by CIM and MIM, respectively, suggesting it might be major gene Ps loci. Only 10 pairs of main QTL interactions for TDU were detected, explained a range of 0.2-5.1% of phenotypic variations for each pair for a total of 22.8%. The QTL on MLG Dlb affecting trichome density were mapped near to Rsc-7 conditioning resistance to SMV （soybean mosaic virus）. This study showed that the genetic mechanism of trichome density was the mixed major gene and polygene inheritance, and also suggested that the causal nature between trichome density and other agronomic traits.

大豆膨胀素基因GmEXPB5和GmEXPB7生物信息学及表达分析

膨胀素(expansin,EXP)通过调控细胞壁的松弛在植物应对环境胁迫过程中起着重要作用。为研究EXP基因在大豆应对非生物胁迫过程中的作用,该文对大豆中的两个EXP基因(GmEXPB5和GmEXPB7)及其蛋白序列进行生物信息学分析,通过实时荧光定量PCR(qRT-PCR)检测基因表达量。结果表明:(1)GmEXPB5和GmEXPB7分别位于大豆第10号和第12号染色体上,编码的蛋白序列长度分别为272和267个氨基酸。GmEXPB5蛋白分子量为29.07 kD,理论等电点为7.51;GmEXPB7蛋白分子量为29.09 kD,理论等电点为8.66。GmEXPB5和GmEXPB7均为稳定的亲水蛋白且定位于细胞壁中。GmEXPB5和GmEXPB7蛋白均含有一段信号肽序列和一个保守的DPBB_1结构域。(2)GmEXPB5蛋白与鹰嘴豆CaEXPB15蛋白亲缘关系最近,GmEXPB7蛋白与密花豆、赤豆和豇豆的EXPB3蛋白有着较近的亲缘关系。(3)GmEXPB5和GmEXPB7在大豆根、茎和叶中均有表达且它们在根和叶中的表达量均显著高于茎中的表达量。(4)GmEXPB5和GmEXPB7在大豆幼苗中可以响应盐、干旱和低温胁迫。(5)GmEXPB5启动子区域含有2种与逆境相关的顺式作用元件(ABRE和ARE);GmEXPB7启动子区域含有5种与逆境相关的顺式作用元件(ABRE、ARE、CGTCA-motif、TC-rich repeats和MBS)。综上所述,GmEXPB5和GmEXPB7能够参与大豆对非生物胁迫的应答。

MicroRNAs Involved in the Pathogenesis of Phytophthora Root Rot of Soybean (Glycine max)

Phytophthora root rot is one of the most prevalent diseases in the world,which can infect the seedlings and plants,with substantial negative impact on soybean yield and quality.MicroRNAs （miRNAs） are a class of post-transcriptional regulators of gene expression during growth and development of organisms.A soybean disease-resistance variety Suinong 10 was inoculated with Phytophthora sojae race No.1,and the specific miRNA resistant expression profile was acquired by microarray for the first time.Different expressional miRNAs have been found after comparing the results of the treated sample with the control sample.Furthermore,the target genes of different expressional miRNAs were predicted.Two miRNAs,cbr-mir-241 and ath-miR854a,regulated the disease-resistance process directly through their targets,some enzymes.Another two miRNAs,gma-miR169a and ath-miR169h,participated in disease-resistance regulation as transcription factors.Similarly,one miRNA,ptc-miR164f,has been reported to regulate the plant development.All of these studies would be served as the foundation for exploring the resistance mechanism.

Genetic Analysis of Combining Abilities and Heterosis for the Contents of Soybean Isoflavone and Its Components Among the Soybean Varieties [Glycine max (L.) Merr.]

The genetic analysis of soybean isoflavone content and its components were carried out based on the NC Ⅱ matingdesign in eight soybean varieties. The results showed that the isoflavone contents and its components of soybean seedare quite differences among the tested materials, the contents of isoflavone and daidzein are controlled not only byadditive effects and but also by non-additive effects, while the content of genistin is dominated by non-additive effects,and genistein, glycitin and daidzin are mainly controlled by additive effects. There are significant differences in thecontents of isoflavone and its components among the combinations derived from different parents. Results also indicatedthat the tested traits are negatively heterosis except for the contens of daidzein and daidzin are positively heterosis basedon the data of the GCA and SCA in average heterosis values. In this research we have a suggestion that soybean varietywith high isoflavone should be used as one of the parents in the breeding program, and it is the best choice that thecombinations crossed between two high isoflavone varieties or a high variety and a low one.

Genetic Analysis of Embryo, Cytoplasm and Maternal Effects and Their Environment Interactions for Isoflavone Content in Soybean [Glycine max (L.) Merr.]

Soybean seed products contain isoflavones （genistein, daidzein, and glycitein） that display biological effects when ingested by humans and animals. These effects are species, dose and age dependent. Therefore, the content and quality of isoflavones in soybeans is a key factor to the biological effect. Our objective was to identify the genetic effects that underlie the isoflavone content in soybean seeds. A genetic model for quantitative traits of seeds in diploid plants was applied to estimate the genetic main effects and genotype x environment （GE） interaction effects for the isoflavone content （IC） of soybean seeds by using two years experimental data with an incomplete diallel mating design of six parents. Results showed that the IC of soybean seeds was simultaneously controlled by the genetic effects of maternal, embryo, and cytoplasm, of which maternal genetic effects were most important, followed by embryo and cytoplasmic genetic effects. The main effects of different genetic systems on IC trait were more important than environment interaction effects. The strong dominance effects on isoflavone from residual was made easily by environment conditions. Therefore, the improvement of the IC of soybean seeds would be more efficient when selection is based on maternal plants than that on the single seed. Maternal heritability （65.73%） was most important for IC, followed by embryo heritability （25.87%） and cytoplasmic heritability （8.39%）. Based on predicated genetic effects, Yudou 29 and Zheng 90007 were better than other parents for increasing IC in the progeny and improving the quality of soybean, The significant effects of maternal and embryo dominance effects in variance show that the embryo heterosis and maternal heterosis are existent and uninfluenced by environment interaction effects.

Correlation Analysis of New Soybean[Glycine max(L.)Merr]Gene Gm15G117700 with Oleic Acid

The fatty acid dehydrogenase gene plays an important role in regulating the oleic acid content in soybean.Genome-wide association study screened out soybean oleic acid related gene Gm15G117700.A fragment size of 693bp was obtained by PCR amplification of the gene and,it was connected by seamless cloning technology to the pMD18T cloning vector.Based on the gene sequence cloned,bioinformatic analysis of gene protein was performed.The overexpression vector of Gm15G117700 and the CRISPR/Cas9 gene editing vector were constructed.The positive plants were obtained by Agrobacterium-mediated transformation of soybean cotyledon nodes and T2 plants were identified by conventional PCR,QT-PCR and Southern blot hybridization.10 copies of high and low oleic acid seeds were selected for QT-PCR to identify the expression content of Gm15G117700 gene in different soybeans,and finally near-infrared spectroscopy analyzer was used to identify the oleic acid quality of soybeans.T2 RT-PCR identification showed that overexpression was reduced by 3.94%,and gene editing was increased by 3.49%.It is determined that the Gm15G117700 gene may belong to a regulatory gene,a minor gene that can promote the conversion to linoleic acid content in soybean oleic acid synthesis.The gene cloning and its functional verification was not reported yet.This is the first report by PCR amplification of soybean Gm15G117700 genes and gene expression vector.Improving the content of oleic acid in soybean lay a foundation for researchers.Therefore;this study clearly identified the function of soybean Gm15G117700 gene and its role played in oleic acid synthesis and metabolism.

干旱响应的大豆G-box结合因子GmDIBL1的克隆和功能鉴定

干旱是典型的非生物逆境之一,严重影响作物的产量品质.鉴定作物抗/耐旱关键基因功能,解析其调控机制,对于培育抗/耐旱作物新种质、保障粮食安全具有重要意义.大豆是对干旱逆境较敏感的作物,挖掘寻找大豆抗/耐旱基因、鉴定其功能、明确其调控机制是大豆抗逆研究领域的热点.通过分析干旱处理的大豆根转录组数据,获得一个受干旱诱导的G-box结合蛋白基因GmDIBL1(Drought Induced B12D Like 1),克隆的cDNA序列包含1个长261 bp的开放阅读框,编码86个氨基酸残基,推导的分子量为10.1 kD,等电点为9.647.GmDIBL1蛋白质序列与其他物种的G-box结合蛋白之间具有较高的保守性,包含B12D结构域(6~73个氨基酸残基)和跨膜螺旋,N-端具有信号肽,C-端构成非胞质区域(34~86氨基酸残基),该蛋白定位于细胞核.GmDIBL1基因在根瘤中的表达水平最高,根中次之.干旱处理12 h后,其表达水平显著升高.在耐旱性较强的‘晋豆21’根中的表达水平显著高于干旱敏感的‘绥农26’中的表达水平.进一步通过在大豆毛状根中超量表达和敲除GmDIBL1基因,鉴定了转基因材料对干旱的响应,证明超量表达GmDIBL1基因显著提高毛状根复合体的抗旱性.这些结果说明GmDIBL1基因是大豆中一个响应干旱胁迫的基因,在大豆抗/耐干旱逆境中具有重要作用.

Isolation and Characterization of Storage Protein Subunit-null-dwarf Mutants in Soybean(Glycine max(L.) Merr.)

Dwarfing is useful to reduce plant height,when breeding high-yielding and non-lodging crops.In this study,a set of natural storage protein subunit-null dwarf mutants of soybean was reported that showed strongly reduced plant stature and deficiency in various 7S and 11S subunits,designated as snd1 mutants.Under normal growth conditions,the snd1 mutants showed a severe dwarf phenotype,with plant height of about 25 cm.Compared with wild-type DN47,the mutant snd1 exhibited no obvious morphological differences at the early stage of development.All the snd1 mutants examined had fewer nodes and shorter than normal internodes;the leaves were similar in shape to normal parents,but were dark-green at the mature stage.The flower size was similar to DN47;however,the flowering period was shorter than in the wild-type.Significant variation was noted for protein content,oil content of the seeds and size of seeds(weight of 100 seeds)among 17 snd1 dwarf lines.Genetic analysis indicated that the dwarfism of snd1 was controlled by a single recessive gene.The snd1 dwarf mutant had markedly different dynamic levels of the endogenous hormones gibberellin(GA),brassinosteroid,indole-3-acetic acid and abscisic acid,at the seedling stage.Exogenous GA3 treatment led to recovery of the plant height phenotype of the snd1 mutant;GA3 at 0.1 mm had the largest effect on enhancing plant height.Using molecular markers,snd1 gene was approximately mapped in an interval of 603 kb between markers Satt166 and Satt561 on chromosome 19.Snd1 mutant provided valuable material for hypoallergenic soybean breeding and the snd1 gene might be a novel gene related to plant height in soybean.

Proteomic Studies of Petal-specific Proteins in Soybean [Glycine Max(L.)Merr.] Florets

A survey of petal-specific proteomes of soybean(Glycine max(L.) Merr[Non-italic].) was conducted comparing protein expression profiles in different petals. Two-dimensional polyacrylamide gel electrophoresis reference maps of protein extracts from standard petals(SP), lateral wings(LW), keel petals(KP), and reproductive organs(RO)(a mixture of stamen and carpel) were obtained. Protein expression in the three petal types was compared using Image Master TM 2 D platinum 6.0 software. This indicated that the proportion of homologous proteins between SP and LW was 59.27%, between SP and KP was 61.48%, and between LW and KP was 60.05%. Within a mass range of 6.5-200.0 ku and pH 4.0-7.0, approximately 590, 646, 544, and 700 protein spots were detected in SP, LW, KP, and RO, respectively. A total of 82 differentially expressed proteins were detected. Sixty-four of these detected spots were differentially expressed and showed more than 2-fold changes in abundance; of these 64 proteins, 26 showed increased expression and 38 showed decreased expression. Among these spots, single organ-specific proteins were also identified.They were ID 49(60.9 ku), ID 45(50.0 ku), and ID 46(40.5 ku) in RO, ID 98(42.0 ku) in SP, and ID 05(29.0 ku) in KP. A total of 14 protein spots from 82 differentially expressed proteins were identified with LC-MS/MS. Further protein identification was conducted using the SwissProt and NCBInr databases. The identified proteins and their putative functions were discussed further. This was the first study reporting the comparison of petal protein profiles of soybean florets using proteomics tools.

Identification of Co-dominant SSR Markers Associated with Genes Controlling α'- and cr-subunit-null β-conglycinin Phenotypes in Soybean (Glycine max (L.) Merr.)

Studies have shown that the three subunits of β-conglycinin are the main potential allergens of soybean sensitive patients.And β-conglycinin has adverse effects on nutrition and food processing.So solation and production of lines with lowerβ-conglycinin content has been the focus of recent soybean breeding projects.Soybean lines with deficiency in one or all subunits of β-congIycinin have been obtained.An effective and rapid system to identify such mutations will facilitate genetic manipulation of the β-conglycinin subunit composition.Here,two segregating F2 populations were developed from crosses between Cgy-1/cgy-1(CC),anα'-lacking line(△α'),and DongNong 47(DN47),a wild-type(Wt)Chinese soybean cultivar with normal globulin components,and Cgy-2/cgy-2(CB),an a-lacking line(△α),and DN47.These populations were used to estimate linkage among the egy-1(conferring α'-null)and cgy-2(α-null)loci and simple sequence repeat(SSR)markers.Seven SSR markers(Sat_038,Satt243,Sat_307,Sat_109,Sat_231,Sat_108 and Sat_190)were determined to co-scgregate with cgy-1,and six SSR markers(Satt650,Satt671,Sat_418,Sat_170,Satt292 and Sat_324)co-segregated with cgy-2.Linkage maps being composed of seven SSR markers and egy-1 locus,and six SSR markers and the cgy-2 locus were then constructed.It assigned that the egy-1 gene to chromosome 10 at a position between Sat_307 and Sat_231,and the cgy-2 gene to chromosome 20 at a position between Satt650 and Satt671.These markers should enable map-based cloning of the egy-1 and cgy-2 genes.For different subunit-deficiency types[α'-null,α-null and(α'+α)-null types],the two sets of SSR markers could also detect of polymorphism between three normal cultivars and seven related mutant lines.The identification of these markers is great significance to the molecular marker-assisted breeding of soybean/9-conglycinin subunits.

Effect of Different Potting Mixtures on Growth and Yield of Soybean （Glycine max L.） and Soil Microbial Activity

Soybean （Glycine max： Fabaceae） is a mycotrophic （mycorrhizal） crop grown commercially for human consumption. Seven different fertilizer mixtures, namely cow dung, coir dust, mush room medium waste, saw dust, compost, decaying leaves and field soil with standard dose of NPK （control） were used for this experiment. The variety used was PM 25. Soil microbial activity was measured using CO2 evolution method. The experiment was carried out as a complete randomized block design with five replicates at the rate of eight plants per replicate. Average number of leaves on 25-day and 45-day old plants, shoot-length, root-length, number of pods per plant, wet weight of pod per plant, dry weight of pod per plant, plant wet weight, plant dry weight and seed dry weight per pod were measured. All management practices were conducted according to recommendations of the Department of Agriculture from seed germination to harvesting. Data were analyzed using SAS program （9.1.3）. Highest number of pods/plant （100, 124, 102, 106） and dry-seed-weight in g/plant （12, 14.8, 12, 12） were recorded in those grown in cow dung, compost, decaying leaves and inorganic mixture （control） whereas the lowest pod production （8.7 pods/plant） and seed dry weight （1.0 g/plant） was recorded in saw dust. Instead of inorganic fertilizer there is possibility to use organic potting mixtures like compost and decaying leaves which gave a significant difference in crop productivity as compared to other treatments. Significantly highest mean microbial activity was observed in potting media filled with coir dust.

Assessment of Genetic Variation in Soybean (<i>Glycine max</i>) Accessions from International Gene Pools Using RAPD Markers: Comparison with the ISSR System

Soybean (Glycine max) is one of the most important crops in the world in terms of total production and usage. It is also among the least diverse species. The main objectives of the present study were to 1) assess the level of genetic variation among soybean (G. max) accessions from different countries using Random Amplified Polymorphic DNA (RAPD) markers and 2) compare Inter Simple Sequence Repeats (ISSR) and RAPD marker systems in detecting polymorphic loci in soybeans (G. max). Genomic DNAs from 108 soybeans (G. max) accessions from 11 different gene pools were analyzed using several ISSR and RAPD primers. The average level of polymorphic loci detected with the RAPD primers was 35%. The soybean accessions from the China, Netherlands, and Canada gene pools were the least genetically variable with 25%, 26%, and 30% of polymorphic loci, respectively. Accessions from Hungary (43%) and France (48%) showed the highest level of polymorphism based on the RAPD analysis. Overall, RAPD data revealed that the accessions from different countries are closely related with 64% genetic distance values below 0.40. The levels of polymorphic loci detected with the RAPD and ISSR marker systems were in general moderate and similar even if they target different regions of the genome. A combination of different marker systems that include RAPD/ISSR, microsatellites (SSR), and SNPs should provide the most accurate information on genetic variation of soybean (G. max) accessions.

中国栽培大豆(Glycine max (L.) Merr.)微核心种质的群体结构与遗传多样性

【目的】评价中国栽培大豆微核心种质的群体结构和遗传多样性水平,为拓宽大豆遗传基础、发掘优异基因、改良大豆品种提供理论依据。【方法】利用大豆20个连锁群上的100个SSR位点,对来自全国28个省补充完善的248份栽培大豆微核心种质进行SSR遗传多样性及群体结构分析;采用PowerMarker Version 3.25软件统计等位变异数、平均等位变异数、多态性信息量(PIC值)及亚群特有等位变异数等参数;基于遗传距离建立了栽培大豆微核心种质的无根Neighbor-Joining树;用Structure2.2软件对微核心种质的群体结构进行评价。【结果】100个SSR位点在248份材料中共检测出等位变异1460个,每个位点变异范围为2—33个,平均为14.6个,每个位点PIC值变异范围为0.158—0.932,平均为0.743。基于模型的群体结构分析显示,依据LnP(D)无法判断最佳K值(群组数),但通过计算系数ΔK发现,K=3为微核心种质的最佳群体结构。结合种质的生态类型及品种类型分析发现,地理来源相同的种质具有聚在一起的倾向,但来源相同的种质也有分在不同组的情况。不同生态类型及品种类型间均存在较多的互补等位变异和特有等位变异。【结论】中国栽培大豆微核心种质具有丰富的遗传多样性,可以用来拓宽大豆品种遗传基础;不同生态类型及品种类型间存在较多的互补及特有等位变异,是种质创新及品种改良的物质基础;栽培大豆微核心种质存在明显的群体结构,为微核心种质在育种中的直接或间接利用提供了理论依据。

大豆(G.max)种子蛋白SBTi位点新类型的发现及甘肃省大豆SBTi位点各等位基因频率的研究

1991年,用PAGE技术检测了甘肃省243份大豆(G.max)品种种子蛋白中SBTi位点的等位基因型。检测结果:Ti^a型品种22份,占品种总数的90.9％,Ti^b21份,占8.6％,未发现Ti^c及ti型。发现一份材料的电泳图谱在与标样上Ti^a、Ti^b、Ti^c对应位置上没有谱带,而在Ti^b位置紧上方,有一条新谱带(Rf=0.79,Ti^b=0.82)。经加胰蛋白酶等措施多次验证,初步认为这很可能是在Ti位点上的一个新的等位基因控制下形成的蛋白谱带。