Identification of QTLs for growth period traits in soybean using association analysis and linkage mapping

The growth period traits of soybean (Glycine max L. Merr.) are quantitatively inherited and crucial for its adaptation to different environments. Association analysis and linkage mapping were used to identify the quantitative trait loci (QTLs) for days to flowering (DF), days from flowering to maturity (DFM) and days to maturity (DM). Considering the effect of sowing date, the phenotypes were evaluated in three or four sowing‐date‐experiments in each year. A total of 96 associations, involving 19 SSRs corresponding to DF, DFM and/or DM, were identified by association mapping. Six, eight and two QTLs were observed relating to DF, DFM and DM by linkage mapping, respectively, and some QTLs were shared by DF, DFM and DM. Four SSRs (Satt150, Satt489, Satt172 and Sat_312) were found to be related to the growth period traits using the two mapping methods. In summary, association analysis and linkage mapping can complement and verify results from both methods to identify QTLs in soybean, and these findings may be useful in facilitating the selection of growth period–related traits via marker‐assisted selection.     

Association analysis using SSR markers to find QTL for seed protein content in soybean

Association analysis studies can be used to test for associations between molecular markers and quantitative trait loci (QTL). In this study, a genome-wide scan was performed using 150 simple sequence repeat (SSR) markers to identify QTL associated with seed protein content in soybean. The initial mapping population consisted of two subpopulations of 48 germplasm accessions each, with high or low protein levels based on data from the USDA’s Germplasm Resources Information Network website. Intrachromosomal LD extended up to 50 cM with r ² > 0.1 and 10 cM with r ² > 0.2 across the accessions. An association map consisting of 150 markers was constructed on the basis of differences in allele frequency distributions between the two subpopulations. Eleven putative QTL were identified on the basis of highly significant markers. Nine of these are in regions where protein QTL have been mapped, but the genomic regions containing Satt431 on LG J and Satt551 on LG M have not been reported in previous linkage mapping studies. Furthermore, these new putative protein QTL do not map near any QTL known to affect maturity. Since biased population structure was known to exist in the original association analysis population, association analyses were also conducted on two similar but independent confirmation populations. Satt431 and Satt551 were also significant in those analyses. These results suggest that our association analysis approach could be a useful alternative to linkage mapping for the identification of unreported regions of the soybean genome containing putative QTL.     

Identification of population-specific QTLs for flowering in soybean

Flowering is an important stage in plant development and crucial for adaptation of plant species to different environments. Two soybean mapping populations were used to identify quantitative trait loci (QTLs) for days to flowering (DF) and days to maturity (DM) by genotyping simple sequence repeat (SSR) markers. Single-factor analysis of variance detected association of phenotypic data with SSR markers in each population. DF QTLs were identified on four chromosomes (chrs.); two QTLs located on chrs. 2 and 13 with Satt041 and Satt206 in the Jinpumkong 2 × SS2-2 population and other two DF QTLs were detected on chrs. 6 and 19 with Satt100 and Satt373 in the Iksannamulkong × SS2-2 population. The major QTLs associated with Satt100 explained 30.3% of maximum phenotypic variation. Especially, all DF QTLs included QTLs for DM, except Satt206 on chr. 13. Moreover, two additional DM QTLs were mapped on chrs. 10 and 11 with Satt243 and Satt359, respectively. DF QTL on chr. 2 with Satt041 was the newly identified QTL only in the Jinpumkong 2 × SS2-2 population and explained 10.3% of the phenotypic variation. The single locus of Satt100 on chr. 6 and Satt373 on chr. 19 were located on soybean genomic regions of the known flowering gene loci E1 and E3, respectively. These population-specific QTLs (Satt100 and Satt373) are the major QTLs for flowering time, putatively, they may be related to maturity QTLs with large effect. Additionally, these QTLs are valuable for marker-assisted approaches and could be widely adopted by soybean breeders.     

Quantitative trait analysis of flowering time in spring rapeseed (B. napus L.)

The inheritance of flowering time trait in spring-type rapeseed (Brassica napus L.) is poorly understood, and the investigations on mapping of quantitative trait loci (QTL) for the trait are only few. We identified QTL underlying variation for flowering time in a doubled haploid (DH) mapping population of nonvernalization-responsive canola (B. napus L.) cultivar 465 and line 86 containing introgressions from Houyou11, a Chinese early-flowering cultivar in Brassica rapa L. Significant genetic variation in flowering time and response to photoperiod were observed among the DH lines from 465/86. A molecular linkage map was generated comprising three types of markers loci. QTL analysis indicated that flowering time is a complex trait and is controlled by at least 4 major loci, localized on four different linkage groups A6, A7, C8 and C9. These loci each accounted for between 9.2 and 12.56 % of the total genotypic variation for first flowering. The published high-density maps for flowering time mapping used different marker systems, and the parents of our crosses have different genetic origins, with either spring-type B. napus or B. rapa. So we cannot determine whether the QTL on the same linkage groups were in the same region or not. There was evidence of additive × additive epistatic effects for flowering time in the DH population. Epistasis existed not only between main-effect QTLs, but also between QTLs with minor effects. Four pair of epistasis effects between minor QTLs explained about 20 % of the genetic variance observed in the DH population. The results indicated that minor QTLs for flowering time should not be ignored. Significant genotypes × environment interactions were also found for the quantitative traits, and with significant change in the ranking of the DH lines in different environments. The results implied that FQ3 was a non-environment-specific QTL and may control flowering time by autonomous pathway. FQ4 were winter-environment-specific QTL and may control flowering time by photoperiod-pathway. Identification of the chromosomal location and effect of the genes influencing flowering time may hasten the development of canola varieties having an optimal time for flowering in target environments such as for high altitude areas, via marker-assisted selection.     

Genetic mapping of quantitative trait loci conditioning salt tolerance in wild soybean (Glycine soja) PI 483463

Salt tolerance in soybean [Glycine max (L.) Merr.] is controlled by major quantitative trait loci (QTL) or single gene(s). Among soybean germplasm, wild soybean plant introduction PI 483463 was reported to have a single dominant gene for salt tolerance. The objective of this study was to genetically map the QTL in a recombinant inbred line (RIL) population derived from a cross between PI 483463 and Hutcheson. Simple sequence repeat (SSR) markers and universal soybean single nucleotide polymorphism (SNP) panel (the USLP 1.0) were utilized for molecular genotyping. The RILs were phenotyped in two independent tests in a greenhouse using a 1–5 scale visual rating method. The results showed that the salt tolerant QTL in PI 483463 was mapped to chromosome 3 in a genomic region between the Satt255 and BARC-038333-10036 markers. The favorable allele inherited from PI 483463 conferred tolerance to salinity and had an additive effect on reducing leaf scorch. A subset of 66 iso-lines was developed from the F₃ families of the same cross and was used for genetic confirmation of the QTL. The integration of recombination events and the salt reaction data indicate that the QTL is located in the region of approximately a 658 kb segment between SSR03_1335 at nucleotide 40,505,992 and SSR03_1359 at nucleotide 41,164,735 on chromosome 3. This narrow region can facilitate further genomic research for salt tolerance in soybean including cloning salt tolerance genes.     

Identification and fine mapping of molecular markers closely linked to fruit spines size <Emphasis Type="Italic">ss</Emphasis> gene in cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis> L.)

Fruit spine size is one of the importantly external quality traits effected the economic value of cucumber fruit. Morphological–cytological observation of the fruit spine size phenotype indicated that large spine formation arises from an increasing of spiny pedestal cell number caused by cell division, and best periods to accurately score fruit spine size trait was 4th day before flowering to 7th day after flowering according the continuous observation. Genetic analysis showed that a single dominant gene determined the fruit spine size trait in cucumber. BC₁ population (189 individuals) of two inbred lines (large spine PI197088 and small spine SA0422) was used for primary mapping of the SS/ss locus with 7 markers covering an interval of 37.1 cM. An F₂ segregating population of 1032 individuals constructed from the same two parents (PI197088 and SA0422) was used to fine mapping of the SS/ss locus. Six new markers linked to the gene were successfully screened for construction of a fine linkage map, in which the SS/ss locus was located in the region flanked by marker SE1 (3 recombinants) and SSR43 (2 recombinants) with a 189 kb physical distance. Markers from this study will be valuable for candidate gene cloning and marker-assisted selection for cucumber breeding.     

QTLs conferring FOV 7 resistance detected by linkage and association mapping in Upland cotton

Fusarium wilt is a worldwide disease that affects cotton production. Molecular markers tightly linked to resistance genes can be used for marker-assisted and/or genomic selection. We performed both family-based linkage mapping and population-based association mapping (AM) to detect quantitative trait loci (QTLs) conferring resistance against Fusarium oxysporum f. sp. vasinfectum race 7 (FOV 7) in Upland cotton. To identify QTLs underlying FOV 7 resistance by linkage mapping, three Upland cotton cultivars/lines, Xuzhou 142, Yumian 21 and Shang 9901, were used to obtain the composite cross population, designated as Xuzhou 142/Yumian 21//Xuzhou 142/Shang 9901. A linkage map containing 185 simple sequence repeat loci and 40 linkage groups was constructed with an average distance of 7.5 cM between adjacent markers. Seven QTLs were detected by linkage mapping, explaining 2.9–6.6 % of the total phenotypic variance. We also performed marker–trait AM with the MLM model (Q + K) in a panel composed of 356 Upland cotton cultivars. In total, 27 loci were significantly associated with FOV 7 resistance at the α = 0.01 level (?log₁₀ P ≥ 2), which were distributed on 16 chromosomes and explained 1.48–12.99 % of phenotypic variation. Three of the 7 QTLs identified by linkage mapping could be detected in AM. We identified the favorable allele for each of the 27 associated loci and investigated the number of favorable alleles in each accession. The results should increase our understanding of the genetic basis of FOV resistance and facilitate future resistance breeding in Upland cotton.     

A consensus genetic map of chickpea (Cicer arietinum L.) based on 10 mapping populations

A consensus genetic map of chickpea (Cicer arietinum L.) was constructed by merging linkage maps from 10 different populations, using STMS (Sequence-tagged Microsatellite Sites) as bridging markers. These populations derived from five wide crosses (C. arietinum × Cicer reticulatum) and five narrow crosses (Desi × Kabuli types) were previously used for mapping genes for several agronomic traits such as ascochyta blight, fusarium wilt, rust resistance, seed weight, flowering time and days to flower. The integrated map obtained from wide crosses consists of 555 loci including, among other markers, 135 STMSs and 33 cross-genome markers distributed on eight linkage groups and covers 652.67 cM. The map obtained from narrow crosses comprises 99 STMSs, 3 SCARs, 1 ASAP, fusarium resistance gene, 5 morphological traits as well as RAPD and ISSR markers distributed on eight linkage groups covering 426.99 cM. Comparison between maps from wide and narrow crosses reflects a general coincidence, although some discrepancies are discussed. Medicago truncatula cross-genome markers were BLASTed against the M. truncatula pseudogenome permitting assignments of chickpea linkage groups LGI, II, III, IV, V and VI on Medicago chromosomes 2, 5, 7, 1, 3 and 4, respectively. A marker detectable on Medicago chromosome 4 were also located on LGVIII, This consensus map is an important progress to assist breeders for selecting suitable markers to be used in marker-assisted selection (MAS).     

运用关联分析定位栽培大豆蛋白11S、7S组分的相关基因位点

大豆贮藏蛋白主要成分是7S和11S球蛋白,大豆贮藏蛋白组分及其亚基组成决定了蛋白质的品质和加工特性。本研究选用134对细胞核SSR标记,对166份栽培大豆微核心种质进行基因分型,运用一般线性回归(general linear model, GLM)和复合线性回归(mixed linear model, MLM)方法进行标记与性状的关联分析,定位大豆蛋白亚基的相关基因。结果表明,2年均检测到的且与蛋白亚基相关联的SSR位点有14个,以MLM方法检测到5个SSR位点(Sat_062、Satt583、Satt291、Satt234和Satt595)与蛋白亚基相关联;7S组分各亚基变异程度较大,是引起11S/7S变异的主要原因;表型变异较大的亚基可能因为相关基因进化中发生重组较多,LD衰减距离较小,导致检测到较少的相关位点。本研究结果对蛋白亚基相关性状的标记辅助选择育种有重要的利用价值。     

Identification of QTL underlying soluble pigment content in soybean stems related to resistance to soybean white mold (<Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis>)

The frequency of soybean white mold (Sclerotinia sclerotiorum, SWM) outbreaks has increased since 1978, and this disease is currently considered to be the second most important cause of soybean yield loss worldwide. We have studied SWM in stems of soybean cultivar Maple Arrow, which shows partial resistance to SWM, in an attempt to identify the quantitative trait loci (QTLs) underlying soluble pigment(s) content, which is associated with SWM resistance. A SWM-susceptible cultivar, Hefeng 25, was crossed with Maple Arrow, and 149 F_5:6 recombinant inbred lines were subsequently advanced through single-seed-descent. A total of 109 simple sequence repeat (SSR) markers were used to construct a genetic linkage map. Three QTLs for soluble pigment content in soybean stems associated with the resistance to SWM, namely, Qsp-1 (Satt502-Sat_159), Qsp-2 (Sat_156-Satt251), and Qsp-3 (Satt525-Satt233), were identified in 2007 and 2008 and located onto linkage groups D1a+q, B1 and A2, respectively. The phenotypic variation (R ²) explained by these QTLs ranged from 6.29 to 15.37%. These three QTLs were not significantly related to known QTLs associated with escape resistant mechanisms. The use of these QTLs in marker-assisted selection may contribute to improved soybean resistance to SWM.     

A linkage map for flowering dogwood (<Emphasis Type="Italic">Cornus florida</Emphasis> L.) based on microsatellite markers

A genetic linkage map of flowering dogwood (Cornus florida L.) was constructed using 94 individuals derived from a cross of two F₁ trees designated 97-6 and 97-7, which were originally from a cross between ‘Appalachian Spring’ and ‘Cherokee Brave’. Out of approximately 800 SSR loci examined, 271 were polymorphic between ‘Appalachian Spring’ and ‘Cherokee Brave’, but were monomorphic between 97-6 and 97-7. These 271 segregating markers were used to build a linkage map for flowering dogwood. Eleven linkage groups were obtained with a log-of-odds (LOD) value of 6.0 using JoinMap® 4.0 software, which matches the chromosome number of flowering dogwood haploid genome. This linkage map consisted of 255 SSR loci, spanned a total of 1,175 centimorgans (cM) with an average internal distance of 4.6 cM. Several larger gaps and slight clustering of markers were present on this linkage map. This is the first linkage map of flowering dogwood and will be a fundamental tool for new gene identification and marker-assisted selection in our flowering dogwood breeding program.     

Molecular mapping of a fertility restorer gene for cytoplasmic male sterility in soybean

In this study, we report the mapping of the Rf locus in soybean by microsatellite simple sequence repeat (SSR) genetic markers. A cross was made between cytoplasmic male sterility (CMS) line JLCMS82A and restorer line JIHUI 1 based on the DNA polymorphisms revealed by 109 SSR markers. A F₂ population derived from a single F₁ plant containing 103 individuals was used for mapping the Rf locus. The Rf gene of JIHUI 1 gametophytically restores male fertility to JLCMS82A. Fertile and semi-fertile DNA bulks and parental DNAs were screened with 219 SSR markers, and Satt215 which was previously mapped to soybean LG J, was found linked to the Rf gene. Five additional polymorphic SSR markers from LG J were used for analysis and a regional linkage map around the Rf locus was established. SSR markers, Sctt011 and Satt547, flanked the Rf locus at 3.6 cM and 5.4 cM, respectively. The availability of these SSR markers will facilitate the selection of restorer lines in hybrid soybean breeding.     

Construction of an integrated genetic map based on maternal and paternal lineages of tea (Camellia sinensis)

Genetic study on important traits of tea is difficult because of its self-incompatibility in nature. Moreover, development of a new variety usually needs more than 20 years, since it takes many years from seedling to matured plants for trait investigation. Genetic map is an essential tool for genetic study and breeding. In this study, we have developed an integrated genetic map of tea (Camellia sinensis) using a segregating F1 population derived from a cross between two commercial cultivars (‘TTES 19’ and ‘TTES 8’). A total of 574 polymorphic markers (including SSR, CAPS, STS, AFLP, ISSR and RAPD), 69 markers with highly significant levels of segregation distortion (P < 0.001) (12.0 %) were excluded from further analyses. Of the 505 mapped markers, there were 265 paternal markers (52.5 %), 163 maternal markers (32.3 %), 65 doubly heterozygous dominant markers (12.9 %), and 12 co-dominant markers (2.4 %). The co-dominant markers and doubly heterozygous dominant markers were used as bridge loci for the integration of the paternal and maternal maps. The integrated map comprised 367 linked markers, including 36 SSR, 3 CAPS, 1 STS, 250 AFLP, 13 ISSR and 64 RAPD that were assigned to 18 linkage groups. The linkage groups represented a total map length of 4482.9 cM with a map density of 12.2 cM. This genetic map has the highest genetic coverage so far, which could be applied to comparative mapping, QTL mapping and marker assisted selection in the future.     

Genetic studies on saline and sodic tolerances in soybean

Salt-affected soils are generally classified into two main categories: saline and sodic (alkaline). Developing and using soybean (Glycine max (L.) Merr) cultivars with high salt tolerance is an effective way of maintaining sustainable production in areas where soybean growth is threatened by salt stress. Early classical genetics studies revealed that saline tolerance was conditioned by a single dominant gene. Recently, a series of studies consistently revealed a major quantitative trait locus (QTL) for saline tolerance located on linkage group N (chromosome 3) around the SSR markers Satt255 and Sat_091; other minor QTLs were also reported. In the case of sodic tolerance, most studies focused on iron deficiency caused by a high soil pH, and several QTLs associated with iron deficiency were identified. A wild soybean (Glycine soja Sieb. & Zucc.) accession with high sodic tolerance was recently identified, and a significant QTL for sodic tolerance was detected on linkage group D2 (chromosome 17). These studies demonstrated that saline and sodic tolerances were controlled by different genes in soybean. DNA markers closely associated with these QTLs can be used for marker-assisted selection to pyramid tolerance genes in soybean for both saline and sodic stresses.     

A quantitative trait locus influencing tolerance to Phytophthora root rot in the soybean cultivar ‘Conrad’

‘Conrad’, a soybean cultivar tolerant to Phytophthora root rot (PRR), and ‘OX760-6-1’, a breeding line with low tolerance to PRR, were crossed. F₂ derived recombinant inbred lines were advanced to F₆ to generate a population through single-seed descent. This population was used to identify quantitative trait loci (QTLs) influencing PRR tolerance in ‘Conrad’. A total of 99 simple sequence repeat (SSR), or microsatellite, markers that were polymorphic and clearly segregated in the F₆ mapping population were used for QTL detection. Based on the data of PRR in the field at two planting locations, Woodslee and Weaver, for the years 2000 and 2001, one putative QTL, designated as Qsatt414-596, was detected using MapMaker/QTL. Qsatt414-596 was flanked by two SSR markers from the linkage group MLG J, Satt414 and Satt596. Satt414 and Satt596 were also detected to be significantly (P < 0.005) associated with PRR using the SAS GLM procedure and were estimated to explain 13.7% and 21.5% of the total phenotypic variance, respectively.     

大豆抗胞囊线虫4号生理小种新品系SSR标记分析

培育抗病品种是大豆胞囊线虫(Soybean Cyst Nematode, SCN)病经济、有效的防治方法。利用130个SSR标记对26份抗SCN 4号生理小种(SCN 4)新品系和15份感病品系进行基因型分析, 旨在明确抗病品系与SCN 4抗性相关联的SSR标记, 提出抗性基因分子标记鉴定方法, 以提高抗病品系在育种中的利用效率。研究表明, Hartwig与晋品系亲本具有不同的SCN 4抗病基因, 其遗传相似系数为0.362。与抗性显著关联的22个SSR位点分布在11个连锁群(LG), 推测LG D1b上分布的SSR标记附近存在1个新的SCN 4抗病基因; 而Satt684、Sat_230、Sat_222、Satt615和Satt231位点, 来自亲本Hartwig等位基因与抗病相关联, 而来自晋品系的等位基因与感病相关联, 在Sat_400、Satt329和Satt557等其他17个SSR位点, 来自Hartwig等位基因与感病相关联, 来自晋品系亲本的等位基因与抗病相关联。利用非连锁不平衡SSR标记Satt684和Sat_400可对供试品系进行有效的抗性辅助选择。     

QTL mapping of agronomically important traits in sorghum (<Emphasis Type="Italic">Sorghum bicolor</Emphasis> L.)

Sorghum is one of the most important cereal crops; it is used to produce feed, sugar, and biofuel. To investigate genetic tradeoffs between grain and stem sugar production, we evaluated plant height, Brix (the percentage of soluble solids in stalk juice), 100-grain weight and flowering time over 3 years in a recombinant inbred line (RIL) population consisting of 189 individuals derived from a cross between the sweet sorghum cultivar ‘Rio’ and grain sorghum ‘BTx623’. We constructed a genetic linkage map (total length, 1418.71 cM; average distance between markers, 11.26 cM), which consisted of 118 simple sequence repeat (SSR) and 8 insertion-deletion (INDEL) markers. A total of 14 QTLs were detected on chromosomes 1, 3, 6, 7, and 9, which included 6 QTLs for plant height; 4 for Brix; and 2 QTLs for each 100-grain weight and flowering time. Eight QTLs were detected at least in 2 years. These results will be useful for future QTL fine mapping and gene mining for these traits, and useful for the improvement of sorghum through molecular marker-assisted selection.     

Molecular mapping of major genes and quantitative trait loci determining flowering time in response to photoperiod in barley

Two major genes (eam8 and eam10) and two quantitative trait loci (QTL) determining flowering time in barley were associated with restriction fragment length polymorphism markers. The loci eam8 and eam10 were found to map in regions of chromosomes 1HL and 3HL, respectively, already estimated from previous classical linkage analyses. While investigating doubled haploid lines of a spring habit barley mapping population, two QTL for flowering time were detected on chromosomes 1HL and 7HS, respectively, when the material was grown under long photoperiod conditions. When growing the same lines under short photoperiod, no QTL were discernible. Allelic and homoeologous relationships with flowering time loci described earlier in barley and other Triticeae species are discussed.     

An SSR-based molecular genetic map of cassava

Summary Microsatellites or simple sequence repeats (SSR) are the markers of choice for molecular genetic mapping and marker-assisted selection in many crop species. A microsatellite-based linkage map of cassava was drawn using SSR markers and a F₂ population consisting of 268 individuals. The F₂ population was derived from selfing the genotype K150, an early yielding genotype from an F₁ progeny from a cross between two non-inbred elite cassava varieties, TMS 30572 and CM 2177-2 from IITA and CIAT respectively. A set of 472 SSR markers, previously developed from cassava genomic and cDNA libraries, were screened for polymorphism in K150 and its parents TMS 30572 and CM 2177-2. One hundred and twenty two polymorphic SSR markers were identified and utilized for linkage analysis. The map has 100 markers spanning 1236.7 cM, distributed on 22 linkage groups with an average marker distance of 17.92 cM. Marker density across the genome was uniform. This is the first SSR based linkage map of cassava and represents an important step towards quantitative trait loci mapping and genetic analysis of complex traits in M. esculenta species in national research program and other institutes with minimal laboratory facilities. SSR markers reduce the time and cost of mapping quantitative trait loci (QTL) controlling traits of agronomic interest, and are of potential use for marker-assisted selection (MAS).     

大豆主茎节数数量性状座定位及遗传效应分析

为了定位控制主茎节数的QTL并明确其遗传效应,利用100对SSR引物,并采用Mapmaker Exp 3.0和复合区间法,研究构建了一张包括3个连锁群的连锁图谱。以‘黑农37’（栽培大豆）×ZYD581（野生大豆）组合的亲本、F₂、F₃为试材,分别在chr1连锁群上定位了一个影响大豆主茎节数的QTL,2007年QTL位于Satt238—Satt242这个区间内,与Satt238的遗传距离是0.01 cM,与Satt242的遗传距离是24.69 cM,其遗传贡献率为17.22%,加性效应为-3.2608;2008年QTL位于Satt238—Satt240之间,与Satt238的遗传距离为0.59 cM,与Satt240的遗传距离为6.01 cM,其遗传贡献率为6.68%,加性效应为-1.4965。2年大豆主茎节数QTL分析表明,在chr1连锁群上Satt238附近确定了1个控制大豆主茎节数QTL位点。