棉花分子标记图谱的构建和一些重要性状的定位

在棉花上,虽然已有若干张不同的分子标记连锁图发表,但在育种中它们仍较难被应用,主要是因为在现有的分子标记连锁图谱中能用于育种的重要农艺性状的DNA分子标记尚不多.为此,本研究选用具有13个质量性状标记的两个陆地棉(Gossypium hirsutum L.)多标记基因系T582和T586作为亲本,用SSR和AFLP的DNA分子标记技术,对F2的作图群体进行DNA分子标记连锁图谱的构建,对棉花重要质量性状基因和数量性状的QTLs进行定位和标记,并建立分子标记连锁群与染色体的对应关系,为棉花分子标记辅助选择育种提供依据.     

ET-ISJ标记的开发及陆地棉遗传图谱构建

根据植物结构基因外显子拼接位点的保守序列,设计扩增外显子的ET-ISJ (exon targeted intron-exon splice junction)标记引物。利用1 280对ET-ISJ引物组合,在陆地棉品种渝棉1号和T586中,筛选获得69对多态性引物组合,占引物组合的5.4%。用多态性ET-ISJ引物组合检测(渝棉1号×T586)F_2:7重组近交系群体,得到70个位点。以70个ET-ISJ标记位点与523个SSR、59个IT-ISJ、29个SRAP和8个形态标记进行连锁分析,构建的遗传连锁图谱包括59个连锁群和673个位点(68个ET-ISJ、510个SSR、58个IT-ISJ、29个SRAP和8个形态标记)。连锁图覆盖3 216.7 cM,占棉花基因组的72.3%,标记间平均长度为4.8 cM。68个ET-ISJ标记分布于20条染色体。研究表明ET-ISJ标记多态性较高、稳定性好,可有效用于棉花与其他植物遗传连锁图谱构建。     

陆地棉重要农艺性状基因的QTL定位研究

本研究利用陆地棉品种新陆中10号与陆地棉品种新陆早7号构建F2作图群体;利用MAPMAKER/EXP（Version3.0b）构建连锁群总长度962.2cm,覆盖率为17.49%,标记数为89个位点。利用复合区间作图法原理对作图群体的果枝始节、单铃重、衣分、籽指、株高、叶主脉、叶次脉等7个农艺性状进行了QTLs筛选,共检测9个稳定的QTLs：其中与株高有关的1个,解释23.1%的表型变异;与籽指有关的QTL共1个,解释18.8%的表型变异;与衣分有关的QTL共2个,分别解释6.5%、7.4%的表型变异;与铃重有关的QTL共2个,分别解释11.6%、14.2%的表型变异;与叶主脉及叶次脉共检测到3个QTLs,解释10.7%～21.9%的表型变异。     

棉花显性无腺体基因的应用及其遗传研究

1986～1992年通过有性杂交,将海_1的显性无腺体性状转育到陆地棉中,培育出既具有优异特性,又具有显性无腺体性状的陆地棉新材料和胞质型显性无腺体不育系、保持系和恢复系。通过等位测验和连锁测验等遗传研究,表明海_1无腺体性状是由位于gl_2点上的一对不完全显性基因Gl_2~e所控制,并带有一对有腺体基因Gl_3和Gl_1,Gl_2~e对Gl_3和Gl_1具有显性上位作用。Gl_2~e与标记系T582的4个和T586的3个标记性状元连锁关系,不在其所属的连锁群上。     

短季棉早熟性的分子标记及QTL定位

以两个陆地棉品种中棉所36×TM-1的207个F2单株为作图群体,筛选出73个多态性引物,25个SSR标记、35个RAPD标记和13个SRAP标记,构建了第一张以研究短季棉为主的包含43个标记,标记间的最小遗传距离为11.8 cM,最大遗传距离为48.9 cM,总长1174.0 cM的遗传连锁图谱,覆盖棉花基因组总长度的23.48%。检测到与短季棉早熟性状相关的12个QTLs,其中有8个QTLs呈簇分布在LG1连锁群上,找到对表型变异的贡献率在30%以上与全生育期、霜前花率和开花期有关的QTL各1个。     

利用陆海杂种BC1群体构建棉花遗传连锁图谱并初步定位产量性状相关的QTL

利用陆地棉推广品种中棉所36和海1配制杂交组合,并用中棉所36为轮回亲本构建回交群体(BC1F1,BC2F1和BC1S1).用亲本和F1对新开发的2102对SSR引物进行多态性筛选,共筛选到317对含有海1显性带的引物,占筛选引物总数的15.08%;最终对其中的275对引物进行了BC1F1群体扩增,获得306个SSR标记差异位点.连锁分析表明(LOD=6.5),有254个标记位点连锁,分布在42个连锁群中,覆盖2252.36 cM,约占棉花基因组的50.05%;平均每个连锁群有6.08个标记,覆盖53.63 cM;标记间平均间距为8.87 cM.利用BC1F1、BC2F1和BC1S1三个不同世代分离群体产量性状数据,共定位16个产量性状QTL,解释表型变异5.77%/～19.86%.其中,衣分6个,铃重6个,籽指4个.有9个增效基因来自陆地棉亲本中棉所36,7个增效基因来自海岛棉亲本海1,说明了表型性状较差的品种同样可能含有可用于性状改良的增效基因.控制衣分的3个QTL可在不同的世代稳定检测到,效应稳定,增效基因均来自高值亲本陆地棉,为进一步分子标记辅助选择奠定了基础.     

甘蓝型油菜产量及相关性状的QTL分析

高产是甘蓝型油菜育种的重要目标之一，产量是多基因控制的数量性状。本文通过QTL作图分析了产量及其相关性状的数量性状位点，以甘蓝型油菜中油821和保604 F₁代小孢子培养获得的DH系为作图群体，构建了由20个连锁群组成的，包括251个分子标记( 2个RFLP标记，72个RAPD标记，91个SSR标记，86个SRAP标记)的遗传连锁图(10个标记没有分配到连锁群中)。图谱的平均图距为6.96 cM，共覆盖油菜基因组1 746.5 cM。在此图谱基础上采取复合区间作图法，检测到与油菜产量及其相关性状有关的QTL共17个。其中控制株高的3个分别位于第4、第9和第10连锁群上，对性状的解释率为9.42%～17.58%；与分枝部位有关的4个分别位于第4、第6和第7连锁群上，其中Bp1 和Bp2 均位于第4连锁群，对性状的解释率为8.13%～15.20%；与主花序有效长有关的3个分别位于第4、第10和第16连锁群上，对性状的解释率为7.49%～23.36%；与一次有效分枝有关的2个分别位于第1、第4连锁群上，对性状的解释率为15.29%～19.58%；与角果总数和千粒重有关的分别位于第4连锁群和第9连锁群上，贡献率分别为17.42%和7.64%；与单株产量有关的3个分别位于第3、第4和第15连锁群，共解释26.60%的表型变异。部分性状的QTL在连锁群上成簇分布,对性状贡献率很大，表现主效QTLs的特点，相应的性状之间也呈显著相关，这表明一因多效或者相关的QTLs之间紧密连锁是性状相关的遗传基础。本研究中与主效QTLs连锁的标记可用于油菜产量性状的分子标记辅助选择。     

棉花表型性状基因的SSR标记定位

本文以两个陆地棉多标记基因系T582和T586,以及杂交获得的F_1、F_2及F_3代作为试验材料,利用11对SSR引物对F_2群体的120个单株的DNA样品进行多态性分析,并利用F_2和F_3群体对F_2群体对应单株的13个表型性状进行基因型的判定,结果得到了3个与表型性状基因连锁的SSR标记,分别是红茎基因(R_1)与J178连锁、遗传距离为24.9cM,簇生铃基因(CL_1)与J236连锁,遗传距离为46.0cM,茸毛基因(T_1)与J252连锁,遗传距离为28.5cM,其中R_1、CL_1、J178和J236在同一连锁群上。红茎和植株茸毛是具有抗虫性能的形态性状,用SSR标记这些性状将有助于提高育种家的育种效率。     

陆地棉衣分差异群体产量及产量构成因素

 以衣分差异较大的陆地棉品种为材料,构建了包含188个F₂单株的作图群体,应用6111对SSR引物对亲本进行了分子标记筛选,结果仅获得了123个多态性位点,其中88个位点构建了总长为666.7 cM、平均距离为7.57 cM的遗传图谱,覆盖棉花基因组的14.9%。通过复合区间作图法对F₂单株和F_2∶3家系进行QTL检测,共鉴定出了18个控制产量及产量构成因素变异的QTLs,包括2个衣分QTLs、4个子棉产量QTLs、4个皮棉产量QTLs、2个衣指QTLs、3个单株铃数QTLs、2个铃重QTLs和1个子指QTL。 解释的表型变异分别为\{6.9%\}～16.9%、5.6%～16.2%、4.8%～15.6%、7.7%～13.3%、8.2%～11.6%、6.1%～7%和6.6%。不同QTLs在相同染色体区段上的成簇分布表明与产量性状相关的基因可能紧密连锁或一因多效。产量及产量构成因素QTLs的遗传方式主要以显性和超显性效应为主。检测到的主效QTLs可以用于棉花产量及产量构成因素的分子标记辅助选择。     

利用置换系检测棉花第22染色体短臂的产量相关性状QTLs

CSB22sh为以陆地棉(Gossypium.hirsutum L.)遗传标准系TM-1为背景的第22染色体短臂被海岛棉(Gossypium.barbadense L.)Pima3-79置换的海陆置换系。TM-1与CSB22sh杂交,构建了104个F2单株的作图群体,应用6748对SSR引物对亲本进行分子标记筛选,获得90个多态性标记位点。其中85个标记位点构建了总长85.24 cM的遗传图谱,标记间平均距离1.0 cM,覆盖棉花基因组的1.8%。通过复合区间作图法对F2:3和F2:4家系的7个产量相关性状(衣分、铃重、子指、株高、第一果枝节位、单株铃数、单株果枝数)进行QTL检测,共检出28个不同QTLs,解释性状表型变异的3.5%～44.8%。仅在一个环境中检测到的QTLs有17个,2个环境同时检测到的QTLs有8个,3个环境同时检测到的QTLs有3个。不同的QTL在相同区段的成簇分布表明,控制不同性状的基因可能紧密连锁或一因多效。检测到的稳定的QTL可以用于相应性状的分子标记辅助选择。     

T1 locus in cotton is the candidate gene affecting lint percentage, fiber quality and spiny bollworm (Earias spp.) resistance

A genetic linkage map of chromosome 6 was constructed by using 270 recombinant inbred lines originated from an upland cotton cross (Yumian 1 × T586) F₂ population. The genetic map included one morphological (T₁) and 18 SSR loci, covering 96.2 cM with an average distance of 5.34 cM between two markers. Based on composite interval mapping (CIM), QTL(s) affecting lint percentage, fiber length, fiber length uniformity, fiber strength and spiny bollworm resistance (Earias spp.) were identified in the t₁ locus region on chromosome 6. The allele(s) originating from T586 of QTLs controlling lint percentage increased the trait phenotypic value while the alleles originating from Yumian 1 of QTLs affecting fiber length, fiber length uniformity, fiber strength and spiny bollworm resistance increased the trait phenotypic value.     

陆地棉重组近交系产量及其构成因素的QTL分析

利用爱字棉1517×德州047重组近交系(recombinant inbred lines, RIL)中G6群体构建的SSR遗传连锁图谱及基于混合线性模型的复合区间作图法对QTL进行定位,并对主效QTL,加性×加性上位性QTL及与环境互作效应进行分析,为利用分子聚合方法提高产量提供理论依据。对2006年、2008年以及2009年的产量性状进行分离分析,检测到24个不同年份的主效QTL,其中相关于单株籽棉、单株皮棉、衣分、子指以及单株铃数的分别检测到1个不同年份稳定存在的主效QTL;对3年的产量性状作环境因子联合分析,检测到14个主效QTL,其中6个与环境互作,检测到20对加加上位性QTL,其中7对与环境互作。不同年份检测的稳定且受环境影响小或不受环境影响的与近处标记紧密连锁的主效QTL可用于分子标记辅助选择,以提高育种的效率。     

Construction of a genetic linkage map and QTL analysis of fiber-related traits in upland cotton (Gossypium hirsutum L.)

A genetic linkage map with 70 loci (55 SSR, 12 AFLP and 3 morphological loci) was constructed using 117 F₂ plants obtained from a cross between two upland cotton cultivars Yumian 1 and T586, which have relatively high levels of DNA marker polymorphism and differ remarkably in fiber-related traits. The linkage map comprised of 20 linkage groups, covering 525 cM with an average distance of 7.5 cM between two markers, or approximately 11.8% of the recombination length of the cotton genome. The present genetic linkage map was used to identify and map the quantitative trait loci (QTLs) affecting lint percentage and fiber quality traits in 117 F_2:3 family lines. Sixteen QTLs for lint percentage and fiber quality traits were identified in six linkage groups by multiple interval mapping: four QTLs for lint percentage, two QTLs for fiber 2.5% span length, three QTLs for fiber length uniformity, three QTLs for fiber strength, two QTLs for fiber elongation and two QTLs for micronaire reading. The QTL controlling fiber-related traits were mainly additive, and meanwhile including dominant and overdominant. Several QTLs affecting different fiber-related traits were detected within the same chromosome region, suggesting that genes controlling fiber traits may be linked or the result of pleiotropy.     

Heritability and predicted selection response of yield components and fibre properties in an inter-specific derived RIL population of cotton

Exploiting genetic variation through inter-specific breeding has improved cotton yield, fibre properties and adaptability. The objectives of this study were to examine heritability and predicted selection response of yield components and fibre properties in a recombinant inbred line (RIL) population from an inter-specific cross between Gossypium hirsutum (Gh) variety Guazuncho 2, and G. barbadense (Gb) line VH8-4602. A population of 93 and 82 RILs was tested in two seasons, with two parents and local controls, Sicot 75 (Gh) and Sipima 280 (Gb) in field experiments. Seed cotton samples hand harvested before and after defoliation were used to measure lint percent, boll weight, 100 seed weight and the lint to measure fibre length, uniformity, short fibre index (SFI), elongation, strength, micronaire, maturity ratio (MR), percent of maturity (PM) and fineness. There was large phenotypic variation for individual traits and transgressive segregation occurred in lint percent, lint weight/seed, fibre no./seed, uniformity, SFI, elongation, MR and PM. Narrow sense heritabilities were moderate for yield components (34.3–41.2%) and for key fibre properties, length, strength, micronaire and fineness (38.3–42.1%), which led to a predicted selection response of 6.7–24.0% for yield components and 3.9–10.9% for key fibre properties under a selection intensity of 10%. Favourable associations were found between key fibre properties, but an adverse association between lint percent and each of these fibre properties. Only five RILs were identified with desirable combinations. The results demonstrated the value of exploiting inter-specific variation to develop cotton germplasm and how breeding strategies can be improved.     

New QTLs for lint percentage and boll weight mined in introgression lines from two feral landraces into Gossypium hirsutum acc TM‐1

Overuse of a small number of Upland cotton cultivars has narrowed cotton's genetic base, leading to major difficulties in developing cotton cultivars with diverse genetic backgrounds that are able to adapt to adverse conditions. To effectively broaden the genetic base, chromosome introgression lines (ILs) were developed, where TM‐1, the genetic standard of Upland cotton, was used as the common recipient and its two feral landraces, TX‐256 and TX‐1046, were used as the donors. A total of 115 ILs, with an average segment length of 11.15 cM, were first developed via intraspecific hybridization by marker‐assisted selection (MAS) in BC₃F₂ generations, spanning 3887.75 cM of the cotton genome. Association analysis showed that 63 markers were found to be associated with boll weight (BW), lint percentage (LP) and seed index. The percent of phenotypic variance explained by 148 QTLs detected was 4.12% on average. Eleven and five new QTLs for BW and LP (one stable QTL identified for LP in all environments) were detected, respectively, which can be used for efficiently pyramiding favourable alleles into one cultivar by MAS.     

Microsatellite markers associated with lint percentage trait in cotton, Gossypium hirsutum

Molecular markers associated with fiber development traits have the potential to play a key role in understanding of cotton fiber development. Seventeen SSRs out of 304 markers tested from MGHES (EST-SSR), JESPR (genomic SSR), and TMB (BAC-derived SSR) collections showed significant linkage associations (using a Kurskal-Wallis non-parametric test) with lint percentage QTL in a set of recombinant inbred cotton lines (RILs) segregating for lint percentage. The permutation test of these potential markers associated with lint percentage QTL(s) determined that 12 SSR markers have stable estimates, exceeding empirically chosen threshold significance values at or above α = 0.01. Interval mapping demonstrated that 9 SSRs with stable critical LOD threshold values at α = 0.01 have significant QTL effect. Multiple QTL-mapping (MQM) revealed that at least, two highly significant fiber development QTLs exist around regions TMB0471 and MGHES–31 (explained about 23–59% of the phenotypic variation of lint percentage) and around markers MGHES–31 and TMB0366 (accounted for 5.4–12.5% phenotypic variation of lint percentage). These markers, in particular fiber-specific EST-SSRs, might be the possible ‘candidate’ loci contributing for fiber development in cotton. BAC-derived SSRs associated with fiber trait are the possible markers that are useful for the identification of physical genomic contigs that contain fiber development genes. Several lint percentage trait associated SSR markers have been located to chromosomes 12, 18, 23, and 26 using deletion analysis in aneuploid chromosome substitution lines. Outcomes of the work may prove useful in understanding and revealing the molecular basis of the fiber development, and the utilization of these markers for development of superior cotton cultivars through marker-assisted selection (MAS) programs. I. Y. Abdurakhmonov and S. Saha contributed equally to the work     

Association analysis of fibre traits in Gossypium arboreum accessions

Advances in the use of diploid Asiatic species in cotton breeding require an understanding of the relatedness and ancestry of diploid cotton accessions, and identification of simple sequence repeat (SSR) markers associated with agronomically important phenotypic traits, for example, fibre quality. Fifty‐six Gossypium arboreum germplasm accessions introduced from nine regions of Africa, Asia and Europe were evaluated for eight fibre characters (lint percentage, lint colour, elongation, micronaire, strength, 50% span length, 2.5% span length and maturity%) and genotyped with 98 SSR markers. When viewed across all accessions most of the SSR markers were polymorphic. Population structure analysis identified six main clusters for the accessions which corresponded to different geographic regions, indicating agreement between genetic and predefined populations. The general linear model method was used to disclose marker–trait associations. Marker–trait associations were investigated by fitting single marker regression models for phenotypic traits on marker band intensities with correction for population structure. This paper illustrates the potential of association mapping in diploid cotton, because existing phenotypic data, a modest number of SSR markers, and a pioneering statistical analysis, identified interesting associations.     

甘蓝型油菜主要脂肪酸组成的QTL定位

应用RAPD、SSR和SRAP技术, 对甘蓝型油菜低芥酸品系APL01与高芥酸品系M083杂交组合的BC1F1群体进行检测, 获得251个分子标记, 构建了19个连锁群组成的分子标记遗传图谱; 应用WinQTLCart 2.0对油菜主要脂肪酸组成进行QTL扫描, 获得与棕榈酸含量相关的QTL 5个, 分别位于N3、N8、N10和N13连锁群, 其中效应值较大的主效QTL qPA8-1和qPA13分别可解释棕榈酸含量表型变异的11.31%和14.47%。获得与硬脂酸含量相关的QTL 3个, 分别位于N1、N8和N16连锁群, 其中效应值较大的主效QTL qST16可解释硬脂酸含量表型变异的12.22%。获得与油酸含量相关的QTL 2个, 位于N8和N13连锁群, 均为主效QTL, 其中qOL8位于N8连锁群的m11e37b~A0226Ba267区间, 可解释油酸含量表型变异的11.73%, qOL13位于N13连锁群的m18e46~m20e25a区间, 可解释表型变异的27.14%。获得与亚油酸含量相关的QTL 3个, 其中主效QTL qLI8-1位于N8连锁群, 可解释亚油酸含量表型变异的13.25%。获得与亚麻酸含量相关的QTL 3个, 效应值均较小, 属微效QTL。获得与廿碳烯酸含量相关的QTL 4个, 分别位于N8、N13和N15连锁群, 其中主效QTL qEI8-1、qEI8-2和qEI13分别可解释廿碳烯酸含量表型变异的12.20%、10.22%和11.14%。获得与芥酸含量相关的QTL 2个, 位于N8和N13连锁群, 均为主效QTL, 其中qER8位于N8连锁群的m11e37b~A0226Ba267区间, 可解释芥酸含量表型变异的16.74%; qER13位于N13连锁群的A0301Bb398~m18e46区间, 可解释芥酸含量表型变异的31.32%。在N8连锁群的分子标记m11e27b附近及N13连锁群的分子标记m18e46附近存在多个主要脂肪酸的主效QTL, 这些标记可用于油菜脂肪酸改良的分子标记辅助选择。     

Genetic association of yield with its component traits in a recombinant inbred line population of cotton

Lint yield of upland cotton (Gossypium hirsutum L.) is determined by its component traits, boll number, boll weight, and lint percentage. Selecting high yielding lines is based on the ability to manipulate component traits. In this study, 188 recombinant inbred lines and two parental lines were grown in 1999 and 2000 at Mississippi State University. Lint yield and its three component traits were measured and analyzed by an extended conditional mixed linear model approach. Boll number unit-area^–1 made the largest contribution to genotypic and genotype × environment (G × E) variations for lint yield. Both boll number and lint percentage, and boll number and boll weight jointly accounted for more than 70% of the genotypic and G × E variations in lint yield. Ninety-nine percent of the genetic and phenotypic variation in lint yield could be explained by the three component traits, indicating that lint yield was mainly dependent on its three component traits. Small phenotypic variation in lint yield could be accounted for by effects of genotype, G × E interactions of boll number or boll number combined with other component trait(s) (Table 5). For boll number unit-area^–1 a wider distribution of genotypic contribution effects was detected than for lint percentage and boll weight in this study. Boll number and boll weight interacted to affect lint yield, indicating that balanced selection for boll weight and boll number is needed in high-yielding line development. Comparative results with other approaches were also discussed in this study.The U.S. Governments right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged