陆地棉低世代群体纤维品质QTL定位及候选基因功能注释

【目的】通过对纤维品质数量性状位点(Quantitative trait loci,QTLs)定位及其基因功能注释,为分子标记辅助育种提供理论依据。【方法】以2个纤维品质有差异的陆地棉品种(系)中棉所49和396289为亲本构建F2群体,以高密度遗传图谱为基础,对3个环境的F2:3家系做包括细度和成熟度等在内的7个纤维品质性状QTLs定位,利用直系同源基因簇(Clusters of orthologous groups,COG)、基因本体(Gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)等数据库对QTLs做基因功能注释。【结果】获得157个与纤维品质有关的QTLs,分布于20条染色体上,A03、A04、D02、A11和D07等有较多性状的QTLs聚集,可能是控制纤维品质性状的关键染色体。共获得13个稳定QTLs,其中qFL-A03-1和qFin-A11-4在3个环境中重复出现,另有9个QTLs则在2个环境中重复出现。通过COG、GO和KEGG对QTLs进行基因功能注释,共获得4 763个候选基因,3个数据库分别注释到2 416、4 188和2 512个基因,在稳定QTLs中共注释到429个基因,其中一些基因可能与纤维品质关系密切。【结论】利用高通量测序获得的高密度遗传图谱有助于获得较多QTLs,有利于纤维品质相关候选基因的筛选及性状的改良,提高育种效率。     

陆地棉遗传图谱构建及产量和纤维品质性状QTL定位

利用3 458对SSR引物筛选陆地棉中棉所35和渝棉1号间的多态性引物, 获得173对。以多态性引物检测(渝棉1号×中棉所35)F₂群体180个单株的标记基因型, 共获得178个标记位点。构建的遗传连锁图谱包括148个标记, 36个连锁群, 总长1 309.2 cM, 标记间平均距离8.8 cM, 覆盖棉花基因组的29.5%。36个连锁群中的28个分别被定位于20条染色体, 8个连锁群未定位于染色体。以渝棉1号×中棉所35的F₂、F_2:3群体的产量、纤维品质性状鉴定结果, 利用区间作图方法, 检测到4个产量性状QTL, 即2个衣分(LP)、1个铃重(BW)、1个籽指(SD); 5个纤维品质性状QTL, 即1个纤维长度(FL)、2个纤维比强度(FS)和2个纤维细度(FF)。LP1、BW、SD、FL和FS1被定位于第7染色体, LP2、FS2、FF1和FF2被分别位于第15、21、9和20染色体。5个纤维品质QTL的有利等位基因均来源于渝棉1号。     

基于两个陆地棉低世代群体定位纤维品质相关QTL

【目的】定位棉花纤维品质性状相关的数量性状位点(Quantitative trait locus,QTL)。【方法】以陆地棉高强纤维品系中棉所679和纤维品质一般的农垦5号为亲本构建包含200个单株的F2群体及对应的F2:3家系群体,对2个群体的纤维长度、断裂比强度等5个纤维品质性状进行检测。用6 688对简单重复序列(Simple sequence repeat, SSR)引物在双亲间筛选,得到149对多态性引物,以F2为作图群体,使用QTL IciMapping软件进行连锁图谱构建,并对F2及F2:3群体进行QTL定位。【结果】根据F2群体基因型信息构建了1张包含119个标记、28个连锁群、总长为1 173.5 cM(centiMorgan)的遗传连锁图谱。分别在F_2、F2:3群体中检测到9个和11个与纤维品质性状相关的QTLs,这些QTLs分布在11个连锁群上。其中F2群体的qFL-D11-1、q BT-D11-1与F2:3群体的qFL-D11-1、q MIC-D11-1均定位在标记DPL0062与HAU0423之间,推测这些位点可能是控制纤维品质性状的重要QTL。【结论】利用多个群体进行QTL定位有益于发现稳定的QTL位点,控制纤维品质性状的基因可能成簇存在,为挖掘纤维品质性状相关基因及分子标记辅助育种奠定基础。     

陆地棉高品质品系纤维品质性状QTL的分子标记及定位

为进一步挖掘利用高品质品系NM03102的优异纤维品质性状的基因,利用陆地棉鲁棉研21作为母本、NM03102为父本构建了F2和F2∶3分离群体。通过7892对SSR引物对亲本进行筛选,获得222对多态性引物,进一步对195个F2群体单株分析得到242个标记位点。其中,182个标记位点连锁构建37个连锁群,共覆盖1661.6 cM,每个连锁群平均包含4.9个标记位点,标记间平均相距9.1 cM,其中35个连锁群被定位到了20条染色体上。利用F2和F2∶3纤维品质数据,通过复合区间作图法,共检测到20个纤维品质性状QTL。其中,1个纤维强度的QTL和1个纤维整齐度的QTL与已有的报道一致,1个纤维强度的QTL和1个麦克隆值的QTL在两世代中稳定存在,这为标记辅助选择奠定了基础。     

新疆棉花纤维品质性状的QTL分析

 以自育高品质中长绒棉品种（新陆中9号提高系）9-1696为母本,与主栽品种中棉所35为父本配置单交组合,筛选出12对SSR引物在F₂群体和B1群体进行纤维长度、整齐度、比强度和伸长率4个纤维品质性状的QTL分析,采用区间作图法（LOD>2.0）,在F₂群体中检测到6个与纤维品质性状连锁的QTL位点。其中,检测到纤维长度、纤维整齐度、伸长率各1个QTL位点,比强度检测到3个QTL。在B₁群体中检测到2个QTL,分别与比强度和纤维长度连锁。     

陆地棉优质纤维QTL的分子标记筛选及优质来源分析

利用陆地棉优质品系7235、渝棉1号做亲本, 以7235 × 渝棉1号的F2与F2:3分离群体为材料, 开展不同来源棉花高强纤维QTL微卫星标记筛选, 为进一步进行优质纤维QTL聚合育种提供基础。通过5 514对SSR引物对亲本进行多态性筛选, 获得117个多态性标记, 用其中80个标记构建了总长为1 147.8 cM的遗传图谱; 应用复合区间作图法分析了该组合的F2单株和F2:3家系纤维品质性状, 共检测到36个纤维品质数量性状基因座(QTL), 其中与纤维长度、比强度、细度、伸长率及整齐度相关的QTL各6、8、7、8和7个, 分别解释各性状表型变异的3.4%~14.4%、5.0%~42.4%、6.5%~11.7%、5.1%~19.4%和6.9%~14.0%。通过分析来源于7235和渝棉1号高强QTL的染色体分布, 表明两亲本在D7、D8、D9染色体上都存在控制纤维比强度的QTL, 其中存在于D7和D8染色体上来自双亲的QTL紧密连锁, 成簇分布在染色体上的一定区间内。而在D7和D9染色体上也发现双亲完全相同的优质QTL, 其优质供体可能来自陆地棉优质品系PD4381。进一步分析了获得的QTL在聚合育种中的应用潜力。     

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。     

新疆主栽陆地棉品种形态性状QTL的标记与定位

为了阐明新疆"矮密早"栽培技术下的高密度、矮化陆地棉形态性状QTL的遗传规律,本研究利用新疆不同陆地棉主栽品种(Gossypium hirsutum L.)构建了3个种内作图群体,进行陆地棉形态性状QTL的分子标记筛选.三个遗传图谱的连锁群长度分别为593.6 cM、830.2 cM和743.1 cM.3个遗传图谱的连锁群覆盖了除棉花Chr.22外的所有染色体.在此基础上鉴定、筛选出果枝始节、株高、叶主脉、叶次脉等15个稳定的QTLs:其中2个果枝始节QTL位于Chr.5和Chr.7上;3个株高QTL分别位于Chr.13、Chr.25和Chr.17上;筛选出叶主脉及叶次脉的QTL共10个,位于Chr.7、Chr.15、Chr.17、Chr.19、Chr.21和Chr.23连锁群17、6上,解释表型变异在6.8%～24.4%之间.对没有分配到连锁群上的标记位点的单标记分析,在LOD值大于2的水平下,共检测出9个与棉株形态性状相关的标记,其中与株高相关标记3个,另外6个标记与叶主脉及叶次脉相关.本研究定位在Chr.15、Chr.21、Chr.23和Chr.25上的棉株形态性状QTL,在染色体水平上的定位与前人报道相同,其它QTL在染色体水平上定位与前人研究不同,可能是新检测出的QTL.     

陆地棉种子物理性状QTL定位

定位棉花种子性状的基因对揭示棉花种子性状的遗传规律,以及明确棉花种子、产量、纤维品质等性状间的遗传关系具有重要意义。以(渝棉1号×T586) F_2:7重组近交系群体构建的遗传连锁图谱,在鉴定270个家系3个环境种子物理性状的基础上,利用MQM作图方法,共检测到34个种子物理性状QTL,包括9个种子重(qSW)、5个短绒重(qFW)、3个短绒率(qFP)、8个种仁重(qKW)、6个种子壳重(qHW)和3个种仁率(qKP)QTL,它们可解释4.6%~80.1%的性状表型变异。9个QTL在2个或3个环境中被检测到,其中包括第12染色体显性光子位点的短绒重与短绒率QTL,以及另外7个微效应QTL。34个QTL分布于15条染色体,其中A染色体组20个,D染色体组14个。有12个染色体区段分布有2个或2个以上的QTL,而且同一染色体区域同一亲本所具有的不同性状QTL的方向大多数与性状表型相关系数的正负一致。     

水稻分蘖相关性状的QTL定位与分析

应用147个SSR标记,对水稻测序品种日本晴(Nipponbare以下简称Nipp) ×广陆矮4号(部分测序,以下简称GLA)的F2群体进行基因检测,构建了全长为1736.6cM、覆盖水稻基因组12条染色体的连锁图,采用QTLCart1.7 统计软件对水稻分蘖相关性状如分蘖数、穗数、有效分蘖数、分蘖率等4个性状的基因座位进行定位分析,检测到5个主效果基因和17个微效基因,同时结合基因组研究的最新成果,对7个与标记间遗传距离为0(QTL POS=0)的QTLs进行生物信息学的分析,尝试用QTL定位预测基因的功能。     

QTL analysis for early-maturing traits in cotton using two upland cotton (Gossypium hirsutum L.) crosses

Making use of the markers linked closely to QTL for early-maturing traits for MAS (Marker-assisted selection) is an effective method for the simultaneous improvement of early maturity and other properties in cotton. In this study, two F₂ populations and their F_2:3 families were generated from the two upland cotton (Gossypium hirsutum L.) crosses, Baimian2 × TM-1 and Baimian2 × CIR12. QTL for early-maturing traits were analyzed using F_2:3 families. A total of 54 QTL (31 suggestive and 23 significant) were detected. Fourteen significant QTL had the LOD scores not only > 3 but also exceeding permutation threshold. At least four common QTL, qBP-17 for bud period (BP), qGP-17a/qGP-17b (qGP-17) for growth period (GP), qYPBF-17a/qYPBF-17b (qYPBF-17) for yield percentage before frost (YPBF) and qHFFBN-17 for height of first fruiting branch node (HFFBN), were found in both populations. These common QTL should be reliable and could be used for MAS to facilitate early maturity. The common QTL, qBP-17, had a LOD score not only > 3 but also exceeding permutation threshold, explaining 12.6% of the phenotypic variation. This QTL should be considered preferentially in MAS. Early-maturing traits of cotton are primarily controlled by dominant and over-dominant effects.     

QTL mapping of fiber quality in an elite hybrid derived-RIL population of upland cotton

Xiangzamian 2 (XZM2) was the most widely cultivated cotton hybrid planted as F₁ hybrids and as selfed F₂ seeds in China before the release of transgenic Bt hybrids. By crossing two parents of XZM2, Gossypium hirsutum cv. Zhongmiansuo12 (ZMS12) and G. hirsutum acc. 8891, and through subsequent selfings, we obtained F₈ and F₉ populations of 180 recombinant inbred lines (RILs). A RIL population was cultivated in two cotton-growing regions in China for 2 years. The purpose of the present research was to detect quantitative trait loci (QTL) for fiber quality and provide information applicable to cotton breeding. A genetic map was constructed mainly using SSR markers. QTL controlling fiber quality traits were determined at the single-locus and two-locus levels, and genotype-by-environment interactions were analyzed. Among the main-effect QTL, a fiber length QTL qFL-D2-1 and a reflectance QTL qFR-D2-1 were simultaneously detected at two growing regions in 2 years, which suggested a high degree of stability in different environments, and might be of particular value for a marker-assisted selection (MAS) program. The results suggested that epistatic effects, as well as additive effects, of QTL play important roles in fiber quality in these RILs. In our research, the phenomenon of QTL clusters was detected in the cotton genome.     

Quantitative analysis and QTL mapping for agronomic and fiber traits in an RI population of upland cotton

Genetic mapping is an essential tool for cotton (Gossypium hirsutum L.) molecular breeding and application of DNA markers for cotton improvement. In this present study, we evaluated an RI population including 188 RI lines developed from 94 F₂-derived families and their two parental lines, ‘HS 46’ and ‘MARCABUCAG8US-1-88’, at Mississippi State, MS, for two years. Fourteen agronomic and fiber traits were measured. One hundred forty one (141) polymorphic SSR markers were screened for this population and 125 markers were used to construct a linkage map. Twenty six linkage groups were constructed, covering 125 SSR loci and 965 cM of overall map distance. Twenty four linkage groups (115 SSR loci) were assigned to specific chromosomes. Quantitative genetic analysis showed that the genotypic effects accounted for more than 20% of the phenotypic variation for all traits except fiber perimeter (18%). Fifty six QTLs (LOD > 3.0) associated with 14 agronomic and fiber traits were located on 17 chromosomes. One QTL associated with fiber elongation was located on linkage group LGU01. Nine chromosomes in sub-A genome harbored 27 QTLs with 10 associated with agronomic traits and 17 with fiber traits. Eight chromosomes in D sub-genome harbored 29 QTLs with 13 associated with agronomic traits and 16 with fiber traits. Chromosomes 3, 5, 12, 13, 14, 16, 20, and 26 harbor important QTLs for both yield and fiber quality compared to other chromosomes. Since this RI population was developed from an intraspecific cross within upland cotton, these QTLs should be useful for marker assisted selection for improving breeding efficiency in cotton line development. Paper number J1116 of the Mississippi Agricultural and Forestry Experiment Station, Mississippi State University, Mississippi State, MS 39762. Mention of trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by USDA, ARS and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

QTL mapping for agronomic and fibre traits using two interspecific chromosome substitution lines of Upland cotton

CS‐B14Sh and CS‐B22Sh are cotton interspecific chromosome substitution (CS)‐B lines, in which a pair of short arms of chromosome 14 and chromosome 22 were introgressed from Gossypium barbadense doubled‐haploid line 3‐79 with the background of Gossypium hirsutum line TM‐1, respectively. These two CS‐B lines were crossed with TM‐1, and segregating progenies (F₂ and F_2:3, respectively) were obtained. Phenotypic data of lint yield, yield‐related traits and fibre‐quality traits were collected from two trials. In the cross CS‐B14SH X TM‐1, QTL for boll weight (BW), lint percentage (LP), fibre upper half mean length (UHML), micronaire reading (MIC), and fibre breaking tensile strength (STR) were repeatedly detected. Alleles from 3‐79 decreased BW and MIC, but increased UHML and STR. In the cross CS‐B22Sh X TM‐1, QTL for BW, LP, UHML, MIC, STR, fibre elongation (EL),seed weight(SW), node of first fruiting branch (NFB) and fibre uniformity index (UI) were repeatedly detected, and alleles from 3‐79 decreased UHML, UI and STR, but increased NFB, SW, MIC and EL. QTL clusters were found in both populations.     

鲁棉研15号纤维品质性状QTL定位研究

 以陆地棉（Gossypium hirsutum L.）杂交种鲁棉研15号的F₂群体为作图群体,利用SSR标记和JoinMap3.0软件构建遗传连锁图谱;利用复合区间作图法分别对随机组成的3个鲁棉研15号的F_2:3家系亚群体进行纤维品质性状QTL定位。构建的遗传连锁图谱包含116个多态位点,25个连锁群,全长892.25 cM,覆盖棉花总基因组的20.05%,平均每个连锁群4.64个标记,标记间平均距离7.76 cM;根据已有图谱的定位结果,19个连锁群与染色体建立了联系。在3个F_2:3家系亚群体中共检测到46个QTL,其中16个为纤维长度（FL）QTL、7个为纤维强度（FS）、12个为麦克隆值（FM）、6个为伸长率（FE）,5个为整齐度指数（FU）。发现在A_h05、A_h08、A_h09、D_h02染色体上QTL有成簇分布的现象,并在3个亚群体中检测到一些受环境影响较小、稳定遗传的QTL。这些QTL可以在今后应用于分子标记辅助选择。     

Dynamic QTL mapping for plant height in Upland cotton (Gossypium hirsutum)

Plant height served as one of model traits to analyse dynamic development. The objective of this research was to investigate quantitative trait loci (QTL) and dynamic QTL for plant height trait using an intraspecific recombinant inbred line (RIL) population and a constructed genetic map in Upland cotton (Gossypium hirsutum L.). Totally, 41 QTL and 23 conditional QTL controlling plant height were detected at two experimental environments, respectively. Four stable QTL were identified simultaneously in both environments. Some QTL identified at the early stage could not be detected at the final stage at plant maturity. Conditional QTL with different genetic effects were identified at certain stages, demonstrating that the expression of QTL had temporal characteristic during plant growth. Therefore, the study of dynamic QTL could unravel temporal genetic patterns controlling complex developmental quantitative traits.     

Identifying QTL for fiber quality traits with three upland cotton (Gossypium hirsutum L.) populations

Cotton fiber quality was quantitative trait, controlled by multiple genes. Identification of stable quantitative trait loci (QTL) effectively contributing to favorable fiber quality traits would provide the key basis for marker-assisted selection used in molecular breeding projects. Three upland cotton F₂ populations were established with a common parent Chinese cultivar Yumian 1 and three American commercial cultivars/lines (Acala Maxxa, CA3084 and TAM94L-25), each of which had unique fiber quality characteristic that was favorable economically. Three whole genome genetic maps were constructed with 323, 302 and 262 SSR loci for population (Yumian 1 × Acala Maxxa), (Yumian 1 × CA3084), and (Yumian 1 × TAM 94L-25) respectively, spanning 1,617.2, 1,639.9 and 1,441.4 cM. Based on these genetic maps and three generation phenotypic data of fiber quality traits (F₂, F_2:3 and F_2:4), 77 QTL were detected, including 19 for fiber length, 14 for fiber uniformity, 17 for micronaire, 10 for fiber elongation, and 17 for fiber strength. Among these QTL, 46 QTL were significant QTL and 31 were putative QTL, including that one QTL (qFL05.1) and four QTL (qFL23.1, qFM06.1, qFM06.2 and qFE25.1) were detected across three and two populations respectively; two QTL qFL10.1 (Yumian 1 × TAM 94L-25) and qFL15.1 (Yumian 1 × Acala Maxxa) were detected in three generations; qFM23.1, qFE18.1 and qFS21.2 detected in population (Yumian 1 × CA3084), qFE10.1, and qFS10.2 detected in population (Yumian 1 × TAM 94L-25), and qFS15.1 detected in population (Yumian 1 × Acala Maxxa), were all detected in two generations. Alleles underlying these stable QTL were valuable candidate gene for fine mapping, cloning, and favorable gene pyramiding projects. Our study also verified that QTL mapping of fiber quality traits using multiple populations with a common parent had higher efficiency compared to single population crossed with two parents and favorable alleles contributed to QTL effect could be conferred by parents with inferior fiber quality traits.     

陆地棉种质资源抗旱性状的关联分析

干旱是导致全世界棉花严重减产、纤维品质下降的重要因素,因此获得高产、优质、耐旱的棉花新品种一直是棉花的育种目标。本研究选取217份陆地棉栽培种组成的自然群体为研究对象,采用全生育期处理组灌水量为对照组50%的干旱胁迫处理,并在处理后期对217份材料的株高、衣分、单铃重等18个性状进行2年2点的表型鉴定,干旱胁迫后,群体间响应差异明显,多个表型性状在对照和处理间表现显著差异。通过BLUP分析表型数据并计算各性状的抗旱系数;全基因组范围选取的214对多态性SSR分子标记扫描群体,共检测到393个多态性位点,基因多样性系数平均值为0.402,范围为0.072～0.631, PIC值平均为0.329,范围为0.070～0.560;群体结构分析表明,该群体可分为2个亚群。用上述SSR标记分别对18个性状的抗旱系数进行关联分析,共关联到76个极显著位点(P<0.01),表型变异解释率为2.930%～7.218%,其中共有14个标记位点能同时被2种或以上性状检测到。研究结果可为后期棉花杂交育种亲本选择及抗旱分子标记辅助育种提供理论基础及参考依据。