棉花陆海渐渗系双交分离群体产量和纤维品质性状的QTL定位

【目的】利用海岛棉染色体片段渐渗系对棉花产量和纤维品质相关QTL(Quantitative trait locus)进行定位。【方法】以陆地棉中棉所45和海岛棉海1高代回交自交获得的4个优质海岛棉染色体片段渐渗系MBI7115、MBI 7412、MBI 7153、MBI 7346为亲本,通过[(MBI 7115×MBI 7412)×(MBI 7153×MBI 7346)]构建双交F_1及F_(1:2)群体。利用微卫星标记(Simple sequence repeat,SSR)对亲本及双交F_1进行基因型分析,并对F_1、F_(1:2)群体的产量和纤维品质相关性状进行QTL定位。【结果】4个渐渗系遗传背景恢复到轮回亲本中棉所45的比例均在97%以上,在2个群体中共检测到41个QTL,分布在11条染色体上。其中与纤维品质相关的有30个,表型贡献率在1.11%~11.80%;与产量相关的有11个,表型贡献率在1.09%~13.57%。【结论】有5个纤维品质相关的QTL能同时在2个群体中检测到且均为新发现的QTL。这一结果为进一步精细定位和开展棉花分子标记辅助育种提供了重要依据。     

陆地棉背景下海岛棉第18染色体片段置换系的培育及相关农艺性状QTL定位

Sub18是以陆地棉遗传标准系TM-1为背景, 含海岛棉3-79第18染色体的置换系材料。本研究以TM-1为受体亲本, 置换系Sub18为供体亲本, 借助分子标记辅助选择技术培育了一套以TM-1为背景, 含海岛棉3-79第18染色体不同长度片段的置换系。这套置换系由45个株系构成, 共78个置换片段。其中27个株系导入单片段, 占总株系的60%; 9个株系导入2个片段, 占20%; 9个株系导入3个及以上片段, 占20%。导入片段总长度为467.6 cM, 约为该染色体遗传长度的4倍, 每个株系内被替换的染色体片段长度不完全相同, 平均遗传长度为5.99 cM, 最短的为0.9 cM, 最长的20.35 cM。其中13个株系表现开放花蕾性状, 涉及的最短导入片段长5.05 cM。对TM-1、Sub18以及培育的45个导入系进行农艺性状调查和QTL联合定位分析, 鉴定出纤维强度(qFS-C18-1)、整齐度(qFU-C18-1)、马克隆值(qFMi-C18-1)、成熟度(qFMa-C18-1)、皮棉重(qLW-C18-1)、籽指 (qSI-C18-1)和衣分 (qLP-C18-1) 7个加性QTL和5个上位性效应QTL。研究结果为进一步精细定位目标QTL、克隆QTL以及重要性状分子设计育种奠定了基础。     

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。     

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.     

利用陆海杂种BC1群体构建遗传连锁图谱并初步定位产量性状相关的QTL

摘要：本研究利用中棉所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2252.36cM,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8.87cM。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。     

利用黄褐棉染色体片段导入系定位产量和纤维品质性状QTL

陆地棉的遗传基础狭窄,阻碍了棉花的遗传改良进程。为有效拓宽陆地棉的遗传基础,本试验利用野生种黄褐棉(AD4)为供体亲本,以综合性状优良的B0011品系为受体亲本(国审棉华杂棉H318的亲本之一),构建了含71个株系的导入系BC5S5群体。基于SLAF-seq的基因分型和多年多点田间试验的综合分析表明,该导入系在产量和纤维性状方面具有很大的变异,共检测到48个QTL,其中包含19个产量和29个纤维构成因素相关的QTL。在At亚组检测到9个性状的32个QTL,在Dt亚组为16个。进一步对QTL加性效应方向分析显示,其中有30个QTL的加性效应为正,18个QTL的加性效应为负。本研究结果为利用黄褐棉重要农艺性状有利等位基因改良陆地棉产量和品质奠定了基础。     

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。     

棉花BC_5F_2代换系的产量及品质相关性状表型分析及QTL定位

本研究利用生产上推广的优良早熟陆地棉栽培品种中棉所36为受体亲本,海岛棉海1为供体亲本,选择培育了一套由303个单株组成的BC5F2代换系。从已构建的BC1F1遗传图谱上以5~10cM为标准挑选391对多态性标记进行分子检测,多数单株含有海岛棉代换片段数为2~10个。对该群体的单株产量、品质性状进行了表型鉴定,存在大量具有优良品质的单株,纤维强度最高的能达到37.8cN/tex,铃重、衣分、纤维长度、整齐度、马克隆值、伸长率及纤维强度超轮回亲本分别为17.82%、44.55%、46.86%、33.33%、74.92%、41.58%、42.57%。采用性状-标记间的单向方差分析,共定位了20个与产量性状和33个与纤维品质性状有关的QTL,其中qUN-14-2、qBW-2-20、qFL-2-20和qMV-1-38这4个QTL存在一定的遗传稳定性。鉴定的QTL大多是微效基因,解释表型变异为3.01%~9.69%,该研究为染色体单片段代换系的精细的分子研究奠定了基础。     

陆地棉抗黄萎病、纤维品质和产量等农艺性状的QTL定位

用一个高抗黄萎病的陆地棉品系5026和一个高感黄萎病的陆地棉品种李8为材料,构建一个RIL群体.用5 300对SSR引物筛选亲本多态,获得115个多态位点并进行标记间连锁分析,构建了一张包括20个连锁群全长560.1 cM的陆地棉品种间分子标记遗传图谱.RIL家系分两份:一份种于病圃,在苗期和成株期分别考查各家系的发病情况;一份种于大田,调查各家系的产量和纤维品质相关性状.用复合区间作图法对抗病、产量和纤维品质等性状进行QTL定位:在苗期检测到了3个抗病QTL,成株期检测到了1个抗病QTL,解释的变异系数范围是7.4%～11.8%;检测到2个纤维长度、2个纤维强度、1个纤维细度、1个整齐度、1个短纤维指数和2个纤维伸长率等9个与纤维品质相关性状的QTL,解释的变异范围是6.7%～15.7%;检测到了2个皮棉产量、1个籽棉产量、2个单株果枝数、1个单株铃数、2个铃重、1个籽指、1个衣分和1个衣指等11个产量构成因素的QTL,解释的变异方差范围是4.1%～10.6%,这些结果为棉花抗病育种同时兼顾产量和品质育种提供了非常有用的信息.     

棉花分子遗传图谱构建和纤维品质性状QTL分析

以陆地棉(Gossypium hirsutum L.)中棉所8号和海岛棉(Gossypium barbadense L.) Pima90-53组配衍生的214个单株的F₂群体为材料,构建了包含110个SSR标记和65个AFLP标记的遗传连锁图谱。该图谱共包括42个连锁群,连锁群长度为4.5~147.3 cM,包括2~22个分子标记,标记间平均距离为11.6 cM,总长为2 030 cM,约占棉花全基因组的40.6%。应用复合区间作图法分析该组合的F₂单株和F_2:3家系纤维品质性状,共得到25个纤维品质数量性状基因座(QTL),其中5个与纤维长度相关s,分布在Chr.21、Chr.15、LG2和LG12上,可解释表型变异的10.2%~35.8%;4个与整齐度相关,分布在Chr.21、LG9、LG18和LG12上,可解释表型变异的12.6%~36.6%;7个与马克隆值相关,分布在Chr.9、LG1、LG9、LG20和LG12上,可解释表型变异的11.5%~26.1%;7个与断裂比强度相关,分布在Chr.21、Chr12、Chr.8、LG1、LG4和LG10上,可解释表型变异的16.5%~52.8%;2个与伸长率相关,分布在Chr.9和Chr.21上,可解释表型变异的18.1%和27.1%。LG9、LG12和Chr.21上存在QTL聚集区。     

QTL Mapping of Important Agronomic Traits on Chromosomes Showing Marker Segregation Distortion in Interspecific Backcross Cotton Populations

[Objective] The purpose of this study was to map QTLs for yield, maturity, and fiber quality traits of cotton interspecific hybrid populations and to mine favorable alleles from the cotton genome for QTL fine mapping, gene cloning, and molecular marker-assisted selection. [Method] Four BC₁F₁ populations, E(E3), E(3E), (E3)E, and (3E)E, were developed by crossing sea island cotton 3-79 ("3") and upland cotton 'Emian 22' ("E"), and with "Emian 22" as the recurrent parent. BC1F2 populations derived from BC1F1 populations were phenotyped to allow mapping of QTLs for yield-, maturity-, and fiber quality-related traits on chromosomes showing segregation distortion. [Result] A normal distribution test indicated that six fiber quality-related traits, except for the micronaire value in E (E3) and the short fiber ratio in all populations, followed a normal distribution pattern; in contrast, earliness-related traits were not normally distributed in all populations, which suggests that genotype-environment interactions have strong effects on maturity-related traits. Using composite interval mapping, 47 QTLs were detected on three chromosomes (2, 16, and 18): 15 in (E3)E, 13 in (3E)E, 12 in E(E3), and 7 in E(3E), with the phenotypic variation explained (PVE) ranging from 8.8%–30.9%. A total of 27 fiber-related QTLs were discovered, with the PVE ranging from 9.0%–30.9%. Five yield-related QTLs (PVE = 8.8%–17.4%) and 13 maturity-related QTLs (PVE = 9.4%–19.4%) were also identified. [Conclusion] Comparison analysis showed that most QTLs in our study were consistent with the MetaQTL, including QTLs for boll weight, fiber length, fiber length uniformity, lint percentage, and micronaire value on chromosome 2; QTLs for fiber length, fiber strength, and fiber length uniformity on chromosome 16; and QTLs for boll weight, date of boll opening, fiber length, fiber strength, fiber length uniformity, and micronaire value on chromosome 18. Several detected QTLs in our study were not found in the MetaQTL, such as QTLs for date of boll opening and lint percentage on chromosome 16 and a QTL for the date from seedling to flowering stages on chromosome 18. These newly identified QTLs may provide novel insights for cotton QTL analysis.     

多环境下陆地棉染色体片段代换系及F_1皮棉产量与纤维品质的表型分析

以1套陆地棉染色体片段代换系为亲本材料,采用株系间随机成对杂交组配F1,在5个环境下鉴定片段代换系亲本及F1的皮棉产量与纤维品质性状表现。结果表明,与轮回亲本(中棉所36)相比,F1在铃重与皮棉产量性状上具有一定的对照优势,亲本与F1的纤维长度与比强度均表现出明显的对照优势,且环境间表现一致。F1铃重、衣分与皮棉产量在各个环境下的中亲优势均值全部为正,且相对较大。除个别环境外,纤维长度、马克隆值与比强度的中亲优势均值也全部为正,但数值相对较小。片段代换系及F1的遗传变异丰富,部分亲本与F1在多个环境下的综合表现优异,其皮棉产量与纤维品质得到同步提高,群体材料的遗传改良与杂种优势利用具有广阔前景。     

Mapping QTL of Fiber Yield and Quality Traits in F2 Populations of Chromosome Segment Substitution Lines from Gossypium hirsutum × Gossypium barbadense

In this study, F₂ populations from two chromosome segment substitution lines MBI7455 and MBI7358, were quantified using SSR to evaluate the parents' genotype and detect QTL related to fiber quality plus yield traits of cotton. Results show that the recurrent parent (CCRI45) hosted 96.70% and 95.60% of chromosome segment substitution in MBI7455 with 12 chromosome segments and in MBI7358 with 16 chromosome substitution segments of Gossypium barbadense, respectively. In the F₂ population, the average rate of chromosome substitution of the recurrent parent (CCRI45) was 96.44%, and the average segments of chromosome substitution of Gossypium barbadense was 13.42, with an average segments of homozygous donor chromosome value of 3.90. Analysis showed 19 fiber quality-related QTL with a phenotypic variance of between 2.52%-13.11% and seven yield traits-related QTL with a range of 2.93%-11.40% phenotypic variance, resulting in a total of 26 QTL. The CSSLs could be used to detect QTL for fiber yield and quality traits, which offer an important foundation for the cotton molecular-assisted breeding.     

Genetic analysis of the fiber quality and yield traits in <Emphasis Type="Italic">G. hirsutum</Emphasis> background using chromosome segments substitution lines (CSSLs) from <Emphasis Type="Italic">Gossypium barbadense</Emphasis>

Developing chromosome segments substitution lines (CSSLs) is an effective method for broadening the cotton germplasm resource, and improving the fiber quality and yield traits. In this study, the 1054 F₂ individual plants and 116 F_2:3 lineages were generated from the two parents of MBI9749 and MBI9915 selected from BC₅F_3:5 lines which originated from hybridization of CCRI36 and Hai1, and advanced backcrossing and repeated selfing. Genotypes of the parents and F₂ population were analyzed. The results showed that 19 segments were introgressed for MBI9749 and 12 segments were introgressed for MBI9915, distributing on 17 linkage groups. The average background recovery rate to the recurrent parent CCRI36 was 96.70% for the two parents. An average of 16.46 segments was introgressed in F₂ population. The average recovery rate of 1054 individual plants was 96.85%, and the mean length of sea island introgression segments was 157.18 cM, accounting for 3.15% of detection length. QTL mapping analysis detected 22 QTLs associated with fiber quality and yield traits in the F₂ and F_2:3 populations. These QTLs distributed on seven chromosomes, and the phenotypic variation was explained ranging from 1.20 to 14.61%. Four stable QTLs were detected in F₂ and F_2:3 populations, simultaneously. We found that eight QTLs were in agreement with the previous research. Six QTL-clusters were identified for fiber quality and yield traits, in which five QTL-clusters were on chromosome20. The results indicated that most of QTL-clusters always improve the fiber quality and have negative additive effect for yield related traits. This study demonstrated that CSSLs provide basis for fine mapping of the fiber quality and yield traits in future, and could be efficiently used for pyramiding favourable alleles to develop the new germplasms for breeding by molecular marker-assisted selection.     

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。     

Quantitative Traits Locus Meta-analysis of Fiber Quality Traits in Cotton

Using bioinformatics methods and meta-analysis with BC₁ map as reference, 92 cotton fiber quality QTL collected from both BC₁ and BC₁F₂ populations constructed previously were used to construct a QTL integrated map for QTL analysis in this study. The five hundred ninety-nine loci were mapped into 26 chromosomes with an average distance between adjacent markers of 5.96 cM and covered 3,571.9 cM. Sixty-three QTL of fiber qualities related were integrated into the new reference map. The fifteen meta-QTL were mapped on 12 chromosomes by the meta-analysis method and also QTL clusters have been discovered on chromosome 9, 16 and 24. The major meta-QTL of Meta-QTL9-1 derived from five QTL on chromosome 9, could explain 17.16% of phenotypic variance. The meta-QTL16-1 derived from ten QTL on chromosome 16, could explain 12.28% of phenotypic variance. And three meta-QTL derived from nine QTLs on chromosome 24, could explain 16.12%,16.69% and 18.27% of phenotypic variance, respectively. On average, one meta-QTL derived from two QTLs on the other chromosomes. The results indicated that these meta-QTL could be used in improving fine QTL mapping and molecular-assisted selection of cotton fiber qualities in breeding.     

QTL Mapping of Boll Weight Trait Based on Recombinant Inbred Lines in Gossypium hirsutum L.

[Objective] The aim of this study was to explore the quantitative trait locus (QTL) related to the boll weight. [Method] A single seed descended population of 137 recombinant inbred lines (RILs) was developed from the cross of upland cotton (Gossypium hirsutum L.) CCRI 36 and G2005, an introgression inbred line introgressed from G. barbadense. Using restriction-site associated DNA sequencing (RAD-seq), a genetic linkage map composed of 6 434 makers, including 6 295 single nucleotide polymorphisms (SNP) and 139 simple sequence repeat (SSR) markers, was developed from the RILs population. [Result] This map spanned 4 071.98 cM with an average distance of 0.63 cM between adjacent markers. QTL mapping was performed by using boll weight data of five environments through WinQTLCart 2.5 software. Thirty-two QTL, with 4.46%-15.84% explained phenotypic variation related boll weight, were detected and found distributing on 15 chromosomes. qBW-A4-1, qBW-A4-2, qBW-A5-2, qBW-D9-1, and qBW-D9-2 were detected in two environments, which explained 5.07%-15.84% of the phenotypic variation. [Conclusion] Major QTLs detected in this study will provide an important reference for analysis of the genetic mechanism of boll weight.     

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。