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.     

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。     

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.     

QTL Mapping of Chlorophyll Content in Gossypium hirsutum and Gossypium barbadense Backcross Inbred Lines

[Objective] The purpose of this study was to map quantitative trait loci (QTL) related to chlorophyll content based on Soil and Plant Analyzer Development (SPAD) readings in cotton. [Method] The 195 BILs (Backcross Inbred Lines) were produced by a cross between Gossypium barbadense Hai 7124 and G. hirsutum CRI 36, using CRI 36 as the recurrent parent for backcrossing with F1 to produce BC1F1, followed by seven generations of selfing. The genetic linkage map was constructed in a previous study. QTLs of chlorophyll SPAD value in the first flowering and boll development stages were identified with inclusive composite interval mapping (ICIM) method of the BIP and MET models in IciMapping 4.1 software, respectively. [Result] In total, nine chlorophyll SPAD reading QTLs were identified on 6 chromosomes. The q-SPAD-A11-1 detected at the first flowering stage overlapped with q-SPAD-A11-2 detected at the boll development stage, contributing 5.08% and 5.75% of the phenotypic variation, respectively. The q-SPAD-D08-2 physical position ranged from 48.71 to 53.65 Mb on chromosome D08, which overlapped with a chlorophyll content QTL detected in a previous study. [Conclusion] The novel stable QTLs, q-SPAD-A11-1 and q-SPAD-A11-2 detected in this study provide an important piece of information for fine mapping chlorophyll content in cotton.     

QTL Mapping of Fiber Quality in Low-Generation Upland Cotton Populations and Functional Annotation of Candidate Genes

[Objective] This article aims to provide a theoretical basis for molecular marker-assisted breeding by quantitative trait loci (QTLs) mapping and gene function annotation of fiber quality. [Method] F₂ populations were constructed using the two cotton cultivars (lines), CCRI 49 and 396289, as parents. Based on high-density genetic maps, the F_2:3 families with three environments were included in the F2 population. The QTL mapping of seven fiber quality traits including fineness and maturity, etc., was performed using the Clusters of orthologous groups (COG), Gene ontology (GO), and Kyoto encyclopedia of genes and genomes (KEGG) databases to annotate QTLs for gene function. [Result] A total of 157 QTLs related to fiber quality were obtained and distributed on 20 chromosomes. QTLs with more traits on A03, A04, D02, A11 and D07 chromosomes clustered and may be the key chromosomes controlling fiber quality traits. A total of 13 stable QTLs were obtained, of which qFL-A03-1 and qFin-A11-4 were repeated in three environments, and nine other QTLs were repeated in two environments. And 4 763 candidate genes were annotated, with 2 416, 4 188, and 2 512 genes being annotated in COG, GO, and KEGG, respectively. Among them, 429 genes were annotated in stable QTLs. Some of these genes may be closely related to fiber quality. [Conclusion] The high-density genetic map obtained by high-throughput sequencing can help to obtain more QTLs, which is beneficial to the screening of candidate genes related to fiber quality and the improvement of fiber traits, and improves the breeding efficiency.     

毛棉苗期抗旱性状的QTL定位

【目的】通过分析毛棉苗期抗旱相关性状的数量性状位点(Quantitative trait locus,QTL),以期检测稳定的主效QTL,促进栽培品种抗旱性状遗传改良及提高抗旱育种效率。【方法】以四倍体野生种毛棉(Gossypium tomentosum)和陆地棉品种中棉所12(CCRI 12)的种间杂种F2及其F2:3家系为研究材料,用于基因型分型的F2有188个系,用于表型分型的F2:3家系有149个株系。分别在干旱胁迫和正常灌水2个环境下调查表型数据。采用复合区间作图法对F2:3家系苗期相关性状抗旱系数进行QTL定位。【结果】对苗期相关性状抗旱系数的QTL定位分析,共得到16个QTL,其中与株高、叶片数、叶绿素含量、脯氨酸含量、丙二醛含量抗旱系数相关的QTL分别有5个、1个、3个、3个、4个,分布在13条染色体上。来自毛棉的5个加性QTL分别为qSHDC-19-1、qSHDC-19-2、qSLNDC-5-1、qMDADC-24-1、qMDADC-24-2,其加性效应值为0.10~0.22,解释变异9.4%~25.8%。【结论】这些与抗旱相关的QTL有助于棉花抗旱分子标记辅助选择。     

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。     

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。     

Evaluation of Chromosome Segment Substitution Lines Related to Fiber Yield and Quality Traits from Gossypium hirsutum×Gossypium barbadense

Three generations of chromosome segment substitution lines lines (BC₄F₃, BC₄F_3:4, and BC₄F_3:5, Anyang, Korla) were used to evaluate phenotypic traits related to fiber yield and quality. The average performance of the traits in the population was near to the control, CCRI45, in the four populations. Seventy-eight percent of the lines showed longer and stronger fibers than the recurrent parent, and 22.59%～53.61% of lines had bigger bolls and higher lint percentages than the recurrent parent. Correlation analysis showed that the yield and fiber quality traits were positively correlated among the four populations. These results indicated that most of the genetic background in the substitution lines has been restored to the recurrent parent with abundant genetic variations. In addition, most of substitution lines had good yield and fiber quality. This study provided a large amount of basic research materials for cotton quantity genetics, gene pyramiding, and gene function analysis, and provided parent materials and new strains that can directly applied for cotton breeding.     

陆地棉产量相关性状的QTL定位

中棉所28和湘杂棉2号分别是以中棉所12×4133和中棉所12×8891配制而成的两个陆地棉强优势杂交种。以其F₂为作图群体,筛选6000多对SSR引物,利用两群体间27个共有多态位点,通过JoinMap 3.0软件整合了一张包含245个多态位点、全长1847.81 cM的遗传图谱。利用Win QTLCart 2.5复合区间作图法分别对两群体8个产量相关性状在F₂和F_2:3中进行QTL定位,在中棉所28群体多环境平均值的联合分析中检测到16个QTL,三环境分离分析中检测到43个QTL;在湘杂棉2号群体分别检测到20个和66个QTL。在A3、D8、D9等染色体上有QTL成簇分布现象,同时在两个群体中发现一些不受环境影响且稳定遗传的QTL。对考察的8个性状在两个群体中发现12对共有QTL,控制果枝数、衣分和籽指的QTL增效基因位点均来源于共同亲本中棉所12。综合分析推测中棉所12的育种价值主要是通过提高后代的结铃性来实现的。研究结果为棉花产量性状的分子设计育种提供了有用的信息。     

海岛棉CSSLs分子评价及纤维品质、产量性状QTL定位

本课题组前期以陆地棉中棉所8号(CCRI8)为轮回亲本, 海岛棉Pima 90-53为供体亲本培育了一套陆地棉中棉所8号为背景的海岛棉染色体片段置换系(CSSLs), 本研究利用SSR标记对该置换系群体BC₃F₅进行基因型检测, 在3个不同环境下(河北保定、青县和新疆轮台)鉴定其纤维品质和产量相关性状并进行QTL定位。该置换系群体包含182个家系, 置换片段数在1~15个之间, 平均为6.6个; 导入片段长度在0.7~83.2 cM之间, 平均长度为16.8 cM; 置换片段总长度20 249.6 cM; 背景回复率在92.3%~99.6%之间, 平均为96.2%。共检测出59个相关的QTL, 其中与纤维品质性状相关的41个, 单个QTL的贡献率为1.27%~26.66%; 与产量性状相关的18个, 单个QTL的贡献率为2.03%~19.38%; 检测到14个稳定的QTL, 其中4个马克隆值和2个纤维伸长率相关的稳定QTL增效基因均来自高值亲本海岛棉Pima 90-53, 2个铃重相关的稳定QTL增效基因来自高值亲本陆地棉中棉所8号。研究结果为深入开展纤维品质和产量性状的QTL精细定位、QTL间互作和分子育种提供了理论依据。     

Improving Fiber Quality Traits of Xinluzao 45 Using Gossypium barbadense Chromosome Segment Substitution Lines

Gossypium barbadense chromosome segment substitution lines could be used as an important germplasm resource for improving fiber quality traits of upland cotton. In this study, we used TM-1 background substitution lines CSSL-122, the Xinjiang upland cotton Xinluzao 45 and hybrid, backcross progeny BC₂F₁ populations comprising 120 individual plants as test materials. Nineteen SSR markers located in Gossypium barbadense chromosomes land inked with fiber length and strength were used to screen out obvious polymorphic primers between the parents. We then traced and detected chromosome segments of Gossypium barbadense in the BC₂F₁ populations. Simultaneously, we compared fiber quality traits of positive plants that contained Gossypium barbadense chromosome segments with non-positive plants in the BC₂F₁ populations. The results showed that two markers, NAU2987 and BNL3145, which were linked with fiber length, could accurately identify the positive plants. In addition, compared with non-positive plants, the increased fiber length and strength of the positive plants were very significant (P < 0.01). Our research suggested that alien Gossypium barbadense chromosome segments significantly improved the fiber quality traits of Xinluzao 45. Thus, the Gossypium barbadense chromosome segment substitution lines will provide a vital theoretical basis and practical reference for improving Upland Cotton fiber quality traits.     

陆地棉转录因子的染色体定位

利用植物转录因子PTFD数据库1 116条陆地棉转录因子DNA序列设计的1 455对SSR引物,筛选陆地棉品种/品系渝棉1号、中棉所35、7235和T586,获得66对多态性引物。它们涉及到27个转录因子家族的64个转录因子,其中渝棉1号与中棉所35间23对多态性引物,渝棉1号与T586间30对多态性引物,渝棉1号与7235间33对多态性引物。以多态性引物检测对应重组近交系群体,共获得93个位点。其中,(渝棉1号×中棉所35)群体23个位点,(渝棉1号×T586)群体32个位点,(渝棉1号×7235)群体38个位点。利用转录因子SSR位点与实验室已定位的SSR位点进行遗传连锁分析,将84个位点定位于23条染色体上,其中32个位点分布于A染色体组,52个位点分布于D染色体组。     

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.     

陆地棉重组自交系群体纤维品质及产量性状遗传变异分析

以陆地棉优质品系0-153和大面积推广的双价转基因抗虫棉品系sGK9708为亲本构建了含有196个F6:8家系的重组自交系群体。在4个环境中,纤维长度、纤维强度、麦克隆值、伸长率、整齐度指数、单株结铃数、铃重、衣分、子指、单株子棉产量及单株皮棉产量等性状均呈正态分布,且存在双向超亲分离。多环境下变异分析表明,各性状家系间及环境间存在极显著差异。除纤维伸长率外,其余各纤维品质及产量性状都具有较高的遗传力,在0.75以上。共有6个家系在纤维长度、细度和强度性状上均超过高亲,可作为优质纤维种质。相关分析显示,有些家系可能打破了纤维品质和产量性状间的负相关。通过聚类分析得出,该群体各系分在了不同的组,表明存在着一定的遗传差异。该群体是进行优异纤维品质相关基础研究的良好材料。     

基于陆地棉背景的海岛棉染色体片段导入系产量性状QTL定位

棉花产量分为籽棉产量和皮棉产量,其中高皮棉产量总是育种的首要目标。皮棉产量由单株铃数、衣分、单铃重等因素组成。其中衣分在各因素中的遗传率最高,同时也是产量育种中重要的选择指标。育种中利用分离群体对单株铃数、铃重等产量性状选择受环境影响较大。利用染色体片段导入系进行铃数、铃重等产量性状的定位,定向改良产量性状,是棉花分子设计育种的有效方法。本研究利用陆地棉TM-1为轮回亲本和海岛棉海7124为非轮回亲本构建了一套陆地棉背景的染色体片段导入系,并在7个环境的田间试验下,鉴定了它们的产量表现,定位了28个与单株铃数、铃重、衣分和籽指相关的QTL。其中,在Dt亚组染色体上鉴定出的产量性状QTL多于在At亚组染色体上鉴定出的。28个QTL中,加性效应为正的16个,加性效应为负的12个,表明海岛棉不同的导入片段效应不同,有的片段可以提高陆地棉产量,有的则降低陆地棉产量。在6个环境下,导入系IL008(特征标记NAU2573和NAU3576)的衣分均显著高于轮回亲本TM-1,因此IL008可以应用于棉花分子育种,定向改良陆地棉的衣分。     

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。     

转Bt抗虫棉与常规棉品种间配合力分析及杂种优势研究

 利用6个抗虫棉品种作母本,4个常规品种作父本,按NCⅡ设计,对24个组合的杂种F₁进行了分析。结果表明,杂种F₁具有明显的优势,所研究的13个性状全部具有中亲优势,9个性状具有高亲优势,11个性状具有竞争优势。配合力分析的10个性状中单株铃数、铃重、纤维长度、纤维伸长率和麦克隆值主要受基因的加性效应控制,而子棉产量和皮棉产量主要受基因的非加性效应影响。鲁棉研17是优良的抗虫棉亲本,而冀棉12 是良好的非抗虫棉亲本,组合鲁棉研17×中棉所12在重要农艺性状上优势明显,相对而言纤维品质的改良较难。