水稻高秆染色体片段代换系Z1377的鉴定及重要农艺性状QTL定位

株高是水稻重要的农艺性状, 往往与产量相关性状密切关联, 在水稻育种中有重要利用价值。本研究以日本晴为受体、缙恢35为供体亲本, 经表型和分子标记双重选择, 鉴定了一个水稻高秆染色体片段代换系Z1377。Z1377共含有18个代换片段, 平均代换长度为2.95 Mb。与日本晴相比, Z1377的株高、倒一节间至倒四节间长、穗长、一次枝梗数、二次枝梗数、粒长、每穗实粒数、总粒数显著增加; 粒宽显著变细, 有效穗数、结实率显著减少, 但仍达86.75%。用日本晴与Z1377杂交构建的次级F₂群体共检测到16个相关QTL, 分布于第2、第3、第4、第5、第6、第7和第9染色体。其中有8个可能与已克隆基因等位, 如GW2、EUI1、ZFP185等, 另8个如qPH3等尚未见报道。Z1377的株高由一个主效QTL (qPH3)和一个微效QTL (qPH5)控制, 其中qPH3的贡献率达28.59%。而且, 在F₂群体中, 高秆和矮秆基本呈现双峰分布, 经卡平方测验, 符合3∶1分离比, 表明高秆对矮秆显性, 并主要由qPH3负责。这将为该主效基因的精细定位和克隆奠定基础, 同时为进一步选育含2~3个代换片段的中高优良染色体片段代换系并应用于育种奠定基础。     

增加穗粒数的水稻染色体代换系Z747鉴定及相关性状QTL定位

增加穗粒数对水稻高产品种培育至关重要。其遗传基础复杂,由多基因控制。水稻染色体片段代换系可以将多基因控制的复杂性状分解,因而是理想的遗传研究材料。本研究通过高代回交和自交结合分子标记辅助选择方法,鉴定了一个以日本晴为受体、西恢18为供体亲本的、含有15个代换片段的增加穗粒数的水稻染色体片段代换系Z747,平均代换长度为4.49 Mb。与受体日本晴相比, Z747的每穗总粒数、一次枝梗数、二次枝梗数、穗长和粒长显著增加,粒宽显著变窄、结实率显著降低,但结实率仍为81%。进一步以日本晴和Z747杂交构建的次级F2群体鉴定出46个相关性状的QTL,分布于水稻1号、2号、3号、5号、6号、9号、11号和12号染色体上。其中qGPP12、qPH-3-1、qPH-3-2等12个QTL可能与已克隆的基因等位, qSPP9等34个可能是新鉴定的QTL。Z747的每穗总粒数由2个具有增加粒数效应的QTL (qSPP3和qSPP5)和1个具有减少粒数效应的QTL (qSPP9)控制。研究结果对主效QTL的精细定位和克隆、以及有利基因的单片段代换系培育有重要意义。     

利用染色体片段代换系定位水稻叶片形态性状QTL

水稻叶片形态是理想株型的重要组成部分,控制叶片形态基因的挖掘对于塑造水稻理想株型,实现水稻超高产目标具有重要意义。本研究利用广陆矮4号为受体亲本,日本晴为供体亲本构建的一套染色体片段代换系,对水稻上三叶(倒一叶、倒二叶和倒三叶)形态性状与单株籽粒产量进行了相关性分析,并开展了相关QTL定位。结果表明,除剑叶宽外,水稻上三叶的叶长、叶宽都与单株产量呈极显著正相关。同时,通过单因素方差分析和Dunnett’s多重比较,在两年间重复检测到20个控制叶形的QTL,其中叶长QTL 13个(8个表现正向效应,5个表现负向效应);叶宽QTL 7个(4个表现正向效应,3个表现负向效应)。这些QTL的鉴定为水稻叶形性状的分子改良提供了重要遗传信息。     

利用染色体片段代换系定位陆地棉株高QTL

以陆地棉中棉所36为轮回亲本和海岛棉海1为供体亲本, 构建染色体片段代换系。为了能检测到稳定的株高QTL,将三个代换系群体(BC₅F₃, BC₅F_3:4和BC₅F_3:5)在5个环境中种植,2009年和2010年分别在河南安阳种植BC₅F₃单株、BC₅F_3:4株行, 2011年分别在河南安阳、辽宁辽阳和新疆石河子种植BC₅F_3:4株系。结果表明,在不同群体环境中株高的超亲比例为53.43%~88.97%。从早期构建的总图距为5088.28 cM, 含有2280个SSR标记位点,覆盖26条染色体的遗传连锁图谱中筛选标记,对408个单株进行的SSR鉴定,结果检测到16个株高QTL,分布在10条染色体上。单个QTL解释的表型变异为7.35%~13.17%。有7个QTL在2个以上环境被检测到。与标记MUSS563紧密连锁的qPH-15-19在一个环境中被检测到,在前人的研究中也有报道。这些结果为进一步精细定位QTL、基因克隆、分子辅助选择等研究奠定基础。     

染色体单片段代换系的构建及应用于QTL精细定位

染色体片段代换系(Chromosome Segment Substitution Lines,CSSLs),又称为代换系(Substitution lines,SLs)或导入系(Introgression lines,ILs)。只含一个代换片段的代换系,即单片段代换系(Single Segment Substitution Lines,SSSLs)是理想的代换系。单片段代换系只有代换片段与受体亲本不同,其它遗传背景与受体亲本完全一致,对代换区段中的QTL进行分析时遗传背景干扰很小,有利于QTL的分析,不少学者利用单片段代换系材料对许多QTL进行了鉴定和精细定位,并克隆了一些重要性状的QTL。本文介绍了染色体单片段代换系构建的原理和利用微卫星标记(SSR）进行单片段代换系鉴定的方法,讨论了代换系构建过程中值得注意的问题,并对用染色体单片段代换系进行QTL精细定位做了展望。     

基于水稻长大粒染色体片段代换系Z66的粒型QTL的鉴定及其聚合分析

水稻籽粒大小是一个复杂的农艺性状,受多基因控制。染色体片段代换系是创造自然变异的有效手段,也是复杂性状研究的理想材料。本研究构建了一个新的水稻长大粒染色体片段代换系Z66, Z66以日本晴的基因组为遗传背景,含有来自R225的12个代换片段,平均代换长度为3.32Mb。然后,以日本晴/Z66创建的次级F₂群体定位出12个控制水稻籽粒大小的QTL,并培育出具有目标QTL的5个新单片段代换系(S1～S5)和4个新双片段代换系(D1～D4)。其中有9个QTL(qGL3、qGL7、qGL10、qGW6、qGW10、qRLW3、qRLW10、qGWT3、qGWT10)可被单片段代换系所验证,表明这些QTL遗传稳定。此外,还利用单片段代换系鉴定到6个新的QTL(qGL9-2、qGW9-2、qRLW6、qRLW7、qRLW9-2、qGWT7)。在这18个QTL中,qGL9-2、qRLW9-1、qRLW9-2、qGW9-2、qGWT9-2可能是新鉴定的QTL。双基因聚合分析表明,不同QTL间聚合产生不同的上位性效应。如qRLW3(a=0.21)和qRLW9-2(a=0.08)聚合...     

利用单片段代换系定位水稻粒形QTL

稻谷粒长、粒宽和长宽比是衡量稻米外观品质的重要指标，稻谷籽粒形状也是影响水稻产量的重要因素。为更好地开展粒形分子育种，对水稻粒形QTL进行了分子定位。以单片段代换系（SSSL）为材料构建分离群体，利用微卫星标记对控制水稻谷粒长和谷粒宽的2个粒形QTL进行分子定位。粒宽QTLGw-8被定位于第8染色体长臂末端微卫星标记RM502与RM447之间，遗传距离均为0．3cM。     

野生稻染色体片段代换系构建及其效应分析

通过回交程序结合分子标记辅助选择构建了一套染色体片段来源于马来西亚普通野生稻的珍汕97B染色体片段代换系。该套染色体片段代换系由105份材料构成,每系含有一个或少数几个导入片段,所有导入片段相互衔接覆盖野生稻全基因组。染色体片段代换系的平均背景回复率为94.6%,平均导入片段长度为41.7cM。利用该群体以及相同亲本的高世代BC3F3群体,共定位到40个QTL影响抽穗期、株高、SPAD值、有效穗数和穗长等农艺性状。该套野生稻染色体片段代换系为发掘和利用野生资源中的优良基因提供重要材料基础。     

利用染色体单片段代换系定位水稻垩白QTL

为了鉴定和定位水稻垩白相关QTL,分析其遗传效应,利用籼稻品种广陆矮4号为受体,粳稻品种日本晴为供体构建的84个染色体单片段代换系群体为试验材料,通过单因素方差分析和Dunnett's多重比较,对代换片段上垩白相关QTL进行鉴定。以P≤0.001为阈值,共检测到36个垩白相关QTL。其中,垩白度QTL共19个,其加性效应值为-6.44~12.86,加性效应百分率为-34.04%~68.02%。垩白粒率相关QTL共17个,其加性效应值为-7.32~3.63,加性效应百分率为-8.07%~4.00%。这些QTL的鉴定为进一步精细定位并克隆相应QTL,提高稻米外观品质奠定了基础。     

基于染色体单片段代换系的水稻粒形QTL定位

水稻的粒形是影响水稻产量和品质的重要因子之一, 是由多基因控制的数量性状。染色体单片段代换系由于减少了个体间遗传背景的干扰, 已经成为鉴定复杂性状QTL的新型遗传材料。本研究以广陆矮4号为受体,日本晴为供体的119个染色体单片段代换系群体为试验材料,通过单因素方差分析和Dunnett’s多重比较,测验单片段代换系与受体亲本之间粒形的差异,鉴定了代换片段上粒形相关的QTL。以P≤0.001为阈值, 共检测到39个粒形相关的QTL。其中,粒长相关的19个,其加性效应值为0.18~1.06 mm,加性效应百分率为2.40%~14.13%;粒宽相关的14个,其加性效应值为0.09~0.31 mm,加性效应百分率为2.71%~9.15%;粒厚相关的6个,其加性效应值为0.05~0.10 mm,加性效应百分率为2.14%~4.46%。这些QTL的鉴定,为进一步精细定位并克隆相应QTL和高产、优质水稻新品种的分子标记辅助选择奠定了基础。     

Quantitative trait loci mapping for flag leaf traits in rice using a chromosome segment substitution line population

To study the genetic basis of rice flag leaf morphology, quantitative genetic analysis was conducted in a population of 37 chromosome segment substitution lines (CSSLs) of indica elite variety ‘Habataki’ in the background of japonica cultivar ‘Sasanishiki’ across three different environments. The CSSLs showed normal distribution with transgressive segregation, indicating that these four traits are controlled by polygenes. Moreover, analyses of variance showed that these traits were highly influenced by the growing environment, which are typical for polygenic quantitative traits. Seven quantitative trait loci (QTLs) on four chromosomes were detected in total: four for flag leaf width, one for flag leaf area and two for flag leaf angle. Two key QTLs, qFLW4 and qFLAG5 controlling flag leaf width and angle, respectively, were identified in all three environments. These QTLs could provide useful information for marker‐assisted selection in improving the performance of plant architecture with regard to leaf angle and area. Moreover, developed CSSLs with these QTLs information are also useful research materials to reveal the importance of leaf morphology in relation to grain yield.     

基于染色体片段置换系群体检测水稻株型性状QTL

株型是由多个形态和生理性状集成的复合性状,它与水稻产量密切相关。挖掘优异株型等位基因或QTL,对水稻超高产育种具有重要意义。本研究利用籼稻昌恢121和粳稻Koshihikari构建的208个染色体片段置换系(chromosome segment substitution lines, CSSLs),在3个环境下,对控制株高、剑叶形态和分蘖数的QTL进行检测,共鉴定到35个株型性状QTL,分布于11条染色体上(除9号染色体以外),解释表型变异2.00%～22.86%。值得关注的是qPH-1-1、qFLW-6和qFLA-3均能在3个环境下被检测到,其中qFLW-6为1个新鉴定到的剑叶宽QTL。对qPH-1-1和qFLA-3位点进行鉴定,验证了这2个位点等位基因的加性效应和环境稳定性。本研究为株型性状QTL的进一步精细定位、克隆及分子辅助聚合育种奠定了基础。     

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.     

利用东乡普通野生稻染色体片段置换系定位产量相关性状QTL

以东乡普通野生稻和日本晴为亲本构建的染色体片段置换系为研究材料, 2019年分别在北京、山东临沂和江西南昌对分蘖数、穗粒数和粒形等11个产量相关性状进行多环境鉴定,结合染色体片段置换系基因型数据定位水稻产量相关性状QTL。3个环境共检测到68个QTL,包括株高4个、穗长5个、分蘖数2个、一次枝梗数7个、一次枝梗粒数8个、二次枝梗数8个、二次枝梗粒数10个、每穗粒数6个、千粒重7个、粒长8个和粒宽3个; LOD值介于2.50～12.66之间,贡献率变幅为4.67%～27.79%,15个QTL的贡献率大于15%;24个QTL与已报道位点/基因位置重叠,44个QTL为新发现位点; 6个QTL在2个环境能被检测到, 1个QTL qTGW2能在3个环境检测到,且是还未报道的新位点。最后,利用BSA法验证了qPH7、qPBPP8-2和qGW10三个QTL的可靠性。本研究将为后续产量相关性状基因克隆以及进一步解析其遗传基础和分子调控机制奠定基础。     

基于染色体片段置换系对水稻粒形及千粒重QTL检测与稳定性分析

粒形及千粒重是水稻产量的重要影响因素,通过挖掘这些性状的优异基因,对水稻超高产育种具有重要意义。本研究利用1套以籼稻恢复系昌恢121为背景亲本,粳稻越光为供体亲本构建的染色体片段代换系为材料,在3个环境下对水稻粒形及千粒重进行QTL检测及稳定性分析,共检测到59个QTL,分布于1号、2号、3号、4号、5号、6号、7号、10号、11号和12号染色体上,贡献率为0.77%～36.26%,其中发现10个QTL多效位点。值得关注的是qGW2-1、qGW2-2、qGW3-1、qGW3-2、qGL3和qGL12这6个QTL能在3个环境中重复检测到,其中qGW3-1为新鉴定的QTL位点。这些结果为进一步开展水稻粒形基因的精细定位、克隆和分子辅助育种奠定了一定的理论基础。     

Construction of high‐throughput genotyped chromosome segment substitution lines in rice (Oryza sativa L.) and QTL mapping for heading date

Chromosome segment substitution lines (CSSLs) provide ideal materials for quantitative trait loci (QTLs) mapping and genetic dissection of complex traits. In this study, we developed a set of CSSL population consisting of 175 lines, which were derived between the recipient ‘Guangluai 4’ and the donor ‘Nipponbare’. Based on 260 molecular markers, we firstly constructed a physical map of core 97 lines. Then, these 97 lines were further genotyped based on resequencing data, and a resequencing‐based physical map was constructed. Compared with the molecular marker‐based physical map, the resequencing‐based physical map of 97 lines contained 367 substituted segments with 252 newly discovered segments. The total size of the 367 substituted segments was 1,074 Mb, which was 2.81 times the size of rice genome. Using the 97 CSSLs as materials, we identified nine QTLs for heading date and three of them were firstly reported. All the QTLs had positive additive effects, ranging from 9.50 to 16.50 days. These CSSLs may greatly help forge a new resource for functional genomics studies and molecular breeding in rice.     

Detection and characterization of epistasis between QTLs on plant height in rice using single segment substitution lines

Hua-jing-xian 74 and its 12 single segment substitution lines (SSSLs) in rice were used as crossing parents to construct a half diallel crossing population. A total number of 91 materials were grown under three planting densities. By analysis of average plant height (PH) over all environments 10 SSSLs were detected with significant additives and 6 SSSLs with significant dominances. These SSSLs were further tested under different densities respectively, indicating that some of single locus effects were sensitive to densities and the conditions under the density of 16.7 cm × 16.7 cm maybe inhibited the expressing of these PH QTLs. Qualitative and quantitative analyses of each four participating genotypes indicated that digenic interactions among these QTLs were prevalent. Of 66 tested interactions, about 42.4% were epistatic (P < 5%). Although some QTLs hadn’t single locus effects, they were possible to form digenic interactions. A significant finding was that the detected epistases were mostly negative. Additionally, these epistases were also found being sensitive to planting densities, the conditions under the density of 10 cm × 16.7 cm perhaps promoted the expressing of epistatic interactions among PH QTLs.     

QTL analysis for soil‐surface roots originating from a New Plant Type rice (Oryza sativa L.)

The introgression line YTH16 harbouring chromosome segments from the New Plant Type cultivar IR65600–87–2‐2–3 with genetic background of an Indica Group rice IR 64 forms soil‐surface roots. To clarify the genetic mechanism, QTL analysis was performed using hybrid populations derived from a cross between IR 64 and YTH16. A total of eight QTLs were detected in the three introgressed segments on chromosomes 2, 5 and 7. Seven chromosome segment lines (CSLs) combining these three QTL regions were selected from the progenies. The two CSLs harbouring a single region (excluding the CSL with a region on chromosome 5) showed high scores and low means of root angle distribution in comparison with IR 64. Four CSLs harbouring two or three regions showed high scores and low means of root angle distribution in comparison with YTH16 and the CSLs harbouring a single QTL region. These results indicated that the soil‐surface rooting of YTH16 was controlled by the three QTLs’ regions and that chr. 5 particularly played a role in supporting the effect with others.