Mapping of QTLs Related to Grain Weight Using Chromosome Single Segment Substitution Lines in Rice

Grain weight, one of the major factors determining rice yield, is a typical quantitative trait control ed by multiple genes. With Guangluai 4 as recipient and Nipponbare as donor, a population of 119 chromosome single segment substitution lines had been developed. Correlation analysis between grain weight and grain shape by SPSS revealed that 1 000-grain weight shared extremely significant posi-tive correlation with grain length and length-width ratio, but no significant correlation with grain width and thickness. The QTL analysis of grain weight was carried out using one-way analysis of variance and Dunnett＇s test. Nineteen stable QTLs re-sponsible for grain weight were identified over two years. Al 19 QTLs were identi-fied on al chromosomes except for chromosome 10 and 12 at a significance level of P≤0.001. Among them, 10 QTLs had a positive effect and were derived from the Nipponbare al ele, the additive effect of these QTLs ranged from 0.49 to 2.74 g, and the contributions of the additive effects ranged from 2.00% to 11.05%. Another 9 QTLs had a negative effect and were al derived from Guangluai 4 al ele, the ad-ditive effect of these QTLs ranged from 0.60 to 2.35 g, and the contributions of the additive effects ranged from 2.40% to 9.84%. The results provide a basis for the fine mapping and gene cloning of novel locus associated with rice grain weight.

Unconditional and Conditional QTL Mapping for Tiller Numbers at Various Stages with Single Segment Substitution Lines in Rice(Oryza sativa L.)

Tiller is one of the most important agronomic traits which influences quantity and quality of effective panicles and finally influences yield in rice. It is important to understand ＂static＂ and ＂dynamic＂ information of the QTLs for tillers in rice. This work was the first time to simultaneously map unconditional and conditional QTLs for tiller numbers at various stages by using single segment substitution lines in rice. Fourteen QTLs for tiller number, distributing on the corresponding substitution segments of chromosomes 1, 2, 3, 4, 6, 7 and 8 were detected. Both the number and the effect of the QTLs for tiller number were various at different stages, from 6 to 9 in the number and from 1.49 to 3.49 in the effect, respectively. Tiller number QTLs expressed in a time order, mainly detected at three stages of 0-7 d, 14-21 d and 35-42 d after transplanting with 6 positive, 9 random and 6 negative expressing QTLs, respectively. Each of the QTLs expressed one time at least during the whole duration of rice. The tiller number at a specific stage was determined by sum of QTL effects estimated by the unconditional method, while the increasing or decreasing number in a given time interval was controlled by the total of QTL effects estimated by the conditional method. These results demonstrated that it is highly effective and accurate for mapping of the QTLs by using single segment substitution lines and the conditional analysis methodology.

Identification of QTLs for Plant Height and Its Components by Using Single Segment Substitution Lines in Rice (Oryza sativa)

QTLs for plant height and its components on the substituted segments of fifty-two single segment substitution lines （SSSLs） in rice were identified through t-test （P〈0.001） for comparison between each SSSL and recipient parent Huajingxian 74. On the 14 substituted segments, 24 QTLs were detected, 10 for plant height, 2 for panicle length, 4 for length of the first internode from the top, 5 for length of the second internode from the top and 3 for length of the third internode from the top, respectively. All these QTLs were distributed on nine rice chromosomes except chromosomes 5, 9 and 11. The additive effect ranged from -4.08 to 3.98 cm, and the additive effect percentages varied from -19.35% to 10.43%.

Identification of Multiple Alleles at the Wx Locus and Development of Single Segment Substitution Lines for the Alleles in Rice

The microsatellite markers 484/485 and 484/W2R were used to identify the multiple alleles at the Wx locus in rice germplasm. Fifteen alleles were identified in 278 accessions by using microsatellite class and G-T polymorphism. Among these alleles, （CT）12-G, （CT）15-G, （CT）16-G, （CT）17-G, （CT）18-G and （CT）21-G have not been reported. Seventy-two single-segment substitution lines （SSSLs） carrying different alleles at the Wx locus were developed by using Huajingxian 74 with the （CT）11-G allele as a recipient and 20 accessions containing 12 different alleles at the Wx locus as donors. The estimated length of the substituted segments ranged from 2.2 to 77.3 cM with an average of 17.4 cM.

QTL Detection for Rice Grain Shape Using Chromosome Single Segment Substitution Lines

Rice grain shape is one of the important factors affecting grain quality and yield,but it is liable to be influenced by genetic backgrounds and environments.The chromosome single segment substitution lines(SSSLs) in rice have been considered as ideal populations to identify the quantitative trait loci(QTLs).In this study,22 QTLs affecting rice grain shape were detected to be distributed on eight chromosomes except chromosomes 6,9,11 and 12 by using SSSLs.Among them,seven QTLs conditioned grain length,six conditioned grain width,five affected grain length-width ratio and four controlled grain thickness.

QTL Detection and Epistasis Analysis for Heading Date Using Single Segment Substitution Lines in Rice(Oryza sativa L.)

Heading date of rice is a key agronomic trait determining cultivated areas and seasons and affecting yield. In the present study, ifve primary single segment substitution lines with the same genetic background were used to detect quantitative trait loci （QTLs） for heading date in rice. Two QTLs, qHD3 and qHD6 on the short arm of chromosome 3 and the short arm of chromosome 6, respectively, were identiifed under natural long-day （NLD）. Nineteen secondary single segment substitution lines （SSSLs） and seven double segments pyramiding lines were designed to map the two QTLs and to evaluate their epistatic interaction between them. By overlapping mapping, qHD3 was mapped in a 791-kb interval between SSR markers RM3894 and RM569 and qHD6 in a 1 125-kb interval between RM587 and RM225. Results revealed the existence of epistatic interaction between qHD3 and qHD6 under natural long-day （NLD）. It was also found that qHD3 and qHD6 had signiifcant effects on plant height and yield traits, indicating that both of the QTLs have pleiotropic effects.

Mapping of Hd-6-2 for Heading Date Using Two Secondary Segregation Populations in Rice

Heading date is one of the most important traits for rice adaption to different cultivation areas and crop seasons. In this study, two single segment substitution lines(SSSLs), W31-41-61-3-11-3-6-7(W31-SSSL) and W32-59-80-2-11-1-10(W32-SSSL) with substituted intervals derived from the donor parents IR66897 B(W31) and IR66167-27-5-1-6(W32), respectively, with Huajingxian 74(HTX74) were found to comprise a gene for extremely late-heading date, and the gene was tentatively designated as Hd-6-2. Two secondary F2 segregating populations were developed by crossing the two heterozygous SSSLs with HJX74 to map Hd-6-2 gene. According to phenotype analysis of the two mapping populations, the late heading date trait was controlled by a major recessive gene. In the segregation population derived from W31-SSSL, Hd-6-2 was mapped on chromosome 6 between PSM677 and RM204 with the genetic distances of 1.3 and 2.7 c M, respectively. In the population of W32-SSSL, the gene for heading date was mapped to the similar region as Hd-6-2 and co-segregated with PSM672. The sequence alignment of Hd3 a in the coding domains and promoter regions of HJX74 and W31-SSSL are completely consistent, whereas there was a great difference between W32-SSSL and HJX74, suggesting that Hd3 a could hardly be the main cause of the heading date variation in W31-SSSL, but it was probably the main reason for the change of heading stage in W32-SSSL.

展颖野生稻株高性状的遗传与SD1基因的等位变异分析

为了探究SSSL-B50和华粳籼74(HJX74)株高差异的原因,利用SSSL-B50与HJX74回交构建F2群体进行株高性状遗传分析与基因定位。结果显示,F1植株表现为高秆;260个单株组成的F2群体株高出现分离,高秆植株与半矮秆植株分离比例符合3∶1(χ^(2)=0.18<3.84),说明SSSL-B50高秆性状为显性性状,受1对显性等位基因控制。利用IciMapping软件对F2进行连锁分析,将SSSL-B50携带的高秆基因定位在1号染色体代换片段上的标记S18和X161之间38.38~39.07 Mb。候选基因筛选分析发现,定位区间内存在着“绿色革命”基因座位SD1。对展颖野生稻、SSSL-B50及HJX74进行SD1基因测序及序列比对,发现HJX74的CDS序列与展颖野生稻、SSSL-B50相比有280 bp的缺失,导致氨基酸编码提前终止。qRT-PCR结果表明,SD1表达量在SSSL-B50茎秆的第2,3,4节均显著高于HJX74。此外,与前人报道结果相比,展颖野生稻SD1基因编码的氨基酸序列发生了2处改变(E100G、Q339R)。研究结果弄清楚了展颖野生稻单片段代换系SSSL-B50高秆性状受SD1调控,同时从展颖野生稻鉴定到了SD1的新等位型SD1Glu。

野生稻单片段代换系苗期耐旱性评价及QTL鉴定

【目的】干旱是影响水稻生产力的主要非生物因素之一,筛选抗旱水稻可有效保障水稻产量。【方法】选取以南方野生稻和展颖野生稻为供体、‘华粳籼74’(HJX74)为受体构建的93份单片段代换系(Single-segment substitution lines,SSSLs)为供试材料,以HJX74为对照,采用20%PEG-6000模拟干旱条件进行苗期耐旱性试验,以4个生长发育性状指标(相对苗高、相对苗干质量、相对根干质量、相对根长)作为评价指标,采用隶属函数法对SSSLs进行排序、筛选相关性状,并筛选与HJX74差异显著的SSSLs,进行QTL鉴定与加性效应分析,初步鉴定与苗期耐旱性状相关的QTL。【结果】4个生长发育性状指标与平均隶属函数值的相关性分析结果均为极显著正相关,可作为SSSLs苗期耐旱性鉴定和评价指标。隶属函数法分析结果显示,93份SSSLs平均隶属函数值范围为0.20~0.71,HJX74平均隶属函数值为0.54;有29个SSSLs的平均隶属函数值> HJX74的平均隶属函数值,其中,M124的隶属函数值最大,表明M124具有较强的耐旱性。通过单因素方差分析,从8个SSSLs(M78-1、M78-2、M124、M107、M151、M103、M130、M115)鉴定出7个相对苗高QTLs(qRSH1-1、qRSH1-2、qRSH2-1、qRSH3-1、qRSH3-2、qRSH5-1、qRSH6-1),分别分布在1、2、3、5、6号染色体,其加性效应为0.05~0.06,表型贡献率为6.97%~9.01%;从3个SSSLs(M79、M145、M148)中鉴定出3个相对根干质量QTLs(qRRDW10-1、qRRDW11-1、qRRDW11-2),分别分布在10、11号染色体,其加性效应为0.12~0.20,表型贡献率为12.51%~19.95%;从2个耐旱SSSLs(M80、X149)中鉴定出1个相对苗干质量QTL(qRSDW5-1),分布在5号染色体,加性效应为0.07,表型贡献率为10.5%。【结论】筛选出13份SSSLs携带苗期耐旱QTLs,为后续苗期耐旱QTLs精细定位与克隆研究奠定基础。

水稻单片段代换系代换片段的QTL鉴定

利用以台中 6 5为受体、窄叶青和低脚乌尖为供体培育的 10个水稻单片段代换系为材料 ,对代换片段上2 1个重要农艺性状的QTL进行了鉴定。 10个代换片段的总长度为 2 30 0 0cM ,占水稻基因组总长度的 12 6 2 %。通过t测验比较单片段代换系与受体亲本的表型差异 ,以P≤ 0 0 0 1为阈值 ,在 10个代换片段上共鉴定出 17个性状的 5 7个QTL ,平均每个性状鉴定出 3.35个 ,这些QTL分布于第 1、3、5、7、8、10和 12染色体上。 5 7个QTL的加性效应百分率在 1 10 %～ 89 73%之间 ,其中 15个QTL的加性效应百分率大于 10 % ,30个QTL在 3%～ 10 %之间 ,12个QTL小于 3%。

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0.18～1.06mm,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0.09～0.31mm,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0.05～0.10mm,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。

籼稻背景的单片段代换系群体的构建

以国内、外12个水稻品种（包括8个籼稻品种和4个粳稻品种）为供体亲本，利用回交和微卫星标记辅助选择相结合的方法构建了一个以籼稻品种“华粳籼74”为遗传背景的单片段代换系（SSSL）群体。该群体由260个不同的SSSL组成，其代换片段分布在水稻的12条染色体上，长度分布于0．2—109．9cM。平均长度为19．9cM；代换片段的总长度为5166．0cM，相当于水稻基因组总长度的3．4倍，代换片段在水稻基因组上的覆盖率为83．8％。这些SSSL的代换片段包含了水稻基因组丰富的等位基因变异，将在基因（QTL）的定位、克隆、功能分析及水稻分子育种中具有重要的利用价值。

水稻恢复基因Rf3和Rf4聚合效应分析

为选育水稻野败型和夜公型细胞质雄性不育的较强恢复系和分析恢复基因Rf3和Rf4的聚合效应,采用5个携带不同供体恢复基因Rf3和4个携带不同供体恢复基因Rf4的单片段代换系两两杂交,配制了11个杂交组合。利用分子标记辅助选择从这11个杂交组合的F2和F3群体中筛选鉴定6个携带基因型Rf3Rf3/Rf4Rf4的双片段聚合系和5个携带Rf3（Rf4）的次级单片段代换系。结果表明,对于野败型不育细胞质,这些双片段聚合系具有不同的恢复性。其中,携带基因型Rf3-4Rf3-4/Rf4-4Rf4-4的DSPL04-01/23-10P1、DSPL04-01/23-10P2和DSPL04-01/23-10P3的恢复性最强,携带基因型Rf3-4Rf3-4/Rf4-2Rf4-2的DSPL11-01/14-10P和DSPL22-01/14-10P的恢复性最弱,携带基因型Rf3-4Rf3-4/Rf4-3Rf4-3的DSPL22-01/27-10P的恢复性居中。5个次级单片段代换系代换片段的总长度为133.9 cM,长度分布范围为24.1～28.5 cM,平均长度为26.8 cM。利用恢复性最强的DSPL04-01/23-10P1及其相应的单片段代换系分别与野败型不育系‘珍汕97A’和夜公型不育系‘Y华农A’杂交,通过考察F1杂种株的花粉育性,研究DSPL04-01/23-10P1中的恢复基因Rf3和Rf4的聚合效应。DSPL04-01/23-10P1较其相应的单片段代换系能够提高F1杂种株的花粉育性,但差异不显著。结论：本研究选育的双片段聚合系中所聚合的来自不同供体的恢复基因Rf3和Rf4之间没有互作效应。

基于SSSL的水稻重要性状QTL的鉴定及稳定性分析

目的单片段代换系(SSSL)是通过高代回交和分子标记辅助选择构建的,只含有来自供体亲本的一个染色体片段,遗传背景与受体亲本相同的品系。本研究的目的是利用SSSL检测不同环境条件下水稻重要性状的QTL。方法以32个SSSL为材料,随机区组试验设计,在2～4个季节中对水稻22个重要性状的QTL进行分析。结果共鉴定出59个QTL,分布于第1、2、3、4、6、7、8、10和11号染色体上。其中的18个QTL能够在2次以上重复检出,稳定性较好的QTL占检出QTL的30.5%,大多数农艺性状的QTL效应较小、稳定性较差。不同的性状,QTL稳定性不同,千粒重、粒长、谷粒长宽比、抽穗天数等性状的QTL较稳定。稳定性好的QTL,不仅具有较大的加性效应,而且受环境影响较小。结论利用单片段代换系可以有效地对水稻重要性状的QTL进行多年多季的稳定性分析。水稻大多数重要农艺性状QTL的不稳定性,反映了水稻生长发育过程的可塑性,可能是通过栽培措施使水稻品种获得高产优质的重要遗传基础。

水稻亚种间单片段代换系的建立

利用微卫星标记辅助选择 ,建立了 2 9个以台中 6 5为受体、低脚乌尖或窄叶青为供体的单片段代换系 ,该代换系群体的代换片段分布于 9条染色体上 ,总长度为 6 43.6 5cM ,平均长度为 2 2 .19cM ,覆盖全基因组 42 7.5 5cM ,覆盖率为2 3.47%。

利用高代回交和分子标记辅助选择建立水稻单片段代换系

以水稻品种华粳籼74为受体,以6个水稻品种为供体,通过高代回交和微卫星标记辅助选择相结合的方法,建立了水稻的一个单片段代换系群体。该群体由86个单片段代换系组成,其中52个在BC3F2中获得,34个在BC3F3中获得。每个单片段代换系只含有来自一个供体的一个染色体代换片段,而遗传背景与华粳籼74相同。这些单片段代换系的代换片段分布于水稻的12条染色体,代换片段的长度为1.5～56.3cM,平均长度为23.0cM。全部代换片段在水稻基因组上的覆盖率为57.1%。

水稻单片段代换系芽期和苗期耐冷性分析及耐冷性QTL鉴定

对籼型两系骨干亲本9311(受体)和粳稻日本晴(供体)构建的单片段代换系群体,在芽期和苗期分别进行低温处理,以芽期处理后的恢复成活率和苗期卷叶程度作为耐冷性的评价指标进行耐冷性评价。结果表明,日本晴在芽期和苗期耐冷性极显著强于9311,不同代换系芽期和苗期耐冷性也存在极显著差异,芽期和苗期耐冷性较强的代换系X724、X732、X733较好地保持了9311的农艺性状,其产量等性状的一般配合力优于9311;鉴定出18个芽期耐冷性QTL和5个苗期耐冷性QTL,分布于12条染色体上的20个代换片段中,第12染色体的RM1261-RM519和RM17区域同时检测出芽期和苗期耐冷性QTL;芽期耐冷性QTLqCTP9、qCTP11.2、qCTP12.1和苗期耐冷性QTLqCTS1.1、qCTS1.2具有较大加性效应,对应代换系耐冷性较强。

水稻粒长基因GS3在聚合育种中的效应

为探索利用水稻粒长基因GS3以及单片段代换系进行谷粒长性状设计育种的可行性,本研究利用以籼稻品种"华粳籼74"为遗传背景且携带谷粒长基因GS3的单片段代换系与携带其它优良基因的单片段代换系杂交进行了分子聚合育种。在F4获得了26份含GS3和其它优良基因的纯合聚合系,谷粒长测定证实这些聚合系都获得了目标谷粒长表型,有效地改良了华粳籼74的外观品质。研究结果表明利用单片段代换系和GS3可以实现谷粒长的设计育种。

利用单片段代换系定位水稻抽穗期QTL

抽穗期是水稻品种的重要农艺性状之一,对抽穗期QTL进行定位并研究其遗传效应在水稻育种中是至关重要的。本研究利用以6个水稻品种为供体的52个单片段代换系为试验材料,通过t测验比较单片段代换系与受体亲本华粳籼74之间抽穗期的差异,对代换片段上的抽穗期QTL进行了鉴定。以P≤0.001为阈值共鉴定出20个抽穗期QTL,这些QTL分布于水稻的10条染色体。QTL加性效应值为–5.9～1.1,加性效应百分率为–7.4%～1.4%。有8个QTL被定位在小于10.0cM的区段内。利用1个单片段代换系与华粳籼74杂交发展的F2群体对qHD-3-1进行了定位。在作图群体中,早抽穗和迟抽穗植株数符合3:1的分离比,早抽穗表现为显性。利用微卫星标记将qHD-3-1定位于3号染色体短臂,PSM304和RM569分别位于其两侧,遗传距离分别为2.4cM和5.1cM。

利用单片段代换系的测交群体定位玉米籽粒性状杂种优势位点

【目的】粒型性状是玉米百粒重的重要组成部分,并具有较强的杂种优势,鉴定玉米粒型性状的杂种优势位点,可以为粒型性状关键杂种优势位点的克隆与分子标记辅助育种提供理论依据。【方法】利用一套中国优良自交系lx9801背景的昌7-2单片段代换系为基础材料,与自交系T7296测交构建包含184个测交种的群体。2013年,在河南浚县和长葛两点对测交群体、单片段代换系群体等材料进行了田间鉴定,完全随机区组设计,3个重复,成熟后连续收获10株对粒长、粒宽、粒厚和百粒重进行考种。利用方差分析和t测验比较每个SSSL×T7296测验种单个性状与对照种T7296×lx9801的显著性差异,对玉米粒型性状的杂种优势位点进行定位。【结果】单片段代换系的两个亲本在粒型性状上表现出明显的差异,昌7-2的粒长和百粒重高于lx9801,而粒厚低于lx9801。测交群体4个籽粒性状均表现出一定的杂种优势,粒长的平均中亲优势在长葛和浚县点分别为19.32%和15.30%,粒宽的中亲优势分别为10.86%和10.07%,粒厚的中亲优势分别为6.23%和4.78%,百粒重的中亲优势分别为20.97%和25.09%。相关分析结果表明,粒长与粒宽、百粒重呈显著正相关,粒宽和粒厚与百粒重也呈显著的正相关,粒宽与粒厚呈负相关关系。在2个环境中定位了13个粒长的杂种优势位点,其中1杂种优势位点同时被检测到。粒宽定位到14个杂种优势位点,在长葛点和浚县点分别检测到1个主效的杂种优势位点。粒厚检测到25个杂种优势位点,在第2、3、7和9染色体上分别定位到1个共同的杂种优势位点。百粒重在2种环境中共检测到24个杂种优势位点,有6个杂种优势位点同时被检测到,其中在第1染色体上检测到2个共同的杂种优势位点h KW1a和h KW1b;在第7染色体上检测一个主效的杂种优势位点h KW7a,在长葛点和浚县的贡献分别为20.12%和11.03%;位于第8染色体的杂种优势位点h KW8b在2种环境中的贡献率分别为18.64%和8.76%。【结论】玉米粒型性状的杂种优势表现依次为粒长>粒宽>粒厚,在2种环境中共检测到75个杂种优势位点,其中11个杂种优势位点被同时检测到。由于粒长与脱籽率显著相关,而且在育种过程中难以对籽粒性状的杂种优势进行预测,因此在育种过程中可以通过分子标记对粒长的杂种优势进行选择,从而选育出脱籽率高的优良玉米杂交种。