基于染色体片段置换系的野生稻粒宽QTL-qGW8.1的精细定位

利用以栽培稻9311为受体、普通野生稻为供体的染色体单片段置换系CSSL182,检测到一个与粒宽相关的QTL。CSSL182与受体亲本9311粒型性状差异显著,且只在8号染色体有一个野生稻导入片段。构建CSSL182/9311的F_2次级分离群体,将粒宽QTL初定位在8号染色体的标记RM447和RM264之间,贡献率达22.49%,将该QTL命名为qGW8.1。随后进一步设计区间内多态性分子标记引物,检测F_2群体的2000株分离个体以及F_(2∶3)群体交换单株,结合后代表型验证,最终将qGW8.1精细定位到8号染色体10 kb区间内。该区间内含有3个候选基因,基因测序发现这3个基因在双亲之间均含有丰富的变异。对双亲子粒颖壳细胞电镜扫描观察发现,CSSL182的颖壳细胞宽度比9311减少16.7%。这一结果表明qGW8.1中来自野生稻的等位基因通过改变颖壳细胞形状影响粒型。     

南瓜遗传连锁图谱的构建及粒宽性状的QTL定位

以印度南瓜纯系大粒材料‘0515-1’和小粒材料‘0460-1-1’为亲本,获得193个南瓜F2单株群体,应用AFLP和SSR分子标记技术进行多态性筛选,构建了含84个标记位点的遗传连锁图谱。结果表明,整个图谱包含12个连锁群,全长683.50cM,标记平均间距为8.13cM。采用复合区间定位分析,共检测到控制南瓜籽粒宽度的4个数量性状位点(QTL),分别位于3个连锁群上,各QTL的贡献率在2.87%～29.68%之间。     

基于水稻染色体片段代换系的耐盐发芽QTL分析

为了检测水稻种子的耐盐相关数量性状位点,也为耐盐遗传机理的研究提供理论基础,本实验以Koshihikari(受体)和Nona Bokra(供体)为亲本构建的全基因组单片段代换系(154个)作为研究群体,采用培养皿培养,在水稻种子的萌发期进行浓度为1％的盐胁迫,以种子的发芽情况为指标,统计发芽率数据,利用分子标记技术,定...     

水稻粒长QTL定位与主效基因的遗传分析

该研究利用短粒普通野生稻矮杆突变体和长粒栽培稻品种KJ01组配杂交组合F_1,构建分离群体F_2;并对该群体粒长进行性状遗传分析,利用平均分布于水稻的12条染色体上的132对多态分子标记对该群体进行QTL定位及主效QTLs遗传分析,为进一步克隆新的主效粒长基因奠定基础,并为水稻粒形育种提供理论依据。结果表明:(1)所构建的水稻杂交组合分离群体F_2的粒长性状为多基因控制的数量性状。(2)对543株F_2分离群体进行QTL连锁分析,构建了控制水稻粒长的连锁遗传图谱,总长为1 713.94 cM,共检测出24个QTLs,只有3个表现为加性遗传效应,其余位点均表现为遗传负效应。(3)检测到的3个主效QTLs分别位于3号染色体的分子标记PSM379～RID24455、RID24455～RM15689和RM571～RM16238之间,且三者对表型的贡献率分别为54.85%、31.02%和7.62%。(4)在标记PSM379～RID24455之间已克隆到的粒长基因为该研究新发现的主效QTL位点。     

基于CSSL的水稻抽穗期QTL定位及遗传分析

抽穗期是水稻（Oryza sativa）品种的重要农艺性状之一,适宜的抽穗期是获得理想产量的前提。鉴定和定位水稻抽穗期基因/QTL,分析其遗传效应对改良水稻抽穗期至关重要。以籼稻品种9311（Oryzasativa ssp.indica‘Yangdao 6’）为受体,粳稻品种日本晴（Oryza sativa ssp.japonica‘Nipponbare’）为供体构建的94个染色体片段置换系群体为材料,以P≤0.01为阈值,对置换片段上的抽穗期QTL进行了鉴定。采用代换作图法共定位了4个控制水稻抽穗期的QTL,分别位于第3、第4、第5和第8染色体;QTL的加性效应值变化范围为–6.4––2.7,加性效应百分率变化范围为–6.4%––2.7%;qHD-3和qHD-8加性效应值较大,表现主效基因特征。为了进一步定位qHD-3和qHD-8,在目标区域加密16对SSR引物,qHD-3和qHD-8分别被界定在第3染色体RM3166–RM16206之间及第8染色体RM4085–RM8271之间,其遗传距离分别为13.9cM和6.4cM。研究结果为利用分子标记辅助选择改良水稻抽穗期奠定了基础。     

利用染色体片段置换系定位水稻落粒性主效QTL

水稻落粒性是与其生产密切相关的重要性状之一。以7个染色体片段置换系为材料,采用重叠群代换作图法对控制落粒性的2个主效QTL进行定位。结果表明,104个SSR标记在亲本间具有多态性,多态率为68.0%;4个置换系的落粒性与亲本日本晴的落粒性相似,表现难落粒。3个置换系与亲本93-11的落粒性相似,表现易落粒;7个染色体片段置换系在第1和第6染色体上检出7个置换片段,其长度分别为23.6、16.5、6.6、9.9、10.4、20.2和7.1 cM;qSH-1-1被定位在第1染色体RM472-RM1387之间,遗传距离约为6.6 cM。qSH-6-1为新发现的落粒性主效QTL,被定位在第6染色体RM6782-RM3430之间,遗传距离约为4.2 cM。利用染色体片段置换系能准确地定位水稻落粒性QTL,qSH-1-1与qSH-6-1的鉴定和初步定位为其进一步的精细定位、图位克隆及分子标记辅助选择奠定了基础。     

利用染色体片段置换系定位水稻落粒性主效QTL

水稻落粒性是与其生产密切相关的重要性状之一。以7个染色体片段置换系为材料, 采用重叠群代换作图法对控制落粒性的2个主效QTL进行定位。结果表明, 104个SSR标记在亲本间具有多态性, 多态率为68.0%; 4个置换系的落粒性与亲本日本晴的落粒性相似, 表现难落粒。3个置换系与亲本93-11的落粒性相似, 表现易落粒; 7个染色体片段置换系在第1和第6染色体上检出7个置换片段, 其长度分别为23.6、16.5、 6.6、 9.9、 10.4、 20.2和7.1 cM; qSH-1-1被定位在第1染色体RM472－RM1387之间, 遗传距离约为6.6 cM。qSH-6-1为新发现的落粒性主效QTL, 被定位在第6染色体RM6782－RM3430之间,遗传距离约为4.2 cM。利用染色体片段置换系能准确地定位水稻落粒性QTL, qSH-1-1与qSH-6-1的鉴定和初步定位为其进一步的精细定位、图位克隆及分子标记辅助选择奠定了基础。     

基于染色体置换系的普通野生稻耐盐性QTL定位

本研究分别利用SSR标记、InDel标记以及简化基因组测序技术对一套以普通野生稻为供体亲本,9311为受体亲本的染色体置换系进行基因型鉴定,并通过对其不同生育期的耐盐性鉴定,共发掘2个与发芽期耐盐相关的QTL,13个与苗期耐盐性相关的QTL。其中与苗期存活率相关的QTL qSSR5.1、苗期耐盐等级相关的QTL qSSG5.1均被定位于同一位点,该位点对苗期存活率、苗期耐盐等级均具有增效作用,贡献率分别为6.36%、8.13%。在此QTL内包含与非生物胁迫相关基因OsDi19-1。经序列比对发现,OsDi19-1启动子区域在两亲本间存在较大差异,且受到盐胁迫时该基因表达量上升。同时,鉴定出水稻芽期耐盐的优异种质资源CSSL72、苗期耐盐的优异种质资源CSSL23、CSSL153,为水稻育种中耐盐性的改良提供了新的种质资源。     

覆盖野生稻基因组的染色体片段替换系的构建及其米质相关数量性状基因座位的鉴定

染色体片段替换系（CSSL）是基因组水平快速初步定位数量性状基因座位（QTL）的良好材料,而水稻的品质性状是多基因控制的数量性状,因此可用替换系鉴定控制水稻品质性状的QTL。本文用分子标记辅助选择技术（MAS）构建了由133个株系组成的以‘特青’（籼稻品种）为轮回亲本,以海南的一种普通野生稻为供体亲本,覆盖绝大部分野生稻基因组的染色体片段替换系。利用这套替换系,初步定位了控制稻米外观和理化品质性状的15个QTL,为今后水稻品质性状QTL的克隆以及稻米品质相关性状的改良提供了依据。     

基于CSSL的水稻抽穗期QTL定位及遗传分析

抽穗期是水稻(Oryza sativa)品种的重要农艺性状之一, 适宜的抽穗期是获得理想产量的前提。鉴定和定位水稻抽穗期基因/QTL, 分析其遗传效应对改良水稻抽穗期至关重要。以籼稻品种9311(Oryza sativa ssp. indica ‘Yangdao 6’)为受体,粳稻品种日本晴(Oryza sativa ssp. japonica ‘Nipponbare’)为供体构建的94个染色体片段置换系群体为材料, 以P≤0.01为阈值, 对置换片段上的抽穗期QTL进行了鉴定。采用代换作图法共定位了4个控制水稻抽穗期的QTL, 分别位于第3、第4、第5和第8染色体; QTL的加性效应值变化范围为–6.4 – –2.7, 加性效应百分率变化范围为–6.4%– –2.7%; qHD-3和qHD-8加性效应值较大, 表现主效基因特征。为了进一步定位qHD-3和qHD-8, 在目标区域加密16对SSR引物, qHD-3和qHD-8分别被界定在第3染色体RM3166–RM16206之间及第8染色体RM4085-RM8271之间, 其遗传距离分别为13.9 cM和6.4 cM。研究结果为利用分子标记辅助选择改良水稻抽穗期奠定了基础。     

陆地棉背景的Pima棉染色体片段置换系创制

染色体片段置换系(CSSLs,chromosome segment substitution lines)基因组内只有1个或少数几个纯合的供体亲本染色体片段,而其余部分与受体亲本相同,是进行QTL分析的理想材料。本研究以陆地棉中棉所8号(CCRI8)为受体亲本,海岛棉Piam90-53为供体亲本在BC3F1-3借助分子标记辅助选择培育了一套182个株系构成的染色体片段置换系。这套置换系置换片段平均长度21.0 c M,总长度19957.8 c M,是棉花基因组总长度4168.7 c M的4.7倍。每个株系内置换片段长度不一,最短为0.7 c M,最长是83.2 c M,导入片段数量为1~11个。CSSLs在纤维品质性状上的分布表现为相对连续的正态分布,部分株系较CCRI8有了明显提高。本研究为进一步开展棉花纤维品质QTL定位以及陆地棉纤维品质育种研究提供了新材料。     

利用CSSLs群体研究稻米粒型QTL的表达稳定性

利用Asominori×IR2 4的染色体片段置换系 (CSSLs)群体 ,对稻米粒长、粒宽和长宽比进行连续两年及 4个地点的QTL表达稳定性分析。结果表明 :3个性状“两年四点”的表现型都为连续分布 ,均存在超亲遗传类型 ;共检测到 13个粒型相关QTL ,其中在 8个环境中都能被重复检测到的QTL有 6个 ,即影响粒长的 qGL 3、控制粒宽的qGW 5a和 qGW 5b以及共同作用于长宽比的qLWR 3、qLWR 5a和 qLWR 5b。这 6个QTL对应置换系的相应性状与背景亲本Asominori的表现型差异在 8个环境中都达到极显著水平 (P <0 0 0 1) ,且同一QTL对应置换系相应性状的表现型在不同环境间呈显著正相关 (r≥ 0 75 ,r0 0 5=0 6 6 6 ) ,说明这 6个QTL表达稳定性较高。由于 qGL 3和 qLWR 3均位于R19 C16 77标记区间 ,qGW 5a和 qLWR 5a位于C2 6 3标记附近 ,qGW 5b和 qLWR 5b被定位在R5 6 9标记附近 ,因此R19、C16 77、C2 6 3和R5 6 9这 4个RFLP标记对优良水稻外观品质的标记辅助选择 (MAS)育种有着重要作用     

水稻粒形性状的遗传及相关基因定位与克隆研究进展

作物育种的首要目标是提高产量。水稻粒形是与水稻产量性状直接相关,与品质性状存在着密切关系的数量性状,其评价指标主要是粒长、粒宽、粒厚、长/宽和长/厚。近年来,水稻粒形的数量遗传研究取得了重要进展,并成功定位克隆了一批控制水稻粒形的基因。文章综述了水稻粒形的经典遗传研究、QTL定位、粒形基因的克隆和功能分析以及在水稻超高产育种中的利用。     

QTL mapping of grain length in rice (Oryza sativa L.) using chromosome segment substitution lines

Chromosome segment substitution (CSS) lines have the potential for use in QTL fine mapping and map-based cloning. The standard t-test used in the idealized case that each CSS line has a single segment from the donor parent is not suitable for non-idealized CSS lines carrying several substituted segments from the donor parent. In this study, we present a likelihood ratio test based on stepwise regression (RSTEP-LRT) that can be used for QTL mapping in a population consisting of non-idealized CSS lines. Stepwise regression is used to select the most important segments for the trait of interest, and the likelihood ratio test is used to calculate the LOD score of each chromosome segment. This method is statistically equivalent to the standard t-test with idealized CSS lines. To further improve the power of QTL mapping, a method is proposed to decrease multicollinearity among markers (or chromosome segments). QTL mapping with an example CSS population in rice consisting of 65 non-idealized CSS lines and 82 chromosome segments indicated that a total of 18 segments on eight of the 12 rice chromosomes harboured QTLs affecting grain length under the LOD threshold of 2.5. Three major stable QTLs were detected in all eight environments. Some minor QTLs were not detected in all environments, but they could increase or decrease the grain length constantly. These minor genes are also useful in marker-assisted gene pyramiding.     

Characterization of a rice variety with high hydraulic conductance and identification of the chromosome region responsible using chromosome segment substitution lines

Background and Aims

The rate of photosynthesis in paddy rice often decreases at noon on sunny days because of water stress, even under submerged conditions. Maintenance of higher rates of photosynthesis during the day might improve both yield and dry matter production in paddy rice. A high-yielding indica variety, ‘Habataki’, maintains a high rate of leaf photosynthesis during the daytime because of the higher hydraulic conductance from roots to leaves than in the standard japonica variety ‘Sasanishiki’. This research was conducted to characterize the trait responsible for the higher hydraulic conductance in ‘Habataki’ and identified a chromosome region for the high hydraulic conductance.

Methods

Hydraulic conductance to passive water transport and to osmotic water transport was determined for plants under intense transpiration and for plants without transpiration, respectively. The varietal difference in hydraulic conductance was examined with respect to root surface area and hydraulic conductivity (hydraulic conductance per root surface area, L_p). To identify the chromosome region responsible for higher hydraulic conductance, chromosome segment substitution lines (CSSLs) derived from a cross between ‘Sasanishiki’ and ‘Habataki’ were used.

Key Results

The significantly higher hydraulic conductance resulted from the larger root surface area not from L_p in ‘Habataki’. A chromosome region associated with the elevated hydraulic conductance was detected between RM3916 and RM2431 on the long arm of chromosome 4. The CSSL, in which this region was substituted with the ‘Habataki’ chromosome segment in the ‘Sasanishiki’ background, had a larger root mass than ‘Sasanishiki’.

Conclusions

The trait for increasing plant hydraulic conductance and, therefore, maintaining the higher rate of leaf photosynthesis under the conditions of intense transpiration in ‘Habataki’ was identified, and it was estimated that there is at least one chromosome region for the trait located on chromosome 4.     

基于CSSLs群体定位和图位克隆水稻长芒基因GAD1-2

水稻是世界上最早驯化的重要粮食作物之一。水稻芒可以保护水稻种子不被鸟琢食,是水稻重要的驯化性状之一。芒在野生稻中普遍存在,对野生稻的生存和传播至关重要,然而在驯化和人工选择过程中该性状逐渐被淘汰。定位和克隆水稻长芒相关基因是研究水稻芒驯化遗传机制的基础。本研究以籼稻恢复系东南恢810为受体、漳浦野生稻为供体构建的146个染色体片段置换系(chromosome segment substitution lines, CSSLs)为研究材料,调查了146个CSSLs株系和双亲的芒长,结果表明在4个置换系中检测到1个控制水稻芒长主效基因GAD1-2,位于水稻第8号染色体;利用重叠代换作图法,将GAD1-2定位在Ind8-10和RM4936标记之间,遗传距离约为4.75 Mb。选择分离群体中的显性单株,利用开发的标记,最终将GAD1-2 基因定位在两个 Indel 标记之间,两者间的物理距离约为27 kb,该区域内只有两个候选基因Os08g0485500和Os08g0485400。经测序和分析表明,Os08g0485500是GAD1-2的候选基因,GAD1-2在保守的ORF区域存在6个碱基缺失,导致丝氨酸和半胱氨酸这两个氨基酸缺失,从而表现长芒的性状;在Os08g0485500基因位点已克隆了1个控制水稻芒长的GAD1基因,推测GAD1-2与GAD1为等位基因。本研究为进一步理解水稻起源演化和水稻芒长发育基因的遗传机制奠定了基础。     

Fine mapping of the region on wheat chromosome 7D controlling grain weight

We report the fine mapping of the previously described quantitative trait loci (QTL) for grain weight QTgw.ipk-7D associated with microsatellite marker Xgwm1002-7D by using introgression lines (ILs) carrying introgressions of the synthetic wheat W-7984 in the genetic background of the German winter wheat variety ‘Prinz’. The BC₄F₃ ILs had a 10% increased thousand grain weight compared to the control group and the recurrent parent ‘Prinz’, and 84.7% of the phenotypic variance could be explained by the segregation of marker Xgwm1002-7D, suggesting the presence of a gene modulating grain weight, which was preliminarily designated gw1. It was possible to delimit the QTL QTgw.ipk-7D to the interval Xgwm295–Xgwm1002, which is located in the most telomeric bin 7DS4-0.61-1.00 in the physical map of wheat chromosome arm 7DS. Furthermore, our data suggest the presence of a novel plant height-reducing locus Rht on chromosome arm 7DS of ‘Prinz’. Larger grain and increased plant height may reflect the pleiotropic action of one gene or may be caused by two linked genes. In general, our data support the concept of using nearly isogenic ILs for validating and dissecting QTLs into single Mendelian genes and open the gateway for map-based cloning of a grain-weight QTL in wheat.     

QTL mapping of protein content in rice using single chromosome segment substitution lines

Protein content (PC) is an important component of rice nutritional quality. In order to better understand the genetic basis of this trait and increase related breeding efficiency, 21 single chromosome segment substitution (SCSS) lines grown in four sites over two growing seasons (regarded as eight environments) were used to associate PC with particular chromosome segments. Segments from 15 chromosomes were found to contain quantitative trait loci (QTLs) for PC in at least one environment. These included segments from chromosome 3 and 8, in which QTLs for PC had not previously been identified. The segment of chromosome 8 in CSSL-48 had the largest positive effect across all environments. The interaction between substitution and environment was highly significant. Some substitutions had large effects in one environment, but no effect in another (i.e. CSSL-08 and CSSL-17), while some substitutions significantly increased PC in one environment but decreased it in another (i.e. CSSL-41 and CSSL-43). By biplot and clustering analysis, the eight environments were grouped into two contrasting environment types, that is, Hainan and Jiangsu. The segment of chromosome 8 in CSSL-48 had PC-enhancing QTLs in both of the environment types. The segments in CSSL-34 had QTLs which increase PC in the Jiangsu environment but have no effect in the Hainan environment. For enhancing PC, CSSL-48 could be explored in breeding for wide adaptation across all environments, while CSSL-12, CSSL-14, CSSL-17, CSSL-41 and CSSL-43, and that in CSSL-34 could be explored in breeding for specific adaptation to the Hainan and Jiangsu environments, respectively. Near isogenic lines are under development to validate the QTLs with large effects in a range of genetic backgrounds relevant to Jiangsu and Hainan breeding programs. Secondary mapping populations are also being developed for further localising the responsible QTLs in CSSL-14, CSSL-34 and CSSL-48.     

稻米外观品质性状遗传与分子定位研究进展

稻米外观品质主要是指稻米的粒形、垩白、透明度和籽粒色泽等,它不仅直接影响到人们的喜好,还与其他品质性状诸如蒸煮食用、加工等密切相关。因此,外观品质对稻米的商品价值有着十分重要的影响。本文从经典遗传与现代分子生物学两个方面对稻米主要外观品质的遗传研究进展进行了较全面的综述,包括粒长、粒宽、长宽比、粒厚、垩白、透明度和籽粒色泽等。综合近年来的遗传研究结果发现,大多数稻米外观品质性状都是由数量基因控制的。利用分子标记技术已将控制外观品质的QTL（qualitative trait locus）定位在不同的染色体上,为下一步的稻米外观品质改良提供了有利条件。     

Identification of quantitative trait loci controlling rice mature seed culturability using chromosomal segment substitution lines

The genetic transformation efficiency of a rice variety is largely determined by its tissue culturability. Establishment of a highly efficient tissue-culture system has greatly accelerated the wide spread application of transgenic japonica varieties. However, such process for indica rice was hampered because this type of variety is recalcitrant to in vitro culture. This study aimed to map the quantitative trait loci (QTLs) for mature seed culturability using a chromosomal segment substitution lines (CSSL) population derived from a cross between an indica variety “Zhenshan 97B” and a japonica variety “Nipponbare”. The CSSLs consist of 139 lines each containing a single or a few introgression segments, and together covering the whole “Nipponbare” genome. Every CSSL was tested by culturing on the two medium systems developed for the respective indica and japonica parental varieties. The performance of culturability was evaluated by four indices: frequency of callus induction (CIF), callus subculture capability (CSC), frequency of plant regeneration (PRF) and the mean plantlet number per regenerated callus (MNR). All four traits displayed continuous variation among the CSSLs. With the culture system for japonica rice, three CIF QTLs, three CSC QTLs, three PRF QTLs and three MNR QTLs were detected. With the culture system for indica variety, six CIF QTLs, two CSC QTLs, three PRF QTLs and six MNR QTLs were identified, and these QTLs distributed on nine rice chromosomes. Two QTLs of CIF and two QTLs of MNR were detected in both the japonica and indica rice culture system. The correlation coefficients of all the four traits varied depending on the culture systems. These results provide the possibilities of enhancing the culturability of indica rice by marker-assisted breeding with those desirable alleles from the japonica. Lina Zhao and Hongju Zhou have contributed equally to this work.