栽培稻种间近等基因系杂种育性研究

 为了研究亚洲栽培稻与非洲栽培稻种间杂种不育的遗传基础，以亚洲栽培稻品种WAB56-104为测验种与来自种间回交组合WAB56-104/CG14//WAB56-104///WAB56-104的14个种间近等基因系测交，对亲本及杂种的花粉及小穗育性研究表明，这14个近等基因系带有来自非洲栽培稻品种CG14的3个互不等位的配子消除位点和2个互不等位的花粉不育基因位点, 其中至少有2个配子消除位点未曾报道过。     

栽培稻种间育性研究进展

杂种不育是影响亚洲栽培稻和非洲栽培稻之间基因交流的主要障碍,栽培稻种间杂交育性表现为数量性状位点控制的复杂遗传模式。杂种不育的遗传基础还需要深入研究。本文综述了育性方面的最新进展并对其实际应用的可能性进行探讨。     

水稻质核互作雄性不育系的微效恢复基因定位和排除方法研究

以232个协青早A/B456///协青早A/B456∥B456植株组成的分离群体为材料,调查花粉可育率和自交结实率,并采用121个在染色体上分布比较均匀的SSR多态性标记进行QTL检测。发现22个SSR标记分别与10个花粉育性位点连锁,分布于第2,5,6,8,10,12染色体上;28个标记分别与16个小穗育性位点连锁,分布于第1,2,4,5,6,8,10和11染色体上;13个标记同时与花粉育性和小穗育性连锁,小穗育性与花粉育性QTL差异是花粉可育度和自交结实率不平行性的遗传基础。各可育位点对花粉育性和小穗育性的提高效应比较小,为微效基因,但每个花粉育性位点的存在都可导致不育系败育不彻底。协青早A中发现1个花粉可育位点Pf5-1,与分子标记Bm55和Rm13紧密连锁,进行分子标记辅助选择,可能排除协青早A的微效恢复基因(可育位点),达到完全不育。协青早A存在8个小穗育性位点,能够提高自交结实率,有助于杂种F1结实率的提高,有利于提高不育系的可恢复性。多数微效恢复基因显示为部分隐性或隐性,是水稻质核互作雄性不育系选育难的重要原因。采用不育系/拟用亲本∥保持系/拟用亲本的方式,观察杂种育性分离,可对拟用亲本的微效恢复基因有所了解,用保持系/部分保持系∥保持系的方式可提高微效恢复基因排除的效率。     

栽培稻(Dryza sativa)杂种不育性的遗传研究 Ⅰ.等基因F_1不育系杂种不育性的双列分析

以栽培稻粳型品种台中65及其5个等基因F_1不育系作双列杂交试验。结果表明,5个等基因系中共带有三组不同位点的F_1不育基因,分别命名为 E2、E3和 E5位点组。E2和E5位点组的作用主要产生染败花粉,而 E3位点组的作用主要产生空败花粉。E3位点组的作用时期早于 E2和 E5位点组。     

栽培稻(Oryza sativa)杂种不育性的遗传研究: Ⅰ.等基因F1不育系杂种不育性的双列分析

 以栽培稻粳型品种台中65及其5个等基因F_1不育系作双列杂交试验。结果表明,5个等基因系中共带有三组不同位点的F_1不育基因,分别命名为 E2、E3和 E5位点组。E2和E5位点组的作用主要产生染败花粉,而 E3位点组的作用主要产生空败花粉。E3位点组的作用时期早于 E2和 E5位点组。     

东南亚与南亚稻属AA基因组种间的遗传多样性差异

选用36对水稻微卫星（SSR）引物,对稻属428份东南亚及南亚AA组种进行遗传多样性分析。试验结果显示选取的SSR标记均具有多态性,多态性位点百分率（P）达100%。36个多态位点共扩增出311个等位基因,每个位点3～17个,平均8.6。Nei基因多样性指数（He）平均为0.650,变幅为0.337（RM455）～0.865（RM169）。东南亚稻属AA组的SSR多样性大于南亚,两地区又以普通野生稻的多样性指数（He）最大。种（类型）间遗传分化东南亚小于南亚,其中以尼瓦拉野生稻与亚洲栽培稻的遗传分化程度最大。特异等位基因的数量、涉及的位点数及频率均表明东南亚及南亚稻属AA组间具有较大的遗传差异,而某些特异位点（如RM161）等位基因所显示的较高频率,则表明该位点较高的鉴别效率。     

水稻粳型亲籼光温敏不育系509S的选育及其杂种优势

水稻籼粳亚种间杂种不育的主要原因在于育性位点上等位基因互作,因此,在籼粳交主要的育性位点上的片段置换,及把籼粳片段相互置换克服生殖障碍进行育种成为可能。本研究将籼稻轮回422S的光温敏不育基因以及籼稻249黄的黄叶形态标记通过回交方法导入到粳稻镇稻88中,同时在S5、S7、S8和S9育性位点上,分别利用4个与育性位点紧密连锁的分子标记RM276、RM455、RM141和RM185进行分子标记辅助选择,最终成功选育出粳稻光温敏不育系509S。该不育系株叶形态表现为粳稻特征,与大量籼稻品种杂交结实率正常,与粳稻杂交结实率反而偏低。选用分布于水稻12条染色体上的SSR标记对509S的遗传背景进行鉴定,实验结果表明不育系509S遗传背景92%为粳稻遗传背景。不育系509S在4个置换的育性位点都为籼稻片段,正是这些籼稻片段的置换使不育系509S与籼稻杂交可育,而与粳稻杂交表现低育,且在不育系与籼稻配制的杂交组合中表现强大的籼粳交杂种优势。该实验结果为籼粳交杂种优势的育种实践利用提供了一条新的可行途径。     

栽培稻种间杂交改良云南粳稻品种研究

为引入非洲栽培稻的有利基因以丰富云南粳稻的遗传基础以达到培育高产、抗病、抗逆云南粳稻品种的目的,利用179个非洲栽培稻品种作母本与6个云南粳稻杂交,并用相应父本回交后形成F1、BC1F1、BC2F1、BC2F2群体,同时对滇粳优1号作轮回亲本的组合在BC2F1中每组合随机用5~10株回交至BC3F1。研究表明,F1及BC1F1的自交结实率为0,种间杂种不育是非洲栽培稻与亚洲栽培稻种间杂交最主要的生殖障碍;但回交至BC2F1自交结实率即达到7.9％, BC3F1的平均结实率为14.6％,在BC2F1选结实率在10％以上的植株自交1次,BC2F2的结实率即为42.8%,变幅15%~80%;表明杂种不育模式符合“单位点孢子体 配子体互作不育”。在BC2F2群体中,10.7％的组合的综合表现优于轮回亲本。大规模育种实践表明,通过两次回交,再自交2~3代,种间杂种不育的障碍基本可得到克服,并可引入非洲栽培稻的有利基因。     

水稻野败胞质低温敏两用系新种质的初步研究

水稻低温敏两用系新资源３６３Ｓ是从三系杂交稻威优６３Ｆ２代分离群体中筛选培育的，具有野败不育胞质。在２４℃以下低温诱导表现稳定不育，２６℃以上高温诱导可育，这两种温度条件下其花粉育性与小穗育性表现一致；在２４－２６℃范围内，随温度升高，可染花粉增加，但小穗仍表现败育；     

利用分子标记辅助选择育成抗螟虫籼稻不育系科龙A

以克螟稻为Bt抗虫基因供体,采用杂交、回交转育方法,结合GUS分子标记辅助选择和田间抗螟虫性选择技术,选育出首个转Bt抗螟虫基因籼型水稻不育系科龙A。经鉴定,科龙A的花器性状和开花习性良好,花粉败育彻底,不育度高,配合力强,高抗螟虫,所配杂种F1代仍保持有高抗螟虫特性。该不育系于2006年8月通过浙江省科技成果鉴定。     

Mapping of Rice Fertility-Restoring Genes for ID-Type Cytoplasmic Male Sterility in a Restorer Line R68

An F2 population derived from the cross Zhong 9NR68 was used to map the fertility-restoring (Rf) gene for ID-type cytoplasmic male sterility (CMS).Two bulks (a fertile bulk and a sterile bulk) were constructed by pooling equal amount of ten highly fertile lines and ten highly sterile lines,respectively.Four hundred and thirteen pairs of simple sequence repeat (SSR) primers,which evenly distributed on 12 chromosomes of rice,were selected for analyzing polymorphisms between the parents and between the two bulks.The primer RM283 on chromosome 1 and the primers RM5756,RM258,RM6100 and RM171 on chromosome 10 were found to be polymorphic between the parents and between the two bulks.These five SSR markers were linked to fertility-restoring genes.A total of 82 excessive sterile lines were selected from Zhong 9NR68 F2 population to estimate the genetic distance between five SSR markers and fertility-restoring genes respectively.The results indicated that one Rf gene was linked to RM283 located on chromosome 1 at a distance of 6.7 cM,and the other Rfgene was mapped to the long arm of chromosome 10 flanked by RM258 and RM6100 at the distances of 8.0 cM and 2.4 cM,respectively.     

两系杂交稻亲本SSR指纹图谱的建立及其在种子纯度鉴定中的应用

利用筛选到的20对SSR引物建立了两系杂交稻32个亲本(24个光温敏不育系和8个恢复系)的DNA指纹图谱,针对所涉及的9个杂交稻组合,获得能在父、母本间表现出多态性的特异SSR标记48个.以杂交稻组合两优932为例,在实验室用一个特异SSR标记(RM302)对200粒种子样本进行了纯度鉴定,结果鉴定纯度为90.50%,与田间种植鉴定结果90.60%(海南鉴定)和91.57%(武汉鉴定)基本一致,表明SSR标记技术适用于构建DNA指纹图谱和种子纯度鉴定.     

水稻印尼水田谷型细胞质雄性不育恢复系R68的恢复基因初步定位

用印尼水田谷型不育系中9A和恢复系R68配组,选取F2的高可育株和极端不育株构建2个基因池,用82个完全不育单株作为定位群体,利用分布于12条染色体的413对SSR引物对双亲和两池进行多态性分析。 位于第1染色体的RM283和位于第10染色体的RM5756、RM258、RM6100、RM171 在亲本、两池间存在多态性,用F2单株验证证明它们与恢复基因连锁。经典遗传分析和分子标记定位研究表明,印尼水田谷型细胞质雄性不育恢复系R68具有2对恢复基因,分别位于第1和第10染色体上。位于第1染色体的恢复基因与分子标记RM283的距离是6.7 cM,位于第10染色体的恢复基因与标记RM5756、RM258、RM6100和RM171间的距离分别是10.4、8.0、2.4和4.2 cM。     

Fine Mapping of C(Chromogen for Anthocyanin)Gene in Rice

Seven residual heterozygous lines(RHLs)displaying different genotypic compositions in the genomic region covering probable locations of C (Chromogen for anthocyanin)gene on the short arm of rice chromosome 6 were selected from the progenies of the indica cross Zhenshan 97B/Milyang 46.Seeds were harvested from each of the seven plants,and the resultant F2:3 populations were used for fine mapping of C gene.It was shown in the populations that the apiculus coloration matched to basal leaf sheath coloration in each plant.By relating the coloration performances of the populations with the genotypic compositions of the RHLS,the C locus was located between rice SSR markers RM314 and RM253.By using a total of 1279 F2:3 individuals from two populations showing coloration segregation.the C locus was then located between RM111 and RM253,with genetic distances of 0.7 cM to RM111 and 0.4 cM to RM253.Twenty-two recombinants found in the two populations were assayed with seven more markers located between RM111 and RM253,including six SSR markers and one marker for the C gene candidate,OsC1.The C locus Was delimited to a 59.3-kb region in which OsC1 was located.     

水稻光温敏雄性不育突变体tms3650的鉴定和基因定位

【目的】雄性不育是水稻杂种优势利用的基础。为进一步解析其调控机制,对一个雄性不育突变体进行了鉴定。【方法】利用⁶⁰Co-γ对籼稻品种93-11进行辐射诱变,从其后代中鉴定到一个雄性不育突变体tms3650。将籼稻明恢63作父本同突变体tms3650杂交构建F₂和F₃群体,采用图位克隆的方法对目的基因进行精细定位。【结果】tms3650突变体其他农艺性状与野生型一致,但花药白绿且瘦小,花粉不能被1%碘-碘化钾溶液染成蓝黑色,穗子包颈,抽穗期延迟。遗传分析表明该突变体表型受一对隐性核基因控制。精细定位结果表明基因位于第3染色体长臂SSR标记RM15927和RM15934之间135.25 kb距离内,且与RM15931标记共分离。陵水冬季南繁鉴定发现,该突变体育性受光温环境影响,说明tms3650突变体的雄性育性在短日低温条件下发生了转育,是一个光温敏雄性不育突变体。【结论】通过将定位位点与已报道的雄性不育基因比较,发现tms3650是一个新的基因位点,暂命名为TMS3650。     

水稻色素原基因C的精细定位

应用籼稻组合珍汕97B/密阳46的衍生材料,针对水稻第6染色体短臂色素原基因C的可能位置,筛选到在C基因周围区间呈不同基因型组合的7个剩余杂合体,收获种子建立F2∶3群体。在各个植株上,稃尖颜色和叶鞘颜色的表现完全相同。通过各个群体颜色表现与原剩余杂合体基因型的比较,将C基因定位于微卫星标记RM314与RM253之间。在该基础上,应用两个分离群体共1279个样本,经标记检测和连锁分析,进一步将C基因定位于RM111和RM253之间, 与RM111和RM253的遗传距离分别为0.7 cM和0.4 cM。最后,应用区间内的另外6个微卫星标记和1个源于C基因候选基因OsC1的标记,检测在RM111 C基因 RM253区间内发生了重组的22个个体,将C基因定位于一个大小为59.3 kb、涵盖C基因候选基因OsC1座位的区间中。     

Molecular Markers and Candidate Genes for Thermo-Sensitive Genic Male Sterile in Rice

The discovery of thermo-sensitive genic male sterility(TGMS) has led to development of a simple and highly efficient two-line breeding system. In this study, genetic analysis was conducted using three F_2 populations derived from crosses between IR68301 S, an indica TGMS rice line, and IR14632(tropical japonica), Supanburi 91062(indica) and IR67966-188-2-2-1(tropical japonica), respectively.Approximately 1:3 ratio between sterile and normal pollen of F_2 plants from the three populations revealed that TGMS is controlled by a single recessive gene. Bulked segregant analysis using simple sequence repeat(SSR) and insertion-deletion(InDel) markers were used to identify markers linked to the tms gene. The linkage analysis based on the three populations indicated that the tms locus was located on chromosome 2 covering the same area. Using IR68301S × IR14632 F_2 population, the results showed that the tms locus was located between SSR marker RM12676 and InDel marker 2gAP0050058. The genetic distance from the tms gene to these two flanking markers were 1.10 and 0.82 cM, respectively.InDel marker 2gAP004045 located between these two markers showed complete co-segregation with the TGMS phenotype. In addition, InDel marker vf0206114052 showed 2.94 cM linked to the tms gene using F_2 populations of IR68301S × Supanburi 91062. These markers are useful tool for developing new TGMS lines by marker-assisted selection. There were ten genes located between the two flanking markers RM12676 and 2gAP0050058. Using quantitative real-time PCR for expression analysis, 7 of the 10 genes showed expression in panicles, and response to temperatures. These genes could be the candidate gene controlling TGMS in IR68301S.     

水稻短根突变体ksr1的遗传分析和基因定位

 从甲基磺酸乙酯诱变的Kasalath突变体库中,在苗期筛选到一个水稻短根突变体 ksr1, 6 d苗龄时该突变体的根长只有野生型的20%左右,遗传分析表明该突变性状由一对隐性核基因控制。利用突变体与粳稻日本晴杂交发展的F2群体对突变基因进行了定位分析,初步定位结果显示目的基因 KSR1 与第4染色体上SSR标记RM1223连锁。在该标记附近进一步发展了8对SSR标记和2对InDel标记,将突变基因定位于InDel标记4 24725K和SSR标记RM17182之间,该区段物理距离为155 kb。     

Fine Mapping of C（Chromogen for Anthocyanin） Gene in Rice

Seven residual heterozygous lines (RHLs) displaying different genotypic compositions in the genomic region covering probable locations of C (Chromogen for anthocyanin) gene on the short arm of rice chromosome 6 were selected from the progenies of the indica cross Zhenshan 97B/Milyang 46. Seeds were harvested from each of the seven plants, and the resultant F_2:3 populations were used for fine mapping of C gene. It was shown in the populations that the apiculus coloration matched to basal leaf sheath coloration in each plant. By relating the coloration performances of the populations with the genotypic compositions of the RHLs, the C locus was located between rice SSR markers RM314 and RM253. By using a total of 1279 F_2:3 individuals from two populations showing coloration segregation, the C locus was then located between RM111 and RM253, with genetic distances of 0.7 cM to RM111 and 0.4 cM to RM253. Twenty-two recombinants found in the two populations were assayed with seven more markers located between RM111 and RM253, including six SSR markers and one marker for the C gene candidate, OsC1. The C locus was delimited to a 59.3-kb region in which OsC1 was located.