豫粳6号保持系与恢复系TR2604间杂种花粉不育基因的定位

阐明BT型杂交粳稻组合间育性差异的遗传基础有助于三系法杂交粳稻组合的选育。根据TR2604与豫粳6号A(B)、9201A(B)后代的花粉育性及小穗育性,明确了豫粳6号A(B)/TR2604 F1不育由双亲间特异性不亲和造成。遗传分析表明豫粳6号A(B)与TR2604 F1花粉不育受单基因S38(t)控制。以352株豫粳6号A/TR2604//TR2604、豫粳6号B/TR2604//豫粳6号B等群体中单株为定位群体,将S38(t)定位于第7染色体上标记RM18和RM234之间,与两标记遗传距离分别为0.43 cM和0.14 cM,两标记间物理距离约为180 kb,相关结果为S38(t)图位克隆工作奠定了基础。     

水稻咪草烟抗性的遗传分析及其紧密连锁分子标记的筛选与应用

抗除草剂水稻的培育及推广能够提高除草效率, 取得巨大经济效益。本研究中金粳818为抗咪草烟资源, 以其与常规粳稻苏垦118杂交产生的F₂群体对抗性基因进行遗传分析与分子定位表明, 其抗性表型受1对显性核基因控制, 位于水稻第2染色体SSR分子标记RM7413和RM7426之间。对该区间候选基因预测和测序发现, 咪草烟靶基因-乙酰乳酸合酶基因(ALS)在重要功能位点发生1个碱基的突变(G变为A), 导致一个氨基酸由丝氨酸(S)突变为天冬酰胺(N), 初步确定ALS是抗性表型的重要候选基因。标记RM7413、RM7426与ALS的物理距离分别为165 kb、1612 kb。以金粳818与南粳9108为亲本, 检测标记RM7413在辅助育种实践中的应用潜力, 对杂交种及其自交后代进行连续表型及标记选择, F₇群体能够稳定遗传抗咪草烟性状, 表明RM7413在粳稻抗咪草烟辅助育种中具有巨大应用潜力。本研究结果为粳稻抗除草剂分子标记辅助改良奠定了基础。     

水稻新质源(CMS-FA)雄性不育恢复基因的遗传研究

发掘水稻新型雄性不育细胞质源CMS-FA,育成系列优质米不育系和系列新质源恢复系,组配成强优势杂交稻组合的基础上研究新质源雄性不育恢复系的恢复基因遗传。采用新质源(CMS-FA)不育系金农1A与恢复系金恢3号杂交获得杂交F₁代种子,种植F₁代,收获自交F₂代种子。用F₁分别与不育系或保持系回交,获得(不育系//不育系/恢复系和不育系/恢复系//保持系)2个测交群体。同时种植P₁、P₂、F₁、F₂、B₁F₁和B₂F₁等群体,考察花粉染色率、套袋结实率和自然结实率,卡平方测验遗传分离适合度。结果表明,不育系与恢复系杂交F₁代正常可育,育性恢复(可育)基因为显性遗传。F₂代分离出可育︰不育适合3︰1,育性恢复(可育)基因为1对显性基因控制。B₁F₁和B₂F₁代2个测交群体的可育︰不育都适合1︰1分离规律,验证了F₂代育性恢复(可育)单基因的遗传模式。暂时确定新质源(CMS-FA)核质互作三系的基因型为不育系S(SS)、保持系F(SS)和恢复系S(FF)。     

水稻抗白叶枯病新基因Xa32（t）的鉴定和初步定位

通过多菌系接种鉴定及抗谱分析,并与目前国际上已知抗白叶枯病基因比较,证明在水稻抗源C4064中含有一个新的抗白叶枯病基因,暂命名为Xa32(t)。应用分离集团分析法(BSA),借助SSR和EST等分子标记,对该基因进行了分子标记定位。通过对F₂分离群体及F₃家系单株进行遗传连锁性检测,发现6个位于水稻第11染色体长臂末端的分子标记RM27256、RM27274、RM2064、ZCK24、RM6293和RM5926与Xa32(t)基因连锁。它们与Xa32(t)基因间的遗传距离分别为2.1、1.0、1.0、0.5、1.5和2.6 cM。其中标记RM6293和RM5926位于染色体近端粒一侧,其他4个标记RM27256、RM27274、RM2064和ZCK24位于基因的另一侧。将Xa32(t)定位在水稻第11染色体长臂末端2.0 cM范围内。     

粳稻恢复系宁恢3-2育性恢复力的遗传

两套粳稻雄性不育系(A)及其保持系(B)分别与宁恢3-2(R)杂交、回交、自交、获得两个组合的 F_1(A/R)、B_2(A//A/R)、B_2′(A//B/R)、F_2和 F_3,以花粉育性和种子育性作为恢复度指标,研究宁恢3-2育性恢复力的遗传。 结果表明,BT 六千辛 A 和 L 平壤3号 A的花粉以染败为主,在光学显微镜下不易区分分离世代植株中可育和不育花粉,     

一个新的水稻花粉半不育性位点的定位分析

利用一套以籼稻珍汕97B为背景的粳稻日本晴染色体片段代换系,鉴定发现1个半不育的代换系。全基因组基因型分析表明,该代换系仅含3个粳稻导入片段,而其他遗传背景与珍汕97B相同。在湖北武汉和海南分别种植其衍生的F₂和F₃分离群体,采用单标记分析和区间作图法分析花粉育性和小穗育性的数量性状位点(QTL),结果表明,该代换系的半不育性是第2染色体上的粳稻导入片段引起的,该片段RM262~RM475区间存在1个新的影响花粉育性的QTL,其贡献率为13.9%。研究结果将为进一步精细定位水稻育性QTL以及鉴定相关功能基因提供重要的试验基础。     

一个水稻开颖不育突变体ohs1(t)的遗传分析及基因定位

我们在明恢86的转基因后代中发现了一个水稻花器官发育突变体,暂命名为开颖不育(open hull sterile 1,ohs1(t)))突变体.ohs1(t)突变体表现颖花开裂,内外稃片变细,内稃微弯向外稃,有雌雄蕊分化,但雌雄蕊较野生犁株的小,大多数花药没有花粉,少数花药中含有不育花粉,雌雄配子均不育.突变体ohs1(t)分别与明恢86、R527、93-11和中花16号杂交后代遗传分析表明,ohs1(t)是一个隐性基因控制的突变体.以ohs1(t)和93-11杂交F2群体中突变个体作为初步定位群体,采用已报道的SSR标记将OHS1(t)初步定位在1号染色体的长臂端RM493和RM5638两个标记间.随后利用已公布的水稻基因组序列(http://rgp.dna.affrc.go.jp/E/index.html)及93-11和日本晴间SSR标记数据库(http://www.gramene.org/),新开发和筛选了SSR和InDel标记,并以ohs1(t))和中花16号杂交F2群体中突变个体作为新定位群体,将OHS1(t)基因进一步定位在NSSR0115和InDel0102之间,遗传距离分别为0.2 cM和0.3 cM,物理距离约66 kb.     

小麦花粉致死基因Ki的SSR标记分析

小麦雄性不育主要是通过花粉的败育表现,其不育材料对小麦杂种优势的利用研究具有重要意义和价值,国外研究表明,某些特定普通小麦品种间杂交F₁表现的花粉部分不育现象,受控于核基因组花粉致死基因Ki,为了筛选小麦花粉致死基因Ki的连锁标记,利用现代分子生物学技术通过定位该基因,克隆出花粉致死基因连锁标记片段,为小麦雄性不育种质材料的转育提供有效的选择标记。对小麦花粉致死基因Ki进行了分子标记定位,以‘中国春’和澳大利亚春小麦品种的BC₁F₁代作为定位群体,利用分离群体分组分析法(BSA)对位于小麦6B染色体上85对SSR引物进行多态性筛选,具有多态性的引物再通过BC₁F₁定位群体进行验证,从中筛选出与目的基因连锁的2个SSR标记Xgwm626和Xgpw4138。运用Mapmaker 3.0软件进行连锁分析。结果表明,Xgwm626和Xgpw4138与Ki基因的遗传距离分别为9.2 cM和6.9 cM,且2个SSR标记位于目的基因两侧,并将Ki定位于小麦6BL染色体上。研究结果为Ki基因的分子标记辅助选择和进一步精细定位奠定了基础。     

粳稻恢复系宁恢3-2育性恢复

两套粳稻雄性不育系(A)及其保持系(B)分别与宁恢3-2(R)杂交、回交、自交、获得两个组合的 F_1(A/R)、B_2(A//A/R)、B_2′(A//B/R)、F_2和 F_3,以花粉育性和种子育性作为恢复度指标,研究宁恢3-2育性恢复力的遗传。 结果表明,BT 六千辛 A 和 L 平壤3号 A的花粉以染败为主,在光学显微镜下不易区分分离世代植株中可育和不育花粉,故不宜用花粉育性作为恢复度指标进行遗传分析。以种子育性为指标,发现两个不育系是配子体不育类型,宁恢3-2对它们的育性恢复力是由一对显性恢复基因控制的。     

大豆质核互作雄性不育系NJCMS3A双亲雄性育性基因的SSR标记

利用大豆质核互作雄性不育系NJMCS3A的质、核供体亲本N21566和N21249构建F₂和BC₁F₁育性分离群体进行雄性育性的遗传分析与基因定位。结果表明, F₁正反交可育,F₂和BC₁F₁的可育株与不育株分离比例经χ²测验分别符合3∶1和1∶1,表明NJCMS3A供体亲本雄性育性由一对基因控制,可育等位基因为显性。该基因可能是NJCMS3A的一个恢复基因。选用793对SSR引物对F₂和BC₁F₁群体分别进行育性基因定位,发现该育性基因位于O连锁群上,在Satt331和Satt477标记之间,与Satt331、CSSR133和Satt477标记距离的次序一致,分别为8.1~10.4 cM、11.4~16.4 cM、13.3~19.2 cM。     

9311与日本晴间一个杂种不育基因的鉴定与定位

以克服亚种间杂种不育来充分发掘亚种间杂种优势是提高水稻单产的一条有效途径。本研究,从一套以日本晴为背景,9311为供体的染色体片段代换系中鉴定出一个系T9424,其与日本晴配置的F₁植株小穗与花粉育性较双亲显著降低,双亲间存在不亲和。重测序结果表明T9424在第1、第4和第5染色体上导入9311片段。日本晴/T9424 F₂群体内单株基因型及育性鉴定结果表明,T9424与日本晴间杂种不育基因位于第5染色体上。利用F₂群体内790株单株将该杂种不育基因定位于第5染色体分子标记PSM8与A14之间110kb的物理区段内。对日本晴/T9424 F₁植株花粉与胚囊育性鉴定结果表明该杂种不育基因同时控制雌、雄配子败育,将该基因暂命名为S39(t)。相关结果有助于加深对水稻亚种间杂种不育现象的认识,为该基因克隆及其育种利用奠定基础。     

Comparative genetic analysis and molecular mapping of fertility restoration genes for WA,Dissi, and Gambiaca cytoplasmic male sterility systems in rice

The genetic relationship among three cytoplasmic male sterility (CMS) systems, consisting of WA, Dissi, and Gambiaca, was studied. The results showed that the maintainers of one CMS system can also maintain sterility in other cytoplasmic backgrounds. The F₁ plants derived from crosses involving A and R lines of the respective cytoplasm and their cross-combination with other CMS systems showed similar pollen and spikelet fertility values, indicating that similar biological processes govern fertility restoration in these three CMS systems. The results from an inheritance study showed that the pollen fertility restoration in all three CMS systems was governed by two independent and dominant genes with classical duplicate gene action. Three F₂ populations, generated from the crosses between the parents of good-performing rice hybrids, that possess WA, Dissi, and Gambiaca CMS cytoplasm, were used to map the Rf genes. For the WA-CMS system, Rf3 was located at a distance of 2.8 cM from RM490 on chromosome 1 and Rf4 was located at 1.6 cM from RM1108 on chromosome 10. For the Dissi-CMS system, Rf3 was located on chromosome 1 at 1.9 cM from RM7466 and Rf4 on chromosome 10 was located at 2.3 cM from RM6100. The effect of Rf3 on pollen fertility appeared to be stronger than the effect of Rf4. In the Gambiaca-CMS system, only one major locus was mapped on chromosome 1 at 2.1 cM from RM576. These studies have led to the development of marker-assisted selection (MAS) for selecting putative restorer lines, new approaches to alloplasmic line breeding, and the transfer of Rf genes into adapted cultivars through a backcrossing program in an active hybrid rice breeding program.     

籼粳亚种间杂种育性相关基因座全基因组分析

利用一套以粳稻品种Asominori为遗传背景、籼稻品种IR24为染色体片段供体的覆盖全基因组的CSSL群体，研究了籼粳亚种间组合Asominori/IR24和02428/IR24杂种小穗低育性的遗传基础。结果发现，Asominori/IR24组合的育性主要受第5染色体上的2个育性位点S-24(t)和S-31(t)及第6染色体上的 S-5位点控制，其中S-31(t)为本研究发现的新育性位点，粳稻品种02428带有该位点的亲和性基因。02428/IR24组合的低育性主要受S-24(t)花粉育性位点的影响。育性基因的表达受遗传背景的影响，在粳稻遗传背景中，S-24(t)位点处在Sⁱ/S^j杂合基因型时可使杂种小穗育性下降70%左右，而S-31(t)和 S-5为杂种半不育位点。在籼粳全基因组杂合遗传背景中，当S-5ⁱ/S-5^j基因型置换成S-5ⁱ/S-5ⁱ基因型后，亚种间杂种小穗育性可平均提高22.5%，接近正常育性水平。在S-5ⁱ/S-5^j遗传背景中，S-24(t)和S-31(t)的Sⁱ/Sⁱ纯合基因型不能改善亚种间杂种的小穗育性。说明S-5位点是影响亚种间小穗育性的关键位点，在亚种间杂交稻育种中，必须首先克服S-5位点造成的育性障碍。提出了等位基因置换法克服水稻籼粳亚种间杂种小穗低育性的技术策略。     

Molecular Mapping of a Pollen Killer Gene S29(t) in Oryza Glaberrima and Co-Linear Analysis with S22 in O. Glumaepatula

Two species in genus Oryza, O. glaberrima and O. glumaepatula, are valuable and potential sources of useful genes of interest for rice improvement. However, the hybrid sterility between O. sativa and these two species is a main reproduction barrier when transferring the favorable traits/genes to mbox{O. sativa.} To overcome it, the nature of hybrid sterility should be understood further. The objective in the report is to map a new hybrid sterility gene as a Mendelian factor from O. glaberrima and analyze the co-linear of hybrid sterility S loci mbox{between} mbox{O. glaberrima} and mbox{O. glumaepatula} via comparative mapping approach. A BC₂F₂ population, derived from a single semi-sterility plant of BC₂F₁ of WAB56-104/ WAB450-11-1-2-P41-HB (WAB450-6) //WAB56-104///WAB56-104 was employed to map this pollen killer in O. glaberrima since WAB450-6 is a progeny of interspecific hybrid between O. sativa and O. glaberrima. A new pollen killer locus, S29(t) in O. glaberrima, was identified and mapped to interval between SSR marker RM7033 (1.1 cM) and RM7562 (1.3 cM) on rice chromosome 2. Comparative mapping indicated that S29(t) closely corresponded to S22 which is also a pollen killer gene in O. glumaepatula and is tightly linked with RFLP marker S910 on the short arm of rice chromosome 2. The good co-linear between S29(t) and S22 implied that there might exist common (orthologous) hybrid sterility loci controlled the reproduction barrier among AA genome species of genus Oryza, which will contribute significantly to our understanding of speciation and operation of hybrid sterility between O. sativa and its AA genome relatives.     

A new gene controlling hybrid sterility between Oryza sativa and Oryza longistaminata

Oryza longistaminata is an excellent gene pool to improve Asian cultivated rice Oryza sativa. However, hybrid sterility between O. longistaminata and O. sativa hinders application of desirable genes in rice production. In order to further understand the nature of interspecific hybrid sterility between these two species, near isogenic lines (NILs) were raised with RD23, an indica cultivar from Thailand, as background and an accession of O. longistaminata as donor. Using two NIL/RD23 F₂ populations, a new pollen sterility gene, denoted as HYBRID STERILITY 44(t) or S44(t) in abbreviation, was mapped between simple sequence repeat markers RM5814 and RM20695 on the long arm of rice chromosome 6. The genetic pattern of S44(t) follows one-locus allelic interaction model. The interaction between S44(t)-sativa and S44(t)-longistaminata leads to partial abortion of male gametes carrying the allele of S44(t)-sativa in the heterozygotes, but the spikelet fertility is not affected by S44(t).     

Mapping three new interspecific hybrid sterile loci between Oryza sativa and O. glaberrima

Hybrid sterility hinders the transfer of useful traits between Oryza sativa and O. glaberrima. In order to further understand the nature of interspecific hybrid sterility between these two species, a strategy of multi-donors was used to elucidate the range of interspecific hybrid sterility in this study. Fifty-nine accessions of O. glaberrima were used as female parents for hybridization with japonica cultivar Dianjingyou 1, after several backcrossings using Dianjingyou 1 as the recurrent parent and 135 BC₆F₁ sterile plants were selected for genotyping and deducing hybrid sterility QTLs. BC₆F₁ plants containing heterozygous target markers were selected and used to raise BC₇F₁ mapping populations for QTL confirmation and as a result, one locus for gamete elimination on chromosome 1 and two loci for pollen sterility on chromosome 4 and 12, which were distinguished from previous reports, were confirmed and designated as S37(t), S38(t) and S39(t), respectively. These results will be valuable for understanding the range of interspecific hybrid sterility, cloning these genes and improving rice breeding through gene introgression.     

水稻温敏核雄性不育系培矮64S广亲和位点S-5的SSR紧密连锁标记

广亲和(WC)是水稻(OryzasativaL.)亚种间杂种优势利用的重要遗传工具。本研究对我国南方大面积推广应用的亚种间两系杂交稻亲本培矮64S的WC基因S-5n进行分子标记定位。从培矮64S//T8/秋光三交F1中选出由263个高育和低育极端类型组成的标记群体,选用来源于CornellSSR连锁图的20个标记和根据GenBank数据库公布的序列合成的9个SSR标记,通过BSA(分离群体分析)法对标记群体进行分析,该S-5位点精确定位于第6染色体上,距SSR标记GXR6和RM276的距离只有0.2cM。培矮64S的S-5n对育性贡献可使小穗结实率从平均43.5%提高到77.5%,贡献率达34.0%,表明它是一个很强的控制广亲和特性的主效基因。本研究所获得的这些紧密连锁标记对分子标记辅助选择培育广亲和水稻品种和基因克隆具有重要的利用价值。     

一个水稻雄性不育突变体的遗传分析和基因定位

ms-np是一个源于自然突变的水稻雄性不育突变体,明显较正常植株矮小,叶色浓绿。小花解剖观察发现,突变体小花花丝细长,花药干瘪,呈白色透明状,但雄性器官的数量和雌性器官正常。碘染证实,突变体的花药壁内没有花粉粒着色,是一个典型的无花粉型雄性不育材料。5个F₂和2个BC₁F₁群体的遗传分析显示,该突变性状受1对隐性基因控制。对组合ms-np/M63衍生F₂不育单株的连锁分析表明,ms-np(t)基因位于水稻第6 染色体微卫星标记RM541和RM343之间,遗传距离分别为15.2 cM和7.9 cM。     

温光敏核不育小麦C412S的育性转换及其APRT基因的表达

以选育的小麦(Triticum aestivum L.)温光敏核不育系C412S为试材,以C412S回交转育时的受体亲本C412为常规品系对照,通过分期播种试验研究了其育性转换特性。用涂抹压片法观察减数分裂和小孢子发育进程,用半定量RT-PCR技术分析不育与可育条件下不同发育时期的幼穗中腺嘌呤磷酸核糖基转移酶基因(APRT)的表达水平。结果表明,通过调整播种期改变雄性发育的温光条件,C412S表现出完全不育—高不育—半不育—正常可育的育性转换特性。C412S花粉败育的高峰在单核小孢子晚期,主要表现圆败型不育。C412S的育性敏感期是从花粉母细胞形成期到成熟花粉期,其中最敏感的时段是花粉母细胞形成期到减数分裂期。与对照相比,C412S的APRT1基因序列有个别碱基变异,但编码氨基酸序列没有变化。在花粉母细胞形成期至单核期,与晚播可育条件相比,早播不育的C412S幼穗中APRT基因转录水平下调,因此认为,其育性转换与幼穗中APRT基因转录水平有一定关系。