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。     

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。     

构建水稻优良恢复系背景的重叠片段代换系及其效应分析

通过回交程序结合分子标记辅助选择构建了一套以优良籼稻恢复系9311为背景、导入片段来源于粳稻日本晴的代换系群体。该套群体由125个代换系组成,每系含有单一或少量导入染色体片段,导入片段间相互重叠或衔接能覆盖粳稻全基因组。代换系的平均背景回复率为98.4%,导入片段平均长度为20.9 cM,纯合和杂合导入片段分别占水稻基因组的1.4%和0.2%。利用该群体,两年共检测到31个QTL影响水稻穗重、穗长、结实率和秃顶等性状;导入片段QTL对穗重和结实率均起减效作用。该套重叠片段代换系将为重要性状的基因定位、功能鉴定以及籼粳杂交育种研究提供极有价值的遗传材料。     

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。     

以珍汕97B和9311为背景的导入系构建及其筛选鉴定

以我国主要推广的优良杂交稻亲本（珍汕97B和9311）为受体亲本,以从国际合作网络所征集的品种资源150多份为供体材料,进行了大规模杂交、3-4次回交,1～2次自交,培育珍汕97B和9311为背景的近等基因导入系近5000多份。通过对导入系材料进行品质、耐盐、耐旱、磷高效、氮高效利用等性状的筛选鉴定,获得了大量目标性状的导入系。通过对品质性状导入系的基因型分析,定位了影响粒型的15个染色体区段（或基因）。同时,构建了一套来源于粳稻日本晴的单片段导入系群体。创建的近等基因导入系（群体）为目标基因的发掘、新品种的培育提供重要的材料基础。     

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...     

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)。     

乙烯与水稻细胞质雄性不育的关系

从幼穗发育的IV到VII期,水稻细胞质雄性不育系(珍汕97A)幼穗和叶片的ACC含量和乙烯释放速率均高于其保持系(珍汕97B)。外施乙烯释放剂乙烯利使保持系花粉可育度明显下降;外施ACC合成酶抑制剂AVG引起两系幼穗ACC含量和乙烯释放速率下降,并使不育系花粉育性得以部分恢复,而在外施AVG的同时再施以乙烯利则AVG的恢复作用消失。     

籼型三系不育系69A选育过程

<正>籼型三系不育系69A(曾用名绣丰A)是用优ⅠB与博B杂交,经系统选育,在F5代挑选优良的早熟株与珍汕97A进行回交转育而成。该不育系的不育性和农艺性状稳定,达到了实用不育系的要求。选育过程69A不育系和保持系的选育共利用了3个亲本:优IB和博B、珍汕97A。2000年利用优IB与博B杂交,经系统选育,在F5代挑选优良的早熟单株与珍汕97A进行回交转育。具体选育过程如下。2000年春季在南昌用采用温汤除雄法以优IB为母本     

豫粳6号B与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,两标记间物理距离为180 kb,相关结果为S38(t)图位克隆工作奠定了基础。     

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。     

水稻第1染色体长臂上微效千粒重QTL qTGW1.2的验证与分解

本文报道了水稻第1染色体长臂上微效千粒重QTL qTGW1.2的验证和分解。针对前期qTGW1.2定位结果, 应用SSR标记检测, 从籼籼交组合珍汕97³/密阳46衍生的1个BC₂F_{7分离群体中, 筛选到杂合区间分别为RM11621-RM297和RM212-RM265的2个单株, 构建了两套BC2F8:9近等基因系, 将qTGW1.2进一步界定在RM212-RM265及其两侧交换区间的区域内。}在此基础上, 筛选出5个在目标区间内分离片段缩小且呈阶梯状排列的单株, 衍生了5套BC₂F₁₀分离群体, 应用Windows QTL Cartographer 2.5进行QTL分析。结果表明, 每套群体均检测到千粒重QTL, 加性效应为0.13~0.38 g, 来自密阳46的等位基因提高千粒重; 经比较各个群体的分离区间, 将qTGW1.2分解为互引连锁的2个QTL, 其中, qTGW1.2a位于RM11730和RM11762之间934 kb的区域内, 呈加性作用, qTGW1.2b位于RM11800和RM11885之间2.1 Mb的区域内, 呈正向超显性。     

Validating a segment on the short arm of chromosome 6 responsible for genetic variation in the hull silicon content and yield traits of rice

Rice is a typical silicon-accumulating plant and the beneficial effect of silicon on rice has long been recognized. In a previous study using 244 recombinant inbred lines (RILs) of an indica rice cross, Zhenshan 97B/Milyang 46 grown in 2003, four QTLs were detected for hull silicon content. QTL qHUS-6 had the largest effect among these, and the same interval also had significant effects on yield traits in the same population. The primary objective of this study was to validate the QTL effect in this region on HUS and yield traits. The same RIL population and another RIL population of lower heterogeneity were grown in 2004. QTL qHUS-6 was found to have significant additive effects on hull silicon content with a consistent direction in the two populations. From a residual heterozygous line selected from RILs of the same cross, 15 F_2:3 lines that differed only in a 2.15-Mb segment extending from RM587 to RM6119 on the short arm of chromosome 6 were derived. In these lines, qHUS-6 displayed a major effect, so did QTLs for yield traits previously detected in the same region. Two more QTLs for HUS detected in 2003, qHUS-1-1 and qHUS-1-2, also had consistent effects in the Zhenshan 97B/Milyang 46 RIL population in 2004. Thus this study verified three candidate regions for fine mapping HUS QTLs and determining the genetic relationship between silicon content and yield traits in rice.     

Breeding of widely compatible restorer lines in rice (Oryza sativa L.) through anther culture

Following anther culture of various F₁ hybrids of indica restorer (R) lines/wide compatible varieties (WCVs) and japonica R lines/WCVs, the homozygous diploid plants (2n) generated were test-crossed with indica WA type cytoplasmic-genetic male sterile (CMS) line Zhanshan 97A (WA), indica testers Nanjing 11 and Nante, japonica BT CMS line Hanfeng A (BT), and japonica testers Balilla and Akihikari to identify widely compatible restorer lines. The results of this study showed that among the diploid pollen plants generated from F₁ hybrids of indica R lines/WCVs, 36.7% and 64.7% possessed normal fertility restoration ability (rate of seed-setting > 80%) to Hanfeng A and Zhanshan 97A, respectively. 71.3% and 32.3% had normal fertility restoration ability to Hanfeng A and Zhanshan 97A, respectively, in diploid pollen plants derived from the japonica R lines/WCVs F₁'s anther culture. Several widely compatible R lines were selected from anther culture of F₁ hybrids of indica R lines/WCVs and japonica R lines/WCVs. These widely compatible R lines derived from diploid pollen plants showed good wide compatibility and restoration ability both to WA and BT type CMS lines. Strong standard heterosis of major agronomic traits and yield traits was observed in F₁ test-crosses of widely compatible R lines with Hanfeng A and Zhanshan 97A. The wide compatibility in widely compatible R lines H17, H158 and H281 was studied by a set of three-way crosses. Segregation of the fertile plants with seed-setting rate > 70% and semi-fertile plants with seed-setting rate > 69.9% agreed with a ratio of 1:1, indicating that wide compatibility in these widely compatible R lines is governed by a pair of major genes. This revised version was published online in July 2006 with corrections to the Cover Date.     

A genetic analysis of low-temperature-sensitive sterility in indica-japonica rice hybrids

Low-temperature-sensitive sterility has become one of the major obstacles in indica–japonica hybrid rice breeding. The objectives of this paper were to evaluate the extent of the fertility reduction and to determine the genetic basis of low-temperature-sensitive sterility. Seventeen varieties were crossed in various ways to produce 21 F₁s including 16 indica-japonica hybrids. Fertility of the F₁s and their parents was examined under both high and low temperature conditions. Considerable reduction in spikelet fertility was observed under low-temperature conditions in the majority of the indica–japonica hybrids having at least one wide compatibility parent. However, the extent of fertility reduction varied greatly, depending on the parental genotypes. Data from five pairs of reciprocal crosses indicated that the cytoplasm had no effect on fertility reduction. The more-or-less bimodal distribution of the fertility segregation of one BC₁F₁ and two F₂: populations under low-temperature conditions suggested that the low-temperature-sensitive sterility was controlled by only one or a few genes. It was also shown that the low-temperature-sensitivity is not related to wide compatibility. We conclude that it is possible to develop indica-japonica hybrids with wide compatibility and also insensitivity to the low-temperature conditions.     

利用籼粳交探讨水稻株高和抽穗期的遗传基础

广亲和基因的发现,克服了籼粳交杂种不育的矛盾,但杂种后代株高过高、抽穗期延迟等问题仍然限制着籼粳亚种间杂种优势的利用。本试验利用一个籼粳交(珍汕97/武育粳2号)双单倍体群体(doublehaploidpopulation,DH系)及其分别与两亲本回交构建的两个回交群体,考查了株高和抽穗期的遗传,将三个群体的表型值和两个回交群体的中亲优势值进行了数量性状位点(quantitativetraitlocus,QTL)检测及效应分析,并对结果进行了比较。2个性状在3个相关群体中一共检测到了21个主效QTL和10个上位性QTL,主效QTL的数目和贡献率都比上位性QTL大,而且,在10个上位性QTL中,发生在2个主效QTL间的互作有3个,主效QTL与背景位点间的互作有6个,2个互补位点间的互作仅有1个,进一步说明了主效QTL在株高和抽穗期遗传中的重要作用。比较加性和非加性QTL的作用,发现加性效应、显性效应和上位性效应是籼粳亚种间杂种株高和生育期变化的共同遗传基础。     

渐渗系IL6的遗传评价和渗入片段的鉴定

 用海岛棉3-79的6号染色体渐渗系(简称IL6)和背景亲本TM-1构建了F₂、F_2：3群体,在2年的田间重复实验中调查了18个重要农艺性状,作遗传评价。并获得F₂群体的分子数据,进行QTL定位和单标记分析。结果表明:亲本IL6中的渐渗片段,组成有3-79的6号染色体,3-79的非6号染色体和大部分的TM-1片段。IL6的产量、品质性状都优于受体亲本TM-1,遗传分析也发现产量、品质性状增效基因位点来自IL6,说明所渗入片段起着关键作用。亲本农艺性状表现和遗传分析结果表明,IL6渗入片段含有总铃数、单株铃数、衣分、果枝数、比强度、麦克隆值等性状的主基因。而QTL定位和单标记分析结果显示,IL6渗入片段含有单株铃数、衣分、果枝数、株高、子指、整齐度等性状的QTLs。     

Molecular mapping of S-c, an F1 pollen sterility gene in cultivated rice

Hybrids between indica and japonica rice varieties usually show partial sterility, and are a major limiting factor in the utilization of heterosis at subspecific level. When studying male-gamete (pollen) abortion, a possibly important cause for sterility, six loci (S-a, S-b, S-c, S-d, S-e and S-f) for F₁ pollen sterility were identified. Here we report genetic and linkage analysis of S-c locus using molecular markers in a cross between Taichung 65, a japonica variety carrying allele S-c ^j, and its isogenic line TISL5, carrying alleleS-c ^j. Our results show that pollen sterility occurring in the hybrids is controlled by one locus. We used 208 RFLP markers, as well as 500 RAPD primers, to survey the polymorphism between Taichung 65 and TISL5. Six RFLP markers located on a small region of chromosome 3, detected different RFLP patterns. Co-segregation analysis of fertility and RFLP patterns with 123 F₂ plants confirmed that the markers RG227, RG391, R1420 were completely linked with the S-c locus. The genetic distances between the markers C730, RG166 and RG369 and the S-c locus were 0.5 cM, 3.4 cM, and 3.4 cM respectively. Distorted F₂ ratios were also observed for these 4 RFLP markers in the cross. This result suggests that the `one locus sporo-gametophytic' model could explain F₁ hybrid pollen sterility in cultivated rice. RG227, the completely linked marker, has been converted to STS marker for marker-assisted selection. This revised version was published online in August 2006 with corrections to the Cover Date.