【作者】 李云伏；

【导师】 孙东发；

【作者基本信息】 华中农业大学 ， 作物遗传育种， 2006， 博士

【摘要】 光温敏雄性不育是一种由遗传因素与光温因子互作所致的雄性不育类型，其育性表现与遗传控制机制较由遗传因素控制的雄性不育更为复杂。本文对新育成的小麦光温敏不育系BS20和BS210的育性转换规律和不育性遗传控制模式及其不育侯选基因进行了研究，主要结果如下：1、分期播种和人工气候箱试验研究表明：不育系BS20和BS210的温度敏感期和光周期敏感期是一致的，即在小麦穗分化的药隔至花粉粒单核期。BS20、BS210都表现为短日低温不育。不育系BS20育性受光照和温度共同作用，温度对育性的影响显著大于光周期的影响，临界温度为10℃-12℃。在临界温度以上，存在一个光敏温度范围，即12℃-14℃，在此温度范围内随着光长的增加，结实率增加。不育系BS210育性主要受温度影响，光长作用不明显，临界温度为10℃-12℃。2、“植物主基因+多基因混合遗传模型”分析表明：不育系BS210的两个组合(BS210／白玉149和BS210／0201)为E模型，即两对主基因+多基因模型。进一步分析表明：两个组合均为E-1模型，是2对加性-显性-上位性主基因+加性-显性多基因混合遗传模型。2对主基因，均为负等加性效应。光温敏不育基因是隐性，但存在部分显性效应。BS210／0201比BS210／白玉149主基因加性和显性效应略高，而且多基因加性效应为正值。两个组合的主基因遗传力高于多基因遗传力。环境对两个组合的结实率影响很大，环境方差分别占表型方差的58.33％和46.67％，BS2101／0201组合的环境方差几乎与主基因方差相当，BS210／白玉149组合的环境方差等于主基因方差的2倍。3、光温敏不育相关基因片段的分离试验表明：以不育系BS210和BS20为材料，在北京(可育环境)和安徽阜阳(不育环境)分别种植，通过DDRT-PCR方法进行不育相关基因的分离和反向Northern杂交验证，分别得到了20条和4条与不育相关的cDNA差异片段，进行了序列的测定和功能的初步分析，筛选出6个候选基因片段，其中5个分别与水稻的DNA修复重组蛋白rad50、小麦穗发育过程中类受体激酶基因、玉米和水稻细胞死亡抑制蛋白基因、水稻调控核编码的叶绿体正常发育及其功能的HAP3基因和一粒小麦磷脂酰丝氨酸脱羧酶基因高度同源，一致性分别为88％、92％、88％、57％和89％，其余1个候选基因片段和其他16个序列为新基因片段。修复蛋白rad50基因和一个新基因片段分别在可育和不育环境下mRNA的5’端编码区相同，但3’端非编码区序列出现长度差异。更多还原

【Abstract】 The thermo-photoperiod sensitive male sterility in plant is controlled by factors ofinheritance and environment. Its performance of fertility and inheritance are muchmore complex than that.of male sterility controlled only by factors of inheritance. Inthis study, the fertility alteration and separation of cDNA fragments genes wereconducted in thermo-photoperiod sensitive male sterile lines BS20 and BS210, andinheritance models were analyzed using parents, F₁ and F₂ in crosses of BS210/Baiyu149 and BS210/O201. Major results are described below:1. Fertility AlterationThe study of fertility alteration was carried out in photo-thermo-controlled growthchambers and with different ecological regions and sow times. The results showed thatthe stage of photo-sensitivity and thermo-sensitivity was coincident from antherchamber formation to mononucleate. BS20 and BS210 showed sterility under theconditions of short day length and low temperature. Fertility alteration of BS20 wasinduced by photoperiod and temperature, but thermo-sensitivity played a leading role,and the critical point of temperature was 10-12℃. Below the critical temperature,BS20 kept the stable sterility, but not affected by the photoperiod. However, above thecritical temperature, the fertility of BS20 improved as the temperature rose. In thetemperature range of 12-14℃the fertility of BS20 improved significantly as thephotoperiod extended. Fertility alteration of BS210 was induced mainly bytemperature, and the critical point of temperature was 10-12℃.2. Inheritance of Photoperiod-temperature Sensitive Male SterilityInheritance of fertility from the crosses of BS210/Baiyu149 and BS210/O201 was investigatedby the mixed major gene plus polygenes inheritance model of quantitative traits. The resultsshowed that the two crosses were controlled by two additive-dominance-epistasis majorgenes+additive-dominance polygenes （E-1 model）. The major sterile genes of BS210/Baiyu149& BS210/O201 were negative equal-additive effect and incomplete dominant. The major additiveand dominant effect of BS210/O201 was little higher than BS210/Baiyu149. The inherited modelsof major genes were almost identical in the crosses with same female parent and different maleparents, while polygenes had obvious differences. The heritabilities of major genes weresignificantly higher than that of polygenes. The heritabilities of major genes and polygenes were different in two crosses. Environment was an important effect factor on fertility of thephotoperiod-temperature sensitive genie male sterility. The variance of environmentwere 58.33% and 46.67% in above two crosses. It almost equaled to or even overthan variance of major genes. Therefore, the photoperiod-temperature sensitivemale sterility was called Environment Sensitive Male Sterile.3. Seperation of Candidate genes controlling Photoperiod-temperature Sensitive MaleSterilityDDRT-PCR was used to analyze the differential expression of genes of thephotoperiod-temperature sensitive genie male sterile lines of winter wheat, BS20 andBS210, which are two core female parents of two-line hybrid wheat. They wereplanted in Beijing （fertile environment） and Fuyang, Anhui （sterile environment） in2002-2003. The materials were collected in five stages, stamens/pistil differentiation,anther chamber formation, pollen mother cell formation, tetrad phase, andmono-nucleate phase. The different cDNA bands were identified with reverse Northernblot hybridization and 24 bands were sequenced. 6 candidate gene fragments werescreened including 8 cDNA sequences. The homology research indicated that the 5gene sequences were identical to the partial mRNA sequences of Oryza sativa（japonica cultivar-group） DNA repair-recombination protein （rad50）, Triticumaestivum partial mRNA for receptor-like kinase with LRR repeats, Zea mays & Oryzasativa cell death suppressor protein llsl, Oryza sativa （japonica cultivar-group） HAP3genes and Triticum monococcum phosphatidylserine decarboxylase, and the identitieswere 88%、92%、88%、57% and 89%, respectively. The sixth candidate gene andother 16 sequences might represent new genes. In two of genes, rad50 gene and anew gene（the sixth candidate gene）, the sequcences of 5’ coding region of mRNA wereidentical while the sequences of 3’ terminal non-coded regions were different betweenfertile environment and sterile environment. The studies of different researchersshowed different results. It maybe means photoperiod-temperature sensitive malesterile wheat involved a complex way of metabolism. The development mechanism ofphotoperiod-temperature sensitive genie male sterility could not yet be revealedcompletely in this paper.更多还原