黑龙江省水稻抽穗期基因的效应研究及育种应用

发布时间：2018-01-16 06:44

本文关键词：黑龙江省水稻抽穗期基因的效应研究及育种应用　出处：《中国科学院研究生院(东北地理与农业生态研究所)》2016年博士论文　论文类型：学位论文

【摘要】：水稻抽穗期(Heading Date, HD)是指水稻从播种到抽穗(开花)所经历的时间,抽穗期是影响水稻种植和生产的最重要因素之一,黑龙江省是我国优质粳稻的主产区和商品粮区,属于北方高纬度寒地稻作区,积温低、日照长。越往北部,积温越低,要求品种熟期越早,造成目前育成的优良品种适应区域较窄,严重的限制了种植区域。抽穗期是多基因控制的复杂数量性状。同时受多种环境因素影响。为了鉴定控制黑龙江省水稻抽穗期的主效和微效基因、及不同基因的效应,我们综合利用连锁分析与关联分析手段对抽穗期基因进行研究,主要结果如下：1.利用第三积温区主栽品种空育131和第二积温区东农422的F7重组白交系群体,发现控制水稻抽穗期主效QTL位点Hd2,该基因在空育131中存在一个L710P氨基酸变异,导致空育131早花。对抽穗期相关基因的表达量研究,发现Hd2是通过Ehdl途径参与水稻抽穗期调控。进一步对黑龙江省395份水稻材料的Hd2基因分型,确定Hd2/OsPRR37是控制黑龙江当地水稻抽穗期的主效基因；2.Hd2定位于细胞核中,可以形成同源二聚体,且依赖于其CCT结构域。利用酵母双杂交的手段,筛选到HD2互作的候选蛋白LTG1和DELLA,并利用多种手段验证了HD2与LTG1和DELLA的互作关系；3.利用本课题组收集的395份黑龙江历年审定水稻品种及育种资源,连续三年调查了抽穗期表型数据,同时从这些材料的基因组重测序数据中,调取与水稻抽穗期相关的基因序列：Hd6、Hdl6、Hd17、NRR、LTG1、DTH2、Ehd4、Ehd3、 Ehd2、SE5、Ehdl、Hd3a、RFT、SE14、SDG724、OsGI、Hd2/OsPRR37, Hd4/Ghd7, Hd5/DTH8/Ghd8等,将抽穗期基因的基因型与表型数据进行关联分析,发现Hdl,Hd2/OsPRR37, Hd4/Ghd7, Hd5/DTH8/Ghd8是控制黑龙江省水稻抽穗期的主效基因,Hd6、Hdl6、Hd17是微效基因,不同抽穗期组合方式决定了水稻材料的地域性分布。4.根据以上鉴定到的控制黑龙江水稻材料的抽穗期基因,通过两种策略进行分子设计育种。一是设计不同抽穗期基因的分子标记,对优良品种进行回交改良。二是利用CRISPR/CAS9介导的基因编辑技术,对抽穗期基因进行定点敲除。期望在优良品种的背景下,创造出一系列抽穗期不同的优良水稻材料,一方面可以扩大优良品种的可种植面积和创造极早熟品种,另一方面创制不同抽穗期基因的多种组合类型的水稻材料,为水稻开花期途径的研究提供基础材料。
[Abstract]:Heading date (HDD) is the time from sowing to heading (flowering) of rice, and heading stage is one of the most important factors that affect the planting and production of rice. Heilongjiang Province is the main producing area and commodity grain area of high quality japonica rice in our country. It belongs to the northern high latitude cold region, the accumulated temperature is low, the sunshine is long. The farther north, the lower the accumulated temperature, and the earlier the maturity of varieties is required. As a result of the current breeding of fine varieties to adapt to a narrower area. The heading stage is a complex quantitative trait controlled by many genes. It is also affected by many environmental factors. In order to identify the main and minor genes that control the heading stage of rice in Heilongjiang Province. And the effects of different genes, we used linkage analysis and correlation analysis to study genes at heading stage. The main results were as follows: 1. Using Gangyu 131, the main cultivar in the third accumulated temperature area, and the F7 recombinant white cross population of Dongnong422 in the second accumulated temperature area, the main effect QTL locus Hd2 was found to control the heading stage of rice. There was a L710P amino acid variation in the gene, which led to the early flowering of empty 131. The expression of genes related to heading stage was studied. It was found that Hd2 was involved in the regulation of heading stage of rice through Ehdl pathway, and the Hd2 genotyping of 395 rice varieties in Heilongjiang Province was further studied. It was confirmed that Hd2/OsPRR37 was the main gene to control the heading stage of local rice in Heilongjiang province. 2. HD2 was located in the nucleus and could form homologous dimer, which was dependent on its CCT domain. The candidate proteins LTG1 and DELLA for HD2 interaction were screened by yeast two-hybrid. The interaction between HD2 and LTG1 and DELLA is verified by various means. 3.The phenotypic data of heading stage were investigated by using 395 rice varieties and breeding resources collected by our research group in Heilongjiang province over the years. Meanwhile, the genome re-sequencing data of these materials were obtained. The sequence of genes related to heading stage of rice was obtained: Hd6, Hdl6, Hd17, NRRG1, DTH2, Ehd4, Ehd3, Ehd2SE5, Ehdl, Ehdl, Ehdl, Ehd3, Ehd2, Ehd3, Ehd2SE5 and Ehdl, respectively. Hd3aI RFT SE14 SDG724, OsGIN Hd2 / OsPRR37, Hd4 / Ghd7, Hd5/DTH8/Ghd8, etc. Genotypic and phenotypic data of heading stage gene were analyzed and the results showed that Hdl1 / Hd2 / OsPRR37, Hd4/Ghd7. Hd5/DTH8/Ghd8 is the main gene that controls the heading stage of rice in Heilongjiang Province. Hd6, Hdl6, Hd17, is a microfunctional gene. Different combination patterns of heading stage determined the regional distribution of rice materials. 4. According to the above identified control of Heilongjiang rice heading stage genes. Molecular design breeding was carried out through two strategies. One was to design molecular markers of genes at different heading stages to improve the backcross of superior varieties, and the other was to use CRISPR/CAS9 mediated gene editing technology. It is expected to create a series of fine rice materials at different heading stages under the background of superior varieties, on the one hand, to expand the acreage of good varieties and to create very early varieties. On the other hand, rice materials with different combination types of heading stage genes were created, which provided basic materials for the study of flowering pathway of rice.
【学位授予单位】：中国科学院研究生院(东北地理与农业生态研究所)
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S511

【参考文献】