【作者】 夏红爱；

【导师】 汪旭东； 杨焕明； 吴先军；

【作者基本信息】 四川农业大学 ， 作物遗传育种， 2002， 硕士

【摘要】 用189对微卫星引物对两优培九亲本材料93-11及培矮64S进行差异筛选，初步获得多态性标记25对，其引物的多态性频率为13％。根据实际需要，选取6对引物在已经推广的13个杂交稻品种中进行多态性分析，分别构建了13个品种的指纹图谱，利用这6对微卫星引物基本上可将13个品种区分开。 利用杂交稻基因组序列信息，结合国际公共数据库中公布的有关水稻数据，利用自编软件SNP＿CROSS．PL分别在不同材料93-11、广陆矮、培矮64S、日本晴的叶绿体基因组和核基因组序列间寻找新型遗传标记(SNPs和InDels)，并对部分多态性位点设计PCR引物进行了验证。具体结果是： 1．对93-11、培矮64S、日本晴三个水稻叶绿体基因组序列比较发现，其差异较小：InDels的变化长度从2bp～69bp不等，SNPs在93-11／培矮64S、培矮64S／日本晴、93-11／日本晴之间分别为57、152和201个。对这些差异分别进行了初步分析。 2．对广陆矮(GLA)与“日本睛”基因组之间的多态性研究发现籼粳亚种在基因组水平的差异主要是由于重复序列(大多为转座子)在基因组中的插入或缺失引起。 3．用培矮64S(PA64S)115万条Reads序列与日本晴第一染色体(约50.8 Mb)比较，找到了SNPs 56979个。 4．用1万个培矮64S(PA64S)与93-11部分基因组Contig(约409Mb)比较得出SNPs4196个，InDels2956个，并对整个基因组的多态性情况进行了估计。，5．针对 93习 叶绿体基困组的 69hP的缺失、32hP的插入，分别 设计了2对引物在不同釉、粳稻材料中进行了生物学验证，结果证明 了该多态性在釉、粳亚种叶绿体基因组中是真实存在的。同时也论证 利用序列比较寻找基因组多态性位点的方法是可行的。更多还原

【Abstract】 189 pairs of Micro-satellite Primers(SSR) were used to screen the polymorphism between 93-11 and Pei’ ai64S,which were the parents of Liang You Pei Jiu. There were 25 pairs of SSR primers,which showed polymorphism between the two varieties. So,the polymorphism rate was 13%. According to necessary,the polymorphisms among 13 widely used hybrid rices were analyzed by 6 pairs of primers and their finger printing maps were constructed. Generally,13 varieties can be identified by the 6,SSR primers.The new polymorphism markers (SNPs & InDels) were searched separately between the chloroplast and nuclear genome sequences using the self-compiled software (SNP_CROSS. PL). The varieties used to mine new polymorphism markers were 93-11,Guanglu’ ai,Pei’ ai64S and Nipponbare. Their sequences were got from BGI (Beijing Genome Institute) and NCBI(National Center of Biotechnology Information). Andwhat’ s more,the partial PCR primers were designed to test polymorphism sites. The detail results were as follows:1. The comparing results among the three chloroplast genome sequence 93-11,Pei’ ai64S,and Nipponbare showed that the chloroplast genome sequences were comparatively conserved:The length of the InDels were changed from 2bp-69bp;there were 57,152 201 SNPs,respectively,among 93-11 & Pei’ ai64S, Pei’ ai64S & Nipponbare, 93-11 & Nipponbare. These differences were analyzed.2. The polymorphism results between Guanglu’ ai and Nipponbare indicated that the insertion and deletion of the repeat sequences,especially transposons,among rice genome were the main reasons which results to the difference between Indica and Japonica varieties in genomic level (about3. 1.15 million Pei’ ai64S reads were compared to the Nipponbare chromosome 1(about 50.8 Mb) and 56979 SNPs were found.4. 10,000 Pel’ ai64S reads were compared to the part sequence of 93-11 genome Contigs and 4196 SNPs & 2956 InDels were found. Consequently, the polymorphism conditions between the two genomes were estimated.5. Two pairs of PCR primers have validated the existence of 69 bp deletion and 32 bp insertion in 93-11 chloroplast. The PCR results among different varieties conformed that the polymorphism sites existed truly between Indica and Japonica varieties,at the same time,these results also proved that it was feasible to search polymorphism sites by sequence comparison.更多还原