栽培稻籼粳亚种基因组的比较研究及药用野生稻的比较核型分析
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摘要
1.利用亚州栽培稻(Oryza sativa L.)的籼稻亚种“广陆矮四号(”Oryza sativa ssp.indica cv Guangluai No.4)的基因组总DNA作为探针,不同的洗脱严谨度条件下对自身和粳稻亚种“日本晴”(Oryza sativa ssp.japonica cv Nipponbare)进行基因组原位杂交(genomic in situ hybridization,GISH)分析。结果发现,杂交信号在两个亚种染色体的分布相似,表明它们亲缘关系较近。通过采用不同洗脱度,杂交信号出现明显不同的带型分布,尤其是在95%的洗脱严谨度下,籼稻和粳稻第1、7、8、9、11染色体上的杂交信号带型明显不一致,说明即使在基因组亲缘关系极近的亚种之间个别染色体同源性程度也较低。这几条同源性相对比较低的染色体可能是进化中比较活跃的染色体,易于发生各种遗传事件,是染色体组进化中的“先行者”,它们的变化可能最终导致了整个基因组水平上的差异并体现在亚种以及种间差异上。
2.利用同时分布在栽培稻(O. sativa)和药用野生稻(O. officinalis)第9和第11染色体上的紧密连锁的RFLP (Restriction Fragment Length Polymorphism)标记为探针,对药用野生稻进行荧光原位杂交(fluorescence in situ hybridization ,FISH)。比较分析了栽培稻和药用野生稻的第9和第11染色体。结果发现,与药用野生稻的常规核型分析不同,本研究中所确定的药用野生稻第9和第11染色体不是遵循染色体长度随染色体编号递减的顺序进行的,而是在比较分子遗传图基础上结合染色体常规核型分析进行的,是比较基因组学细胞水平上的研究,有助于在同一个属中对不同种作比较研究。
3.基于AA、CC基因组中高度重复序列C0t-1DNA杂交带型,对栽培稻、药用野生稻进一步进行了比较核型分析,并构建了相应的杂交带型模式图。研究发现栽培稻和药用野生稻第9和第11染色体着丝粒与端粒均有信号分布,染色体中部则明显不同。这说明在进化的过程中中高度重复序列在如维持染色体稳定的着丝粒与端粒等重要结构区域受到的选择压力大,保守性较强,其它区域则变化要快,这可能是形成种特异性染色体的重要原因之一。
1. Genomic in situ hybridization (GISH) procedure was adopted for genome analysis between Oryza sativa ssp. indica cv Guangluai No.4 and Oryza sativa ssp.japonica cv Nipponbare under several different post-hybridization washing stringencies with genomic DNA from Oryza sativa ssp. indica cv Guangluai No.4 as probes . The signals on the chromosomes of the two subspecies were similar showed that the relationship of these two AA genomes is very close. While under different post-hybridization washing stringencies, signal bands were some different. Especially under the 95% post-hybridization washing stringency , the signal bands on chromosome 1,7,8,9,and 11 of the two subspecies were obviously different. The results indicate that even in the subspecies genomes which have close relationship, the special chromosomes have low homology, these chromosomes maybe make genome evolution. The changes on these chromosomes may induce the differences in the whole genome, and incarnated in the differentiation on subspecies and species.
2. Fluorescence in situ hybridization was adopted for comparative karyotype analysis of chromosome 9 and chromosome 11 between O. sativa and O. officinalis with the Restriction Fragment Length Polymorphisms (RFLPs) as probes .These RFLPs based on the comparative Restriction Fragment Length Polymorphism map between O. sativa and O. officinalis..The results showed that chromosome 9 and 11 ascertained in this research did not followed the general rule that the length of chromosome decreases with the chromosome number increases. It is obviously different from the general karyotype analysis. The results are based on the comparative genetic maps and integrated the general karyotype analysis. It is in the comparative genomics and the cellular level, and will facilitate the comparative analysis on different species in the same genus.
3. Karyotype analysis of O. sativa, and O. officinalis were made according to C0t-1DNA sigal bands, and the C0t-1DNA signal bands mode chart were constructed. It was observed the signals existed on telomeres and centromeres in chromosome 9 and chromosome 11 of O. officinalis , while few signals on other parts of chromosome arms . It indicated that the moderate and highly repetitive sequences in these regions were conservative, while moderate and highly repetitive sequences in other regions changed quickly .These regions were important for preserving the stability of chromosome . It may be one of the most important reasons why the specific chromosome formed.
2.利用同时分布在栽培稻(O. sativa)和药用野生稻(O. officinalis)第9和第11染色体上的紧密连锁的RFLP (Restriction Fragment Length Polymorphism)标记为探针,对药用野生稻进行荧光原位杂交(fluorescence in situ hybridization ,FISH)。比较分析了栽培稻和药用野生稻的第9和第11染色体。结果发现,与药用野生稻的常规核型分析不同,本研究中所确定的药用野生稻第9和第11染色体不是遵循染色体长度随染色体编号递减的顺序进行的,而是在比较分子遗传图基础上结合染色体常规核型分析进行的,是比较基因组学细胞水平上的研究,有助于在同一个属中对不同种作比较研究。
3.基于AA、CC基因组中高度重复序列C0t-1DNA杂交带型,对栽培稻、药用野生稻进一步进行了比较核型分析,并构建了相应的杂交带型模式图。研究发现栽培稻和药用野生稻第9和第11染色体着丝粒与端粒均有信号分布,染色体中部则明显不同。这说明在进化的过程中中高度重复序列在如维持染色体稳定的着丝粒与端粒等重要结构区域受到的选择压力大,保守性较强,其它区域则变化要快,这可能是形成种特异性染色体的重要原因之一。
1. Genomic in situ hybridization (GISH) procedure was adopted for genome analysis between Oryza sativa ssp. indica cv Guangluai No.4 and Oryza sativa ssp.japonica cv Nipponbare under several different post-hybridization washing stringencies with genomic DNA from Oryza sativa ssp. indica cv Guangluai No.4 as probes . The signals on the chromosomes of the two subspecies were similar showed that the relationship of these two AA genomes is very close. While under different post-hybridization washing stringencies, signal bands were some different. Especially under the 95% post-hybridization washing stringency , the signal bands on chromosome 1,7,8,9,and 11 of the two subspecies were obviously different. The results indicate that even in the subspecies genomes which have close relationship, the special chromosomes have low homology, these chromosomes maybe make genome evolution. The changes on these chromosomes may induce the differences in the whole genome, and incarnated in the differentiation on subspecies and species.
2. Fluorescence in situ hybridization was adopted for comparative karyotype analysis of chromosome 9 and chromosome 11 between O. sativa and O. officinalis with the Restriction Fragment Length Polymorphisms (RFLPs) as probes .These RFLPs based on the comparative Restriction Fragment Length Polymorphism map between O. sativa and O. officinalis..The results showed that chromosome 9 and 11 ascertained in this research did not followed the general rule that the length of chromosome decreases with the chromosome number increases. It is obviously different from the general karyotype analysis. The results are based on the comparative genetic maps and integrated the general karyotype analysis. It is in the comparative genomics and the cellular level, and will facilitate the comparative analysis on different species in the same genus.
3. Karyotype analysis of O. sativa, and O. officinalis were made according to C0t-1DNA sigal bands, and the C0t-1DNA signal bands mode chart were constructed. It was observed the signals existed on telomeres and centromeres in chromosome 9 and chromosome 11 of O. officinalis , while few signals on other parts of chromosome arms . It indicated that the moderate and highly repetitive sequences in these regions were conservative, while moderate and highly repetitive sequences in other regions changed quickly .These regions were important for preserving the stability of chromosome . It may be one of the most important reasons why the specific chromosome formed.
引文
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