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1、中南民族大学硕士学位论文摘要1.为研究大颖野生稻、高杆野生稻和宽叶野生稻基因组的差异，比较它们之间进化关系，采用荧光原位杂交技术，对这三种CCDD野生稻进行基因组分析。利用栽培稻和药用野生稻基因组DNA为探针，对大颖野生稻体细胞染色体进行FISH分析，并结合不同的洗脱严谨度对染色体上杂交信号进行检测，结果显示栽培稻基因组 DNA在不同的洗脱严谨度下都分布在大颖野生稻染色体上，其中C组染色体上的杂交信号较D组染色体多，说明A、C基因组亲缘关系较A、D基因组亲缘关系近；利用药用野生稻基因组DNA为探针，分别对大颖野生稻、高杆野生稻和宽叶野生稻进行FISH分析，结合不同的洗脱严谨度观察染色体上杂交信
2、号的变化，结果显示：药用野生稻基因组DNA在低严谨洗脱度下，就将大颖野生稻的C、D组染色体区分开，高杆野生稻和宽叶野生稻在高洗脱严谨度下，C、D组染色体才被区分开，说明高杆野生稻和宽叶野生稻在基因组水平上差异不明显，而大颖野生稻与二者在基因组水平上差异较大。2.利用栽培稻和药用野生稻Cot-1DNA（中高度重复序列）为探针，分别对大颖野生稻体细胞染色体进行FISH分析，结合不同的洗脱严谨度，观察染色体上杂交信号，结果显示：栽培稻Cot-1DNA在80%的洗脱严谨度下，杂交信号大部分集中在大颖野生稻的C组染色体着丝粒、近着丝粒和端粒区域，而在D组染色体这三个区域的杂交信号与C组染色体相比较少，说
3、明大颖野生稻的D基因组中高度重复序列跟A基因组的中高度重复序列有一定的同源性，但C基因组与A基因组的中高度重复序列同源性更高；利用药用野生稻Cot-1DNA为探针，对大颖野生稻体细胞染色体进行FISH分析，在很高的洗脱严谨度（ 80%）下，大颖野生稻的C、D组染色体才被区分开；利用药用野生稻Cot-1DNA为探针，对高杆野生稻体细胞染色体进行FISH分析，也发现在高洗脱严谨度下，C、D组染色体才被分开，但其C组染色体上杂交信号较大颖野生稻C组染色体弱，说明高杆野生稻和大颖野生稻在Cot-1DNA水平上差异明显。3.利用栽培稻着丝粒DNA为探针，分别对药用野生稻、宽叶野生稻和高杆野生稻体细胞染色
4、体进行FISH分析，结果显示：药用野生稻几乎所有的染色体着丝粒区域都有杂交信号分布，相比之下，宽叶野生稻和高杆野生稻只有个别染色体上着丝粒区域有杂交信号，但杂交结果显示：宽叶野生稻有杂交信号的染色体有8条，而高杆野生稻有杂交信号的染色体有16条，说明宽叶野生稻和高杆野生稻的着丝粒 DNA有一定程度的差异，基于二者着丝粒DNA的差I利用 FISH技术从细胞水平对三种 CCDD野生稻的比较分析异，可以考虑把宽叶野生稻和高杆野生稻划分为同一个物种下的不同亚种处理。关键词：荧光原位杂交，Cot-1DNA，洗脱严谨度，基因组II中南民族大学硕士学位论文ABSTRACT1. In order to stu
5、dy genome difference of O. grandiglumis, O. alta and O. latifolia,analyze the genome of these three CCDD wild rice with FISH technique. With O.sativa and O. officinalis genomic DNA for probe respectively, O.grandiglumissomatic chromosomes on fluorescence in situ hybridization(FISH) combineddifferenc
6、e stringency to detect the hybridization signal on chromosomes.The resultsshow that O. sativa genomic DNA is distributed in the C and D genome,But thehybridization signal is much more on the C genome than those on the Dgenome,which illustrates A and C genome have closer relations than A and D genome
7、With O. officinalis genomic DNA for probe, O. grandiglumis, O. alta and O. latifoliasomatic chromosomes on fluorescence in situ hybridization(FISH) combineddifference stringency to detect the hybridization signal on chromosomes.The resultsshow that O. officinalis genomic DNA can distinguish the C an
8、d D genome of O.grandiglumis under the lower stringency,while C and D genome of O. alta and O.latifolia can be distinguished under the higher stringency,which shows O. alta and O.latifolia have difference with O. grandiglumis in the genome level.2. With O.sativa and O. officinalis Cot-1 DNA for prob
9、e respectively, O.grandiglumissomatic chromosomes on fluorescence in situ hybridization combined differencestringency to detect the hybridization signal on chromosomes. The results show thatCot-1 DNA of O. sativa mainly lie on the centromere, near centromere and telomerearea of C genome of O.grandig
10、lumis under the 80% stringency, while the signals ofthese region on D genome is less, which shows that the medium and high repeatsequence on D genome of O.grandiglumis have a certain homology with the mediumand high repeat sequence of A genome, but the homology between medium and highrepeat sequence
11、 of C and A ge nome is larger. With Cot-1 DNA of O. officinalis forprobe, O.grandiglumis somatic chromosomes on fluorescence in situ hybridization, Cand D genome of O.grandiglumis can only be separated under higher stringency(80%). With Cot-1 DNA of O. officinalis for probe, O.alta somatic chromosom
12、es onfluorescence in situ hybridization, also found that the C and D genome of O.alta canbe distinguished only under the higher stringency, however, the signals on the Cgenome of O.alta is weaker than those on the C genome of O.grandiglumis, whichillustrates that O.alta have difference with O.grandi
13、glumis in the Cot-1 DNA level.III利用 FISH技术从细胞水平对三种 CCDD野生稻的比较分析3. With centromere DNA of O.sativa for probe, O. officinalis, O. latifolia and O. altasomatic chromosomes on fluorescence in situ hybridization, the results display thatnearly every chromosomes of O. officinalis have hybridization signal
14、s, whilehybridization signals only distributed on a few chromosomes of O. latifolia and O.alta, and there are 8 chromosomes with signals for O. latifolia,while there are 16chromosomes with signals for O. alta, which display that the centomere DNA of O.latifolia and O. alta have a certain distincts. According to the centromere DNAdifference of O. latifolia and O. alta,we can consider them to be classified thedifference subspecies under the same specie.Keyword: FISH, Cot-1DNA, Stringency, GenomeIV中南民族大学硕士学位论文缩写符号缩写FISHGISHSA-APBCIPNBTDAPIEDTAbpkbMbhmins
