滇山茶复合多倍体基因组原位杂交研究
其他题名The Genomic In Situ Hybridization Studies on the Complex Poliploid of Camellia reticulata Lindl
杜宁
学位类型硕士
导师顾志建
2004
学位授予单位中国科学院昆明植物研究所
学位授予地点中国科学院昆明植物研究所
学位专业植物学
关键词基因组原位杂交 滇山茶复合多倍体 起源
摘要本论文分两部分。首先,对滇山茶(Gamelllia reticulata Lindl.)及其近缘种的相关研究和荧光原位杂交技术进行综述。分别从滇山茶的栽培历史,系统位置,分类学现状、地理分布、主要的形态学以及细胞学特征进行了综述,同时对原位杂交的原理,发展历史及其应用进行综述。其次,叙述了开展此实验的研究背景以及基因组荧光原位杂交的应用。同时利用基因组荧光原位杂交技术对滇山茶(C.reticulata)的六倍体的起源进行研究讨论,并对实验过程中,影响实验成功与否的因素讨论。本研究利用山茶(Camellia japonia L)、怒江山茶(Camellia saluenensisStapf ex Beans)、滇山茶(C.reticulata)二倍体的基因组总DNA作探针,以滇山茶(C.reticulata)六倍体减数分裂(n=45,3个基因组)染色体为靶DNA进行荧光原位杂交。根据杂交信号的分布及多少,进行讨论。主要研究结果和结论如下:怒江山茶(C.saluenensis)作为探针进行杂交,14个二价体完全杂交,21个二价体部分杂交,10个二价体没有杂交信号;滇山茶(C.reticulata)二倍体作为探针进行杂交,15个二价体完全杂交,16个二价体部分杂交,14个二价体没有杂交;山茶(C.japonica)作为探针,3个二价体进行杂交,为同源的染色体,15个二价体部分杂交为异源染色体,27个二价体没有杂交。根据以上杂交结果:说明滇山茶(C.reticulata)六倍体染色体一个基因组中有14对同源染色体与怒江山茶(C.saluenensis)同源;另外有1个基因组成分与滇山茶(C.reticulata)二倍体同源。证明滇山茶(c.reticulata)二倍体,怒江山茶(C.saluenensis),是滇山茶(C.reticulata)异源六倍体的亲本。从三种探针杂交结果看:在滇山茶(C.reticulata)45个二价体中,有32(14+15+3)个同源二价体,J还有13对同源染色体没有杂交,这充分说明虽然滇山茶(C.reticulata)六倍体(x=15,n=45)由三个基因组组成,但它的亲本种(或是供体种)不只3种,肯定是3种以上。根据部分杂交结果证明慎山茶(C.reticulata)六倍体的基因组还含有山茶(C.japonica)的基因组成分。同时进一步推测滇山茶多倍复合体的其他亲本种为西南山茶Camelliapitardii Coh.Stuat,猴子木Camellia yunnanensis Cohen Stuart
其他摘要This thesis included two parts. Part 1 reviewed the research history and previous studies about the cultured history, systematic position, the taxonomy, the distribution, morphological and cytological characteristics of Camellia reticulata and its related species. And it also summarized the principle, application and prospects of ISH (in situ hybridization) more detailedly. In Part 2, summarized the foundation of the experiment and the principle, application of the GISH(genomic in situ hybridization). And the gDNA of C. japonica、C. saluenensis、diploid of C. reticulata as probe separatly, GISH (genomic in situ hybridization) on hexaploid of C. reticulata was investigated. Origin of C. reticulata was further discussed. Moreover, the factors deciding whether the experiment succeeds or not were summed up. The gDNA of C. japonica、C. saluenensis、diploid of C. reticulatabeing probe separately was located on the genomes of the hexaploid of C. reticulata, Comparison of the results was analyzed about the number of the signals. The main results and conclusion were as follows: C. japonica being probe: three bivalents have whole signals, fifteen bivalents have part of signals, and twenty-senven bivalents has no signal C. saluenensis being probe: fourteen bivalents have whole signals, twenty-one bivalents have part of signals, and ten bivalents has no signal Diploid of C. reticulata being probe: fifteen bivalents have whole signals, sixteen bivalents have part of signals, and fourteen bivalents has no signal Based on the results of GISH, In one genome of hexaploid of C. reticulata, fourteen pairs of chromosomes are homologous with C. saluenensis. And the other genome is diploid of C. reticulata. It is concluded that C. saluenensis and diploid of C. reticulata are the progenitors of hexaploid of C. reticulata. The other side, we can find thirty-two bivalents are homologous in fourty-five bivalents, and the other thireen bivalents has no signal. It shows that the hexaploid of C. reticulata consists of not three parents but three genomes. Finally , Camelliapitardii Coh. Stuat and Camellia yunnanensisCohen Stuart may be the parents of the hexaploid of C. reticulata.
页数57
语种中文
文献类型学位论文
条目标识符http://ir.kib.ac.cn/handle/151853/732
专题昆明植物所硕博研究生毕业学位论文
推荐引用方式
GB/T 7714
杜宁. 滇山茶复合多倍体基因组原位杂交研究[D]. 中国科学院昆明植物研究所. 中国科学院昆明植物研究所,2004.
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