四种植物的细胞学和分子细胞遗传学研究
其他题名Studies of cytology and molecular cytogenetics of four different plants
薛瑞娟
学位类型硕士
导师顾志建
2009-01-13
学位授予单位中国科学院昆明植物研究所
学位授予地点昆明植物研究所
学位专业植物学
关键词核型 甜菜树 山柚子科 猪腰豆 居群 重楼属植物 凌云重楼 滇重楼 显微分离技术 荧光原位杂交(fish) 端粒重复序列
摘要本文通过常规核型分析、染色体显微分离技术和荧光原位杂交技术对四种植物进行了细胞学和分子细胞遗传学的研究。从甜菜树植物简单的核型分析到猪腰豆植物种内不同地理居群的核型研究,再到通过染色体荧光原位杂交技术对两种重楼属植物进行的分子细胞遗传学研究,由窄到宽,由浅入深,不同层次地展开了植物染色体变异与演化的研究。 1.甜菜树[Yunnanopilia longistaminea (W. Z. Li) C. Y. Wu et D. Z. Li]的核型研究 研究了产于我国云南省红河流域山柚子科甜菜树属甜菜树(Y.longistaminea)的核型,其核型公式为2n=2x=20=6m+12sm+2st。根据该结果,甜菜树属的染色体数目与已报道的山柚子科其它属植物的染色体数目一致,说明该科各属植物没有很大的变异和分化。且对其核型的分析发现,甜菜树在同源染色体的相对长度、同源染色体臂比值方面分化和变异的程度都比较低。甜菜树属的核型为3A型,对称性较高,属于比较原始的核型。研究结果为重新思考甜菜树的分类地位提供了依据。 2.猪腰豆[Whitfordiodendron filipes (Dunn) Dunn]四个地理居群的核型研究 对豆科猪腰豆属一国产猪腰豆种(W. filipes)的四个地理居群进行了核型分析,染色体数目都为2n=16。四个居群(永平、河口、文山和思茅居群)的核型公式依次为:2n=2x=16=13m+3sm、2n=2x=16=8m+8sm、2n=2x=16=7m+9sm和2n=2x=16=7m+9sm。永平居群与河口居群的核型类型为2A,文山居群与思茅居群的核型类型为2B。本实验的四个居群分别采集于云南省的永平、河口、文山和思茅,遍及云南省的西部、西南及东南地区。可以看出,在本文中较小的研究范围内,猪腰豆的核型只有细微的差异,比较稳定。 3.两种重楼属植物(Paris L.)的染色体荧光原位杂交 通过染色体显微分离技术和荧光原位杂交技术对延龄草科(Trilliaceae)重楼属的两种植物凌云重楼[Paris cronquistii (Takht.) H.]与滇重楼[Paris polyphylla Smith var. yunnnanensis (Franch.) Hand.-Mazz.]进行了分子细胞遗传学研究。 凌云重楼(Paris cronquistii)的研究主要有三个方面。首先对凌云重楼进行了核型研究,并结合已有的研究结果,将凌云重楼的核型归纳为十四种细胞型。其次通过染色体显微切割技术将凌云重楼B染色体分离并扩增,以其为探针,与凌云重楼中期染色体进行荧光原位杂交,结果不仅B染色体上有杂交信号,A染色体上也有,表明B染色体与A染色体同源,B染色体起源于A染色体。最后,以端粒重复序列为探针,与凌云重楼B染色体进行了荧光原位杂交,结果在两种B染色体的端部都有明显的杂交信号,说明了B染色体在染色体结构组成上与A染色体的相似性。 另外,本文以端粒重复序列为探针,在滇重楼间期和中期染色体上进行了荧光原位杂交定位,结果在间期和中期染色体上都有强烈的杂交信号,且端粒重复序列在中期染色体上的杂交信号都分布于染色体端部,其他部位如着丝粒和染色体臂上则没有杂交信号。根据该结果,认为滇重楼染色体在进化过程中比较保守,极少发生染色体融合、臂倒位、易位或重组等染色体结构变异。
其他摘要In this sduty, conventional karyotype analysis, chromosome microdissection techniques and fluorescence in situ hybridization (FISH) had been applied to studies of cytology and molecular cytogenetics of four different plants. Firstly, a karyotype study was performed on Yunnanopilia longistaminea. Secondly, the karyotypes of four geographical populations of Whitfordiodendron filipes were studied. Thirdly, the molecular cytogenetics was studied in two species of Paris by chromosome microdissection techniques and FISH. The origin and evolution of plants were studied at different levels. 1. The karyotype study of Yunnanopilia A karyotype study was performed on Yunnanopilia, a genus of Opiliaceae from the Red River Valley, South-Central Yunnan Plateau, China. The karyotype formula is: 2n=2x=20=6m+12sm+2st. The chromosome number of Yunnanopilia is consistent with the existing chromosome numbers of other genera of Opiliaceae. Based on the result, there was little differentiation in the karyotypes among genera of Opiliaceae. By analyzing the karyotype of Yunnanopilia, it can be concluded that the differentiation degree of Yunnanopilia is low both in relative length of homologous chromosome and arm ratio of homologous chromosome.The karyotype belongs to 3A type which is relatively symmetrical and original. The result provided evidence for reconsidering the taxology status of the Yunnanopilia longistaminea. 2. The karyotype studies of four geographical populations of Whitfordiodendron filipes Karyotype studies were performed on four geographical populations of Whitfordiodendron filipes, a species of Whitfordiodendron Elmer (Leguminosae) from China. The chromosome numbers of four populations are all 2n=16. The karyotype formulas of four populations (Yongping, Heko, Wenshan and Simao) are 2n=16=13m+3sm, 2n=16=8m+8sm, 2n=16=7m+9sm and 2n=16=7m+9sm. The karyotypes of Yongping population and Hekou population both belong to 2A type. Karyotypes of other two populations both belong to 2B type. The four populations in this study were collected respectively from Yongping, Hekou, Wenshan and Simao, which locate the west, southwest and southeast of Yunnan province. This study revealed that the karyotype variation in the four populations of W. filipes is not apparent. 3. The FISH of two species in Paris L. Molecular cytogenetic studies were performed on two species (Paris cronquistii and Paris polyphylla) of Paris in Trilliaceae by chromosome microdissection techniques and FISH. Paris cronquistii was studied from three aspects. Firstly, the karyotype study of Paris croquistii was performed. Combining with existent studies, the karyotype of Paris cronquistii was reduced to fourteen cellular types. Secondly, B chromosome was microdissected from a metaphase cell of Paris cronquistii and then was amplified by PCR. Fluorescence in situ hybridization (FISH) was carried out in metaphase chromosomes of Paris cronquistii with B chromosome as the probe. Both the B chromosome and A chromosomes showed fluorescent signals. Therefore, it can be concluded that B chromosome was homologous with A chromosomes and probably originated from A chromosomes. Thirdly, we located the telomere repeated sequences in B chromosome of Paris cronquistii by FISH. Signals showed on the telomeres of both two types of B chromosome. Therefore, it can be concluded that B chromosome is similar as A chromosomes in the composement of chromosome structure. In addition, the telomere repeated sequences were located in mitosis interphase and metaphase chromosomes of Paris polyphylla by FISH. The results showed that strong signals were found just in telomere of metaphase chromosomes but not in other areas of metaphase chromosomes. Based on the result, it can be inferred that chromosomes of Paris polyphylla were conservative in the process of evolution and chromosome structure variations (such as chromosome fusion, arm reversion, transposal, recombination and so on) rarely happened.
页数70
语种中文
文献类型学位论文
条目标识符http://ir.kib.ac.cn/handle/151853/342
专题昆明植物所硕博研究生毕业学位论文
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薛瑞娟. 四种植物的细胞学和分子细胞遗传学研究[D]. 昆明植物研究所. 中国科学院昆明植物研究所,2009.
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