中国种子植物染色体数据库研制和一些重要类群的细胞学研究
杨静
学位类型博士
导师顾志建 ; 孙卫邦
2010-05
学位授予单位中国科学院研究生院
学位专业植物学
摘要植物的细胞学研究是对植物自身一项重要的生物学特征的探索,也可以在一定程度上揭示植物的系统发生和遗传物质分配等问题。在植物细胞学研究蓬勃进展的情况下,各个植物类群的染色体资料日益增多,但这些资料通常是几个种、几个种的发表在各个研究期刊上,零碎分散,在细胞学研究领域少见从整个分类群的细胞学统计数据的高度上作出的研究结论。随着科研资料的日益繁杂,现代科学领域对科研数据库的需要渐显迫切。在植物染色体研究蓬勃发展和中国缺乏数字化的植物染色体研究统计数据和染色体图谱的情况下,本研究启动了中国种子植物染色体数据库的研制工作,旨在建立和完善一个收纳中国种子植物染色体研究资料的数据库。基于数据库的统计资料,我们选取了裸子植物类群对其染色体的变异和演化特征做了探讨。论文还选取了一些重要的植物类群开展了系统进化上和亲本基因组鉴定上的细胞学研究工作。第一部分:中国种子植物染色体数据库和裸子植物类群的染色体特征探讨:1 中国种子植物染色体数据库:运用Microsoft Office 2003的Microsoft Access 2003数据库软件建立了数据库的框架。数据库包含数据类别19项,分别是:每种植物的科属种及拉丁名;原植物图、有丝分裂中期图、核型图;该植物的形态特征和分布;染色体数目和基数;核型公式;核型描述;材料产地;文献和图片来源。数据库可以按照科和种的方式提取生成数据界面,方便查阅和共享。目前已查找和整理输入了裸子植物类群(Gymnosperm)所有染色体研究资料共120种。涵盖10个科29个属。被子植物类群(Angiosperm),木兰科(Magnoliaceae)10个属61个种、山茶科(Theaceae)3个属80个种,共141个种。2 裸子植物类群的染色体变异与演化特征探讨:在数据库的统计数据基础上,我们分析了裸子植物的染色体特征。裸子植物具有悠久的演化历史,其化石记录可上溯至泥盆纪。然而,这一类群的植物已过了其在地球上的繁盛期,除了现存的部分外,多数已经绝灭。裸子植物的染色体具有以下3个特点:一是染色体大,由本数据库所列的每个种的染色体中期图即可看出。二是数目相对稳定,裸子植物大多数在科的范围内都具有一致的染色体基数。三是核型变异相对小,一个属甚至科内的染色体核型变异都局限在很小的范围内。裸子植物类群中染色体的变异主要以Robertson易位为主,在环境条件艰苦复杂的区域,在有些植物类群中容易导致染色体变异的产生。与一般植物染色体的常见变异形式相反,在裸子植物中存在从高不对称性到低不对称的变异形式。第二部分:一些重要植物类群的细胞学研究:3 十齿花目三个种的核形态学研究及其系统学意义:研究了核子木属(Perrottetia)核子木(Perrottetia racemosa)、十齿花属(Dipentodon)十齿花(Dipentodon sinicus)和瘿椒树属(Tapiscia)瘿椒树(Tapiscia sinensis)的核形态, 首次报道了十齿花和核子木的染色体数目。它们的核型公式分别为: 2n = 34 = 22sm + 12st (十齿花)、2n = 20 = 11m + 9sm (核子木)和2n = 30 = 22m(2SAT) + 8sm (瘿椒树)。核型不对称系数为: 十齿花(3B)、核子木(2A)、瘿椒树(2A)。近期的分子研究揭示这三个属有相近的亲缘关系,并在分子系统树中将十齿花科(Dipentodontaceae)和瘿椒树科(Tapisciaceae)放置于新成立的十齿花目(Huerteales)中。目前十齿花目有三个科:十齿花科(Dipentodontaceae)、瘿椒树科(Tapisciaceae)和非杨料科(Gerrardinaceae)。同时久有争议的核子木属被移置入十齿花科(Dipentodontaceae)内。这三个种分别具有不同的染色体基数和独立的核形态学特征,核子木与十齿花的染色体基数各不相同, 且后者具较高的核型不对称性和不同的分裂间期及分裂前期的细胞核形态, 针对将核子木归入十齿花科(Dipentodontaceae)的处理, 我们认为还需要更多的证据支持。4 对紫花含笑和灰岩含笑的杂种F1代基因组原位杂交(GISH)分析:以基因组为探针的荧光原位杂交技术即基因组原位杂交技术(GISH)被广泛应用于对杂交后代的种质鉴定中。研究对木兰科(Magnoliaceae)含笑属(Michelia)两种具有较高观赏价值的紫花含笑(M. crassipes)和灰岩含笑(M. calcicola)的杂交F1代开展了以亲本全基因组为探针的双色GISH研究。研究结果显示,紫花含笑和灰岩含笑的杂种F1代的染色体数目为侠义木兰科和含笑属的恒定染色体数目2n=38,染色体基数为x=19,核型公式为2n=38=32m+6sm,核型不对称性系数为1B型。从数据库木兰科含笑属的染色体统计数据来看,含笑属植物的染色体的特征为以较高对称性的m染色体(中部着丝粒染色体)为主,sm染色体(近中部着丝粒染色体)的变异范围在4个到18个之间,核型不对称性系数从1A到2B。该杂种F1代的细胞学特征从属于含笑属核型变异范围以内。从GISH图谱上可见,紫花含笑和灰岩含笑的亲本DNA探针在F1代的所有染色体上都存在交叉的杂交信号,在第1和第7号染色体上存在明显的父母本信号差异,证明了该杂交F1代确实含有亲本的染色体组分。
资助项目Cytology study can reveal important biological features of plants and answers to a certain degree in phylogeny and distribution of genetic materials and so forth. By hard working of cytologists, chromosome data of plants have been increased to a great abundance, but yet disorderly distributed in different magazines, which made researches based on the whole chromosome data of one taxon rarely launched. Scientific databases have become increasingly indispensable as researching data growing daily. As Cytological studies are booming in China, in order to fill the absence of digital and statistical data of plant chromosome researches and chromosome atlas, we started to develop a Chinese Seed Plants Chromosome Database, aiming to construct a database and start to record published chromosome data of Chinese seed plants. Based on this database, we chose the part of gymnosperms and gave a discussion to the features of its chromosomes’ evolution and variation. Cytological experiments have been applied to some important phyto-groups for phylogeny research and germplasm identification.Part I: The Chinese Seed Plants Chromosome Database and Discussion on the features of Gymnosperms chromosomes,1  The Chinese Seed Plants Chromosome Database,The frame of database was constructed by Microsoft Access 2003. 19 items of data were included in, they are: Chinese and Latin names of family, genus and species; plant pictures, mitosis metaphase and karyotype figures; morphological characteristics and distributions of the plant; chromosome numbers and basic numbers; karyotype formula; karyotype description; origin of the plant material; literature and the source of photos. In this database, data can be checked and shared easily by extracted out in species sorted interface or family sorted interface. 120 species in 29 genera and 10 families of Gymnospers have been collected and input to the database. In Angiosperms, 61 species in 10 genera of family Magnoliaceae and 80 species in 3 genera of family Theaceae have been collected and input to the database.2 Discussion on the features of evolution and variation of Gymnosperms chromosomes,By data collection of the database, we analyzed chromosome features of the group Gymnosperm. Plants of Gymnosperm had been through a long historical evolution on earth, fossil records of which originated from the late Devonian period. Once an authoritative and major classification level in the plant kingdom, most Gymnosperms have been extinct unless conifers, cycads, Ginkgo and Getales. Three main features of Gymnosperm chromosomes are: relatively large chromosome, which can be recognized from figures in the database; constant chromosome numbers, in most families of Gymnosperm the basic chromosome number keeps a certain value; comparatively low variation, karyotype under family level differs a little. The variation of chromosomes in Gymnosperm is dominated by Robertsonian changes. Contrary to common variation type in Angiosperms, the variation from high unsymmetric karyotype to low unsymmetric karyotype was found in existence in Gymnosperm.Part II: cytology research on some important phyto-groups,3 Karyomorphology of three species in the order Huerteales and their phylogenetic implications,The karyomorphology of three species in Dipentodon (Dipentodontaceae), Perrottetia (Celastraceae), and Tapiscia (Tapisciaceae), namely Dipentodon sinicus, Perrottetia racemosa, and Tapiscia sinensis, was investigated in the study. Recent molecular research has discovered close relationships among these three genera, which has led to the establishment of the order Huerteales with Perrottetia being placed in Dipentodontaceae. Herein we report the chromosome numbers of D. sinicus and P. racemosa for the first time, and present their karyotype formulas as 2n = 34 = 22sm + 12st (D. sinicus), 2n = 20 = 11m + 9sm (P. racemosa), and 2n = 30 = 22m(2SAT) + 8sm (T. sinensis). Asymmetry of their karyotypes is categorized to be Type 3B in D. sinicus, Type 2A in P. racemosa, and Type 2A in T. sinensis. Each of the species shows special cytological features. Compared with Perrottetia, Dipentodon has a different basic chromosome number, a higher karyotype asymmetry, and different karyomorphology of its interphase nuclei, mitotic prophase, and metaphase. Thus, on the basis of these results, we have reservations regarding the suggestion of placing Dipentodon and Perrottetia together in the family Dipentodontaceae.4 Genomic analyses of intergeneric hybrids between Michelia crassipes and M. calcicola by GISH,Genomic in situ hybridization (GISH) is becoming the method of choice for identifying parental chromosomes in interspecific hybrids. Interspecific F1 hybrid between Michelia crassipes and M. calcicola, tow highly ornamental species in Michelia of Magnolicaceae, has been analized by double-colored GISH with its parents’ genome as the probe. Research gave the results that the chromosome number of the F1 hybrid is 2n=38 as the same of species in Michelia and other genera in Magnoliaceae, the basic chromosome is x=19, the karyotype formula is 2n=38=32m+6sm, and the asymmetry of karyotype is 1B type. Based on chromosome data of Michelia in our database, the karyotype of this genus is featured mostly by metacentric chromosomes and submetacentric chromosomes. In Mechelia, the variation range of submetacentric chromosomes is 4 to 18 and of the karyotype asymmetry is 1A to 2B type. Both the karyotype and karyotype asymmetry type of F1 hybrid is among the variation range of Michelia. The figure of GISH showed that all the 38 chromosomes of F1 hybrid have crossing parental signals, and signal on the no.1 and no.7 chromosome showed differences, which proved that both the parental genome have been transmitted to and recombinated in F1 hybrid.
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文献类型学位论文
条目标识符http://ir.kib.ac.cn/handle/151853/16042
专题昆明植物所硕博研究生毕业学位论文
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杨静. 中国种子植物染色体数据库研制和一些重要类群的细胞学研究[D]. 中国科学院研究生院,2010.
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