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1、.蚇羇羀莇蝿袀艿莆葿肅膅莅薁袈肁莄蚃肄羇蒄螆袇芅蒃蒅虿膁蒂薈袅肇蒁螀蚈肃蒀蒀羃罿葿薂螆芈蒈蚄羁膄蒈螆螄肀薇蒆羀羆薆薈螂芄薅蚁羈芀薄袃螁膆薃薃肆肂膀蚅衿羈腿螇肅芇膈蒇袇膃芇蕿肃聿芆蚂袆羅芅螄蚈莃芅薄袄艿芄蚆螇膅芃螈羂肁节蒈螅羇芁薀羁芆莀蚂螃膂荿螅罿肈荿蒄螂羄莈蚇羇羀莇蝿袀艿莆葿肅膅莅薁袈肁莄蚃肄羇蒄螆袇芅蒃蒅虿膁蒂薈袅肇蒁螀蚈肃蒀蒀羃罿葿薂螆芈蒈蚄羁膄蒈螆螄肀薇蒆羀羆薆薈螂芄薅蚁羈芀薄袃螁膆薃薃肆肂膀蚅衿羈腿螇肅芇膈蒇袇膃芇蕿肃聿芆蚂袆羅芅螄蚈莃芅薄袄艿芄蚆螇膅芃螈羂肁节蒈螅羇芁薀羁芆莀蚂螃膂荿螅罿肈荿蒄螂羄莈蚇羇羀莇蝿袀艿莆葿肅膅莅薁袈肁莄蚃肄羇蒄螆袇芅蒃蒅虿膁蒂薈袅肇蒁螀蚈肃蒀蒀羃罿葿薂螆芈
2、蒈蚄羁膄蒈螆螄肀薇蒆羀羆薆薈螂芄薅蚁羈芀薄袃螁膆薃薃肆肂膀蚅衿羈腿螇肅芇膈蒇袇膃芇蕿肃聿芆蚂袆羅芅螄蚈莃芅薄袄艿芄蚆螇膅芃螈羂肁节蒈螅羇芁薀羁芆莀蚂螃膂荿螅罿肈荿蒄螂羄莈蚇羇羀莇蝿袀艿莆葿肅膅莅薁袈肁莄蚃肄羇蒄螆袇芅蒃蒅虿膁蒂薈袅肇蒁螀蚈肃蒀蒀羃罿葿薂螆芈蒈蚄羁膄蒈螆螄肀薇蒆羀羆薆薈螂芄薅蚁羈芀薄袃螁膆薃薃肆肂膀蚅衿羈腿螇肅芇膈蒇袇膃芇蕿肃聿芆蚂袆羅芅螄蚈莃芅薄袄艿芄蚆螇膅芃螈羂肁节蒈螅羇芁薀羁芆 粪便DNA论文：越冬白头鹤种群遗传多样性的初步研究【中文摘要】白头鹤是IUCN红色物种名录中的濒危物种,需要加强种群保护遗传学的研究。本文以粪便为研究材料,通过高质量粪便DNA样品的提取,对白头鹤个
3、体(来自3个不同地区即升金湖、菜子湖、鄱阳湖自然保护区)的线粒体基因组控制区序列进行测定和群体遗传分析。(1)以1103bp左右长度的线粒体控制区DNA序列进行分析,获得38条线粒体控制区序列,序列比对后共检测到12个变异位点,定义12种单倍型,其中2种为安徽(菜子湖、升金湖)和鄱阳湖两个种群所共有,10种为各个种群所特有。从Genebank下载两条白头鹤线粒体控制区序列,增加了2种单倍型。白头鹤两个种群总单倍型多样性(h)为0.784,核苷酸多样性()为0.00128,核苷酸差异均数(k)为1.414。(2)安徽种群(菜子湖、升金湖)和鄱阳湖种群的单倍型多样性分别为0.802、0.679,核
4、苷酸多样性分别为0.00143、0.00071,核苷酸差异均数分别为1.572、0.786。相对于鄱阳湖种群,安徽种群(菜子湖、升金湖)的单倍型多样性、核苷酸多样性、核苷酸差异均数相对较高。中性检验和歧点分布检验(SSD=0.00879,P=0.16)均表明白头鹤在历史上发生过种群扩张事件,时间约在17916年前。分子变异分(Analysis of Molecular Variance, AMOVA)的结果表明白头鹤的遗传变异99.40%发生在种群内,种群间的遗传差异只有0.6%。种群遗传分化不明显Fst=0.00600(P>0.05)。从系统树上也可以进一步说明这两个种群间没有出现显著
5、的遗传分化,两个种群间存在基因交流,存在共享单倍型,这种单倍型的存在可能源于古老单倍型,也可能是群体近期分歧或者群体间存在一些基因流所产生的。【英文摘要】Hooded Crane Grus monacha is a threatened species on the Red List of IUCN and their population is still in decline due to habitat loss. It is necessary to study the genetic diversity for effective conservation of the bird.
6、In this study, faeces samples from three lakes, Shengjin Lake, Caizi Lake and Poyang Lake were collected for genetic analysis. The results show as fellow.(1) A total of 1103 base pairs bp of the mitochondrial control region from Hooded crane collected from two lakes in Anhui(Caizi Lake, Shengjin Lak
7、e) and Poyang Lake in Jiangxi Province were sequenced.38 control region sequences were obtained. In 38 control region sequences,12 variable sites were determined and 12 haplotypes were identified, of which two haplotypes were shared by all populations, 10 haplotypes were unique for single population
8、. Combining with the two sequences obtained from GeneBank,2 haplotypes were identified. Haplotypes diversity(h) of all populations was 0.784, nucleotides diversity() was 0.00128, and average number of nucleotide differences(k) was 1.414.(2) Haplotypes diversity(h) of populations in Anhui and Poyang
9、were 0.802 and 0.679, nucleotide diversity () were 0.00143 and 0.00071 respectively, while average number of nucleotide differences(k)were 1.572 and 0.786 respectively. In these populations, haplotypes diversity, nucleide diversity and average number of nucleotide differences were higher in the Anhu
10、i population than in the Poyang population. Neutrality test and mismatch distribution analysis(SSD=0.00879, P=0.16) revealed a significant population expansion sign in the Hooded crane population, which happened in about 17916 year ago. Analysis of molecular variance(AMOVA) revealed that 99.4% of th
11、e variation was within populations, while among populations was only 0.6%. The genetic variance was not significant (Fst=0.00600, P>0.05), as was also indicated in the phylogenetic tree. Gene flow was identified between the two populations, which shared haplotypes. The haplotypes may result from
12、the presence of ancient haplotypes, or differences between groups in the near future, or gene flow among populations.【关键词】粪便DNA 白头鹤 遗传多样性 线粒体【英文关键词】Faecal DNA Grus monacha genetic diversity mitochondrial DNA【目录】越冬白头鹤种群遗传多样性的初步研究 摘要 3-4 Abstract 4-5 目录 6-8 第一章 引言 8-17 1 濒危水鸟保护遗传学的研究概况 8-12 1.1 遗传多样性对
13、于濒危物种种群维持的意义 8-9 1.2 保护遗传学的研究方法 9-12 2 白头鹤种群现状及保护遗传学概况 12-16 2.1 白头鹤的种群现状 12-13 2.2 白头鹤保护遗传学研究概况 13-15 2.3 越冬白头鹤种群保护中存在的问题 15-16 3 本研究的目的、意义 16-17 第二章 研究方法 17-23 1 材料和方法 17-21 1.1 实验材料 17-18 1.2 实验方法 18-21 2 数据分析 21-23 2.1 数据整理 21-22 2.2 序列变异和单倍型 22 2.3 中性检验和种群扩张 22 2.4 种群遗传结构 22-23 第三章 研究结果 23-39 3
14、.1 基因组DNA的PCR扩增结果 23-25 3.2 基因组测序结果 25-28 3.3 线粒体控制区序列变异及遗传多态性 28-39 第四章 讨论 39-45 4.1 白头鹤种群遗传多样性分析 39-41 4.2 白头鹤种群历史动态分析 41 4.3 白头鹤两地理单元遗传多样性分析及保护对策 41-42 4.4 硫氰酸胍改进法提取粪便DNA的优点 42-43 4.5 粪便DNA提取的影响因素及消除措施 43-45 第五章 结论 45-46 参考文献 46-59 致谢 59-60 攻读学位期间发表的论文目录 60 螈肀膁薀螇膂莆蒆螆袂腿莂螅肄蒅莈螅膇芈蚆螄袆蒃薂螃罿芆蒈螂肁蒁莄袁膃芄蚃袀袃肇
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