【作者】 王晓明；

【导师】 马泽芳； 金煜；

【作者基本信息】 东北林业大学 ， 野生动植物保护与利用， 2003， 硕士

【摘要】 本文克隆并测定了13只东北虎mtDNA D-loop区381bp的序列，并对这些序列和GenBank中注册的2个东北虎D-loop区序列和32个虎cyt b基因全序列(包含4个亚种)进行了分析，研究了cyt b基因和D-loop区序列的特点及其在亚种识别中的价值。研究发现，测得的东北虎D-Loop区381bp的序列中共发现79个变异位点，共有(?)个单倍型；D-Loop区中存在着两种序列：一是与其他物种同源性很高的“正常序列”，称为“短序列”；二是带有22bp的插入片断的“非正常序列”，称为“长序列”。11个“短序列”间同源性在99.5％～100％；4个“长序列”间的同源性在98.5％～99.2％；“短序列”和“长序列”间的同源性为75.5-81.6％。上述发现表明，在选择亚种识别遗传标记时，应区别对待D-loop区的两种序列；本文所研究的区域因为两种序列间的差异很大，所以不适合用作系统进化的分析。在32个虎mtDNA cyt b 1140bp的序列中，共发现14个位点存在变异，占总序列的1.23％，个体间序列的同源性在99.3～100％，平均为99.6％，没有发现缺失或插入。在cyt b基因上共检出10种单倍型，各亚种均有特有的标志性碱基，亚种间无共享单倍型。这表明mtDNA cyt b基因非常适合虎亚种识别；本文提出了cyt b基因上识别东北虎、孟加拉虎、印支虎、苏门答腊虎4个亚种的特异性单倍型。更多还原

【Abstract】 A sequence of 381bp in D-loop region of 13 Amur tiger mtDNA were cloned and sequenced. 2 sequences of Amur tiger D-loop region and 32 sequences of cyt b gene of 4 tiger subspecies in GenBank were also combined to our data in the analysis of characteristics of the two regions for evaluating their significance of subspecies identification. The result showed that 79 polymorphic sites forming 9 haplotypes were found within the 381bp region in D-loop; Two types of sequences were discovered in D-loop region of Amur tiger, Short Sequence that is highly homologous to other species, and Long Sequence that contains a 22bp insertion. The homology among 11 Short Sequences is 99.5%~100%, whilst that among 4 Long Sequences is 98.5%~99.2%. The homology between is Short Sequence and Long Sequence is75.5-81.6%. It indicated that the Long and Short sequence should be considered respectively when they become candidates for identifying subspecies; This region in D-loop is not suitable for phylogenetic analysis in case of their extremely large divergence. In the 32 full length cyt b sequences, 14 polymorphic sites forming 10 haplotypes were found, occupying 1.23% of the whole sequence. The homology among individuals is between 99.3 ~ 100%, averagely 99.6%, neither insertion nor deletion is found. All subspecies have their specific base at certain sites. These results showed that cyt b gene is very suitable for identifying subspecies of tiger. We proposed specific haplotypes on cyt b gene for specifically identifying Amur tiger, Bengal tiger, Indo-China tiger and Sumatran tiger.更多还原