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Studies of population genetics on four mangrove tree species in Okinawa, Japan

日本冲绳四种红树树种的群体遗传学研究

基本信息

批准号：
09660169
负责人：
BABA Shigeyuki
金额：
$ 2.11万
依托单位：
University of the Ryukyus
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

In this studies, attempts have been made to analyze genetical variation of four mangrove tree species, Bruguiera gymnorrhiza (Rhizophoraceae). Kandelia candel (Rhizophoraceae), Rhizophora stylosa (Rhizophoraceae) and Sonneratia alba (Sonneratiaceac), distributed in Okinawa, Japan, and adding one species, Lumnitzera racemosa (Combretaceae) for DNA analysis. Leaves as samples were collected from trees of four species, and stocked in a deep-freezer before experiment. Polyacrylamide gel electrophoresis (PAGE) of a vertical slab-format method was used for analyzing isozyme loci of four mangroves. After the electrophoresis, 23 enzyme species were stained. Fine zymogrames were obtained five enzyme species for Rhizophora stylosa and five enzyme species for Bruguiera gymnorrhiza. Polymorphism was found only peroxidase (POD) for Bruguiera gymnorrhiza, but diaphorase (DIA), esterase (EST) and glucokinase GR) for Sonneratia alba.DNA was extracted from leaves of Rhizophora stylosa. Both CTAB (Cetyl Trimethyl Ammmonia Bromide) and Ice Cold Buffer methods were applied for DNA extraction. The Ice Cold Buffer method was better than the CTAB method for DNA extraction. To obtain mitochondrial DNA fragments, 18 restriction enzyme species were applied. For amplification of chloroplast DNA, 23 primers were used. Mitokondorial and genom DNA fragments were amplified and analyzed. Since genetical variations were small in studied mangrove species collected in Okinawa, primers applied were not so efficiently worked. However, we could obtain evidence that micro-satellites are one of efficient ways for gentical variation analysis of Rhizophora stylosa in Ryukyu islands.

在这项研究中，尝试分析四种红树树种木榄（红树科）的遗传变异。秋茄（Rhizophoraceae）、红海桑（Rhizophoraceae）和海桑（Sonneratiaceac），分布于日本冲绳，并添加一种种，Lumnitzera racemosa（Combretaceae）进行DNA分析。从四种树上采集叶子作为样品，并在实验前储存在深冰箱中。采用垂直平板法的聚丙烯酰胺凝胶电泳（PAGE）对四种红树林的同工酶位点进行了分析。电泳后，23种酶被染色。获得了红树5种酶和木榄5种酶的精细酶谱。木榄仅检测到过氧化物酶(POD)，而白海桑(Sonneratia alba)的心肌黄酶(DIA)、酯酶(EST)和葡萄糖激酶GR)存在多态性。从红树叶中提取DNA。 CTAB（十六烷基三甲基溴化铵）和冰冷缓冲液方法均用于 DNA 提取。冰冷缓冲液法比 CTAB 法提取 DNA 效果更好。为了获得线粒体 DNA 片段，使用了 18 种限制性内切酶。为了扩增叶绿体 DNA，使用了 23 个引物。扩增并分析了线粒体和基因组 DNA 片段。由于在冲绳收集的研究红树物种中遗传变异很小，因此所使用的引物效果不那么有效。然而，我们可以获得证据表明微卫星是琉球群岛红树遗传变异分析的有效方法之一。