Genetic mapping in Japanese coniferous tree species using microsatellite DNA markers

使用微卫星 DNA 标记对日本针叶树种进行遗传图谱分析

基本信息

批准号：
13460070
负责人：
SHIRAISHI Susumu
金额：
$ 9.28万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13460070/
关键词：
forest tree breeding DNA molecular marker microsatelliteDNA SSR linkage map Cryptomeria japonica Chamaecyparis obtusa Pinus thunbergii TAC アカシア PIMA ジーンマッピンギ

项目摘要

To establish an efficient selection system of SSR (simple sequence repeats) DNA regions, TAC (triplex affinity capture) that is a new method instead of a molecular hybridization method, a modified PIMA (PCR-based isolation of microsatellite arrays) and so on was investigated. The results were as follows:1.Using model DNAs in which [CT]-repeats ([CT]_<10>,[CT]_<15>,[CT]_<20>, and [CT]_<25>) were inserted, an optimization of TAC was executed on pH and temperature conditions of the reaction. In pH 6.4 -7.0, highly repeated DNA fragments were selected efficiently.2.The original PIMA method was modified for the reliable screening of the recombinant colony in which SSR was inserted.3.The modified TAC and PIMA were applied into actual screening of SSRs from the genomes of Cryptomeria japonica, Pinus thunbergii, Chamaecyparis obtuse, and Acacia mangium. In C.japonica, About 300 SSR fragments were selected, and SCAR (sequence characterized amplified region) markers were designed based on their sequences.4.The materials of families for constructing linkage-maps were made in C.japonica (a haploid-endospermic family of Sawara-1), P.thunbergii(a haploid-endospermic family of Sima-64), and C.obtuse (a cross-pollinated family between Aira-32 and Satsuma-8).

为了建立一个有效的SSR选择系统（简单序列重复）DNA区域，TAC（Trilex亲和力捕获），它是一种新方法，而不是一种分子杂交方法，一种修饰的PIMA（基于PCR的微磷灰石阵列分离）等等，因此调查。结果如下：1。使用模型DNA，其中[ct] - repeats（[ct] _ <10>，[ct] _ <15>，[ct] _ <20>和[ct] _ <25 >）插入，在反应的pH和温度条件下对TAC进行了优化。在pH 6.4 -7.0中，有效地选择了高度重复重复的DNA片段。2。对原始PIMA方法进行了修改，以可靠地筛选插入SSR的重组菌落的可靠筛选。3。修改后的TAC和PIMA被应用于SSR的实际筛选中来自japonica，Pinus thunbergii，chamaecyparis钝的基因组和杂种。在japonica中，选择了约300个SSR片段，并根据其序列设计了疤痕（序列表征了放大区域）标记。4。用于构建连锁图的家族的材料是在C.Japonica中制成的（单倍倍性粘膜 - 内膜体验Sawara-1的家族），P.Thunbergii（Sima-64的单倍体 - 式经典家族）和C.Obtuse（Aira-32和Satsuma-8之间的一个交叉授粉家庭）。