Construction of a new improvement system using DNA molecular technique in the forest tree species

利用DNA分子技术构建森林树种新改良体系

基本信息

批准号：
09556032
负责人：
SHIRAISHI Susumu
金额：
$ 8.45万
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：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09556032/
关键词：
DNA molecular marker breeding system cultivar diagnosis clone analysis parentage diagnosis MuPS analysis chloroplast DNA marker SCAR 林木育種品種識別育種戦略父性遺伝

项目摘要

For constructing an effective improvement system of forest tree species using DNA molecular technique, we developed a new DNA molecular marker and tried to apply the actual breeding.(1) A new DNA molecular marker for identification of cultivars/clones : In Acacia mangium and Cryptomeria japonica (Sugi), more than 18 SCAR (sequence characterized amplified regions) markers were designed from RAPD (random amplified polymorphic DNA) fragments in each species. By combining these SCAR markers, a MuPS (multiplex-PCR of SCAR markers) system was constructed using Multiplex-PCR.This system possesses the discrimination power to identify ca. 260,000 genotypes.(2) Survey of the intra-specific chloroplast DNA polymorphism in Pinus thunbergii : BESS (base excision sequence scanning), CFLP (cleavase fragment length polymorphism), and SSR (simple sequence repeats) analysis of the chloroplast DNA revealed the eight mutations in the intergenic spacer regions. These chloroplast DNA markers were useful for ascent the pollen parent of progenies.(3) Application MuPS system into the classification of Sugi cultivars and plus tree clones : We determined the MuPS types of 77 cultivars and 589 plus tree clone of Kyushu and constructed the data base of their MuPS types. As the result of MuPS analysis, quite a few plus tree clones originated from some cultivars or some unknown cutting clones. These duplicated clones should be eliminated from the plus trees population of Kyushu region.

为了使用DNA分子技术构建有效的森林树种的改进系统，我们开发了一种新的DNA分子标记，并试图应用实际的育种。（1）一种新的DNA分子标记物，用于鉴定品种/克隆：杂种杂种和cryptptomeria in Acacia in Japonica（SUGI），超过18个疤痕（表征放大区域的序列）标记是根据每种物种中RAPD（随机扩增的多态性DNA）片段设计的。通过结合这些疤痕标记，使用多重PCR构建了MUP（疤痕标记的多重PCR）系统。该系统具有识别CA的歧视能力。 260,000个基因型。（2）对Pinus thunbergii中特异性内叶绿体DNA多态性的调查：BESS（碱基切除序列扫描），CFLP（切割酶片段长度多态性）和SSR（简单的序列重复）揭示了八个80基因间间隔区的突变。这些叶绿体DNA标记物可用于上升的后代花粉母体。（3）将MUPS系统应用于Sugi品种和加上树克隆的分类：我们确定了77种品种的MUPS类型和589个Plus Tree Clone Clone of Kyushu并构建了数据他们的MUPS类型的基础。作为MUPS分析的结果，相当多的树克隆起源于某些品种或一些未知的切割克隆。这些重复的克隆应从九州地区的普通树木种群中消除。