An evaluation system of resistance ability of Japanese black pine clones against pine wood nematode based on parentage diagnoses using DNA molecular markers

基于DNA分子标记亲子鉴定的黑松无性系抗松材线虫能力评价体系

基本信息

批准号：
12556024
负责人：
SHIRAISHI Susumu
金额：
$ 7.1万
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：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

To establish the evaluation system in commercial forests of the strongness of resistance Japanese black pine (Pinus thunbergii) clones against pine wood nematode, a parentage diagnosis was developed as a basic technique for the system. Since chloroplast DNA (cpDNA) is paternally inherited in coniferous species, useful cpDNA markers were developed for the determination of the paternal parent of the progenies. The results were as follows ;1. For screening sequence polymorphisms of cpDNA, 18,569 bp-sequence constructed with 52 non-coding regions (13,360 bp) and coding regions (5,209 bp) were sequenced in 16 clones. The length is one seventh of the whole cpDNA. As the results, seven variations in five non-coding regions were recognized. Two mutations of them were single nucleotide polymorphism (SNP), three were simple sequence repeat polymorphism (SSR), one was insertion, and one was insertion / deletion (indel).2. To evaluate the length polymorphism in cpDNA SSR, twelve SSR regions were investigated. Eleven variations were recognized in the five regions.3. Chloroplast DNA markers inherited from its paternal parent were actually applied into the parentage diagnosis of the seeds produced in seed orchards. The DNA types of the embryo and endosperm isolated from a seed were analyzed separately and the maternal parent and paternal parent were detected from their cpDNA types. The contribution of each clone was estimated from the parentage diagnosis data. The result indicated that a few clones product the next generation (seeds) in seed orchards. This suggests a serious problem that must be overcome in seed orchard management.4. An evaluation system of resistance ability of 16 clones is available by using these cpDNA markers and nuclear SSRs together.

为了在商业森林中建立评估系统的耐药性强度日本黑松（Pinus thunbergii）针对松树线虫的克隆，开发了一个亲子症诊断作为该系统的基本技术。由于叶绿体DNA（cpDNA）是在针叶性物种中遗传的，因此开发了有用的cpDNA标记来确定后代父母的父母。结果如下; 1。对于CPDNA的筛选序列多态性，在16个克隆中测序了由52个非编码区（13,360 bp）构建的18,569个BP序列（13,360 bp）和编码区（5,209 bp）。长度是整个cpDNA的第七个。作为结果，认识到五个非编码区域的七个变化。它们的两个突变是单核苷酸多态性（SNP），三个是简单的序列重复多态性（SSR），一个是插入，一个是插入 /缺失（Indel）.2。为了评估CPDNA SSR的长度多态性，研究了十二个SSR区域。在五个区域中确认了11种变化。3。实际上，从其父母父母继承的叶绿体DNA标记实际上被应用于种子果园中产生的种子的父母诊断中。分别分析了从种子中分离的胚胎的DNA类型和胚乳的类型，并从其CPDNA类型中检测到母亲父母和父母。每个克隆的贡献是从父母诊断数据中估计的。结果表明，几个克隆产品是种子果园中的下一代（种子）。这表明在种子果园管理中必须克服一个严重的问题4。通过将这些CPDNA标记和核SSR一起使用16个克隆的电阻能力的评估系统。