Molecular analysis of the mechanisms for controlling tannin accumulation of pollination-constant nonastringent (PCNA)-type persimmons in Chinese and Japanese cultivars to clarify different origins between them.

对中国和日本品种授粉恒定非涩性（PCNA）型柿子的单宁积累控制机制进行分子分析，以阐明它们之间的不同起源。

基本信息

批准号：
16380025
负责人：
YONEMORI Keizo
金额：
$ 10.05万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

项目摘要

The analysis was performed to identify key genes for regulating tannin accumulation into fruits of pollination constant non-astringent (PCNA)-type of persimmon originated in Japan and China. For this purpose, we conducted the following three experiments:1)We constructed genomic library from closely related diploid species, D. lotus, and a seed clone was screened from the library with a probe tightly linked to the trait of natural astringency-loss of Japanese-type PCNA fruits. Then, we constructed ca. 300 kbp contig of D. lotus. Due to the sequence analysis of this contig, we found that the sequences are very homologous to those of D. kaki and it can use for exploring the gene of persimmon (D. kaki). In addition, primer sets designed from the sequences of this contig were able to be applicable to distinguish Japanese-type PCNA from others. We could make a universal primer set to select PCNA persimmon.2)We conducted the expression analysis of mRNA to identify the genes involved in tannin accumulation of persimmon. For this purpose, suppression subtractive hybridization (SSH) analysis was performed between astringency-removed fruits with ethanol treatment on the tree and non-treated fruits. According to this analysis, we were able to obtain several candidates for the genes involved in tannin accumulation into the fruits.3)As for investigation of a dominant gene conferring the trait of Chinese-type PCNA, we looked for molecular markers by AFLP method combined with bulked segregant analysis (BSA) using progenies derived from a Chinese PCNA 'Luo Tian Tian Shi'. By this analysis, we found three markers closely linked to the Chinese-PCNA trait. Using a marker among them, we screened genomic library of `Luo Tian Tian Shi' and obtained a genomic clone as the seed for constructing contig for the survey of the gene.

进行了分析，以鉴定将单宁积累调节成授粉恒定非群体（PCNA） - 柿子型的关键基因，起源于日本和中国。为此，我们进行了以下三个实验：1）我们从密切相关的二倍体物种D. lotus构建了基因组文库，并从文库中筛选了一个种子克隆，并与日本型PCNA果实的自然误射性特征密切相关。然后，我们构建了Ca。 300 kbp的D.莲花。由于该重叠群的序列分析，我们发现序列与D. kaki的序列非常同源，并且可以用于探索柿子的基因（D. kaki）。此外，根据该重叠群的序列设计的底漆集可以适用于将日本型PCNA与其他pCNA区分开。我们可以制作一个通用底漆来选择PCNA柿子。2）我们进行了mRNA的表达分析，以鉴定柿子单宁积累的基因。为此，在树上对乙醇处理和未处理的水果进行了与气的水果和未处理的水果之间进行抑制减法杂交（SSH）分析。根据这一分析，我们能够获得几个候选单宁积聚到水果中的候选物。3）对于赋予中国型PCNA特征的主要基因的研究，我们通过AFLP方法与使用Bumbed Seggregant分析（BSA）相结合的AFLP方法来寻找分子标记，使用了来自中国pcna shian pcna theian pcna'luo'luo'lu o'lu'lu'lu'通过此分析，我们发现三个标记与中国pcna性状密切相关。使用其中的标记，我们筛选了“ Luo Tian Tian Shi”的基因组文库，并获得了基因组克隆作为构造基因调查的重叠群的种子。