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CELLULAR AND MOLECULAR APPROACHES TO THE BREEDING BARRIER IN PLANTS, APOMIXIS.

植物无融合生殖育种障碍的细胞和分子方法。

基本信息

批准号：
09460004
负责人：
ADACHI Taiji
金额：
$ 3.07万
依托单位：
Osaka Prefecture University (1999)University of Miyazaki (1997-1998)
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-09460004/
关键词：
Apomixis Guineagrass Bahiagrass RT-PCR Simplified DD Method Adventitious embryony バヒア・グラス RT・PCR法

项目摘要

Cytological Approach: At first, the evaluation of the apomictic ratio was performed in facultative apomict species, Guineagrass and Bahiagrass by means of Nomarski's differential interference contrast mirography (DIC). Then, virietal differences of the capability to form adventitious embyos in tissue culture were compared with that of apomictic ratios. There was some corelations among them. In situ hybridization experiment was also carried out at mRNA levels in each developmental stages of zygotic embryo by using DNA probes concerning to the adventitious embryo formation in alfalfa and rice. To now, unfortunately, technical problems still remain to be resolved.Molecular Approach: By checking the stage at which apospory is determined in Guineagrass, some candidate of cDNA were isolated from developmentally selected samples by means of simplified Differential Display Methods (DD). The gene expression of the candidates were confirmed by Northern hybridization. And then, homology search of candidate, A2-134 was performed with the BLAST-Version 2.1 PIR database. Database search for sequence homology unvailed a relation to the RD22 gen of Arabidopsis thaliana with 50.9% similarity in nucleotide sequence. The RD22 protein regulates the expression of seed-specific genes and probably also genes in responcse to dehydration stress.

细胞学方法：首先，通过Nomarski的微分干涉显微照相术(DIC)对兼性无融合生殖物种、几内亚草和巴伊亚草进行无融合生殖比率的评估。然后，将组织培养中形成不定胚的能力的虚拟差异与无融合生殖比率进行比较。它们之间存在一些相关性。利用苜蓿和水稻不定胚形成的DNA探针，在合子胚各发育阶段的mRNA水平上进行了原位杂交实验。不幸的是，到目前为止，技术问题仍有待解决。分子方法：通过检查几内亚草中无孢子确定的阶段，通过简化的差异显示方法(DD)从发育选择的样品中分离出一些候选cDNA。候选者的基因表达通过Northern杂交得到证实。然后，利用BLAST-Version 2.1 PIR数据库对候选A2-134进行同源性搜索。数据库搜索序列同源性，未发现与拟南芥 RD22 基因的核苷酸序列相似性为 50.9%。 RD22 蛋白调节种子特异性基因的表达，也可能调节响应脱水胁迫的基因的表达。