GENE TRANSFORMATION EXPERIMENT OF THE SELF-INCOMPATIBILITY RELATED GENES

自交不亲和性相关基因的基因转化实验

• 批准号: 05556002
• 负责人: HINATA Kokichi
• 金额: $ 10.62万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05556002/
• 关键词: SELF-INCOMPATIBILITY; BRASSICA CAMPESTRIS; PYRUS PILIFOLIA; SLG; SRK; S-RNASE; GENE-MANIPULATION; TRANSGENIC PLANT; S遺伝子; RNase; アブラナ科; ニホンナシ; クローニング; 二十世紀ナシ; SELF-INCOMPATIBILITY; BRASSICA CAMPESTRIS; PYRUS PILIFOLIA; SLG; SRK; S-RNASE; GENE-MANIPULATION; TRANSGENIC PLANT; S遺伝子; RNase; アブラナ科; ニホンナシ; クローニング; 二十世紀ナシ

Objectives : To understand the molecular mechanism of self-incompatibility and to assess a possibility of molecular breeding of this character, molecular characterization and gene manipulation was conducted on the S-related genes in sporophytic self-incompatible plant : Brassica campestris and in gemetophytic self-incompatible plant : Pyrus pilifolia.Results : In B.campestris, SLG8, SLG9, SLG12, SRK8, SRK9, SRK12, SRA1, SRA2, SRA3, SRA4 and SRBs were isolated from cDNA or genomic DNA libraries and sequenced. Dendrograms constructed on the basis of synonymous and nonsynonymous substitutions indicated that SRA differentiated first, followed by SRB and SLG/SRK.When the SLG8 genomic clone was introduced into B.campestris some of transformed plants changed to self-compatible. Furthermore, the antisense SLG gene was transformed into the same species. Analysis of a transformant showed that the self-incompatibility was broken down by the action of the introduced antisense gene. These results i … More ndicated the self-incompatibility could be controlled, at least partly, by gene manipulation. The genes encoding 3 RNases were cloned from the style of a self-incompatible cultivar, Nijisseiki (S2S4), and its self-compatible mutant, Osa-Nijisseiki (S2S4sm, sm means stylar part mutant), of Japanese pear. For Nijisseiki, cDNAs coding for two S-RNases (S2-RNase and S4-RNase) and an RNase unrelated to self-incompatibility (non-S-RNase) were cloned from the stylar cDNA library. However, the cDNA coding for S4-RNase was neither amplified by PCR nor cloned from the library of Osa-Nijisseiki, suggesting that the mutation of self-incompatible Nijisseiki to self-compatible Osa-Nijisseiki is due to a failure of expression of S4-RNase. The S3-gene promoter of Petunia inflata was used in constructing the vectors for the Japanese pear S-genes. The binary vectors of the Agrobacterium were transformed in the petunia resulting in shoot regeneration. In vitro cultures of the Japanese pear were also extended. Less

目的：了解自我兼容性的分子机制并评估该特征分子繁殖的可能性，分子表征和基因操纵是对孢子型自我不合别植物中与S相关的基因进行的：Brassica Campestris和gemetphy campestris和gemetphoys pofysephophy timpophy timpophy timpophy timpophy timpophy timpophy timpophy timpophy torcamibletic plastibaltic plastibaltic plastiby植物：pyrus pilfgolia。从cDNA或基因组DNA库中分离出SRK8，SRK9，SRK12，SRK12，SRA1，SRA2，SRA3，SRA4和SRB并进行了测序。基于同义和非同步取代构建的树状图表明，SRA首先区分了，然后是SRB和SLG/SRK。当SLG8基因组克隆引入b.campestris中时，一些转化的植物变成了自我兼容。此外，反义SLG基因被转化为同一物种。对转化剂的分析表明，引入的反义基因的作用分解了自我兼容性。这些结果i…更加谨慎，至少可以通过基因操纵来控制自我兼容性。编码3个RNass的基因是从自兼容的品种Nijisseiki（S2S4）的风格中克隆的，其自我兼容的突变体Osa-Nijisseiki（S2S4SM，SM含义SM含义stylar Part突变体），日本梨。对于nijisseiki，从Stylar cDNA库中克隆了两个S-RNase（S2-RNase和S4-RNase）和与自我不相容性（非S-RNase）无关的RNase编码的cDNA。然而，编码为S4-RNASE的cDNA既不是由PCR扩展，也没有从OSA-Nijisseiki图书馆中克隆，这表明自我不相容的Nijisseiki对自我兼容的Osa-Nijisseiki的突变是由于S4-RNASE表达的失败。矮牵牛的S3基因启动子用于构建日本珍珠S-Genes的矢量。农杆菌的二元载体在矮牵牛中转化，导致芽再生。日本珍珠的体外培养物也扩展了。较少的