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Study on mitotic and meiotic homologous recombination in plant and cultured cells of rice

水稻植株和培养细胞有丝分裂和减数分裂同源重组的研究

基本信息

批准号：
15580001
负责人：
NIIZEKI Minoru
金额：
$ 2.37万
依托单位：
Hirosaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15580001/
关键词：
homologous recombination RiLIM15 Alternative splicing Meiosis southern blot Exon Intron gene family 相同染色体組換え遺伝子 mRNA cDNA ソマクローナル変異培養細胞 OsRAD51 相同染色体組換えイネゲノムモチ・ウルチ分離比

项目摘要

1.Analysis of occurrence of MCO in cultured cell of rice : Calli were induced from F_1 seed (Wxwx) of glutinous rice (wxwx) as a seed parent and non-glutious rice(WxWx) as a pollen parent. After regeneration of rice plamts, it was found a dominant homologous plant(WxWx) which might be segregated by the MCO.2.Analysis of structure and expression pattern of RiLIM15 in rice : In order to clarify the relationships between occurrence of MCO and expression of RiLIM15 gene in cultured cells and panicles investigated on the aspects of structure and expression of RiLIM15. Two RiLIM15 homologous RiLIM15A and RiLIM15B were isolated. In exon region, the DNA sequences for two genes were highly homologous but not intron regions. The amino acid sequences these two genes were highly homologous, sugggesting that they may be nearly equivalent in function. Analysis of cDNA sequences that eight patterns of deletions of DNA sequences in RiLIM15 cDNA clones derived from both meiotic and cultured cells. They may be resulted from alternative splicing. The putative mRNA with these deletion may translate different functions of proteins.3.Analysis of RAD51 in rice : RT-PCR was performed using total RNA extracted from the cultured cells, young meiotic panicles, mature leaves and X-ray treated seedlings. Expression of the RAD51 gene was found to occur in cultured cells, young meiotic panicles and X-ray treated seedlings, but not mature leaves. The gene possibly participates in DNA repariring of MCO.4.Application to substantial plant breeding : In nine generation of progenies regenerated from calli of cv.Akihikari it was observed that a line is 3-4 early heading date and almost the same number of panicle as the parent cultivar and significantly higher thousand kernel weight than the parent cultivar.

1.水稻离体培养细胞中MCO发生的分析：以糯性水稻（WxWx）为种子亲本，非糯性水稻（WxWx）为花粉亲本，F_1代种子（WxWx）诱导出愈伤组织。水稻植株再生后，发现一个显性同源植株（WxWx），可能是由MCO分离的。2.水稻RiLIM 15基因的结构和表达模式分析：为了阐明MCO的发生与RiLIM 15基因在培养细胞和幼穗中的表达之间的关系，从RiLIM 15基因的结构和表达方面进行了研究。分离到两个RiLIM 15同源物RiLIM 15 A和RiLIM 15 B。在外显子区，两个基因的DNA序列高度同源，但在内含子区没有同源性。这两个基因的氨基酸序列高度同源，提示它们可能在功能上几乎相同。对RiLIM 15细胞减数分裂和培养细胞中8种DNA序列缺失模式的cDNA序列进行分析。它们可能是可变剪接的结果。3.水稻RAD 51基因的分析：以水稻离体培养细胞、幼穗、成熟叶片和经X射线处理的幼苗为材料，提取总RNA，进行RT-PCR分析。RAD 51基因的表达被发现发生在培养的细胞，年轻的减数分裂穗和X射线处理的幼苗，但不成熟的叶片。该基因可能参与MCO的DNA修复。4.在植物育种上的应用：在9个世代的秋光愈伤组织再生后代中，观察到一个株系的抽穗期早3-4天，穗数与亲本基本相同，千粒重显著高于亲本。