一个水稻CCCH型锌指蛋白基因OsZF142功能的分析
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摘要
锌指蛋白转录因子是植物中最丰富的一类转录因子,调控植物各个时期的生长发育和环境胁迫下基因的表达。本研究通过RT-PCR和转基因方法分析了一个新的水稻CCCH型锌指蛋白基因OsZF142的表达特征、启动子活性及其生物学功能。RT-PCR分析发现,OsZF142在水稻根、茎、叶、花和种子等器官中倾向于发育的种子中表达,且倾向于胚中优势表达,不在胚乳中表达,表明OsZF142基因是一个水稻种子胚乳不表达、胚优势表达锌指蛋白基因。进一步利用OsZF142基因启动子与GUS构建的融合表达载体转基因分析发现,OsZF142基因的启动子序列具有类似的启动子活性,转基因水稻植株种子的胚乳中没有检测到GUS活性,而胚中检测到较强的GUS活性,根、茎、叶等器官和组织中也检测到相对较弱的GUS活性,进一步表明锌指基因OsZF142的启动子是一个胚乳不表达、胚优势表达启动子。对正反义OsZF142转基因日本晴水稻植株与野生型日本晴植株比较观察发现,反义转基因日本晴植株在形态表型上与野生型日本晴差异不明显;但正义OsZF142转基因水稻植株普遍表现生长发育变异,主要表现在:水稻整个生长发育延迟(包括种子萌发及幼苗生长延迟、主茎出叶及分蘖变慢、分蘖能力受到抑制、抽穗延迟、叶片延缓衰老等),水稻穗颈变短和植株变矮等一系列异常的多效性表型,暗示了OsZF142蛋白是一个水稻生长发育的负调控因子。
Zinc-finger protein transcription factors arc one of the most abundant transcription factors in plants, and they widely regulate gene expression in different growth phases and environmental stress. By RT-PCR and transgene techniques, we analyzed expression profile, promoter activity and gene function of OsZF142, a novel CCCH-type zinc finger protein gene in rice (Oryza sativa L). The OsZF142 transcripts were detected in roots, culms, leaves, flowers and immature seed in rice plant using RT-PCR. The results were found that the OsZF142 gene was preferentially expressed in developmental seed and its spatial expression profile was preferentially expressed in rice embryo, but not in rice endosperm. A full-length of the OsZF142 gene promoter was cloned and a fusion expression vector was constructed with gus gene. Analysis of transgenic rice showed expression pattern of gus gene in these transgenic plants was similar to that of the RT-PCR analysis of the OsZF142 transcript. It suggested that the OsZF142 promoter activity was preferentially in rice embryo, but not in rice endosperm. The comparation analysis was conducted between wild type plants and transgenic plants carrying the sense and antisese OsZF142 gene in Nipponbare rice.The transgenic plants with the antisense OsZF142 gene were not remarkable difference with the wild type. But the transgenic plants with the sense OsZF142 gene indicated significance mutation in rice growth and development. They included that the whole growth phase of rice was delayed, such as the delay of seed germination and seedling growth, the slowness of leaves growth and tillering in main culm, the restraint of the tiller ability, the lateness of heading, the postponement of leaves senescence, shorter statue and uppermost internode, and so on, which indicated that the sense OsZF142 had a pleiotropic impact on rice growth and development. Together, these results demonstrated that OsZF142 protein was a negative regulation factor for the growth and development of rice.
Zinc-finger protein transcription factors arc one of the most abundant transcription factors in plants, and they widely regulate gene expression in different growth phases and environmental stress. By RT-PCR and transgene techniques, we analyzed expression profile, promoter activity and gene function of OsZF142, a novel CCCH-type zinc finger protein gene in rice (Oryza sativa L). The OsZF142 transcripts were detected in roots, culms, leaves, flowers and immature seed in rice plant using RT-PCR. The results were found that the OsZF142 gene was preferentially expressed in developmental seed and its spatial expression profile was preferentially expressed in rice embryo, but not in rice endosperm. A full-length of the OsZF142 gene promoter was cloned and a fusion expression vector was constructed with gus gene. Analysis of transgenic rice showed expression pattern of gus gene in these transgenic plants was similar to that of the RT-PCR analysis of the OsZF142 transcript. It suggested that the OsZF142 promoter activity was preferentially in rice embryo, but not in rice endosperm. The comparation analysis was conducted between wild type plants and transgenic plants carrying the sense and antisese OsZF142 gene in Nipponbare rice.The transgenic plants with the antisense OsZF142 gene were not remarkable difference with the wild type. But the transgenic plants with the sense OsZF142 gene indicated significance mutation in rice growth and development. They included that the whole growth phase of rice was delayed, such as the delay of seed germination and seedling growth, the slowness of leaves growth and tillering in main culm, the restraint of the tiller ability, the lateness of heading, the postponement of leaves senescence, shorter statue and uppermost internode, and so on, which indicated that the sense OsZF142 had a pleiotropic impact on rice growth and development. Together, these results demonstrated that OsZF142 protein was a negative regulation factor for the growth and development of rice.
引文
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