拟南芥细胞核-叶绿体双定位蛋白Sig6调控子叶叶绿体发育的机理研究

批准号:
32000185
项目类别:
青年科学基金项目
资助金额:
24.0 万元
负责人:
胡芬红
依托单位:
学科分类:
植物光合与固氮
结题年份:
2023
批准年份:
2020
项目状态:
已结题
项目参与者:
胡芬红
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中文摘要
高等植物子叶的叶绿体由黄化质体在光照条件下转绿而成,在此转化过程中需要大量细胞核基因进行精准调控。拟南芥Sig6一直被认为是定位于叶绿体并调控质体基因转录的σ因子,然而我们发现它在黑暗生长的黄化苗中还定位于细胞核。缺失核定位Sig6的突变体子叶呈黄化表型,而缺失叶绿体定位Sig6的突变体子叶叶绿体则发育正常。这说明核定位而不是叶绿体定位的Sig6调控了子叶黄化质体的转绿过程,但具体作用机制不清楚。本项目拟通过RNA-seq和qPCR等方法阐明Sig6在细胞核内调控哪些基因的表达;利用ChIP-seq等手段研究Sig6调控基因表达的方式;以前期酵母双杂交筛库获得的Sig6候选互作蛋白为研究对象,通过生物化学和遗传学等策略阐明这些蛋白与Sig6互作的方式和它们协同调控相关基因表达的分子机制。相关研究对于我们深入理解Sig6调控高等植物子叶黄化质体转化为叶绿体的分子机制和网络具有重要的科学意义。
英文摘要
In higher plants, chloroplasts in cotyledons develop from etioplasts under light condition. The transition from etioplasts to chloroplasts is precisely regulated by nuclear-encoding genes. The sigma factor 6 has been demonstrated to localize in the chloroplast to regulate transcription of the plastid-encoded genes. However, we found that Sig6 can be localized in the nucleus, which is critical for cotyledon greening of etiolated seedlings. It will be important to elucidate how the nuclear-localized Sig6 regulates chloroplast development in cotyledons. In this project, we will identify the genes regulated by nuclear-localized Sig6 using RNA-seq and qPCR assays, investigate how the nuclear-localized Sig6 regulate genes expression via ChIP-seq assay, and confirm the interaction between Sig6 and the candidate interacting proteins that were identified by Y2H system. Biochemical and genetics approaches will be employed to explore how Sig6 and the candidate interacting proteins coordinately regulate gene expression. This project will focus on the direct targets of Sig6 and the molecular and cellular determinants responsible for Sig6 function. More globally, our results will provide a fundamental insight into the mechanism by which early chloroplast development is regulated in etiolated cotyledons.
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