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政府計畫
計畫名稱(中文) 調控早期胚胎發育與種子萌發基因的分子機制研究
計畫名稱(英文) Molecular Mechanic Study of Genes Involve in Early Embryogenesis and Seed Germination
文獻分類 其它
計畫編號 MOST105-2311-B001-074-MY3
計畫主辦人 趙光裕
計畫主管機關 科技部
計畫執行機構 中央研究院植物暨微生物學研究所
全程計畫年 2017年 ~ 2018年
中文關鍵字 阿拉伯芥;⽣⾧素;吉⾙素;胚胎發育;種⼦萌發;rDNA 變種;核醣體RNA; NOT 複合體;微絲束(F-actin bundle);微絲結合蛋⽩(F-actin binding protein);胞質分裂 (cytokinesis)
英文關鍵字 Arabidopsis;auxin;gibberellin;embryogenesis;seed germination;rDNA variant;rRNA Processing;nucleolus;NOT complex;F-actin bundle;F-actin binding protein; cytokinesis
中文摘要 種?是?等植物?命起始器官及許多動物主?來源,研究參與胚胎發育及種?萌發基因功能, 不僅可作為探究植物發育過程分?調控機制的重要材料,更可作為對增進種?產量與品質的重 要依據。利?擬南芥進?正向遺傳學突變分析時獲得數個參與種?胚胎發育的基因,基因突變 造成胚胎發育受阻或是種?能無法萌發。NHP247 突變造成??素於胚胎發育過程分布異常致 使胚胎發育受阻,胞質分裂時細胞板形成亦發?異常。NHP247 是?微絲結合蛋?能促進微絲 束的形成,NHP247 可能透過與微絲束的作?參與囊胞 (vesicles) 於胞質分裂時細胞板的形 成。NHP231 突變造成成熟種籽無法萌發,NHP231 是阿拉伯芥NOT11 蛋?並與NOT 複合體 中的NOT10 及 NTO1 作?。將nhp231 同源種?進?基因表現分析發現許多參與GA 調控種 ?發育的基因表現降低;?參與ABA 調控種?休眠的許多基因表現。進?雙重突變實驗發現 LEC1, SPT, NCED, PIF5, 及 DELLA 基因發?突變可讓nhp231 與not10 的種?恢復萌發。 NHP148 是?位於核仁 SAS10/C1D 蛋?可與nucleolins、MPP10 及Jumonji14 作?,NHP148 突變致使核醣體RNA processing 受阻,進?影響正常核仁的形成與作?。本計畫主要以遺傳 學及分?細胞?物學實驗,對這三個基因如何參與胚胎發育及種?萌發進?系統性且廣泛深? 的分?作?模式及調控機制探究,期望將結果發表於重要期刊並作為未來進?步研究課題之依據。
英文摘要 Seeds and fruits constitute the most important source of food supply around the world and in most cases their production is the result of successful double fertilization and following embryogenesis. Therefore, a better understanding of embryogenesis and seed development will not only have a great impact to grain yield, but also may provide a great potential to develop and generate beneficial merchandise. Our previous genetic screening of T-DNA mutants with defects in gametophytic development or embryogenesis has successfully identified more than 20 novel mutants. Several T-DNAs confirmed as single T-DNA inserted mutants by segregation analyses and fully complemented by introducing the original genomic sequence for each of the genes will be further studied their roles during embryogenesis or seed germination in this proposal. The mutant nhp247 showed a severe disturbance in the distribution and transport of auxin to result in embryo lethal. Further investigation revealed cell plate formation in nhp247-2 was significantly disturbed that was consistent with by the localization of NHP247 to the proximal region of the developing cell plate during cytokinesis. The defect of cell divisions in nhp247 and special localization of NHP247 during cell division imply that NHP247 is required to organize the formation of the cell plate. Interestingly, we found NHP247 binds to F-actin and has F-actin bundling activity in vitro. All these suggest NHP247, a F-actin bundling protein, is involved in phragmoplast formation during cytokinesis. NHP231 encodes NOT11 ortholog in Arabidopsis and its null mutation causes defective seed germinate under normal condition. The interaction between NHP231 with other components of NOT complex (NOT10 and NTO1 N-terminus) in cytoplasm of Arabidopsis protoplast by the BiFC assay confirmed that NHP231 is the NOT11 ortholog in Arabidopsis. Microarray analyses from seeds of nhp231/- revealed down- and up-regulated of genes participating in GA signaling repression and ABA-dependent seed dormancy maintained, respectively. We have generated double mutation of nhp231 and not10 with all these available mutants of the genes shown up-regulated from our array data. Mutations in LEC1, SPT, NCED, PIF5, and DELLA could fully or partially rescue two mutant alleles of nhp231 and not10. Disruption of NHP148 caused aberrant processing of precursor rRNAs at the 5’ External Transcribed Spacer, repression of the major rDNA variant, and embryonic lethality. Strikingly, overexpressed NHP148 resulted in multiple nucleoli, presumably owing to ectopic activation and translocation of rDNA loci. NHP148 encodes a nucleolus-localized SAS10/C1D family protein and interacted with Small Subunit Processome components nucleolins, and Arabidopsis MPP10 ortholog, and histone demethylase Jumonji14. Our results uncovering the dual role of THAL in transcription and processing events critical for proper rRNA biogenesis and nucleolar organization during reproduction are the first to define the function of SAS10/C1D family members in plants. We will try to compel evidence of their important roles in seed maturation (NHP231) and embryogenesis (NHP247 and NHP148) with diverse genetic, molecular and cellular approaches. We believe that the study of these genes will not only advance our current knowledge into diverse level of regulation in plant reproduction, but also generate high quality publications.
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