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  2. 農業暨自然資源學院
  3. 生物科技學研究所
Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/36256
標題: 鐵炮百合花藥發育和花粉萌發生長的三個專一性基因之分子調控分析
The analysis for molecular regulation of three specific genes involved in the lily anther development and pollen germination and tube growth
作者: 徐思維
Hsu, Ssu-Wei
關鍵字: Desiccation;乾燥;Abscisic acid;Pollen;離層酸;花粉
出版社: 生物科技學研究所
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摘要: 
在百合花粉成熟期和花粉管生長期間，Rop基因會受乾燥和離層酸訊息傳遞調控，當把未成熟的花藥/花粉進行乾燥處理和外加不同濃度的離層酸處理時，LLP-Rop1基因會降低其mRNA的累積，而LLP12-2，一個Rop-interactive CRIB motif-containing (RIC)基因則會增加其mRNA的累積；利用離層酸合成抑制劑norflurazon處理，也顯示離層酸會抑制LLP-Rop1，且會誘導LLP12-2基因表現。LLP-Rop1大量表現時，會抑制花粉管延長和造成管尖膨大的現象，CFP-LLP-Rop1分布在花粉管尖端的原生質膜上，Fluorescence resonance energy transfer (FRET)分析顯示，在花粉管中，活化的LLP-Rop1會和LLP12-2發生交互作用，但此交互作用可能不會受花粉乾燥的影響。當LLP12-2大量表現時，會抑制花粉萌發和花粉管生長，利用鈣離子流入過程的抑制劑LaCl3和EGTA及actin去極化藥物LatB處理，顯示LLP12-2會調控鈣離子流入的過程，而與actin的聚集無關。進一步以蛋白激酶抑制劑staurosporine或蛋白磷酸酶抑制劑okadaic acid處理，顯示LLP12-2調控鈣離子流入的過程與對staurosporine敏感的蛋白激酶和對okadaic acid敏感的蛋白磷酸酶無關。更進一步利用LLP12-2突變體來探討LLP12-2蛋白質的功能，顯示LLP12-2蛋白的N端扮演effector domain的功能而影響花粉萌發和花粉管生長

Here, we report unique desiccation-associated abscisic acid (ABA) signaling transduction through which the Rop (Rho GTPase of plants) gene is regulated during the stage of pollen maturation. Accumulation of the LLP-Rop1 transcript decreased its level of accumulation while LLP-12-2, a Rop-interactive CRIB motif-containing (RIC) transcript increased either by premature drying of developing anther/pollen or by the exogenous application of various concentrations of ABA during pollen maturation and tube growth. Application of norflurazon, an ABA biosynthesis inhibitor, also resulted in the downregulation of the LLP-Rop1 gene while the LLP-12-2 was upregulated by ABA. LLP-Rop1 overexpression inhibited tube elongation, caused tube expansion and the formation of a ballooned tip. CFP-LLP-Rop1 was localized to the cytoplasm having a greater intensity along the tube plasma membrane. Fluorescence resonance energy transfer (FRET) analysis of lily pollen tubes coexpressing CFP-LLP-Rop1 and YFP-LLP-12-2 demonstrated that LLP-12-2 is a target RIC protein of active LLP-Rop1, but the interaction between LLP-Rop1 and LLP-12-2 proteins is probably irrelevant of dehydration in the dried pollen. Overexpression of LLP12-2 inhibits pollen germination and tube growth. Studies with the germination inhibitors, the Ca2+ influx blocking agents LaCl3 and EGTA and an actin-depolymerizing drug, latrunculin B (LatB), suggested that LLP12-2 is involved in modulating calcium influx in the cytoplasm, but not actin assembly. When pollen germination and tube elongation were further examined by LaCl3 or EGTA, each with the addition of either staurosporine or okadaic acid, the results revealed that the involvement of LLP12-2 in modulation of calcium influx in the cytoplasm is independent of staurosporine-sensitive protein kinases and okadaic acid-sensitive phosphatases. The deletion mutant analysis of LLP12-2 indicated that the N-terminal region of LLP12-2 acts as an effector domain which regulates pollen germination and tube growth.
URI: http://hdl.handle.net/11455/36256
其他識別: U0005-2107201018420000
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