Molecular Cloning and Expression of a Sweet Potato Cysteine Protease SPCP1 from Senescent Leaves

文化大學機構典藏 CCUR > 環境規劃暨生物資源學院 > 農學院 > 園藝暨生物技術學系 > 期刊論文 > Item 987654321/18872

jsp.display-item.identifier=請使用永久網址來引用或連結此文件: https://irlib.pccu.edu.tw/handle/987654321/18872

题名:	Molecular Cloning and Expression of a Sweet Potato Cysteine Protease SPCP1 from Senescent Leaves
其它题名:	從甘藷老化葉片分子選殖及表現半胱胺酸蛋白分解酶
作者:	陳顯榮黃冠中陳巍珊蘇正庭侯文琪林耀輝
贡献者:	園藝系
关键词:	半胱胺酸蛋白分解酶 Ethephon 葉片老化 SPCP1 甘藷
日期:	2009-04
上传时间:	2011-01-19 14:19:41 (UTC+8)
摘要:	本研究報告從甘藷老化葉片分離出一條全長的cDNA稱之SPCP1，SPCP1其ORF含有1020個核苷酸（339氨基酸）且與不同物種包括赤楊、阿拉伯芥、紫雲英、油菜、胡蘿蔔、棉花、大麥、鳶尾、苜蓿、菸草、水稻、蓖麻、及白花三葉草等類似木瓜酶的半胱胺酸蛋白分解酶胺基酸序列具有高相似性（約58%至74%）。半定量RT-PCR及西方墨點雜交結果顯示SPCP1在自然老化及黑暗、ethephon、及ABA誘導的老化葉片中會被活化及增加表現量，然而於成熟的綠色葉片、莖、及根中SPCP1並無顯著增加。葉片老化過程中葉綠素開始降解的時間比SPCP1基因表現早。1 mM ethephon處理對於甘藷懸浮培養細胞亦會誘導SPCP1的表現，Evan blue染色結果顯示ethephon處理濃度高至2 mM時對懸浮培養細胞亦無顯著影響，然而10 mM濃度時大部份的細胞會死亡。依據這些實驗結果我們結論甘藷SPCP1為有功能的老化相關基因，其mRNA及蛋白質的表現於天然的或誘導的老化葉片及懸浮培養細胞皆顯著增加，SPCP1的功能與角色可能與老化過程中葉綠素開始降解及細胞死亡無關。 In this report a full-length cDNA, SPCP1, was isolated from senescent leaves of sweet potato (Ipomoea batatas (L.) Lam). SPCP1 contained 1020 nucleotides (339 amino acids) in the open reading frame, and exhibited high amino acid sequence homologies (ca. 58% to 74%) with papain-like cysteine proteases of Alnus glutinosa, Arabidopsis thaliana, Astragalus sinicus, Brassica napus, Daucus carota, Gossypium hirsutum, Hordeum vulgare, Iris hollandica, Medicago truncatula, Nicotiana tabacum, Oryza sativa, Ricinus communis, Trifolium repens. Semi-quantitative RT-PCR and Western blot hybridization showed that SPCP1 gene expression was enhanced significantly in natural senescent leaves and in dark-, ethephon-, and ABA-induced senescent leaves, whereas, was almost not detected in mature green leaves, stems, and roots. Initiation of chlorophyll degradation is earlier than the SPCP1 gene expression during leaf senescence. SPCP1 expression was also induced in sweet potato suspension cells treated with 1 mM ethephon. Evan blue staining showed that suspension cells were not significantly affected by ethephon treatment up to 2 mM, however, most of the cells died when treated with 10 mM ethephon. In conclusion, sweet potato SPCP1 is likely a functional, senescence-associated gene and its expression levels were significantly enhanced at mRNA and protein levels in natural and induced senescent leaves and suspension cells. The physiological role and function of SPCP1 were likely not in association with initiation of chlorophyll degradation and cell death during senescence.
關聯:	Botanical Studies 50卷2期 P.159-170
显示于类别:	[園藝暨生物技術學系] 期刊論文

文件中的档案:

档案	描述	大小	格式	浏览次数
index.html		0Kb	HTML	490	检视/开启

在CCUR中所有的数据项都受到原著作权保护.

数据加载中.....