ETHYLENE-REGULATED EXPRESSION OF A CARNATION CYSTEINE PROTEINASE DURING FLOWER PETAL SENESCENCE
ETHYLENE-REGULATED EXPRESSION OF A CARNATION CYSTEINE PROTEINASE DURING FLOWER PETAL SENESCENCE
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DOI：
10.1007/bf00020397
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发表时间：
1995-06-01
影响因子：
5.1
通讯作者：
WOODSON, WR
中科院分区：
文献类型：
--
作者：
JONES, ML;LARSEN, PB;WOODSON, WR
以香石竹（Dianthus caryophyllus L.）花瓣由植物激素乙烯调节，并且与相当大的分解代谢活性包括蛋白质的损失有关。在本文中，我们提出的半胱氨酸蛋白酶的分子克隆，并表明其表达受乙烯和花瓣衰老相关。用5 ′-特异性引物和3 ′-非特异性引物从反转录的花柱RNA中扩增出1-氨基环丙烷-1-羧酸合酶cDNA，通过聚合酶链反应扩增出1600 bp的cDNA。发现克隆产物（pDCCP 1）的核苷酸序列与几种半胱氨酸蛋白酶而不是ACC合酶具有显着的同源性。428个氨基酸的单个开放阅读框显示与其它植物半胱氨酸蛋白酶共享显著的同源性，包括与来自拟南芥的半胱氨酸蛋白酶的大于70%的同一性。半胱氨酸蛋白酶活性位点的氨基酸在pDCCP 1肽中是保守的。RNA凝胶印迹分析表明，pDCCP 1的表达量随着乙烯产生和花瓣衰老的开始而显著增加。增加pDCCP 1的表达也与其他衰老的花器官，包括子房和花柱乙烯的产生。pDCCP 1转录积累在花瓣中处理与外源乙烯在3小时内和治疗的花与2，5-降冰片二烯，乙烯作用的抑制剂，防止pDCCP 1在花瓣中的表达增加。pDCCP 1表达的时间和空间模式表明半胱氨酸蛋白酶在花衰老过程中蛋白质损失中的作用。
The senescence of carnation (Dianthus caryophyllus L.) flower petals is regulated by the phytohormone ethylene and is associated with considerable catabolic activity including the loss of protein. In this paper we present the molecular cloning of a cysteine proteinase and show that its expression is regulated by ethylene and associated with petal senescence. A 1600 bp cDNA was amplified by polymerase chain reaction using a 5'-specific primer and 3'-nonspecific primer designed to amplify a 1-aminocyclopropane-1-carboxylate synthase cDNA from reverse-transcribed stylar RNA. The nucleotide sequence of the cloned product (pDCCP1) was found to share significant homology to several cysteine proteinases rather than ACC synthase. A single open reading frame of 428 amino acids was shown to share significant homology with other plant cysteine proteinases including greater than 70% identity with a cysteine proteinase from Arabidopsis thaliana. Amino acids in the active site of cysteine proteinases were conserved in the pDCCP1 peptide. RNA gel blot analysis revealed that the expression of pDCCP1 increased substantially with the onset of ethylene production and senescence of petals. Increased pDCCP1 expression was also associated with ethylene production in other senescing floral organs including ovaries and styles. The pDCCP1 transcript accumulated in petals treated with exogenous ethylene within 3 h and treatment of flowers with 2,5-norbornadiene, an inhibitor of ethylene action, prevented the increase in pDCCP1 expression in petals. The temporal and spatial patterns of pDCCP1 expression suggests a role for cysteine proteinase in the loss of protein during floral senescence.