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利用GUS报告基因对激素和胁迫诱导的拟南芥ACC合酶基因AtACS4、AtACS5和AtACS7的启动子活性进行分析。

The GUS reporter-aided analysis of the promoter activities of Arabidopsis ACC synthase genes AtACS4, AtACS5, and AtACS7 induced by hormones and stresses.

作者信息

Wang Ning Ning, Shih Ming-Che, Li Ning

机构信息

Department of Plant Biology and Ecology, Nankai University, Tianjin 300071, China.

出版信息

J Exp Bot. 2005 Mar;56(413):909-20. doi: 10.1093/jxb/eri083. Epub 2005 Feb 7.

Abstract

Ethylene biosynthesis in higher plants is regulated developmentally and environmentally. To investigate the regulation of ACC synthase gene expression, the promoters of Arabidopsis ACS genes, AtACS4, AtACS5, and AtACS7, were fused to a GUS reporter gene, and the recombinant transgenes were introduced into Arabidopsis to produce three groups of AtACS::GUS transgenic plants. Histochemic and fluorometric study of these transgenic plants revealed that promoters of AtACS4, AtACS, and AtACS7 are all active in dark-germinated seedlings. AtACS5 has the highest promoter activity in leaves of 2-week-old light-grown seedlings among the three AtACS genes studied. In the mature leaves, AtACS4 and AtACS7 genes are expressed in both veins and areoles, whereas AtACS5 is expressed at a higher level in the areoles and epidermal cells surrounding trichomes. The promoter activities of all these AtACS genes are found in the reproductive organs. AtACS5 and AtACS7 are highly expressed in petals, sepals, carpels, stamens, cauline leaves, inflorescence stems, and siliques, while AtACS4 expression is undetectable in the petals of open flowers. All three AtACS genes are expressed in root tissue. In the 2-week-old light-grown Arabidopsis, the AtACS4 promoter is responsive to the plant hormones IAA, ethylene, and ABA, and to darkness and wounding; the AtACS5 promoter to IAA, ABA, salt, high temperature, and wounding; and the AtACS7 promoter to GA3, ethylene, and ABA, and to darkness and salt. Low-temperature treatment abolishes the darkness-induced AtACS7 gene expression, but not that of AtACS4. Each AtACS gene has a unique expression profile during growth and development. It appears that at any developmental stage or any growth period of Arabidopsis, there is always a member of AtACS multigene family that is actively expressed.

摘要

高等植物中的乙烯生物合成受发育和环境因素调控。为了研究ACC合酶基因表达的调控机制,将拟南芥ACS基因AtACS4、AtACS5和AtACS7的启动子与GUS报告基因融合,构建重组转基因并导入拟南芥,获得了三组AtACS::GUS转基因植株。对这些转基因植株进行组织化学和荧光分析发现,AtACS4、AtACS5和AtACS7的启动子在黑暗萌发的幼苗中均有活性。在所研究的三个AtACS基因中,AtACS5在2周龄光生幼苗的叶片中启动子活性最高。在成熟叶片中,AtACS4和AtACS7基因在叶脉和小叶中均有表达,而AtACS5在小叶和毛状体周围的表皮细胞中表达水平较高。所有这些AtACS基因的启动子活性在生殖器官中均有发现。AtACS5和AtACS7在花瓣、萼片、心皮、雄蕊、茎生叶、花序茎和角果中高度表达,而在开放花朵的花瓣中未检测到AtACS4的表达。所有三个AtACS基因在根组织中均有表达。在2周龄光生拟南芥中,AtACS4启动子对植物激素IAA、乙烯和ABA以及黑暗和创伤有响应;AtACS5启动子对IAA、ABA、盐、高温和创伤有响应;AtACS7启动子对GA3、乙烯和ABA以及黑暗和盐有响应。低温处理消除了黑暗诱导的AtACS7基因表达,但不影响AtACS4的表达。每个AtACS基因在生长发育过程中都有独特的表达模式。在拟南芥的任何发育阶段或生长时期,AtACS多基因家族中似乎总有一个成员在活跃表达。

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