棉花油菜素类固醇合成酶基因GhDWF1的克隆和表达分析.pdfVIP

中国农业科学 2007,40(7):1337-1344 Scientia Agricultura Sinica 棉花油菜素类固醇合成酶基因 GhDWF1 的克隆和表达分析 罗 明，周建平，肖月华，李先碧，侯 磊，肖忠意，裴 炎 （西南大学生物技术中心，重庆 400716 ） 摘要：【目的】研究油菜素类固醇物质（BRs）在棉花纤维生长发育过程中的作用。【方法】通过棉花 EST 序 列的筛选和整合，从陆地棉徐州142纤维中克隆了一个BRs合成酶基因 GhDWF1。【结果】其cDNA序列全长1 849 bp， 包含一个 1 692 bp 的开放阅读框，推导编码 563 个氨基酸，与水稻、玉米、豌豆、番茄和拟南芥的 BRs 合成酶 DWARF1/DIMINUTO有较高的同源性，具有该类蛋白的典型结构。GhDWF1 基因在开花当天的胚珠和纤维中表达最强， 在根、幼茎和纤维发育的各个时期表达较高。与徐州142野生型相比，无绒无絮突变体胚珠中表达量较低。【结论】 暗示 GhDWF1 基因的表达与棉花纤维的生长发育有密切关系。 关键词：棉花；DWARF1 基因；纤维；油菜素类固醇；植物固醇 Cloning and Expression Analysis of A Brassinosteroid Biosynthetic Enzyme Gene, GhDWF1, from Cotton (Gossypium hirsuturm L.) LUO Ming, ZHOU Jian-ping, XIAO Yue-hua, LI Xian-bi, HOU Lei, XIAO Zhong-yi, PEI Yan (Center of Biotechnology, Southwest University, Chongqing 400716) Abstract: 【Objective 】Brassinosteroids (BRs) are an important class of plant steroidal hormones that are essential in a wide variety of physiological processes. The present study determines the effects of BRs on development of cotton fibers. 【Method 】 Through screening cotton EST data base and the candidated ESTs, a key gene (GhDWF1) involved in the upstream biosynthetic pathway of BRs has been cloned from developing fibers of upland cotton (Gossypium hirsutum L.) cv. XuZhou142. 【Result 】The full length of the cloned cDNA is 1 849 bp, including a 37 bp 5′-untranslated region, an ORF of 1 692 bp and a 120 bp 3′-untranslated region. The cDNA encodes a polypeptide of 563 amino acid residues with a predicted molecular mass of 65 kD. The deduced amino acid sequence has high homology with the BR biosynthetic enzyme, DWARF1/DIMINUTO, from rice, maize, pea, tomato and Arabidopsis . Furthermore, the typical conserved structure, such as