棉花倍半萜合成酶基因GhTPS1的克隆及棉铃虫取食诱导表达谱分析
Molecular cloning of the sesquiterpene synthase gene GhTPS1 ofGossypium hirsutum and its induction by the feedingactivity of cotton bollworm larvae
黄欣蒸 武 娟 徐琦霞 张永军 郭予元
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作者单位：1. 西北农林科技大学植物保护学院杨陵712100;
中文关键词：棉铃虫，萜烯合成酶，GhTPS1，表达谱
英文关键词：Helicoverpa armigera，terpene synthase，GhTPS1，expression pattern
中文摘要： 棉花被植食性昆虫取食后大量释放的特异性萜烯挥发物，能够有效吸引天敌昆虫进行寄主搜索和定位。
本文从陆地棉(中棉12) Gossypium hirsutum 叶片中克隆获得一个倍半萜合成酶基因的全长cDNA，命名为GhTPS1(GenBank 登录号:JQ365627)。该基因编码545 个氨基酸的蛋白，预测分子量为63. 3 ku，等电点为5. 92。氨基酸序列比对分析表明该基因与其它被子植物倍半萜合成酶基因一致性为34% ～ 60% ，其中与欧洲葡萄( － ) －germacrene D synthase 一致性最高(60% )。聚类分析表明GhTPS1 属于由被子植物倍半萜合成酶基因组成的TPSa 亚家族。采用实时荧光定量PCR 检测了GhTPS1 mRNA 在棉铃虫Helicoverpa armigera (Hübner)幼虫为害棉花不同时间的表达谱，结果表明接虫24 h 该基因表达量显著上调。
英文摘要：Cotton plants attacked by herbivorous insects emit large amounts of characteristic volatile terpenoids that are
highly attractive to predatory insects and trigger specific behavioral reactions in these，such as detection and location of the
chemical source． Here，we identified a novel sesquiterpene synthase gene GhTPS1 ( GenBank accession no． JQ365627)
from leaves of Gossypium hirsutum cv． Zhongmian12． GhTPS1 encodes a protein comprised of 545 amino acid residues
with a calculated molecular mass of 63． 3 ku and an isoelectric point of 5． 92． Multiple sequence alignment revealed that
the deduced amino acid sequence has 34% —60% identity with the sesquiterpene synthases of other angiosperm species，
and shared the highest identity with that of ( － ) － germacrene D synthase from Vitis vinifera． Phylogenetic analysis
clustered GhTPS1 into the TPSa subfamily，which is predominantly comprised of angiosperm sesquiterpene synthase genes．
We used real-time quantitative PCR to further investigate the temporal expression of GhTPS1 in G． hirsutum leaves
induced by the feeding activity of Helicoverpa armigera ( Hübner ) larvae． The results indicate that the GhTPS1
transcription level significantly increased after 24 hours of larval feeding activity．
Key words Helicoverpa armigera，terpene synthase，GhTPS1，expression pattern