杨树因伤诱导型启动子的克隆及功能分析

摘要/Abstract

摘要：

机械损伤或病虫害侵扰引起植物一系列防御相关基因的激活.其中一些是因伤诱导表达的.Bradshaw等1989年报道了杨树Win3基冈家族中的一员.其编码产物类似于白薯和豆类胰蛋白酶抑制剂.并且是因伤诱导表达的。为了更全面的了解这一基因启动子在因伤介导表达中的作用.探讨其在基因上程用于控制外源基因表达的可能性，本文分别从欧洲黑杨(P.Nigra)和美洲黑杨(P.Deltoides)中扩增出win3基因启动子wlNP(705bp)和WIDP(791 bp).这两个启动子与来自大肠杆菌的GUS基因融合，通过根癌农杆菌Ti质粒转化系统引入烟草中.证实了WINP和WIDP控制的GUS基因的表达不仅在损伤部位，而且具有系统的因伤诱导效应.对8株转基因烟草的组织化学分析表明.其表达模式与以前的报道一致。无论是在受损伤组织还是完整组织中.WINP的伤诱导活性总比WIDP高。这两个启动子需要进一步改造以增强其活性。

关键词: 因伤诱导型启动子, Gus报告基因, 转基因植物

Abstract:

Mechanical injury or pathogen and insect invasion usually involves a seties of defence-related gene expression in plants, Some of these genes show distinct wound-inducibility.One of the wound-inducible win3 multigene families from hybrid poplar, encoding a protein similar to sweet potato sporamins and legume trypsin inhibitors, was previously isolated by Bradshaw et al.(1989).To further understand the role of its upstream regulatory region in wound-mediated response, and its possibility to be used in geNetic engineering for the control of the expression of foreign genes, the win3 gene promoter WINP(705bp)and WIDP(791bp)were amplified from P.nigra and P.deltoides, respectively.Both promoters were fused to the E.coli β-glucuronidase(GUS)reporter gene, and introduced into tobacco plants by virtue of Agrobacte rium-Ti system.This study demonstrated that GUS expression responded to wounding locally and remotely in transgenic tobacco under the WIN(D)P control, and the biochemical and histochemical assays of GUS activity for 8 transgenic lines gave the similar expression pattern as reported by Hollick et al. (1995).It was also observed that the responsibility to wounding of WINP was always stronger than that of WIDP, both on the injured or uninjured sites.Further modification of both promoters is necessary to increase their ability to enhance the expression thus to be used in genetic engineering to obtain wound-inducible expression of foreign gene(s).

Key words: Wound-inducible promoter, GUS reporter gene, Transgenic plant

李强,卢孟柱,田颖川,陈颖,韩一凡. 杨树因伤诱导型启动子的克隆及功能分析[J]. 林业科学, 1999, 35(4): 111-118.

Qiang Li,Meng-Zhu Lu,Ying-Chuan Tian,Ying Chen,Yi-Fan Han. CLONING AND CHARACTERIZATION OF WOUND-INDCIBLE PROMOTERS FROM POPUL US[J]. Scientia Silvae Sinicae, 1999, 35(4): 111-118.

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