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作 者 ：黄真池,罗齐军,欧阳乐军,曾富华*

期 刊 ：西北植物学报 2013年 5期 页码：892~897

关键词：辣椒；prp1-1；Rs-AFP2；疫霉菌；

Keywords:pepper, prp1-1, Rs-AFP2, Phytophthora capsici,

摘 要 ：病原物诱导型启动子能精确控制抗病基因在侵染位点的表达,是抗病基因工程的有效工具。prp1-1是来自马铃薯谷胱甘肽巯基转移酶基因启动子的一个273 bp的片段,能够快速准确地启动被侵染位点抗病基因的表达；Rs-AFP2是具有对致病性丝状真菌的广谱抗性。该研究构建prp1-1调控Rs-AFP2基因表达的载体,经农杆菌介导转化法导入辣椒。逆转录PCR检测发现,转基因辣椒只在受到疫霉菌孢子侵染时,才由prp1-1启动Rs-AFP2基因的转录。用疫霉菌孢子灌根接种转基因辣椒T₁代植株,35株T₁代辣椒中有29株表现出明显的疫霉菌抗性。另将23株T₁代辣椒种于人工气候箱,发现其形态和发育特征与相同条件下的非转基因植株无明显区别。研究表明,prp1-1调控Rs-AFP2的诱导表达达到了增强辣椒疫霉菌抗性的目的,而且避免了负面效应的发生。

Abstract:Pathogen inducible promoters are helpful for genetic engineering on disease resistance by restricting expression to infection sites.A 273 bp fragment from the promoter of potato glutathione S transferase gene,named as prp1-1,could mediate rapid and strictly localized transcriptional activation at infection sites.Rs-AFP2 exerts antifungal activity against a broad spectrum of plant pathogenic filamentous fungi.Rs-AFP2 gene under the control of pathogen inducible promoter prp1-1 was introduced into pepper by Agrobacterium tumefaciens-mediated transformation.Reversed transcript PCR results showed that the Rs-AFP2 gene was induced to transcribe only after inoculation with the zoospores of Phytophthora capsici.Tested by root-irrigating with the zoospores of P.capsici,29 of 35 Rs-AFP2 transgenic T₁ lines enhanced resistance markedly.Other 23 T₁ lines were planted in soil of growth cabinet.And no effect on plant morphology was observed at the Rs-AFP2 transgenic T₁ lines.The results suggest the inducible expression of Rs-AFP2 derived by prp1-1 could enhance pepper disease resistance to P.capsici and avoid the adverse effect.

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