2006 Expression of the lipid transfer protein Ace-AMP1-小麦转基因

Abstract To enhance fungal disease resistance,wheat plants (cv.Bobwhite)were engineered to constitutively express the potent antimicrobial protein Ace -AMP1from Allium cepa ,driven by a maize ubiquitin promoter along with its first intron.The bar gene was used for selection of putative transformants on medium containing phosphinothricin (PPT).Transgene inheritance,integration and stability of expression were confirmed over two generations by PCR,Southern,northern and western blot analyses,respectively.The levels of Ace -AMP1in different transgenic lines correlated with the transcript levels of the transgene.Up to 50%increase in resistance to Blumeria graminis f.sp.tritici was detected in detached leaf assays.In ears of transgenic wheat inoculated with Neovossia indica ,Ace -AMP1inten-sified expression of defense-related genes.Elevated levels of salicylic acid and of transcripts of phenyl-alanine ammonia lyase (PAL ),glucanase (PR 2)and chitinase (PR 3)in the transgenic plants indicated manifestation of systemic acquired resistance (SAR).

Keywords Triticum ÆTransgenic wheat ÆAce -AMP1ÆBlumeria ÆNeovossia ÆPR genes

Introduction

Wheat is one of the most important grain crops for human consumption.The rusts,Karnal bunt,powdery mildew and Fusarium head blight can reduce yield by 30–50%.At present,these diseases are controlled through breeding,crop husbandry practices and the application of fungicides.In addition,genetic engi-neering may complement conventional breeding programmes to develop resistant cultivars (Bieri et al.2003;Clausen et al.2000;Oldach et al.2001).

The large (17,000Mb)and complex (hexaploid)genome,genotype-dependent tissue culture re-sponses,transgene silencing (Anand et al.2003)and the lack of an efficient gene delivery system are major hurdles for efficient genetic transformation of wheat;it was the last major cereal to be genetically transformed (Vasil et al.1993)and its transformation efficiency is much lower than that of other cereals.All this has retarded the application of recombinant DNA technology in the genetic improvement of this crop.In recent years,particle bombardment and Agrobacterium tumefaciens -mediated transformation (ATMT)have been used for engineering agronomi-cally useful traits into wheat.However,evidence for successful transformation of wheat using ATMT remains rare.It has been demonstrated that

S.Roy-Barman ÆB.B.Chattoo (&)

Department of Microbiology and Biotechnology Centre,Faculty of Science,The M.S.University of Baroda,Vadodara 390002,India

e-mail:chattoo@msubiotech.ac.in

C.Sautter

Plant Biotechnology,Institute for Plant Sciences,Swiss Federal Institute of Technology (ETH),Zurich CH 8092,Switzerland

Transgenic Res (2006)15:435–446DOI 10.1007/s11248-006-0016-1

ORIGINAL PAPER

Expression of the lipid transfer protein Ace -AMP1in transgenic wheat enhances antifungal activity and defense responses

Subhankar Roy-Barman ÆChristof Sautter ÆBharat B.Chattoo

Received:9November 2005/Accepted:10March 2006ÓSpringer Science+Business Media B.V.2006