Purification of multiple glutathione transferases involved in herbicide detoxification from wheat (Triticum aestivum L.) treated with the safener fenchlorazole-ethyl

被引:87
作者
Cummins, I
Cole, DJ
Edwards, R [1 ]
机构
[1] Univ Durham, Dept Biol Sci, Crop Protect Grp, Durham DH1 3LE, England
[2] Rhone Poulenc Agr Ltd, Ongar CM5 0HW, Essex, England
基金
英国生物技术与生命科学研究理事会;
关键词
D O I
10.1006/pest.1997.2308
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Multiple isoenzymes of glutathione transferase (GST), purified from the shoots of wheat seedlings treated with the herbicide safener fenchlorazole-ethyl, could be resolved into polar and hydrophobic types using hydrophobic interaction chromatography. Both types of GSTs could also be resolved based on their affinities for S-hexyl-glutathione-agarose. A minor proportion of the GSTs could be eluted from the column with a salt wash and this loosely bound fraction contained polypeptides which were recognized by an antiserum raised against the theta-type maize GST ZmGST I-II. The major proportion could only be recovered using S-hexyl-glutathione and these GSTs were characterized in detail. These isoenzymes catalyzed the glutathione conjugation of xenobiotics, including herbicides, and showed additional activities as glutathione peroxidases. Each GST was composed of two subunits, with four distinct classes of subunit being determined. A 25-kDa polypeptide, termed Triticum aestivum GST 1 (TaGST 1), was the most abundant subunit and could be resolved into two variants, TaGST la and TaGST Ib by reversed-phase HPLC. This GST subunit was recognised by an antiserum raised against the maize GST ZmGST V-VI, which is a tau-type GST. In addition to TaGST 1, two 26-kDa polypeptides, TaGST 2 and TaGST 3, and a 24-kDa polypeptide, TaGST 4, could be resolved by a combination of hydrophobic interaction chromatography, SDS-PAGE, and reversed-phase HPLC. In the shoots of untreated wheat seedlings the major isoenzyme was TaGST la-lb, while in the shoots of fenchlorazole-ethyl-treated plants the heterodimers TaGST 1-2, TaGST 1-3, and TaGST 1-4 also accumulated. Significantly, only the safener-inducible TaGST 1-2, TaGST 1-3, and TaGST 1-4 isoenzymes catalyzed the detoxification of fenoxaprop-ethyl and this may help to explain why fenchlorazole-ethyl enhances the glutathione-mediated metabolism and also the tolerance of wheat toward this herbicide. All isoenzymes were active in detoxifying the herbicides metolachlor and fluorodifen, bur only TaGST 1-2 and TaGST 1-3 showed any activity toward atrazine. (C) 1997 Academic Press.
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页码:35 / 49
页数:15
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