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Plant and Cell Physiology Advance Access originally published online on October 18, 2007
Plant and Cell Physiology 2007 48(12):1702-1712; doi:10.1093/pcp/pcm142
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Plastidial Glutathione Reductase from Haynaldia villosa is an Enhancer of Powdery Mildew Resistance in Wheat (Triticum aestivum)

Ya-Ping Chen1,2,4, Li-Ping Xing1,4, Guo-Jiang Wu2, Hua-Zhong Wang1,3, Xiu-E Wang1, Ai-Zhong Cao1 and Pei-Du Chen1,*

1 The National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, PR China
2 South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, PR China
3 The Key Laboratory of Gene Engineering, School of Chemistry and Life Sciences, Tianjing Normal University, Tianjing 300074, PR China

*Corresponding author: E-mail, pdchen{at}njau.edu.cn; Fax, +86-25-8439-5344.


  Abstract

A full-length cDNA (Hv-GR) whose transcript accumulation increased in response to infection by Blumeria graminis DC.f.sp. tritici (Bgt) was isolated from Haynaldia villosa. Southern analysis revealed a single copy of Hv-GR present in H. villosa. This gene encodes a glutathione reductase (GR) with high similarity to chloroplastic GRs from other plant species. Chloroplastic localization of Hv-GR was confirmed by targeting of the green fluorescent protein (GFP)–Hv-GR fusion protein to chloroplasts of epidermal guard cells. Following inoculation with Bgt, transcript accumulation of Hv-GR increased in a resistant line of wheat, but no significant change was observed in a susceptible line. In vivo function of Hv-GR in converting oxidized glutathione (GSSG) to the reduced form (GSH) was verified through heterologous expression of Hv-GR in a yeast GR-deficient mutant. As expected, overexpression of this gene resulted in increased resistance of the mutant to H2O2, indicating a critical role for Hv-GR in protecting cells against oxidative stress. Moreover, overexpression of Hv-GR in a susceptible wheat variety, Triticum aestivum cv.Yangmai 158, enhanced resistance to powdery mildew and induced transcript accumulation of other pathogenesis-related genes, PR-1a and PR-5, through increasing the foliar GSH/GSSG ratio. Therefore, we concluded that a high ratio of GSH to GSSG is required for wheat defense against Bgt, and that chloroplastic GR enzymes might serve as a redox mediator for NPR1 activation.

Keywords: Chloroplastic glutathione reductase - Disease resistance - Haynaldia villosa - Powdery mildew - Triticum aestivum

Abbreviations: AOS, active oxygen species; Bgt, Blumeria graminis DC.f.sp. tritici; DI, disease index; EST, expressed sequence tag; GFP, green fluorescent protein; GR, glutathione reductase; GUS, β-glucuronidase; NPR1, non-expressor of pathogenesis-related genes 1;; ORF, open reading frame;; PR, pathogenesis-related; RT–PCR, reverse transcription–PCR; SSH, suppression subtractive hybridization

4These authors contributed equally to this work.

(Received August 4, 2007; Accepted October 14, 2007)
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