Differential regulation of wheat quinone reductases in response to powdery mildew infection.
At least two types of quinone reductases are present in plants: (1) the zeta-crystallin-like quinone reductases (QR1, EC 1.6.5.5) that catalyze the univalent reduction of quinones to semiquinone radicals, and (2) the DT-diaphorase-like quinone reductases (QR2, EC 1.6.99.2) that catalyze the divalent reduction of quinones to hydroquinones. QR2s protect cells from oxidative stress by making the quinones available for conjugation, thereby releasing them from the superoxide-generating one electron redox cycling, catalyzed by QR1s. Two genes, putatively encoding a QR1 and a QR2, respectively, were isolated from an expressed sequence tag collection derived from the epidermis of a diploid wheat Triticum monococcum L. 24 h after inoculation with the powdery mildew fungus Blumeria graminis (DC) EO Speer f. sp. tritici Em. Marchal. Northern analysis and tissue-specific RT-PCR showed that TmQR1 was repressed while TmQR2 was induced in the epidermis during powdery mildew infection. Heterologous expression of TmQR2 in Escherichia coli confirmed that the gene encoded a functional, dicumarol-inhibitable QR2 that could use either NADH or NADPH as an electron donor. The localization of dicumarol-inhibitable QR2 activity around powdery mildew infection sites was accomplished using a histochemical technique, based on tetrazolium dye reduction.
Greenshields
David L
DL
Department of Biology, University of Saskatchewan, Saskatoon, SK, S7N 5E2, Canada.
Liu
Guosheng
G
Selvaraj
Gopalan
G
Wei
Yangdou
Y
D000595:Amino Acid Sequence; D001203:Ascomycota / Q000472:pathogenicity; D001483:Base Sequence; D018744:DNA, Plant / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D008954:Models, Biological; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D010935:Plant Diseases / Q000235:genetics* / Q000382:microbiology*; D011808:Quinone Reductases / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014908:Triticum / Q000201:enzymology* / Q000235:genetics* / Q000382:microbiology
D018744:DNA, Plant; D011994:Recombinant Proteins; D011808:Quinone Reductases