ABSTRACT
When comparing the responses of two wheat (Triticum aestivum L.) genotypes, the drought-tolerant Plainsman V and the drought-sensitive Cappelle Desprez, to reduced amounts of irrigation water, we found differences in ascorbate metabolism: both ascorbate oxidation and transcription levels of enzymes processing ascorbate were changed. Relative transcript levels of ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) isoenzymes, predicted to localize in distinct subcellular organelles, showed different transcriptional changes in the two genotypes. Among APX coding mRNAs, expression levels of two cytosolic (cAPX I, II) and a thylakoid-bound (tAPX) variants increased significantly in Plainsman V while a cytosolic (cAPX I) and a stromal (sAPX II) APX coding transcripts were found to be higher in Cappelle Desprez after a 4-week-long water-deficit stress. Examining the MDARs, two cytosolic isoforms (cMDAR I, II) displayed significant up-regulation of mRNA levels in the sensitive genotype, whereas only one of them (cMDAR II) did in the tolerant cultivar. We found an up-regulated chloroplastic DHAR (chlDHAR) mRNA only in the sensitive Cappelle Desprez. However, increased expression levels of a cytosolic GR (cGR) and a chloroplastic GR (chlGR) were detected only in the tolerant Plainsman V. After 4 weeks of reduced irrigation, a significantly lower ascorbate/dehydroascorbate ratio was detected in leaves of the sensitive Cappelle Desprez than in the tolerant Plainsman V. Our results indicate that more robust transcription of ascorbate-based detoxification machinery may prevent an adverse shift of the cellular redox balance.
Subject(s)
Ascorbic Acid/metabolism , Multigene Family , Triticum/genetics , Water/metabolism , Droughts , Gene Expression Regulation, Plant , Isoenzymes/genetics , Isoenzymes/metabolism , Lipid Peroxidation , Oxidation-Reduction , Oxidative Stress , Oxidoreductases/genetics , Oxidoreductases/metabolism , RNA, Messenger/metabolism , Stress, Physiological , Transcription, Genetic , Triticum/metabolismABSTRACT
Total glutathione S-transferase (GST, EC 2.5.1.18) and glutathione peroxidase (GPOX) activity were measured spectrophotometrically in Triticum aestivum cv. MV Emese and cv. Plainsman (drought tolerant) and cv. GK Elet and Cappelle Desprez (drought-sensitive) flag leaves under control and drought stress conditions during the grain-filling period, in order to reveal possible roles of different GST classes in the senescence of flag leaves. Six wheat GSTs, members of 3 GST classes, were selected and their regulation by drought and senescence was investigated. High GPOX activity (EC 1.11.1.9) was observed in well-watered controls of the drought-tolerant Plainsman cultivar. At the same time, TaGSTU1B and TaGSTF6 sequences, investigated by real-time PCR, showed high-expression levels that increased with time, indicating that the gene products of these genes may play important roles in monocarpic senescence of wheat. Expression of these genes was also induced by drought stress in all of the four investigated cultivars, but extremely high transcript amounts were detected in cv. Plainsman. Our data indicate genotypic variations of wheat GSTs. Expression levels and early induction of two senescence-associated GSTs under drought during grain filling in flag leaves correlated with high yield stability.
