Identification, Characterization, and Stress Responsiveness of Glucose-6-phosphate Dehydrogenase Genes in Highland Barley
G6PDH provides intermediate metabolites and reducing power (nicotinamide adenine dinucleotide phosphate, NADPH) for plant metabolism, and plays a pivotal role in the cellular redox homeostasis. In this study, we cloned five <i>G6PDH</i> genes (<i>HvG6PDH1</i> to <i>HvG6...
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2020-12-01
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author | Ruijun Feng Xiaomin Wang Li He Shengwang Wang Junjie Li Jie Jin Yurong Bi |
author_facet | Ruijun Feng Xiaomin Wang Li He Shengwang Wang Junjie Li Jie Jin Yurong Bi |
author_sort | Ruijun Feng |
collection | DOAJ |
description | G6PDH provides intermediate metabolites and reducing power (nicotinamide adenine dinucleotide phosphate, NADPH) for plant metabolism, and plays a pivotal role in the cellular redox homeostasis. In this study, we cloned five <i>G6PDH</i> genes (<i>HvG6PDH1</i> to <i>HvG6PDH5</i>) from highland barley and characterized their encoded proteins. Functional analysis of <i>HvG6PDHs</i> in <i>E. coli</i> showed that <i>HvG6PDH1</i> to <i>HvG6PDH5</i> encode the functional G6PDH proteins. Subcellular localization and phylogenetic analysis indicated that HvG6PDH2 and HvG6PDH5 are localized in the cytoplasm, while HvG6PDH1, HvG6PDH3, and HvG6PDH4 are plastidic isoforms. Analysis of enzymatic activities and gene expression showed that <i>HvG6PDH1</i> to <i>HvG6PDH4</i> are involved in responses to salt and drought stresses. The cytosolic <i>HvG6PDH2</i> is the major isoform against oxidative stress. <i>HvG6PDH5</i> may be a house-keeping gene. In addition, <i>HvG6PDH1</i> to <i>HvG6PDH4</i> and their encoded enzymes responded to jasmonic acid (JA) and abscisic acid (ABA) treatments, implying that JA and ABA are probably critical regulators of <i>HvG6PDH</i>s (except for <i>HvG6PDH5</i>). Reactive oxygen species analysis showed that inhibition of cytosolic and plastidic G6PDH activities leads to increased H<sub>2</sub>O<sub>2</sub> and O<sub>2</sub><sup>−</sup> contents in highland barley under salt and drought stresses. These results suggest that G6PDH can maintain cellular redox homeostasis and that cytosolic HvG6PDH2 is an irreplaceable isoform against oxidative stress in highland barley. |
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spelling | doaj.art-f4a9e99780c14399a93df4040b38a4de2023-11-21T01:31:23ZengMDPI AGPlants2223-77472020-12-01912180010.3390/plants9121800Identification, Characterization, and Stress Responsiveness of Glucose-6-phosphate Dehydrogenase Genes in Highland BarleyRuijun Feng0Xiaomin Wang1Li He2Shengwang Wang3Junjie Li4Jie Jin5Yurong Bi6Ministry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaMinistry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaMinistry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaMinistry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaMinistry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaMinistry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaMinistry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaG6PDH provides intermediate metabolites and reducing power (nicotinamide adenine dinucleotide phosphate, NADPH) for plant metabolism, and plays a pivotal role in the cellular redox homeostasis. In this study, we cloned five <i>G6PDH</i> genes (<i>HvG6PDH1</i> to <i>HvG6PDH5</i>) from highland barley and characterized their encoded proteins. Functional analysis of <i>HvG6PDHs</i> in <i>E. coli</i> showed that <i>HvG6PDH1</i> to <i>HvG6PDH5</i> encode the functional G6PDH proteins. Subcellular localization and phylogenetic analysis indicated that HvG6PDH2 and HvG6PDH5 are localized in the cytoplasm, while HvG6PDH1, HvG6PDH3, and HvG6PDH4 are plastidic isoforms. Analysis of enzymatic activities and gene expression showed that <i>HvG6PDH1</i> to <i>HvG6PDH4</i> are involved in responses to salt and drought stresses. The cytosolic <i>HvG6PDH2</i> is the major isoform against oxidative stress. <i>HvG6PDH5</i> may be a house-keeping gene. In addition, <i>HvG6PDH1</i> to <i>HvG6PDH4</i> and their encoded enzymes responded to jasmonic acid (JA) and abscisic acid (ABA) treatments, implying that JA and ABA are probably critical regulators of <i>HvG6PDH</i>s (except for <i>HvG6PDH5</i>). Reactive oxygen species analysis showed that inhibition of cytosolic and plastidic G6PDH activities leads to increased H<sub>2</sub>O<sub>2</sub> and O<sub>2</sub><sup>−</sup> contents in highland barley under salt and drought stresses. These results suggest that G6PDH can maintain cellular redox homeostasis and that cytosolic HvG6PDH2 is an irreplaceable isoform against oxidative stress in highland barley.https://www.mdpi.com/2223-7747/9/12/1800abiotic stressesglucose-6-phosphate dehydrogenasehighland barleyreactive oxygen speciesredox homeostasis |
spellingShingle | Ruijun Feng Xiaomin Wang Li He Shengwang Wang Junjie Li Jie Jin Yurong Bi Identification, Characterization, and Stress Responsiveness of Glucose-6-phosphate Dehydrogenase Genes in Highland Barley Plants abiotic stresses glucose-6-phosphate dehydrogenase highland barley reactive oxygen species redox homeostasis |
title | Identification, Characterization, and Stress Responsiveness of Glucose-6-phosphate Dehydrogenase Genes in Highland Barley |
title_full | Identification, Characterization, and Stress Responsiveness of Glucose-6-phosphate Dehydrogenase Genes in Highland Barley |
title_fullStr | Identification, Characterization, and Stress Responsiveness of Glucose-6-phosphate Dehydrogenase Genes in Highland Barley |
title_full_unstemmed | Identification, Characterization, and Stress Responsiveness of Glucose-6-phosphate Dehydrogenase Genes in Highland Barley |
title_short | Identification, Characterization, and Stress Responsiveness of Glucose-6-phosphate Dehydrogenase Genes in Highland Barley |
title_sort | identification characterization and stress responsiveness of glucose 6 phosphate dehydrogenase genes in highland barley |
topic | abiotic stresses glucose-6-phosphate dehydrogenase highland barley reactive oxygen species redox homeostasis |
url | https://www.mdpi.com/2223-7747/9/12/1800 |
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