Dehydrin CaDHN2 Enhances Drought Tolerance by Affecting Ascorbic Acid Synthesis under Drought in Peppers

Peppers (Capsicum annuum L.), as a horticultural crop with one of the highest ascorbic acid contents, are negatively affected by detrimental environmental conditions both in terms of quality and productivity. In peppers, the high level of ascorbic acid is not only a nutrient substance but also plays a role in environmental stress, i.e., drought stress. When suffering from drought stress, plants accumulate dehydrins, which play important roles in the stress response. Here, we isolated an SK3-type DHN gene CaDHN2 from peppers. CaDHN2 was located in the nucleus, cytoplasm, and cell membrane. In CaDHN2-silenced peppers, which are generated by virus-induced gene silencing (VIGS), the survival rate is much lower, the electrolytic leakage is higher, and the accumulation of reactive oxygen species (ROS) is greater when compared with the control under drought stress. Moreover, when CaDHN2 (CaDHN2-OE) is overexpressed in Arabidopsis, theoverexpressing plants show enhanced drought tolerance, increased antioxidant enzyme activities, and lower ROS content. Based on yeast two-hybrid (Y2H), GST-pull down, and bimolecular fluorescence complementation (BiFC) results, we found that CaDHN2 interacts with CaGGP1, the key enzyme in ascorbic acid (AsA) synthesis, in the cytoplasm. Accordingly, the level of ascorbic acid is highly reduced in CaDHN2-silenced peppers, indicating that CaDHN2 interacts with CaGGP1 to affect the synthesis of ascorbic acid under drought stress, thus improving the drought tolerance of peppers. Our research provides a basis for further study of the function of DHN genes.

[1]  W. Ye,et al.  GhIMP10D, an inositol monophosphates family gene, enhances ascorbic acid and antioxidant enzyme activities to confer alkaline tolerance in Gossypium hirsutum L. , 2023, BMC Plant Biology.

[2]  S. Tiwari,et al.  Wheat derived glucuronokinase as a potential target for regulating ascorbic acid and phytic acid content with increased root length under drought and ABA stresses in Arabidopsis thaliana. , 2023, Plant science : an international journal of experimental plant biology.

[3]  Lijun Qin,et al.  Heterologous expression of HsPstS gene reducing arsenic accumulation and improving As-tolerance in transgenic tobaccos by enhancing CAT activity. , 2023, Journal of plant physiology.

[4]  N. Samarah,et al.  Effect of drought stress on wheat (Triticum durum) growth and metabolism: insight from GABA shunt, reactive oxygen species and dehydrin genes expression. , 2022, Functional plant biology : FPB.

[5]  D. Papaefthimiou,et al.  Over-expression of GGP1 and GPP genes enhances ascorbate content and nutritional quality of tomato. , 2022, Plant physiology and biochemistry : PPB.

[6]  A. Kadıoğlu,et al.  Increased dehydrin level decreases leaf rolling grade by altering the reactive oxygen species homeostasis and abscisic acid content in maize subjected to osmotic stress , 2022, 3 Biotech.

[7]  M. MacDonald,et al.  Ascorbic Acid Preconditioning Effect on Broccoli Seedling Growth and Photosynthesis under Drought Stress , 2022, Plants.

[8]  A. Xiong,et al.  AgGMP encoding GDP-D-mannose pyrophosphorylase from celery enhanced the accumulation of ascorbic acid and resistance to drought stress in Arabidopsis , 2022, PeerJ.

[9]  S. Graether,et al.  The Disordered Dehydrin and Its Role in Plant Protection: A Biochemical Perspective , 2022, Biomolecules.

[10]  Li-jing Chen,et al.  AnDHN, a Dehydrin Protein From Ammopiptanthus nanus, Mitigates the Negative Effects of Drought Stress in Plants , 2021, Frontiers in Plant Science.

[11]  Y. Magata,et al.  Inhibition of cryoaggregation of phospholipid liposomes by an Arabidopsis intrinsically disordered dehydrin and its K-segment. , 2021, Colloids and surfaces. B, Biointerfaces.

[12]  Gezi Li,et al.  Exogenous ascorbic acid application alleviates cadmium toxicity in seedlings of two wheat (Triticum aestivum L.) varieties by reducing cadmium uptake and enhancing antioxidative capacity , 2021, Environmental Science and Pollution Research.

[13]  Z. Gong,et al.  The CaCIPK3 gene positively regulates drought tolerance in pepper , 2021, Horticulture research.

[14]  A. Nayak,et al.  Combined application of ascorbic acid and endophytic N-fixing Azotobacter chroococcum Avi2 modulates photosynthetic efficacy, antioxidants and growth-promotion in rice under moisture deficit stress. , 2021, Microbiological research.

[15]  Jun-Peng Niu,et al.  Effects of exogenous ascorbic acid on seed germination and seedling salt-tolerance of alfalfa , 2021, PloS one.

[16]  Sung Chul Lee,et al.  Pepper E3 ligase CaAIRE1 promotes ABA sensitivity and drought tolerance by degradation of protein phosphatase CaAITP1. , 2021, Journal of experimental botany.

[17]  H. Yasmin,et al.  Foliar application of ascorbic acid enhances salinity stress tolerance in barley (Hordeum vulgare L.) through modulation of morpho-physio-biochemical attributes, ions uptake, osmo-protectants and stress response genes expression , 2021, Saudi journal of biological sciences.

[18]  Yuan Meng,et al.  CaDHN3, a Pepper (Capsicum annuum L.) Dehydrin Gene Enhances the Tolerance against Salt and Drought Stresses by Reducing ROS Accumulation , 2021, International journal of molecular sciences.

[19]  V. Rao,et al.  Arabidopsis F-box protein At1g08710 interacts with transcriptional protein ADA2b and imparts drought stress tolerance by negatively regulating seedling growth. , 2020, Biochemical and biophysical research communications.

[20]  Shushen Yang,et al.  Retraction Note: The specific MYB binding sites bound by TaMYB in the GAPCp2/3 promoters are involved in the drought stress response in wheat , 2020, BMC plant biology.

[21]  C. Merodio,et al.  Functional characterization of VviDHN2 and VviDHN4 dehydrin isoforms from Vitis vinifera (L.): An in silico and in vitro approach. , 2020, Plant physiology and biochemistry : PPB.

[22]  S. Hou,et al.  The LRR-RLK Protein HSL3 Regulates Stomatal Closure and the Drought Stress Response by Modulating Hydrogen Peroxide Homeostasis , 2020, Frontiers in Plant Science.

[23]  Bo Gyeong Kim,et al.  Thermal processing under oxygen-free condition of blueberry puree: Effect on anthocyanin, ascorbic acid, antioxidant activity, and enzyme activities. , 2020, Food chemistry.

[24]  Hao Liu,et al.  Effect of K-/S- segments on subcellular localization and dimerization of wheat dehydrin WZY1-2 , 2020, Plant signaling & behavior.

[25]  D. Vertommen,et al.  Interplay of Structural Disorder and Short Binding Elements in the Cellular Chaperone Function of Plant Dehydrin ERD14 , 2020, Cells.

[26]  M. Hara,et al.  Cryoprotective activity of Arabidopsis KS-type dehydrin depends on the hydrophobic amino acids of two active segments. , 2020, Archives of biochemistry and biophysics.

[27]  Sung Chul Lee,et al.  Pepper Novel Serine-Threonine Kinase CaDIK1 Regulates Drought Tolerance via Modulating ABA Sensitivity , 2020, Frontiers in Plant Science.

[28]  Jian Li,et al.  The Apple microR171i-SCARECROW-LIKE PROTEINS26.1 Module Enhances Drought Stress Tolerance by Integrating Ascorbic Acid Metabolism1 , 2020, Plant Physiology.

[29]  Z. khazaei,et al.  Ameliorative effects of ascorbic acid on tolerance to drought stress on pepper (Capsicum annuum L) plants , 2020, Physiology and Molecular Biology of Plants.

[30]  C. Ulrichs,et al.  Ascorbic Acid Induces the Increase of Secondary Metabolites, Antioxidant Activity, Growth, and Productivity of the Common Bean under Water Stress Conditions , 2020, Plants.

[31]  Shaoliang Chen,et al.  Heterologous Expression of Three Ammopiptanthus mongolicus Dehydrin Genes Confers Abiotic Stress Tolerance in Arabidopsis thaliana , 2020, Plants.

[32]  N. Smirnoff,et al.  Deficiency of GDP-l-galactose phosphorylase, an enzyme required for ascorbic acid synthesis, reduces tomato fruit yield , 2020, Planta.

[33]  Sung Chul Lee,et al.  Roles of pepper bZIP transcription factor CaATBZ1 and its interacting partner RING-type E3 ligase CaASRF1 in modulation of ABA signaling and drought tolerance. , 2019, The Plant journal : for cell and molecular biology.

[34]  J. Wen,et al.  Dehydrin MtCAS31 promotes autophagic degradation under drought stress , 2019, Autophagy.

[35]  Qing-hui Yu,et al.  A small heat shock protein CaHsp25.9 positively regulates heat, salt, and drought stress tolerance in pepper (Capsicum annuum L.). , 2019, Plant physiology and biochemistry : PPB.

[36]  Sung Chul Lee,et al.  A pepper RING‐type E3 ligase, CaASRF1, plays a positive role in drought tolerance via modulation of CaAIBZ1 stability , 2019, The Plant journal : for cell and molecular biology.

[37]  Sung Chul Lee,et al.  Roles of pepper bZIP protein CaDILZ1 and its interacting partner RING-type E3 ligase CaDSR1 in modulation of drought tolerance. , 2018, The Plant journal : for cell and molecular biology.

[38]  J. Wen,et al.  MtCAS31 Aids Symbiotic Nitrogen Fixation by Protecting the Leghemoglobin MtLb120-1 Under Drought Stress in Medicago truncatula , 2018, Front. Plant Sci..

[39]  Jung-Hyun Kim,et al.  Pepper CaREL1, a ubiquitin E3 ligase, regulates drought tolerance via the ABA-signalling pathway , 2017, Scientific Reports.

[40]  G. Richard Strimbeck.  Hiding in plain sight: the F segment and other conserved features of seed plant SKn dehydrins , 2017, Planta.

[41]  Tingting Zhu,et al.  Ascorbic acid metabolism during sweet cherry (Prunus avium) fruit development , 2017, PloS one.

[42]  Jarosław Gocławski,et al.  Automated image analysis for quantification of reactive oxygen species in plant leaves. , 2016, Methods.

[43]  Jian‐Kang Zhu.  Abiotic Stress Signaling and Responses in Plants , 2016, Cell.

[44]  Supratim Basu,et al.  Plant adaptation to drought stress , 2016, F1000Research.

[45]  J. Danielsson,et al.  Membrane-Induced Folding of the Plant Stress Dehydrin Lti301[OPEN] , 2016, Plant Physiology.

[46]  M. Strömvik,et al.  De Novo Regulatory Motif Discovery Identifies Significant Motifs in Promoters of Five Classes of Plant Dehydrin Genes , 2015, PloS one.

[47]  A. Covarrubias,et al.  Dissecting the cryoprotection mechanisms for dehydrins , 2014, Front. Plant Sci..

[48]  S. Graether,et al.  Disorder and function: a review of the dehydrin protein family , 2014, Front. Plant Sci..

[49]  S. Graether,et al.  A dehydrin-dehydrin interaction: the case of SK3 from Opuntia streptacantha , 2014, Front. Plant Sci..

[50]  S. Graether,et al.  The Protection of Membranes from Cold-Stress: A Structural Study of the Intrinsically Disordered Dehydrin Bound to Micelles and Liposomes , 2014 .

[51]  Do Young Hyeon,et al.  The Arabidopsis NAC Transcription Factor ANAC096 Cooperates with bZIP-Type Transcription Factors in Dehydration and Osmotic Stress Responses[W] , 2013, Plant Cell.

[52]  Jiangli Dong,et al.  Overexpression of MtCAS31 enhances drought tolerance in transgenic Arabidopsis by reducing stomatal density. , 2012, The New phytologist.

[53]  W. Kim,et al.  Constitutive expression of CaRma1H1, a hot pepper ER-localized RING E3 ubiquitin ligase, increases tolerance to drought and salt stresses in transgenic tomato plants , 2012, Plant Cell Reports.

[54]  Jianhua Zhang,et al.  Ascorbic acid and reactive oxygen species are involved in the inhibition of seed germination by abscisic acid in rice seeds , 2011, Journal of experimental botany.

[55]  K. Miura,et al.  The Arabidopsis GTL1 Transcription Factor Regulates Water Use Efficiency and Drought Tolerance by Modulating Stomatal Density via Transrepression of SDD1[W][OA] , 2010, Plant Cell.

[56]  L. Kay,et al.  Quaternary dynamics and plasticity underlie small heat shock protein chaperone function , 2010, Proceedings of the National Academy of Sciences.

[57]  Ai-guo Qin,et al.  [Relative expression of SOD and CAT mRNA and activities of SOD and CAT in grafted cucumber leaves under NaCl stress]. , 2008, Ying yong sheng tai xue bao = The journal of applied ecology.

[58]  S. Mahboob,et al.  The effect of water stress on the antioxidant content, protective enzyme activities, proline content and lipid peroxidation in wheat seedling. , 2008, Pakistan journal of biological sciences : PJBS.

[59]  P. Tompa,et al.  Chaperone Activity of ERD10 and ERD14, Two Disordered Stress-Related Plant Proteins1[OA] , 2008, Plant Physiology.

[60]  Desen Ke,et al.  Effects of superoxide radicals on ACC synthase activity in chilling-stressed etiolated mungbean seedlings , 2007, Plant Growth Regulation.

[61]  A. Tunnacliffe,et al.  LEA proteins prevent protein aggregation due to water stress. , 2005, The Biochemical journal.

[62]  Peter Tompa,et al.  The role of structural disorder in the function of RNA and protein chaperones , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[63]  D. Robertson.  VIGS vectors for gene silencing: many targets, many tools. , 2004, Annual review of plant biology.

[64]  M. Alsheikh,et al.  Ion Binding Properties of the Dehydrin ERD14 Are Dependent upon Phosphorylation* , 2003, Journal of Biological Chemistry.

[65]  David C Baulcombe,et al.  Virus-induced gene silencing in plants. , 2003, Methods.

[66]  A. Able.  Role of reactive oxygen species in the response of barley to necrotrophic pathogens , 2003, Protoplasma.

[67]  P. Kasraie,et al.  The effect of ascorbic acid and bio fertilizers on basil under drought stress. , 2022, Brazilian journal of biology = Revista brasleira de biologia.

[68]  N. Jambunathan.  Determination and detection of reactive oxygen species (ROS), lipid peroxidation, and electrolyte leakage in plants. , 2010, Methods in molecular biology.

[69]  J. Svensson,et al.  A dehydrin gene in Physcomitrella patens is required for salt and osmotic stress tolerance. , 2006, The Plant journal : for cell and molecular biology.

[70]  Ch. R. Allagulova,et al.  Induction of expression of the dehydrin gene TADHN and accumulation of abscisic acid in wheat plants in hypothermia , 2005, Doklady Biochemistry and Biophysics.

[71]  Jian-Kang Zhu,et al.  Salt and drought stress signal transduction in plants. , 2002, Annual review of plant biology.

[72]  S. Paik,et al.  Structural changes in α‐synuclein affect its chaperone‐like activity in vitro , 2000, Protein science : a publication of the Protein Society.