GhWRKY6 Acts as a Negative Regulator in Both Transgenic Arabidopsis and Cotton During Drought and Salt Stress

Drought and high salinity are key limiting factors for cotton production. Therefore, research is increasingly focused on the underlying stress response mechanisms of cotton. We first identified and cloned a novel gene encoding the 525 amino acids in cotton, namely GhWRKY6. qRT-PCR analysis indicated that GhWRKY6 was induced by NaCl, PEG 6000 and ABA. Analyses of germination rate and root length indicated that overexpression of GhWRKY6 in Arabidopsis resulted in hypersensitivity to ABA, NaCl, and PEG 6000. In contrast, the loss-of-function mutant wrky6 was insensitive and had slightly longer roots than the wild-type did under these treatment conditions. Furthermore, GhWRKY6 overexpression in Arabidopsis modulated salt- and drought-sensitive phenotypes and stomatal aperture by regulating ABA signaling pathways, and reduced plant tolerance to abiotic stress through reactive oxygen species (ROS) enrichment, reduced proline content, and increased electrolytes and malondialdehyde (MDA). The expression levels of a series of ABA-, salt- and drought-related marker genes were altered in overexpression seedlings. Virus-induced gene silencing (VIGS) technology revealed that down-regulation of GhWRKY6 increased salt tolerance in cotton. These results demonstrate that GhWRKY6 is a negative regulator of plant responses to abiotic stress via the ABA signaling pathway.

[1]  Ting Zhao,et al.  Overexpression of VaWRKY14 increases drought tolerance in Arabidopsis by modulating the expression of stress-related genes , 2018, Plant Cell Reports.

[2]  Xiyan Yang,et al.  A novel cotton WRKY gene, GhWRKY6-like, improves salt tolerance by activating the ABA signaling pathway and scavenging of reactive oxygen species. , 2018, Physiologia plantarum.

[3]  Fuguang Li,et al.  Genome-Wide Study of YABBY Genes in Upland Cotton and Their Expression Patterns under Different Stresses , 2018, Front. Genet..

[4]  Chao-jun Zhang,et al.  A Phi-Class Glutathione S-Transferase Gene for Verticillium Wilt Resistance in Gossypium arboreum Identified in a Genome-Wide Association Study , 2018, Plant & cell physiology.

[5]  Haitao Shi,et al.  Alcohol dehydrogenase 1 (ADH1) confers both abiotic and biotic stress resistance in Arabidopsis. , 2017, Plant science : an international journal of experimental plant biology.

[6]  Chao-jun Zhang,et al.  Genome-wide analysis of WOX genes in upland cotton and their expression pattern under different stresses , 2017, BMC Plant Biology.

[7]  D. Ankerst,et al.  Interactions between temperature and drought in global and regional crop yield variability during 1961-2014 , 2017, PloS one.

[8]  Zhou Du,et al.  agriGO v2.0: a GO analysis toolkit for the agricultural community, 2017 update , 2017, Nucleic Acids Res..

[9]  J. Flexas,et al.  Ectopic Expression of CDF3 Genes in Tomato Enhances Biomass Production and Yield under Salinity Stress Conditions , 2017, Front. Plant Sci..

[10]  I. Szarejko,et al.  The Role and Regulation of ABI5 (ABA-Insensitive 5) in Plant Development, Abiotic Stress Responses and Phytohormone Crosstalk , 2016, Front. Plant Sci..

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

[12]  Anagh Ray,et al.  ABI3 mediates dehydration stress recovery response in Arabidopsis thaliana by regulating expression of downstream genes. , 2016, Plant science : an international journal of experimental plant biology.

[13]  Ning Wang,et al.  GhWRKY25, a group I WRKY gene from cotton, confers differential tolerance to abiotic and biotic stresses in transgenic Nicotiana benthamiana , 2016, Protoplasma.

[14]  Prakash P. Kumar,et al.  Plant hormone-mediated regulation of stress responses , 2016, BMC Plant Biology.

[15]  Wei-Hua Wu,et al.  Arabidopsis WRKY6 Transcription Factor Acts as a Positive Regulator of Abscisic Acid Signaling during Seed Germination and Early Seedling Development , 2016, PLoS genetics.

[16]  Wei Zhang,et al.  WRKY1 regulates stomatal movement in drought-stressed Arabidopsis thaliana , 2016, Plant Molecular Biology.

[17]  M. Pinedo,et al.  On the role of a Lipid-Transfer Protein. Arabidopsis ltp3 mutant is compromised in germination and seedling growth. , 2015, Plant signaling & behavior.

[18]  Yanchen Tian,et al.  A wheat salinity-induced WRKY transcription factor TaWRKY93 confers multiple abiotic stress tolerance in Arabidopsis thaliana. , 2015, Biochemical and biophysical research communications.

[19]  Wenya Guo,et al.  LTP3 contributes to disease susceptibility in Arabidopsis by enhancing abscisic acid (ABA) biosynthesis. , 2015, Molecular plant pathology.

[20]  R. Hu,et al.  Overexpression of Cotton RAV1 Gene in Arabidopsis Confers Transgenic Plants High Salinity and Drought Sensitivity , 2015, PloS one.

[21]  K. Yamaguchi-Shinozaki,et al.  ABA-dependent and ABA-independent signaling in response to osmotic stress in plants. , 2014, Current opinion in plant biology.

[22]  Xiaoyan Ma,et al.  NRGA1, a putative mitochondrial pyruvate carrier, mediates ABA regulation of guard cell ion channels and drought stress responses in Arabidopsis. , 2014, Molecular plant.

[23]  Xiaobo Chen,et al.  The cotton WRKY transcription factor GhWRKY17 functions in drought and salt stress in transgenic Nicotiana benthamiana through ABA signaling and the modulation of reactive oxygen species production. , 2014, Plant & cell physiology.

[24]  M. Margis-Pinheiro,et al.  Genome-wide annotation of the soybean WRKY family and functional characterization of genes involved in response to Phakopsora pachyrhizi infection , 2014, BMC Plant Biology.

[25]  K. Nam,et al.  A subset of Arabidopsis RAV transcription factors modulates drought and salt stress responses independent of ABA. , 2014, Plant & cell physiology.

[26]  N. Suzuki,et al.  ROS as key players in plant stress signalling. , 2014, Journal of experimental botany.

[27]  R. Oelmüller,et al.  WRKY transcription factors , 2014, Plant signaling & behavior.

[28]  Yule Liu,et al.  Development of Agrobacterium-Mediated Virus-Induced Gene Silencing and Performance Evaluation of Four Marker Genes in Gossypium barbadense , 2013, PloS one.

[29]  L. Holm,et al.  Defense-related transcription factors WRKY70 and WRKY54 modulate osmotic stress tolerance by regulating stomatal aperture in Arabidopsis , 2013, The New phytologist.

[30]  A. Molina,et al.  Disease resistance or growth: the role of plant hormones in balancing immune responses and fitness costs , 2013, Front. Plant Sci..

[31]  I. Szarejko,et al.  Open or Close the Gate – Stomata Action Under the Control of Phytohormones in Drought Stress Conditions , 2013, Front. Plant Sci..

[32]  Søren Lindemose,et al.  Structure, Function and Networks of Transcription Factors Involved in Abiotic Stress Responses , 2013, International journal of molecular sciences.

[33]  Xiaoyan Zhang,et al.  Lipid transfer protein 3 as a target of MYB96 mediates freezing and drought stress in Arabidopsis , 2013, Journal of experimental botany.

[34]  Gang Liang,et al.  Activated expression of WRKY57 confers drought tolerance in Arabidopsis. , 2012, Molecular plant.

[35]  Manoj Prasad,et al.  NAC proteins: regulation and role in stress tolerance. , 2012, Trends in plant science.

[36]  I. Somssich,et al.  Arabidopsis WRKY33 Is a Key Transcriptional Regulator of Hormonal and Metabolic Responses toward Botrytis cinerea Infection1[W] , 2012, Plant Physiology.

[37]  K. Shinozaki,et al.  NAC transcription factors in plant abiotic stress responses. , 2012, Biochimica et biophysica acta.

[38]  Diqiu Yu,et al.  The role of WRKY transcription factors in plant abiotic stresses. , 2012, Biochimica et biophysica acta.

[39]  K. Shinozaki,et al.  AP2/ERF family transcription factors in plant abiotic stress responses. , 2012, Biochimica et biophysica acta.

[40]  Jun Liu,et al.  Signal transduction during cold, salt, and drought stresses in plants , 2011, Molecular Biology Reports.

[41]  Xiaoping Zhou,et al.  WRKY22 transcription factor mediates dark-induced leaf senescence in Arabidopsis , 2011, Molecules and cells.

[42]  K. Shinozaki,et al.  ABA-mediated transcriptional regulation in response to osmotic stress in plants , 2011, Journal of Plant Research.

[43]  J. Msanne,et al.  Characterization of abiotic stress-responsive Arabidopsis thaliana RD29A and RD29B genes and evaluation of transgenes , 2011, Planta.

[44]  P. Verslues,et al.  Arabidopsis decuple mutant reveals the importance of SnRK2 kinases in osmotic stress responses in vivo , 2011, Proceedings of the National Academy of Sciences.

[45]  M. Zhang,et al.  ABO3, a WRKY transcription factor, mediates plant responses to abscisic acid and drought tolerance in Arabidopsis. , 2010, The Plant journal : for cell and molecular biology.

[46]  Diqiu Yu,et al.  Overexpression of OsWRKY72 gene interferes in the abscisic acid signal and auxin transport pathway of Arabidopsis , 2010, Journal of Biosciences.

[47]  Q. Shen,et al.  WRKY transcription factors. , 2010, Trends in plant science.

[48]  R. Mittler,et al.  The zinc finger network of plants , 2008, Cellular and Molecular Life Sciences.

[49]  U. Sonnewald,et al.  Specific Roles of α- and γ-Tocopherol in Abiotic Stress Responses of Transgenic Tobacco1[W][OA] , 2007, Plant Physiology.

[50]  Liangjiang Wang,et al.  The WRKY transcription factor superfamily: its origin in eukaryotes and expansion in plants , 2005, BMC Evolutionary Biology.

[51]  K. Shinozaki,et al.  A combination of the Arabidopsis DREB1A gene and stress-inducible rd29A promoter improved drought- and low-temperature stress tolerance in tobacco by gene transfer. , 2004, Plant & cell physiology.

[52]  Jian-Kang Zhu,et al.  Regulation of Abscisic Acid Biosynthesis1 , 2003, Plant Physiology.

[53]  E. Grotewold,et al.  MYB transcription factors in Arabidopsis. , 2002, Trends in plant science.

[54]  Julian I. Schroeder,et al.  Guard cell abscisic acid signalling and engineering drought hardiness in plants , 2001, Nature.

[55]  K. Shinozaki,et al.  Arabidopsis basic leucine zipper transcription factors involved in an abscisic acid-dependent signal transduction pathway under drought and high-salinity conditions. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[56]  T. Eulgem,et al.  The WRKY superfamily of plant transcription factors. , 2000, Trends in plant science.

[57]  C. Kim,et al.  The Arabidopsis thaliana SOS2 gene encodes a protein kinase that is required for salt tolerance. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[58]  S. Clough,et al.  Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[59]  Wang,et al.  Differential responses of abaxial and adaxial guard cells of broad bean to abscisic acid and calcium , 1998, Plant physiology.

[60]  M. Ishitani,et al.  Genetic analysis of osmotic and cold stress signal transduction in Arabidopsis: interactions and convergence of abscisic acid-dependent and abscisic acid-independent pathways. , 1997, The Plant cell.

[61]  M. Delseny,et al.  Two different Em-like genes are expressed in Arabidopsis thaliana seeds during maturation , 1993, Molecular and General Genetics MGG.

[62]  Q. Shen,et al.  WRKY transcription factors: key components in abscisic acid signalling. , 2012, Plant biotechnology journal.

[63]  C. Abdelly,et al.  [Proline, a multifunctional amino-acid involved in plant adaptation to environmental constraints]. , 2012, Biologie aujourd'hui.

[64]  Kazuo Shinozaki,et al.  Transcriptional Regulation of ABI3- and ABA-responsive Genes Including RD29B and RD29A in Seeds, Germinating Embryos, and Seedlings of Arabidopsis , 2005, Plant Molecular Biology.