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Journal of Integrative Agriculture  2015, Vol. 14 Issue (7): 1229-1241    DOI: 10.1016/S2095-3119(14)60872-8
Crop Genetics · Breeding · Germplasm Resources Advanced Online Publication | Current Issue | Archive | Adv Search |
Overexpression of a maize SNF-related protein kinase gene, ZmSnRK2.11, reduces salt and drought tolerance in Arabidopsis
 ZHANG Fan, CHEN Xun-ji, WANG Jian-hua, ZHENG Jun
1、College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, P.R.China
2、Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
3、Institute of Nuclear Technology and Biotechnology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, P.R.China
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摘要  Sucrose non-fermenting-1 related protein kinase 2 (SnRK2) is a unique family of protein kinases associated with abiotic stress signal transduction in plants. In this study, a maize SnRK2 gene ZmSnRK2.11 was cloned and characterized. The results showed that ZmSnRK2.11 is up-regulated by high-salinity and dehydration treatment, and it is expressed mainly in maize mature leaf. A transient expression assay using onion epidermal cells revealed that ZmSnRK2.11-GFP fusion proteins are localized to both the nucleus and cytoplasm. Overexpressing-ZmSnRK2.11 in Arabidopsis resulted in salt and drought sensitivity phenotypes that exhibited an increased rate of water loss, reduced relative water content, delayed stoma closure, accumulated less free proline content and increased malondialdehyde (MDA) content relative to the phenotypes observed in wild-type (WT) control. Furthermore, overexpression of ZmSnRK2.11 up-regulated the expression of the genes ABI1 and ABI2 and decreased the expression of DREB2A and P5CS1. Taken together, our results suggest that ZmSnRK2.11 is a possible negative regulator involved in the salt and drought stress signal transduction pathways in plants.

Abstract  Sucrose non-fermenting-1 related protein kinase 2 (SnRK2) is a unique family of protein kinases associated with abiotic stress signal transduction in plants. In this study, a maize SnRK2 gene ZmSnRK2.11 was cloned and characterized. The results showed that ZmSnRK2.11 is up-regulated by high-salinity and dehydration treatment, and it is expressed mainly in maize mature leaf. A transient expression assay using onion epidermal cells revealed that ZmSnRK2.11-GFP fusion proteins are localized to both the nucleus and cytoplasm. Overexpressing-ZmSnRK2.11 in Arabidopsis resulted in salt and drought sensitivity phenotypes that exhibited an increased rate of water loss, reduced relative water content, delayed stoma closure, accumulated less free proline content and increased malondialdehyde (MDA) content relative to the phenotypes observed in wild-type (WT) control. Furthermore, overexpression of ZmSnRK2.11 up-regulated the expression of the genes ABI1 and ABI2 and decreased the expression of DREB2A and P5CS1. Taken together, our results suggest that ZmSnRK2.11 is a possible negative regulator involved in the salt and drought stress signal transduction pathways in plants.
Keywords:  maize       salt and drought stresses       ZmSnRK2.11       SnRK2  
Received: 15 July 2014   Accepted:
Fund: 

This work was supported by the National High Technology R&D Program of China (2012AA10A306), the National Natural Science Foundation of China (31330056) and the Xinjiang High-Tech Research Projects, China (201011109).

Corresponding Authors:  ZHENG Jun, Tel: +86-10-82105863, Mobile: +86-13683230661, E-mail: zhengjun02@ caas.cn     E-mail:  zhengjun02@caas.cn
About author:  ZHANG Fan, Tel: +86-10-82105866, E-mail: zhangfan_cau@126.com

Cite this article: 

ZHANG Fan, CHEN Xun-ji, WANG Jian-hua, ZHENG Jun. 2015. Overexpression of a maize SNF-related protein kinase gene, ZmSnRK2.11, reduces salt and drought tolerance in Arabidopsis. Journal of Integrative Agriculture, 14(7): 1229-1241.

Alexandrov N N, Brover V V, Freidin S, Troukhan M E,Tatarinova T V, Zhang H, Swaller T J, Lu Y P, Bouck J,Flavell R B. 2009. Insights into corn genes derived fromlarge-scale cDNA sequencing. Plant Molecular Biology,69, 179-194

Anderberg R J, Walker-Simmons M 1992. Isolation of a wheatcDNA clone for an abscisic acid-inducible transcript withhomology to protein kinases. Proceedings of the NationalAcademy of Sciences of the United States of America, 89,10183-10187

Bateman A, Coin L, Durbin R, Finn R D, Hollich V, Griffiths-Jones S, Khanna A, Marshall M, Moxon S, SonnhammerE L L, Studholme D J, Yeats C, Eddy S R. 2004. The Pfamprotein families database. Nucleic Acids Research, 32,D138-D141.

Chakraborty K, Sairam R K, Bhattacharya R C. 2012. Differentialexpression of salt overly sensitive pathway genesdetermines salinity stress tolerance in Brassica genotypes.Plant Physiology Biochemistry, 51, 90-101

Coello P, Hey S J, Halford N G. 2010. The sucrose nonfermenting-1-related (SnRK) family of protein kinases:potential for manipulation to improve stress toleranceand increase yield. Journal of Experimental Botany, 62,883-893

Diedhiou C J, Popova O V, Dietz K J, Golldack D. 2008.The SNF1-type serine-threonine protein kinase SAPK4regulates stress-responsive gene expression in rice. BMCPlant Biology, 8, 49.

Earley K W, Haag J R, Pontes O, Opper K, Juehne T, Song K,Pikaard C S. 2006. Gateway-compatible vectors for plantfunctional genomics and proteomics. The Plant Journal,45, 616-629

Fujii H, Verslues P E, Zhu J K. 2007. Identification of twoprotein kinases required for abscisic acid regulation ofseed germination, root growth, and gene expression inArabidopsis. The Plant Cell, 19, 485-494

Fujita Y, Nakashima K, Yoshida T, Katagiri T, Kidokoro S,Kanamori N, Umezawa T, Fujita M, Maruyama K, IshiyamaK, Kobayashi M, Nakasone S, Yamada K, Ito T, ShinozakiK, Yamaguchi-Shinozaki K. 2009. Three SnRK2 proteinkinases are the main positive regulators of abscisic acidsignaling in response to water stress in Arabidopsis. Plantand Cell Physiology, 50, 2123-2132

Gomez-Cadenas A, Verhey S D, Holappa L D, Shen Q,Ho T H, Walker-Simmons M K. 1999. An abscisic acidinducedprotein kinase, PKABA1, mediates abscisic acid suppressed gene expression in barley aleurone layers.Proceedings of the National Academy of Sciences of theUnited States of America, 96, 1767-1772

Halford N G, Hardie D G. 1998. SNF1-related protein kinases:global regulators of carbon metabolism in plants? PlantMolecular Biology, 37, 735-748

Hrabak E M. 2003. The Arabidopsis CDPK-SnRK superfamilyof protein kinases. Plant Physiology, 132, 666-680

Huai J, Wang M, He J, Zheng J, Dong Z, Lv H, Zhao J, WangG. 2008. Cloning and characterization of the SnRK2 genefamily from Zea mays. Plant Cell Reports, 27, 1861-1868

Kim T H, Maik B. 2010. Guard cell signal transduction network:advances in understanding abscisic acid, CO2, and Ca2+signaling. Annual Review of Plant Biology, 61, 561-591

Kobayashi Y, Yamamoto S, Minami H, Kagaya Y, Hattori T. 2004.Differential activation of the rice sucrose nonfermenting1-related protein kinase 2 family by hyperosmotic stressand abscisic acid. The Plant Cell, 16, 1163-1177

Kulik A, Wawer I, Krzywinska E, Bucholc M, DobrowolskaG. 2011. SnRK2 protein kinases-key regulators ofplant response to abiotic stresses. OMICS: A Journal ofIntegrative Biology, 15, 859-872

Li J, Wang X Q, Watson M B, Assmann S M. 2000. Regulation ofabscisic acid-induced stomatal closure and anion channelsby guard cell AAPK kinase. Science, 287, 300-303

Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki K, Shinozaki K. 1998. Two transcriptionfactors, DREB1 and DREB2, with an EREBP/AP2 DNAbinding domain separate two cellular signal transductionpathways in drought-and low-temperature-responsive geneexpression, respectively, in Arabidopsis. The Plant Cell,10, 1391-1406

Mao X, Zhang H, Tian S, Chang X, Jing R. 2010. TaSnRK2.4,an SNF1-type serine/threonine protein kinase of wheat(Triticum aestivum L.), confers enhanced multistresstolerance in Arabidopsis. Journal of Experimental Botany,61, 683-696

McLoughlin F, Galvan-Ampudia C S, Julkowska M M, CaarlsL, van der Does D, Lauriere C, Munnik T, Haring M A,Testerink C. 2012. The Snf1-related protein kinasesSnRK2.4 and SnRK2.10 are involved in maintenance of rootsystem architecture during salt stress. The Plant Journal,72, 436-449

Merlot S, Mustilli A C, Genty B, North H, Lefebvre V, Sotta B,Vavasseur A, Giraudat J. 2002. Use of infrared thermalimaging to isolate Arabidopsis mutants defective in stomatalregulation. The Plant Journal, 30, 601-609

Murashige T, Skoog F. 1962. A revised medium for rapid growthand bio assays with tobacco tissue cultures. PhysiologiaPlantarum, 15, 473-497

Mustilli A C. 2002. Arabidopsis OST1 protein kinase mediatesthe regulation of stomatal aperture by abscisic acid andacts upstream of reactive oxygen species production. ThePlant Cell, 14, 3089-3099

Podell S, Gribskov M. 2004. Predicting N-terminal myristoylationsites in plant proteins. BMC Genomics, 5, 37.

Sairam R, Deshmukh P, Saxena D. 1998. Role of antioxidantsystems in wheat genotypes tolerance to water stress.Biologia Plantarum, 41, 387-394

Savouré A, Hua X J, Bertauche N, Van Montagu M, VerbruggenN. 1997. Abscisic acid-independent and abscisic aciddependentregulation of proline biosynthesis following coldand osmotic stresses in Arabidopsis thaliana. Molecular andGeneral Genetics, 254, 104-109

Shan G, Embrey S K, Schafer B W. 2007. A highly specificenzyme-linked immunosorbent assay for the detection ofCry1Ac insecticidal crystal protein in transgenic WideStrikecotton. Jouranl of Agricultural and Food Chemistry, 55,5974-5979

Shukla V, Mattoo A K. 2008. Sucrose non-fermenting 1-relatedprotein kinase 2 (SnRK2): A family of protein kinasesinvolved in hyperosmotic stress signaling. Physiology andMolecular Biology of Plants, 14, 91-100

Steven J. Clough, Bent A F. 1998. Floral dip a simplified methodfor Agrobacterium-mediated transformation of Arabidopsisthaliana. The Plant Journal, 16, 735-743

Swamy B P M, Ahmed H U, Henry A, Mauleon R, Dixit S, VikramP, Tilatto R, Verulkar S B, Perraju P, Mandal N P, Variar M,Robin S, Chandrababu R, Singh O N, Dwivedi J L, Das SP, Mishra K K, Yadaw R B, Aditya T L, Karmakar B, et al.2013. Genetic, physiological, and gene expression analysesreveal that multiple QTL enhance yield of rice mega-varietyIR64 under drought. PlOS ONE, 8, e62795.

Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecularevolutionary genetics analysis (MEGA) software version 4.0.Molecular Biology and Evolution, 24, 1596-1599

Tian S, Mao X, Zhang H, Chen S, Zhai C, Yang S, Jing R.2013. Cloning and characterization of TaSnRK2.3, a novelSnRK2 gene in common wheat. Journal of ExperimentalBotany, 64, 2063-2080

Umezawa T, Sugiyama N, Mizoguchi M, Hayashi S, Myouga F,Yamaguchi-Shinozaki K, Ishihama Y, Hirayama T, ShinozakiK. 2009. Type 2C protein phosphatases directly regulateabscisic acid-activated protein kinases in Arabidopsis.Proceedings of the National Academy of Sciences of theUnited States of America, 106, 17588-17593

Vlad F, Rubio S, Rodrigues A, Sirichandra C, Belin C, RobertN, Leung J, Rodriguez P L, Laurière C, Merlot S. 2009.Protein phosphatases 2C regulate the activation of theSnf1-related kinase OST1 by abscisic acid in Arabidopsis.The Plant Cell, 21, 3170-3184

Wang M, Yuan F, Hao H, Zhang Y, Zhao H, Guo A, Hu J,Zhou X, Xie C G. 2013. BolOST1, an ortholog of OpenStomata 1 with alternative splicing products in Brassicaoleracea, positively modulates drought responses in plants.Biochemical and Biophysical Research Communications,442, 214-220

Ying S, Zhang D F, Li H Y, Liu Y H, Shi Y S, Song Y C, Wang T Y,Li Y. 2011. Cloning and characterization of a maize SnRK2protein kinase gene confers enhanced salt tolerance intransgenic Arabidopsis. Plant Cell Reports, 30, 1683-1699

Yoshida R, Hobo T, Ichimura K, Mizoguchi T, Takahashi F, Aronso J, Ecker J R, Shinozaki K. 2002. ABA-activatedSnRK2 protein kinase is required for dehydration stresssignaling in Arabidopsis. Plant and Cell Physiology, 43,1473-1483

Zhang H, Mao X, Jing R, Chang X, Xie H. 2011. Characterizationof a common wheat (Triticum aestivum L.) TaSnRK2.7gene involved in abiotic stress responses. Journal ofExperimental Botany, 62, 975-988

Zhang H, Mao X, Wang C, Jing R. 2010. Overexpression of acommon wheat gene TaSnRK2.8 enhances tolerance todrought, salt and low temperature in Arabidopsis. PLoSOne, 5, e16041.

Zhao J, Sun Z, Zheng J, Guo X, Dong Z, Huai J, Gou M, He J,Jin Y, Wang J, Wang G. 2009. Cloning and characterizationof a novel CBL-interacting protein kinase from maize. PlantMolecular Biology, 69, 661-674

Zheng J, Zhao J, Tao Y, Wang J, Liu Y, Fu J, Jin Y, Gao P,Zhang J, Bai Y, Wang G. 2004. Isolation and analysisof water stress induced genes in maize seedlings bysubtractive PCR and cDNA macroarray. Plant MolecularBiology, 55, 807-823
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