Abstract
Aims
To examine heavy metal-induced regulatory mechanisms at the transcriptional level, a cell wall-associated receptor kinase (WAK) gene, OsWAK11 and its upstream promoter region (−946/+28) were isolated from Oryza sativa. OsWAK11 expression in response to abiotic stress was examined using a β-glucuronidase (GUS) gene fusion.
Methods
Semi-quantitative RT-PCR was used to analyze expression of the OsWAK11 gene. Histochemical detection of GUS was conducted by X-gluc staining methods, and fluorometric measurements of GUS activity were made with 4-methyl umbelliferyl glucuronide (MUG) substrate.
Results
The WAK promoter (−946/+28) responded to aluminum chloride, sodium chloride, and copper (II) sulfate with 3.0-, 2.2-, or 6.4-fold induction of GUS activity, respectively. Sodium nitroprusside and wounding treatment stimulated GUS activity. A histochemical analysis revealed strong GUS staining in the hypocotyls, cotyledons, first leaf, and petiole of cotyledons in transgenic tobacco seedlings. Strong GUS staining was also observed in the stigma and ovary of mature flowers, but not in the stamens.
Conclusion
OsWAK11 expression is regulated by aluminum, sodium, and copper. The GUS expression observed in transgenic tobacco carrying WAK11 promoter demonstrated significant tissue-specificity. The OsWAK11 promoter was strongly upregulated in response to metals and wounding.
Similar content being viewed by others
References
Ahn YO, Kim SH, Lee J, Kim H, Lee HS, Kwak SS (2012) Three Brassica rapa metallothionein genes are differentially regulated under various stress conditions. Mol Biol Rep 39(3):2059–2067
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dyebinding. Anal Biochem 72:248–254
Butt A, Mousley C, Morris K, Beynon J, Can C, Holub E, Greenberg JT, Buchanan-Wollaston V (1998) Differential expression of a senescence-enhanced metallothionein gene in Arabidopsis in response to isolates of Peronospora parasitica and Pseudomonas syringae. Plant J 16(2):209–221
Choi D, Kim HM, Yun HK, Park JA, Kim WT, Bok SH (1996) Molecular cloning of a metallothionein-like gene from Nicotiana glutinosa L. and its induction by wounding and tobacco mosaic virus infection. Plant Physiol 112(1):353–359
Coupe SA, Taylor JE, Roberts JA (1995) Characterisation of an mRNA encoding a metallothionein-like protein that accumulates during ethylene-promotedabscission of Sambucus nigra L. leaflets. Planta 197:442–447
Decreux A, Messiaen J (2005) Wall-associated kinase WAK1 interacts with cell wall pectins in a calcium-induced conformation. Plant Cell Physiol 46:268–278
Eriksson M, Moseley JL, Tottey S, Del Campo JA, Quinn J, Kim Y, Merchan S (2004) Genetic dissection of nutritional copper signaling in chlamydomonas distinguishes regulatory and target genes. Genetics 168(2):795–807
Fradin EF, Zhang Z, Ayala JCJ, Castroverde CD, Nazar RN, Robb J, Liu CM, Thomma BP (2009) Genetic dissection of Verticillium wilt resistance mediated by tomato Ve1. Plant Physiol 150(1):320–332
Garcia L, Welchen E, Gonzalez DH (2014) Mitochondria and copper homeostasis in plants. Mitochondrion. doi:10.1016/j.mito.2014.02.011
Gens JS, Fujiki M, Pickard BG (2000) Arabinogalactan protein and wall-associated kinase in a plasmalemmal reticulum with specialized vertices. Protoplasma 212(1–2):115–134
Giuliano G, Hoffman NE, Ko K, Scolnik PA, Cashmore AR (1988) A light-entrained circadian clock controls transcription of several plant genes. EMBO J 7(12):3635–3642
He ZH, Fujiki M, Kohorn BD (1996) A cell wall-associated, receptor-like protein kinase. J Biol Chem 27:19789–19793
He ZH, He D, Kohorn BD (1998) Requirement for the induced expression of a cell wall associated receptor kinase for survival during the pathogen response. Plant J 14(1):55–63
He ZH, Cheeseman I, He D, Kohorn BD (1999) A cluster of five cell wall-associated receptor kinase genes, Wak1-5, are expressed in specific organs of Arabidopsis. Plant Mol Biol 39:1189–1196
Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database: 1999. Nucleic Acids Res 27(1):297–300
Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general-method for transferring genes into plants. Science 227:1229–1231
Hou XW, Tong HY, Selby J, Dewitt J, Peng X, He ZH (2005) Involvement of a cell wall-associated kinase, WAKL4, in Arabidopsis mineral responses. Plant Physiol 139:1704–1716
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6(13):3901–3907
Kanneganti V, Gupta AK (2008) Wall associated kinases from plants—an overview. Physiol Mol Biol Plants 14:109–118
Kanneganti V, Gupta AK (2011) RNAi mediated silencing of a wall associated kinase, OsiWAK1 in Oryza sativa results in impaired root development and sterility due to anther indehiscence. Physiol Mol Biol Plants 17:65–77
Kaur R, Singh K, Singh J (2013) A root-specific wall-associated kinase gene, HvWAK1, regulates root growth and is highly divergent in barley and other cereals. Funct Integr Genomics 13(2):167–177
Kohorn BD (2001) WAKs; cell wall associated kinases. Curr Opin Cell Biol 13(5):529–533
Kohorn BD, Kobayashi M, Johansen S, Friedman HP, Fischer A, Byers N (2006a) Wall-associated kinase 1 (WAK1) is crosslinked in endomembranes, and transport to the cell surface requires correct cell-wall synthesis. J Cell Sci 119:2282–2290
Kohorn BD, Kobayashi M, Johansen S, Riese J, Huang LF, Koch K, Fu S, Dotson A, Byers N (2006b) An Arabidopsis cell wall-associated kinase required for invertase activity and cell growth. Plant J 46(2):307–316
Kohorn BD, Johansen S, Shishido A, Todorova T, Martinez R, Defeo E, Obregon P (2009) Pectin activation of MAP kinase and gene expression is WAK2 dependent. Plant J 60:974–982
Kropat J, Tottey S, Birkenbihl RP, Depege N, Huijser P, Merchant S (2005) A regulator of nutritional copper signaling in Chlamydomonas is an SBP domain protein that recognizes the GTAC core of copper response element. Proc Natl Acad Sci U S A 102:18730–18735
Lally D, Ingmire P, Tong HY, He ZH (2001) Antisense expression of a cell wall-associated protein kinase, WAK4, inhibits cell elongation and alters morphology. Plant Cell 13:1317–1331
Lejeune A, Constant S, Delavault P, Simier P, Thalouarn P, Thoiron S (2006) Involvement of a putative Lycopersicon esculentum wall-associated kinase in the early steps of tomato-Orobanche ramosa interaction. Physiol Mol Plant Pathol 69:3–12
Lescot M, Déhais P, Thijs G, Marchal K, Moreau Y, Van de Peer Y, Rouzé P, Rombauts S (2002) PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res 30(1):325–327
Li H, Zhou SY, Zhao WS, Su SC, Peng YL (2009) A novel wall-associated receptor-likeprotein kinase gene, OsWAK, plays important roles in rice blast disease resistance. Plant Mol Biol 69:337–346
Liu Y, Liu DC, Zhang HY, Gao HB, Guo XL, Fu XD, Zhang A (2006) Isolation and characterisation of six putative wheat cell wall-associated kinases. Funct Plant Biol 33(9):811–821
Lü S, Gu H, Yuan X, Wang X, Wu AM, Qu L, Liu JY (2007) The GUS reporter-aided analysis of the promoter activities of a rice metallothionein gene reveals different regulatory regions responsible for tissue-specificand inducible expression in transgenic Arabidopsis. Transgenic Res 16(2):177–191
Meier S, Ruzvidzo O, Morse M, Donaldson L, Kwezi L, Gehring C (2010) The Arabidopsis wall associated kinase-like 10 gene encoded a functional guanylyl cyclase is coexpressed with pathogen defense related genes. PLoS ONE 5:1–17
Nishiuchi T, Shinshi H, Suzuki K (2004) Rapid and transient activation of transcription of the ERF3 gene by wounding in tobacco leaves. J Biol Chem 279(53):55355–55361
Palmieri MC, Sell S, Huang X, Scherf M, Werner T, Durner J, Lindermayr C (2008) Nitric oxide-responsive genes and promoters in Arabidopsis thaliana: a bioinformatics approach. J Exp Bot 59:177–186
Qi XT, Zhang YX, Chai TY (2007) Characterization of a novel plant promoter specifically induced by heavy metal and identification of the promoter regions conferring heavy metal responsiveness. Plant Physiol 143:50–59
Quinn JM, Merchant S (1995) Two copper-responsiveelements associated with the Chlamydomonas Cyc6 gene function as targets for transcriptional activators. Plant Cell 7:623–638
Quinn JM, Barraco P, Eriksson M, Merchant S (2000)Coordinate copper-and oxygen-responsiveCyc6 and Cpx1 expression in chlamydomonas is mediated by the same element. J Biol Chem 275:6080–6089
Quinn JM, Kropat J, Merchant S (2003) Copper response element and Crr1-dependentNi2+-responsive promoter for induced, reversible gene expression in Chlamydomonas reinhardtii. Eukaryotic Cell 2(5):995–1002
Rushton PJ, Reinstädler A, Lipka V, Lippok B, Somssich IE (2002) Synthetic plant promoters containing defined regulatory elements provide novel insights into pathogen-and wound-induced signaling. Plant Cell 14(4):749–762
Rushton PJ, Somssich IE, Ringler P, Shen QXJ (2010) WRKY transcription factors. Trends Plant Sci 15:247–258
Schenk PM, Kazan K, Wilson I, Anderson JP, Richmond T, Somerville SC, Manners JM (2000) Coordinated plant defense responses in Arabidopsis revealed by microarray analysis. Proc Natl Acad Sci USA 97:11655–11660
Shiu SH, Bleecker AB (2001) Plant receptor-like kinase gene family: diversity, function, and signaling. Sci Signal 113:re22
Shiu SH, Bleecker AB (2003) Expansion of the receptor-like kinase/Pelle gene family and receptor-like proteins in Arabidopsis. Plant Physiol 132(2):530–543
Simpson SD, Nakashima K, Narusaka Y, Seki M, Shinozaki K, Yamaguchi-ShinozakiK (2003) Two different novel cis-acting elements of erd1, a clpA homologous Arabidopsis gene function in induction by dehydration stress and dark-induced senescence. Plant J 33(2):259–270
Sivaguru M, Ezaki B, He ZH, Tong H, Osawa H, Baluska F, Vokmann D, Matsumoto H (2003) Aluminum-induced gene expression and protein localization of a cell wall-associated receptor kinase in Arabidopsis. Plant Physiol 132:2256–2266
Snowden KC, Richards KD, Gardner RC (1995) Aluminum-inducedgenes: induction by toxic metals, low calcium, and wounding and pattern of expression in root tips. Plant Biol 107(2):341–348
Steinwand BJ, Kieber JJ (2010) The role of receptor-likekinases in regulating cell wall function. Plant Physiol 153:467–478
Stuart GW, Searle PF, Palmiter RD (1985) Identification of multiple metal regulatory elements in mouse metallothionein-Ipromoter by assaying synthetic sequences. Nature 317:828–831
Verica JA, He ZH (2002) The cell wall-associatedkinase (WAK)and WAK-likekinase gene family. Plant Physiol 129:455–459
Verica JA, Chae L, Tong HY, Ingmire P, He ZH (2003) Tissue-specificand development regulated expression of a cluster of tandemly arrayed cell wall-associated kinase-likekinase genes in Arabidopsis. Plant Physiol 133:1732–1746
Wagner TA, Kohorn BD (2001) Wall-associatedkinases are expressed throughout plant development and are required for cell expansion. Plant Cell 13:303–318
Wang N, Huang HJ, Ren ST, Li JJ, Sun Y, Sun DY, Zhang SQ (2012) The rice wall-associated receptor-likekinase gene OsDEES1 plays a role in female gametophyte development. Plant Physiol 160:696–707
Wei J, Theil EC (2000) Identification and characterization of the iron regulatory element in the ferritin gene of a plant (soybean). J Biol Chem 275:17488–17493
Yamasaki H, Hayashi M, Fukazawa M, Kobayashi Y, Shikanai Y (2009) SQUAMOSA promoter binding protein-like7is a central regulator for copper homeostasis in Arabidopsis. Plant Cell 21:347–361
Yi SY, Sun AQ, Sun Y, Yang JY, Zhao CM, Liu J (2006) Differential regulation of Lehsp23.8 in tomato plants: analysis of a multiple stress-induciblepromoter. Plant Sci 171(3):398–407
Zhang H, Li L (2013) SQUAMOSA promoter binding protein-like7 regulated microRNA408 is required for vegetative development in Arabidopsis. Plant J 74:98–109
Zhang SB, Chen C, Li L, Meng L, Singh J, Jiang N, Deng XW, He ZH, Lemaus PG (2005) Evolutionary expansion, gene structure, and expression of the rice wall-associated kinase gene family. Plant Physiol 139:1107–1124
Zhang HX, Lian CL, Shen ZG (2009) Proteomic identification of small, copper-responsive proteins in germinating embryos of Oryza sativa. Ann Bot 103:923–930
Zhao H, Butler E, Rodgers J, Spizzo T, Duesterhoeft S, Eide D (1998) Regulation of zinc homeostasis in yeast by binding of the ZAP1 transcriptional activator to zinc-responsive promoter elements. J Biol Chem 273:28713–28720
Acknowledgments
This work was financially supported by the National Natural Science Foundation of China (31201146, J1210056), the Opening Foundation of State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science (Y052010017), the Doctoral Program Foundation of Institutions of Higher Education of China (20120097130004), the Fundamental Research Funds for the Central Universities (KYZ201316), and the Innovative Research Team Development Plan of the Ministry of Education of China (IRT1256).
Author information
Authors and Affiliations
Corresponding authors
Additional information
Responsible Editor: Philip John White.
Rights and permissions
About this article
Cite this article
Hu, W., Lv, Y., Lei, W. et al. Cloning and characterization of the Oryza sativa wall-associated kinase gene OsWAK11 and its transcriptional response to abiotic stresses. Plant Soil 384, 335–346 (2014). https://doi.org/10.1007/s11104-014-2204-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11104-014-2204-8