Regulation of OsPR10a Promoter Activity by Phytohormone and Pathogen Stimulation in Rice
#These authors contributed equally to this work
Received date: 2020-06-15
Accepted date: 2020-10-10
Online published: 2021-09-28
OsPR10a is one of the well known pathogenesis-related genes in rice, and is induced by multiple plant hormones and pathogens. However, the underlying transcriptional regulation mechanisms in response to different signals and their crosstalks are still largely unknown. In order to find new players participated in the activation of OsPR10a, we systematically analyzed the basal expression patterns as well as the expression responses of a 2.5 kb OsPR10a promoter in rice transgenic plants after phytohormone and pathogen stimulations. In agreement with the native gene expression, the OsPR10a promoter can drive glucuronidase (GUS) gene expressing in spots of leaf cells, leaf trichomes, lemmas and paleae, germinating embryos, calli and root tips. The leaf expression of OsPR10a::GUS was dramatically increased upon jasmonic acid (JA) and cytokinin (CK) treatments, or challenges of the pathogen Magnaporthe grisea and Xanthomonas oryzae pv. oryzae. Thus, the OsPR10a promoter reported here can faithfully reflect its native gene expression. The effects of several JA and CK responsive OsWRKY genes on the regulation of OsPR10a promoter were then inspected by luciferase transient expression assay, and the JA inducible OsWRKY10 transcription factor was found as a new positive regulator of OsPR10a. However, the key transcription factors of JA and CK signaling pathways, OsMYC2 and B-type response regulators, were not responsible for the activation of OsPR10a promoter. Our findings provided new insights into the regulation of OsPR10a expression during plant-hormone/pathogen interactions, and the OsPR10a reporter system can be useful to unravel novel regulators from both pathogen and host.
Key words： OsPR10a promoter; β-glucuronidase; phytohormone; pathogen; OsWRKY10; rice
Ersong Zheng, Xuming Wang, Rumeng Xu, Feibo Yu, Chao Zheng, Yong Yang, Yang Chen, Jianping Chen, Chengqi Yan, Jie Zhou . Regulation of OsPR10a Promoter Activity by Phytohormone and Pathogen Stimulation in Rice[J]. Rice Science, 2021 , 28(5) : 442 -456 . DOI: 10.1016/j.rsci.2021.07.005
| [1] | Agrawal G K, Rakwal R, Jwa N S. 2000. Rice (Oryza sativa L.) OsPR1b gene is phytohormonally regulated in close interaction with light signals. Biochem Biophy Res Commun, 278(2): 290-298. |
| [2] | Agarwal P, Agarwal P K. 2014. Pathogenesis related-10 proteins are small, structurally similar but with diverse role in stress signaling. Mol Biol Rep, 41(2): 599-611. |
| [3] | Andrade L B S, Oliveira A S, Ribeiro J K C, Kiyota S, Vasconcelos I M, de Oliveira J T, de Sales M P. 2010. Effects of a novel pathogenesis-related class 10 (PR-10) protein from Crotalaria pallida roots with papain inhibitory activity against root-knot nematode Meloidogyne incognita. J Agric Food Chem, 58(7): 4145-4152. |
| [4] | Brederode F T, Linthorst H J M, Bol J F. 1991. Differential induction of acquired resistance and PR gene expression in tobacco by virus infection, ethephon treatment, UV light and wounding. Plant Mol Biol, 17(6): 1117-1125. |
| [5] | Breiteneder H, Pettenburger K, Bito A, Valenta R, Kraft D, Rumpold H, Scheiner O, Breitenbach M. 1989. The gene coding for the major birch pollen allergen Betv1, is highly homologous to a pea disease resistance response gene. EMBO J, 8(7): 1935?1938. |
| [6] | Breiteneder H, Hoffmann-Sommergruber K, O’Riordain G, Susani M, Ahorn H, Ebner C, Kraft D, Scheiner O. 1995. Molecular characterization of Api g 1, the major allergen of celery (Apium graveolens), and its immunological and structural relationships to a group of 17-kDa tree pollen allergens. Eur J Biochem, 233(2): 484-489. |
| [7] | Brown R L, Kazan K, McGrath K C, Maclean D J, Manners J M. 2003. A role for the GCC-box in jasmonate-mediated activation of the PDF1.2 gene of Arabidopsis. Plant Physiol, 132(2): 1020-1032. |
| [8] | Cai Q, Yuan Z, Chen M J, Yin C S, Luo Z J, Zhao X X, Liang W Q, Hu J P, Zhang D B. 2014. Jasmonic acid regulates spikelet development in rice. Nat Commun, 5: 3476. |
| [9] | Chen S B, Tao L Z, Zeng L R, Vega-Sanchez M E, Umemura K, Wang G L. 2006. A highly efficient transient protoplast system for analyzing defence gene expression and protein-protein interactions in rice. Mol Plant Pathol, 7(5): 417-427. |
| [10] | Choi C, Hwang S H, Fang I R, Kwon S I, Park S R, Ahn I, Kim J B, Hwang D J. 2015. Molecular characterization of Oryza sativa WRKY6, which binds to W-box-like element 1 of the Oryza sativa pathogenesis-related (PR) 10a promoter and confers reduced susceptibility to pathogens. New Phytol, 208(3): 846-859. |
| [11] | Choi J, Huh S U, Kojima M, Sakakibara H, Paek K H, Hwang I. 2010. The cytokinin-activated transcription factor ARR2 promotes plant immunity via TGA3/NPR1-dependent salicylic acid signaling in Arabidopsis. Dev Cell, 19(2): 284-295. |
| [12] | Dhatterwal P, Basu S, Mehrotra S, Mehrotra R. 2019. Genome wide analysis of W-box element in Arabidopsis thaliana reveals TGAC motif with genes down regulated by heat and salinity. Sci Rep, 9(1): 1681. |
| [13] | Dou S J, Guan M L, Li L Y, Liu G Z. 2014. Pathogenesis-related genes in rice. Chin Sci Bull, 59: 245?258. (in Chinese with English abstract) |
| [14] | Durrant W E, Dong X. 2004. Systemic acquired resistance. Annu Rev Phytopathol, 42: 185-209. |
| [15] | Farmer E E, Almeras E, Krishnamurthy V. 2003. Jasmonates and related oxylipins in plant responses to pathogenesis and herbivory. Curr Opin Plant Biol, 6(4): 372-378. |
| [16] | Fernandez-Calvo P, Chini A, Fernandez-Barbero G, Chico J M, Gimenez-Ibanez S, Geerinck J, Eeckhout D, Schweizer F, Godoy M, Franco-Zorrilla J M, Pauwels L, Witters E, Puga M I, Paz-Ares J, Goossens A, Reymond P, de Jaeger G, Solano R. 2011. The Arabidopsis bHLH transcription factors MYC3 and MYC4 are targets of JAZ repressors and act additively with MYC2 in the activation of jasmonate responses. Plant Cell, 23(2): 701-715. |
| [17] | Feys B J, Parker J E. 2000. Interplay of signaling pathways in plant disease resistance. Trends Genet, 16(10): 449-455. |
| [18] | Fitzgerald H A, Canlas P E, Chern M S, Ronald P C. 2005. Alteration of TGA factor activity in rice results in enhanced tolerance to Xanthomonas oryzae pv. oryzae. Plant J, 43(3): 335-347. |
| [19] | Gutierrez-Alcala G, Calo L, Gros F, Caissard J C, Gotor C, Romero L C. 2005. A versatile promoter for the expression of proteins in glandular and non-glandular trichomes from a variety of plants. J Exp Bot, 56: 2487-2494. |
| [20] | Hashimoto M, Kisseleva L, Sawa S, Furukawa T, Komatsu S, Koshiba T. 2004. A novel rice PR10 protein, RSOsPR10, specifically induced in roots by biotic and abiotic stresses, possibly via the jasmonic acid signaling pathway. Plant Cell Physiol, 45(5): 550-559. |
| [21] | Heyl A, Schmulling T. 2003. Cytokinin signal perception and transduction. Curr Opin Plant Biol, 6(5): 480-488. |
| [22] | Huang L F, Lin K H, He S L, Chen J L, Jiang J Z, Chen B H, Hou Y S, Chen R S, Hong C Y, Ho S L. 2016. Multiple patterns of regulation and overexpression of a ribonuclease-like pathogenesis- related protein gene,OsPR10a, conferring disease resistance in rice and Arabidopsis. PLoS One, 11(6): e0156414. |
| [23] | Hwang S H, Lee I A, Yie S W, Hwang D J. 2008. Identification of an OsPR10a promoter region responsive to salicylic acid. Planta, 227(5): 1141-1150. |
| [24] | Hwang S H, Kwon S I, Jang J Y, Fang I L, Lee H, Choi C, Park S, Ahn I, Bae S C, Hwang D J. 2016. OsWRKY51, a rice transcription factor, functions as a positive regulator in defense response against Xanthomonas oryzae pv. oryzae. Plant Cell Rep, 35(9): 1975?1985. |
| [25] | Ito Y, Kurata N. 2006. Identification and characterization of cytokinin-signalling gene families in rice. Gene, 382: 57-65. |
| [26] | Iwai T, Seo S, Mitsuhara I, Ohashi Y. 2007. Probenazole-induced accumulation of salicylic acid confers resistance to Magnaporthe grisea in adult rice plants. Plant Cell Physiol, 48(7): 915-924. |
| [27] | Jameson P E. 2000. Cytokinins and auxins in plant-pathogen interactions: An overview. Plant Growth Regul, 32(2): 369-380. |
| [28] | Jefferson R A, Kavanagh T A, Bevan M W. 1987. GUS fusions: Beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J, 6(13): 3901-3907. |
| [29] | Jia Y L, Valent B, Lee F N. 2003. Determination of host responses to Magnaporthe grisea on detached rice leaves using a spot inoculation method. Plant Dis, 87(2): 129-133. |
| [30] | Jiang C J, Shimono M, Sugano S, Kojima M, Liu X Q, Inoue H, Sakakibara H, Takatsuji H. 2013. Cytokinins act synergistically with salicylic acid to activate defense gene expression in rice. Mol Plant Microbe Interact, 26(3): 287-296. |
| [31] | Jwa N S, Kumar Agrawal G, Rakwal R, Park C H, Prasad Agrawal V. 2001. Molecular cloning and characterization of a novel jasmonate inducible pathogenesis-related class 10 protein gene, JIOsPR10, from rice (Oryza sativa L.) seedling leaves. Biochem Biophys Res Commun, 286(5): 973-983. |
| [32] | Jwa N S, Agrawal G K, Tamogami S, Yonekura M, Han O, Iwahashi H, Rakwal R. 2006. Role of defense/stress-related marker genes, proteins and secondary metabolites in defining rice self-defense mechanisms. Plant Physiol Biochem, 44: 261-273. |
| [33] | Kauffman H E, Reddy A P K, Hsieh S P Y, Merca S D. 1973. An improved technique for evaluating resistance of rice varieties to Xanthomonas oryzae. Plant Dis Rep, 57(6): 537-541. |
| [34] | Kim S G, Kim S T, Wang Y M, Yu S, Choi I S, Kim Y C, Kim W T, Agrawal G K, Rakwal R, Kang K Y. 2011. The RNase activity of rice probenazole-induced protein1 (PBZ1) plays a key role in cell death in plants. Mol Cells, 31(1): 25-31. |
| [35] | Kim S T, Cho K S, Yu S, Kim S G, Hong J C, Han C D, Bae D W, Nam M H, Kang K Y. 2003. Proteomic analysis of differentially expressed proteins induced by rice blast fungus and elicitor in suspension-cultured rice cells. Proteomics, 3(12): 2368-2378. |
| [36] | Kim S T, Kim S G, Hwang D H, Kang S Y, Kim H J, Lee B H, Lee J J, Kang K Y. 2004. Proteomic analysis of pathogen-responsive proteins from rice leaves induced by rice blast fungus, Magnaporthe grisea. Proteomics, 4(11): 3569-3578. |
| [37] | Kim S T, Kim S G, Kang Y H, Wang Y M, Kim J Y, Yi N, Kim J K, Rakwal R, Koh H J, Kang K Y. 2008. Proteomics analysis of rice lesion mimic mutant (spl1) reveals tightly localized probenazole- induced protein (PBZ1) in cells undergoing programmed cell death. J Proteome Res, 7(4): 1750-1760. |
| [38] | Lee M W, Qi M, Yang Y. 2001. A novel jasmonic acid-inducible rice myb gene associates with fungal infection and host cell death. Mol Plant Microbe Interact, 14(4): 527-535. |
| [39] | Liu J J, Ekramoddoullah A K M. 2006. The family 10 of plant pathogenesis-related proteins: Their structure, regulation, and function in response to biotic and abiotic stresses. Physiol Mol Plant Pathol, 68(1): 3-13. |
| [40] | Lorenzo O, Chico J M, Sanchez-Serrano J J, Solano R. 2004. JASMONATE-INSENSITIVE1 encodes a MYC transcription factor essential to discriminate between different jasmonate-regulated defense responses in Arabidopsis. Plant Cell, 16(7): 1938?1950. |
| [41] | Maleck K, Levine A, Eulgem T, Morgan A, Schmid J, Lawton K A, Dangl J L, Dietrich R A. 2000. The transcriptome of Arabidopsis thaliana during systemic acquired resistance. Nat Genet, 26(4): 403-410. |
| [42] | McGee J D, Hamer J E, Hodges T K. 2001. Characterization of a PR-10 pathogenesis-related gene family induced in rice during infection with Magnaporthe grisea. Mol Plant Microbe Interact, 14(7): 877-886. |
| [43] | Midoh N, Iwata M. 1996. Cloning and characterization of a probenazole-inducible gene for an intracellular pathogenesis- related protein in rice. Plant Cell Physiol, 37(1): 9-18. |
| [44] | Mitsuhara I, Iwai T, Seo S, Yanagawa Y, Kawahigasi H, Hirose S, Ohkawa Y, Ohashi Y. 2008. Characteristic expression of twelve rice PR1 family genes in response to pathogen infection, wounding, and defense-related signal compounds (121/180). Mol Genet Genom, 279(4): 415-427. |
| [45] | Moons A, Prinsen E, Bauw G, Van Montagu M. 1997. Antagonistic effects of abscisic acid and jasmonates on salt stress-inducible transcripts in rice roots. Plant Cell, 9(12): 2243-2259. |
| [46] | Park C J, Kim K J, Shin R, Park J M, Shin Y C, Paek K H. 2004. Pathogenesis-related protein 10 isolated from hot pepper functions as a ribonuclease in an antiviral pathway. Plant J, 37(2): 186-198. |
| [47] | Peng Y, Bartley L E, Chen X W, Dardick C, Chern M, Ruan R, Canlas P E, Ronald P C. 2008. OsWRKY62 is a negative regulator of basal and Xa21-mediated defense against Xanthomonas oryzae pv. oryzae in rice. Mol Plant, 1(3): 446-458. |
| [48] | Pfaffl M W. 2001. A new mathematical model for relative quantification in real-time RT-PCR. Nucl Acids Res, 29(9): e45. |
| [49] | Raines T, Blakley I C, Tsai Y C, Worthen J M, Franco-Zorrilla J M, Solano R, Schaller G E, Loraine A E, Kieber J J. 2016. Characterization of the cytokinin-responsive transcriptome in rice. BMC Plant Biol, 16(1): 260. |
| [50] | Rakwal R, Agrawal G K, Yonekura M. 2001a. Light-dependent induction of OsPR10 in rice(Oryza sativa L.) seedlings by the global stress signaling molecule jasmonic acid and protein phosphatase 2A inhibitors. Plant Sci, 161(3): 469-479. |
| [51] | Rakwal R, Agrawal G K, Agrawal V P. 2001b. Jasmonate, salicylate, protein phosphatase 2A inhibitors and kinetin up-regulate OsPR5 expression in cut-responsive rice(Oryza sativa). J Plant Physiol, 158(10): 1357-1362. |
| [52] | Reymond P, Farmer E E. 1998. Jasmonate and salicylate as global signals for defense gene expression. Curr Opin Plant Biol, 1(5): 404-411. |
| [53] | Ryu H S, Han M, Lee S K, Cho J I, Ryoo N, Heu S, Lee Y H, Bhoo S H, Wang G L, Hahn T R, Jeon J S. 2006. A comprehensive expression analysis of the WRKY gene superfamily in rice plants during defense response. Plant Cell Rep, 25(8): 836-847. |
| [54] | Sakai H, Aoyama T, Oka A. 2000. Arabidopsis ARR1 and ARR2 response regulators operate as transcriptional activators. Plant J, 24(6): 703-711. |
| [55] | Schindler U, Beckmann H, Cashmore A R. 1992. TGA1 and G-box binding factors: Two distinct classes of Arabidopsis leucine zipper proteins compete for the G-box-like element TGACGTGG. Plant Cell, 4(10): 1309-1319. |
| [56] | Schweizer F, Fernandez-Calvo P, Zander M, Diez-Diaz M, Fonseca S, Glauser G, Lewsey M G, Ecker J R, Solano R, Reymond P. 2013. Arabidopsis basic helix-loop-helix transcription factors MYC2, MYC3, and MYC4 regulate glucosinolate biosynthesis, insect performance, and feeding behavior. Plant Cell, 25(8): 3117-3132. |
| [57] | Sels J, Mathys J, De Coninck B M A, Cammue B P A, De Bolle M F C. 2008. Plant pathogenesis-related (PR) proteins: A focus on PR peptides. Plant Physiol Biochem, 46(11): 941-950. |
| [58] | Serna L, Martin C. 2006. Trichomes: Different regulatory networks lead to convergent structures. Trends Plant Sci, 11(6): 274-280. |
| [59] | Shimono M, Sugano S, Nakayama A, Jiang C J, Ono K, Toki S, Takatsuji H. 2007. Rice WRKY45 plays a crucial role in benzothiadiazole-inducible blast resistance. Plant Cell, 19(6): 2064?2076. |
| [60] | Takeuchi K, Gyohda A, Tominaga M, Kawakatsu M, Hatakeyama A, Ishii N, Shimaya K, Nishimura T, Riemann M, Nick P, Hashimoto M, Komano T, Endo A, Okamoto T, Jikumaru Y, Kamiya Y, Terakawa T, Koshiba T. 2011. RSOsPR10 expression in response to environmental stresses is regulated antagonistically by jasmonate/ethylene and salicylic acid signaling pathways in rice roots. Plant Cell Physiol, 52(9): 1686-1696. |
| [61] | Talbot N J, Ebbole D J, Hamer J E. 1993. Identification and characterization of MPG1, a gene involved in pathogenicity from the rice blast fungus Magnaporthe grisea. Plant Cell, 5(11): 1575-1590. |
| [62] | Thaler J S, Humphrey P T, Whiteman N K. 2012. Evolution of jasmonate and salicylate signal crosstalk. Trends Plant Sci, 17(5): 260-270. |
| [63] | Tsai Y C, Weir N R, Hill K, Zhang W J, Kim H J, Shiu S H, Schaller G E, Kieber J J. 2012. Characterization of genes involved in cytokinin signaling and metabolism from rice. Plant Physiol, 158(4): 1666-1684. |
| [64] | van Loon L C. 1985. Pathogenesis-related proteins. Plant Mol Biol, 4: 111-116. |
| [65] | van Loon L C, Pierpoint W S, Boller T, Conejero V. 1994. Recommendations for naming plant pathogenesis-related proteins. Plant Mol Biol Rep, 12(3): 245-264. |
| [66] | van Loon L C, van Strien E A. 1999. The families of pathogenesis- related proteins, their activities, and comparative analysis of PR-1 type proteins. Physiol Mol Plant Pathol, 55(2): 85-97. |
| [67] | van Loon L C, Rep M, Pieterse C M. 2006. Significance of inducible defense-related proteins in infected plants. Annu Rev Phytopath, 44: 135-162. |
| [68] | Wu Q, Hou M M, Li L Y, Liu L J, Hou Y X, Liu G Z. 2011. Induction of pathogenesis-related proteins in rice bacterial blight resistant gene Xa21-mediated interactions with Xanthomnas oryzae pv. oryzae. J Plant Pathol, 93(2): 455-459. |
| [69] | Xie Y R, Chen Z Y, Brown R L, Bhatnagar D. 2010. Expression and functional characterization of two pathogenesis-related protein 10 genes from Zea mays. J Plant Physiol, 167(2): 121-130. |
| [70] | Xiong M, Meng S, Qiu J H, Shi H B, Shen X L, Kou Y J. 2020. Putative phosphatase UvPsr1 is required for mycelial growth, conidiation, stress response and pathogenicity in Ustilaginonidea virens. Rice Sci, 20(6): 529-536. |
| [71] | Xu Z M, Li L H, Gao X Q, Yuan Z J, Li Xin, Tian X D, Wang L L, Qu S H. 2020. Comparative transcriptome analysis of transgenic rice line carrying the rice blast resistance gene Pi9. Chin J Rice Sci, 34(3): 245?255. (in Chinese with English abstract) |
| [72] | Yamamoto T, Yoshida Y, Nakajima K, Tominaga M, Gyohda A, Suzuki H, Okamoto T, Nishimura T, Yokotani N, Minami E, Nishizawa Y, Miyamoto K, Yamane H, Okada K, Koshiba T. 2018. Expression of RSOsPR10 in rice roots is antagonistically regulated by jasmonate/ethylene and salicylic acid via the activator OsERF87 and the repressor OsWRKY76, respectively. Plant Direct, 2(3): e00049. |
| [73] | Yang B, Bogdanove A. 2013. Inoculation and virulence assay for bacterial blight and bacterial leaf streak of rice. In: Yang Y. Rice Protocols. Methods in Molecular Biology (Methods and Protocls). Vol 956. Totowa: Humana Press: 249?255. |
| [74] | Yoshida S, Forno D A, Cock J H, Gomez K A. 1976. Laboratory Manual for Physiological Studies of Rice. Manila, the Philippines: International Rice Research Institute. |
| [75] | Yu F B, Zhou J, Wang X M, Yang Y, Yu C L, Cheng Y, Yan C Q, Chen J P. 2014. Expression characteristics of rice OsPR1b gene promoter. Lett Biotechnol, 25(4): 24?30. (in Chinese with English abstract) |
| [76] | Zeevaart J A D, Creelman R A. 1988. Metabolism and physiology of abscisic acid. Ann Rev Plant Physiol Plant Mol Biol, 39(1): 439-473. |
| [77] | Zhang J, Peng Y L, Guo Z J. 2008. Constitutive expression of pathogen-inducible OsWRKY31 enhances disease resistance and affects root growth and auxin response in transgenic rice plants. Cell Res, 18(4): 508-521. |
| [78] | Zhang Y, Su J B, Duan S, Ao Y, Dai J R, Liu J, Wang P, Li Y G, Liu B, Feng D R, Wang J F, Wang H B. 2011. A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes. Plant Methods, 7(1): 30. |
| [79] | Zhou J, Yang Y, Wang X M, Yu F B, Yu C L, Chen J, Cheng Y, Yan C Q, Chen J P. 2013. Enhanced transgene expression in rice following selection controlled by weak promoters. BMC Biotechnol, 13: 29. |
| [80] | Zhou J, Yu F B, Wang X M, Yang Y, Yu C L, Liu H J, Cheng Y, Yan C Q, Chen J P. 2014. Specific expression of DR5 promoter in rice roots using a tCUP derived promoter-reporter system. PLoS One, 9(1): e87008. |
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