Abstract
Although strawberry is an economically important fruit crop worldwide, production of strawberry is limited by its susceptibility to a wide range of pathogens and the lack of major commercial cultivars with high levels of resistance to multiple pathogens. The objective of this study is to ectopically express the Arabidopsis thaliana NPR1 gene (AtNPR1) in the diploid strawberry Fragaria vesca L. and to test transgenic plants for disease resistance. AtNPR1 is a key positive regulator of the long-lasting broad-spectrum resistance known as systemic acquired resistance (SAR) and has been shown to confer resistance to a number of pathogens when overexpressed in Arabidopsis or ectopically expressed in several crop species. We show that ectopic expression of AtNPR1 in strawberry increases resistance to anthracnose, powdery mildew, and angular leaf spot, which are caused by different fungal or bacterial pathogens. The increased resistance is related to the relative expression levels of AtNPR1 in the transgenic plants. In contrast to Arabidopsis plants overexpressing AtNPR1, which grow normally and do not constitutively express defense genes, the strawberry transgenic plants are shorter than non-transformed controls, and most of them fail to produce runners and fruits. Consistently, most of the transgenic lines constitutively express the defense gene FvPR5, suggesting that the SAR activation mechanisms in strawberry and Arabidopsis are different. Nevertheless, our results indicate that overexpression of AtNPR1 holds the potential for generation of broad-spectrum disease resistance in strawberry.
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References
Barney DL, Davis BB, Fellman JK (1992) Strawberry production: overview. Alternative agricultural enterprises. http://www.cals.uidaho.edu/edcomm/pdf/CIS/CIS0931.pdf
Brunings AM, Moyer C, Peres N, Folta KM (2010) Implementation of simple sequence repeat markers to genotype Florida strawberry varieties. Euphytica 173:63–75. doi:10.1007/s10681-009-0112-4
Cao H, Bowling SA, Gordon SA, Dong X (1994) Characterization of an Arabidopsis mutant that is nonresponsive to inducers of systemic acquired resistance. Plant Cell 6:1583–1592. doi:10.1105/tpc.6.11.1583
Cao H, Li X, Dong X (1998) Generation of broad-spectrum disease resistance by overexpression of an essential regulatory gene in systemic acquired resistance. Proc Natl Acad Sci USA 95:6531–6536. doi:10.1073/pnas.95.11.6531
Chang S, Puryear J, Cairney J (1993) A simple and efficient method for isolating RNA from pine trees. Plant Mol Biol Rep 11:113–116. doi:10.1007/BF02670468
Chern MS, Fitzgerald HA, Yadav RC, Canlas PE, Dong X, Ronald PC (2001) Evidence for a disease-resistance pathway in rice similar to the NPR1-mediated signaling pathway in Arabidopsis. Plant J 27:101–113. doi:10.1046/j.1365-313x.2001.01070.x
Chern M, Fitzgerald HA, Canlas PE, Navarre DA, Ronald PC (2005) Overexpression of a rice NPR1 homolog leads to constitutive activation of defense response and hypersensitivity to light. Mol Plant Microbe Interact 18:511–520. doi:10.1094/MPMI-18-0511
Durrant WE, Dong X (2004) Systemic acquired resistance. Annu Rev Phytopathol 42:185–209. doi:10.1146/annurev.phyto.42.040803.140421
Epstein AH (1966) Angular leaf spot of strawberry. Plant Dis 50:167
Essghaier B, Fardeau ML, Cayol JL, Hajlaoui MR, Boudabous A, Jijakli H, Sadfi-Zouaoui N (2009) Biological control of grey mould in strawberry fruits by halophilic bacteria. J Appl Microbiol 106:833–846. doi:10.1111/j.1365-2672.2008.04053.x
Fan W, Dong X (2002) In vivo Interaction between NPR1 and transcription factor TGA2 leads to salicylic acid-mediated gene activation in Arabidopsis. Plant Cell 14:1377–1389. doi:10.1105/tpc.001628
Feng W, Guan Z, Whidden A (2012) Strawberry industry overview and outlook. Food and Resource Economics Department—University of Florida. http://www.cals.uidaho.edu/edcomm/pdf/CIS/CIS0931.pdf. Accessed 08 Aug 2014
Fitzgerald HA, Chern MS, Navarre R, Ronald PC (2004) Overexpression of (At)NPR1 in rice leads to a BTH- and environment-inducible lesion-mimic/cell death phenotype. Mol Plant Microbe Interact J 17:140–151
Friedrich L, Lawton K, Dietrich R, Willits M, Cade R, Ryals J (2001) NIM1 overexpression in Arabidopsis potentiates plant disease resistance and results in enhanced effectiveness of fungicides. Mol Plant Microbe Interact J 14:1114–1124. doi:10.1094/MPMI.2001.14.9.1114
Göllner K, Schweizer P, Bai Y, Panstruga R (2008) Natural genetic resources of Arabidopsis thaliana reveal a high prevalence and unexpected phenotypic plasticity of RPW8-mediated powdery mildew resistance. New Phytol 177:725–742. doi:10.1111/j.1469-8137.2007.02339.x
Howard CM, Maas JL, Chandler CL, Albregts EE (1992) Anthracnose of strawberry caused by the Colletotrichum complex in Florida. Plant Dis 76:976–981
Jordan VWL, Hunter T (1972) The effects of glass cloche and coloured polyethylene tunnels on microclimate, growth, yield and disease severity of strawberry plants. J Hortic Sci 47:419–426
Karimi M, Inzé D, Depicker A (2002) GATEWAY vectors for Agrobacterium-mediated plant transformation. Trends Plant Sci 7:193–195
Kennedy BW, King TH (1962) Angular leaf spot of strawberry caused by Xanthomonas fragariae sp. Phytopathology 52:873–875
Le Henanff G, Heitz T, Mestre P, Mutterer J, Walter B, Chong J (2009) Characterization of Vitis vinifera NPR1 homologs involved in the regulation of pathogenesis-related gene expression. BMC Plant Biol 9:54–67. doi:10.1186/1471-2229-9-54
Le Henanff G, Kieffer-Mazet F, Miclot AS, Heitz T, Mestre P, Bertsch C, Chong J (2011) Vitis vinifera VvNPR1.1 is the functional ortholog of AtNPR1 and its overexpression in grapevine triggers constitutive activation of PR genes and enhanced resistance to powdery mildew. Planta 234:405–417. doi:10.1007/s00425-011-1412-1
Lin WC et al (2004) Transgenic tomato plants expressing the Arabidopsis NPR1 gene display enhanced resistance to a spectrum of fungal and bacterial diseases. Transgenic Res 13:567–581. doi:10.1007/s11248-004-2375-9
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−∆∆CT. Methods 25:402–408. doi:10.1006/meth.2001.1262
Maas JL, Gouin-Behe C, Hartung JS, Hokanson SC (2000) Sources of resistance for two differentially pathogenic strains of Xanthomonas fragariae in Fragaria genotypes. HortScience 35:4
Makandar R, Essig JS, Schapaugh MA, Trick HN, Shah J (2006) Genetically engineered resistance to Fusarium head blight in wheat by expression of Arabidopsis NPR1. Mol Plant Microbe Interact J 19:123–129. doi:10.1094/MPMI-19-0123
Malnoy M, Jin Q, Borejsza-Wysocka EE, He SY, Aldwinckle HS (2007) Over-expression of the apple MpNPR1 gene confers increased disease resistance in Malus x domestica. Mol Plant Microbe Interact J 20:1568–1580. doi:10.1094/MPMI-20-12-1568
Meur G, Budatha M, Srinivasan T, Rajesh Kumar KR, Dutta Gupta A, Kirti PB (2008) Constitutive expression of Arabidopsis NPR1 confers enhanced resistance to the early instars of Spodoptera litura in transgenic tobacco. Physiol Plant 133:765–775. doi:10.1111/j.1399-3054.2008.01098.x
Mou Z, Fan W, Dong X (2003) Inducers of plant systemic acquired resistance regulate NPR1 function through redox changes. Cell 113:935–944. doi:10.1016/s0092-8674(03)00429-x
Oosumi T et al (2006) High-efficiency transformation of the diploid strawberry (Fragaria vesca) for functional genomics. Planta 223:1219–1230. doi:10.1007/s00425-005-0170-3
Peres N (2011) 2011 Florida plant disease management guide: strawberry, March 2011. https://edis.ifas.ufl.edu/pdffiles/PG/PG05600.pdf. Accessed 08 Aug 2014
Potlakayala SD, DeLong C, Sharpe A, Fobert PR (2007) Conservation of non-expressor of pathogenesis-related genes1 function between Arabidopsis thaliana and Brassica napus. Physiol Mol Plant Pathol 71:174–183. doi:10.1016/j.pmpp.2008.01.003
Potter D et al (2007) Phylogeny and classification of Rosaceae. Plant Syst Evol 266:5–43. doi:10.1007/s00606-007-0539-9
Roberts PD, Jones JB, Chandler CK, Stall RE, Berger RD (1996) Survival of Xanthomonas fragariae on strawberry in summer nurseries in Florida detected by specific primers and nested polymerase chain reaction. Plant Dis 80:1283–1288
Sehringer B, Zahradnik HP, Deppert WR, Simon M, Noethling C, Schaefer WR (2005) Evaluation of different strategies for real-time RT-PCR expression analysis of corticotropin-releasing hormone and related proteins in human gestational tissues. Anal Bioanal Chem 383:768–775. doi:10.1007/s00216-005-0067-9
Shaner G, Finney RE (1977) The effect of nitrogen fertilization on the expression os slow mildewing resistance in Knox wheat. Phytopathology 67:1051–1056. doi:10.1094/Phyto-67-1051
Sreenivasaprasad S, Talhinhas P (2005) Genotypic and phenotypic diversity in Colletotrichum acutatum, a cosmopolitan pathogen causing anthracnose on a wide range of hosts. Mol Plant Pathol 6:361–378. doi:10.1111/j.1364-3703.2005.00291.x
Subramaniam R, Desveaux D, Spickler C, Michnick SW, Brisson N (2001) Direct visualization of protein interactions in plant cells. Nature 19:769–772. doi:10.1038/90831
Verberne MC, Brouwer N, Delbianco F, Linthorst HJ, Bol JF, Verpoorte R (2002) Method for the extraction of the volatile compound salicylic acid from tobacco leaf material. Phytochem Anal 13:45–50. doi:10.1002/pca.615
Wally O, Jayaraj J, Punja ZK (2009) Broad-spectrum disease resistance to necrotrophic and biotrophic pathogens in transgenic carrots (Daucus carota L.) expressing an Arabidopsis NPR1 gene. Planta 231:131–141. doi:10.1007/s00425-009-1031-2
Yuan Y et al (2007) Functional analysis of rice NPR1-like genes reveals that OsNPR1/NH1 is the rice orthologue conferring disease resistance with enhanced herbivore susceptibility. Plant Biotechnol J 5:313–324. doi:10.1111/j.1467-7652.2007.00243.x
Zhang Y, Fan W, Kinkema M, Li X, Dong X (1999) Interaction of NPR1 with basic leucine zipper protein transcription factors that bind sequences required for salicylic acid induction of the PR-1 gene. PNAS 96:6523–6528. doi:10.1073/pnas.96.11.6523
Zhang X, Francis MI, Dawson WO, Graham JH, Orbović V, Triplett EW, Mou Z (2010) Over-expression of the Arabidopsis NPR1 gene in citrus increases resistance to citrus canker. Eur J Plant Pathol 128:91–100. doi:10.1007/s10658-010-9633-x
Acknowledgments
Funding for this research was provided by Florida Strawberry Research and Education Foundation. The authors are grateful to the doctoral fellowship from CAPES (Brazilian National Council for the Improvement of Higher Education—Grant Procs. BEX 5640/10-5). The authors also thank Dr. Sixue Chen (University of Florida) for access to the HPLC equipment.
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Silva, K.J.P., Brunings, A., Peres, N.A. et al. The Arabidopsis NPR1 gene confers broad-spectrum disease resistance in strawberry. Transgenic Res 24, 693–704 (2015). https://doi.org/10.1007/s11248-015-9869-5
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DOI: https://doi.org/10.1007/s11248-015-9869-5