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Broad-spectrum disease resistance to necrotrophic and biotrophic pathogens in transgenic carrots (Daucus carota L.) expressing an Arabidopsis NPR1 gene

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Abstract

The development of transgenic plants highly resistant to a range of pathogens using traditional signal gene expression strategies has been largely ineffective. Modification of systemic acquired resistance (SAR) through the overexpression of a controlling gene such as NPR1 (non-expressor of PR genes) offers an attractive alternative for augmenting the plants innate defense system. The Arabidopsis (At) NPR1 gene was successfully introduced into ‘Nantes Coreless’ carrot under control of a CaMV 35S promoter and two independent transgenic lines (NPR1-I and NPR1-XI) were identified by Southern and Northern blot hybridization. Both lines were phenotypically normal compared with non-transformed carrots. Northern analysis did not indicate constitutive or spontaneous induction in carrot cultures of SAR-related genes (DcPR-1, 2, 4, 5 or DcPAL). The duration and intensity of expression of DcPR-1, 2 and 5 genes were greatly increased compared with controls when the lines were treated with purified cell wall fragments of Sclerotinia sclerotiorum as well as with 2,6-dichloroisonicotinic acid. The two lines were challenged with the necrotrophic pathogens Botrytis cinerea, Alternaria radicina and S. sclerotiorum on the foliage and A. radicina on the taproots. Both lines exhibited 35–50% reduction in disease symptoms on the foliage and roots when compared with non-transgenic controls. Leaves challenged with the biotrophic pathogen Erysiphe heraclei or the bacterial pathogen Xanthomonas hortorum exhibited 90 and 80% reduction in disease development on the transgenic lines, respectively. The overexpression of the SAR controlling master switch in carrot tissues offers the ability to control a wide range of different pathogens, for which there is currently little genetic resistance available.

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Abbreviations

SAR:

Systemic acquired resistance

NPR1:

Non-expressor of PR genes

SA:

Salicylic acid

PR:

Pathogenesis related

ISR:

Induced systemic resistance

JA:

Jasmonic acid

INA:

2, 6-Dichloroisonicotinic acid

References

  • Baranski R, Klocke E, Nothnagel T (2008) Chitinase CHIT36 from Trichoderma harzianum enhances resistance of transgenic carrot to fungal pathogens. J Phytopathol 156:513–521

    Article  CAS  Google Scholar 

  • Cao H, Bowling SA, Gordon AS, Dong XN (1994) Characterization of an Arabidopsis mutant that is nonresponsive to inducers of systemic acquired-resistance. Plant Cell 6:1583–1592

    Article  CAS  PubMed  Google Scholar 

  • Cao H, Glazebrook J, Clarke JD, Volko S, Dong XN (1997) The Arabidopsis NPR1 gene that controls systemic acquired resistance encodes a novel protein containing ankyrin repeats. Cell 88:57–63

    Article  CAS  PubMed  Google Scholar 

  • Cao H, Li X, Dong XN (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

    Article  CAS  PubMed  Google Scholar 

  • Chen WP, Punja ZK (2002) Transgenic herbicide- and disease-tolerant carrot (Daucus carota L.) plants obtained through Agrobacterium-mediated transformation. Plant Cell Rep 20:929–935

    Article  CAS  Google Scholar 

  • Chern MS, Fitzgerald HA, Yadav RC, Canlas PE, Dong XN, 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

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Clarke JD, Liu YD, Klessig DF, Dong XN (1998) Uncoupling PR gene expression from NPR1 and bacterial resistance: characterization of the dominant Arabidopsis cpr6-1 mutant. Plant Cell 10:557–569

    Article  CAS  PubMed  Google Scholar 

  • Davis RM, Raid RN (2002) Compendium of umbelliferous crop diseases. American Phytopathological Society Press, St. Paul

    Google Scholar 

  • Delaney TP, Friedrich L, Ryals JA (1995) Arabidopsis signal-transduction mutant defective in chemically and biologically induced disease resistance. Proc Natl Acad Sci USA 92:6602–6606

    Article  CAS  PubMed  Google Scholar 

  • Endah R, Beyene G, Kiggundu A, van den Berg N, Schluter U, Kunert K, Chikwamba R (2008) Elicitor and Fusarium-induced expression of NPR1-like genes in banana. Plant Physiol Biochem 46:1007–1014

    Article  CAS  PubMed  Google Scholar 

  • Fan WH, Dong XN (2002) In vivo interaction between NPR1 and transcription factor TGA2 leads to salicylic acid-mediated gene activation in Arabidopsis. Plant Cell 14:1377–1389

    Article  CAS  PubMed  Google Scholar 

  • Farrar JJ, Pryor BA, Davis RM (2004) Alternaria diseases of carrot. Plant Dis 88:776–784

    Article  Google Scholar 

  • Ferrari S, Plotnikova JM, De Lorenzo G, Ausubel FM (2003) Arabidopsis local resistance to Botrytis cinerea involves salicylic acid and camalexin and requires EDS4 and PAD2, but not SID2, EDS5 or PAD4. Plant J 35:193–205

    Article  CAS  PubMed  Google Scholar 

  • Fitzgerald HA, Chern MS, Navarre R, Ronald PC (2004) Overexpression of (At)NPR1 in rice leads to a BTH- and environment-induced lesion-mimic/cell death phenotype. Mol Plant Microbe Interact 17:140–151

    Article  CAS  PubMed  Google Scholar 

  • 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 14:1114–1124

    Article  CAS  PubMed  Google Scholar 

  • Glazebrook J (2005) Contrasting mechanisms of defense against biotrophic and necrotrophic pathogens. Annu Rev Phytopathol 43:205–227

    Article  CAS  PubMed  Google Scholar 

  • Govrin EM, Levine A (2002) Infection of Arabidopsis with a necrotrophic pathogen, Botrytis cinerea, elicits various defense responses but does not induce systemic acquired resistance (SAR). Plant Mol Biol 48:267–276

    Article  CAS  PubMed  Google Scholar 

  • Guo XM, Stotz HU (2007) Defense against Sclerotinia sclerotiorum in Arabidopsis is dependent on jasmonic acid, salicylic acid, and ethylene signaling. Mol Plant Microbe Interact 20:1384–1395

    Article  CAS  PubMed  Google Scholar 

  • Heil M, Hilpert A, Kaiser W, Linsenmair KE (2000) Reduced growth and seed set following chemical induction of pathogen defence: does systemic acquired resistance (SAR) incur allocation costs? J Ecol 88:645–654

    Article  CAS  Google Scholar 

  • Jayaraj J, Punja ZK (2007) Combined expression of chitinase and lipid transfer protein genes in transgenic carrot plants enhances resistance to foliar fungal pathogens. Plant Cell Rep 26:1539–1546

    Article  CAS  PubMed  Google Scholar 

  • Jayaraj J, Wan A, Rahman M, Punja ZK (2008) Seaweed extract reduces foliar fungal diseases on carrot. Crop Protect 27:1360–1366

    Article  Google Scholar 

  • Jayaraj J, Rahman M, Wan A, Punja ZK (2009) Enhanced resistance to foliar fungal pathogens in carrot by application of elicitors. Ann Appl Biol 155:71–80

    Article  CAS  Google Scholar 

  • Kauffman HE, Reddy APK, Hsieh SPY, Merca SD (1973) Improved technique for evaluating resistance of rice varieties to Xanthomonas oryzae. Plant Dis Rep 57:537–541

    Google Scholar 

  • Kora C, McDonald MR, Boland GJ (2003) Sclerotinia rot of carrot: an example of phenological adaptation and bicyclic development by Sclerotinia sclerotiorum. Plant Dis 87:456–470

    Article  Google Scholar 

  • Lin WC, Lu CF, Wu JW, Cheng ML, Lin YM, Yang NS, Black L, Green SK, Wang JF, Cheng CP (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

    Article  CAS  PubMed  Google Scholar 

  • 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 19:123–129

    Article  CAS  PubMed  Google Scholar 

  • Malnoy M, Jin Q, Borejsza-Wysocka EE, He SY, Aldwinckle HS (2007) Overexpression of the apple MpNPR1 gene confers increased disease resistance in Malus × domestica. Mol Plant Microbe Interact 20:1568–1580

    Article  CAS  PubMed  Google Scholar 

  • Melchers LS, Stuiver MH (2000) Novel genes for disease-resistance breeding. Curr Opin Plant Biol 3:147–152

    Article  CAS  PubMed  Google Scholar 

  • Meur G, Budatha M, Gupta AD, Prakash S, Kirti PB (2006) Differential induction of NPR1 during defense responses in Brassica juncea. Physiol Mol Plant Pathol 68:128–137

    Article  CAS  Google Scholar 

  • Meur G, Budatha M, Srinivasan T, Kumar KRR, Gupta AD, 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

    Article  CAS  PubMed  Google Scholar 

  • Mou Z, Fan WH, Dong XN (2003) Inducers of plant systemic acquired resistance regulate NPR1 function through redox changes. Cell 113:935–944

    Article  CAS  PubMed  Google Scholar 

  • Oliver RP, Ipcho SVS (2004) Arabidopsis pathology breathes new life into the necrotrophs vs. biotrophs classification of fungal pathogens. Mol Plant Pathol 5:347–352

    Article  CAS  Google Scholar 

  • Pieterse CM, Van Loon L (2004) NPR1: the spider in the web of induced resistance signaling pathways. Curr Opin Plant Biol 7:456–464

    Article  CAS  PubMed  Google Scholar 

  • Pieterse CMJ, Van Wees SCM, Van Pelt JA, Knoester M, Laan R, Gerrits N, Weisbeek PJ, Van Loon LC (1998) A novel signaling pathway controlling induced systemic resistance in Arabidopsis. Plant Cell 10:1571–1580

    Article  CAS  PubMed  Google Scholar 

  • Punja ZK (2005) Transgenic carrots expressing a thaumatin-like protein display enhanced resistance to several fungal pathogens. Can J Plant Pathol 27:291–296

    CAS  Google Scholar 

  • Punja ZK, Raharjo SHT (1996) Response of transgenic cucumber and carrot plants expressing different chitinase enzymes to inoculation with fungal pathogens. Plant Dis 80:999–1005

    CAS  Google Scholar 

  • Punja ZK, Jayaraj J, Wally O (2007) Carrot. In: Pua EC, Davies DR (eds) Transgenic crops. Springer, Berlin, pp 277–294

    Chapter  Google Scholar 

  • Quilis J, Penas G, Messeguer J, Brugidou C, Segundo BS (2008) The Arabidopsis AtNPR1 inversely modulates defense responses against fungal, bacterial, or viral pathogens while conferring hypersensitivity to abiotic stresses in transgenic rice. Mol Plant Microbe Interact 21:1215–1231

    Article  CAS  PubMed  Google Scholar 

  • Shah J (2003) The salicylic acid loop in plant defense. Curr Opin Plant Biol 6:365–371

    Article  CAS  PubMed  Google Scholar 

  • Shah J, Tsui F, Klessig DF (1997) Characterization of a salicylic acid-insensitive mutant (sai1) of Arabidopsis thaliana, identified in a selective screen utilizing the SA-inducible expression of the tms2 gene. Mol Plant Microbe Interact 10:69–78

    Article  CAS  PubMed  Google Scholar 

  • Silverman P, Seskar M, Kanter D, Schweizer P, Metraux JP, Raskin I (1995) Salicylic-acid in rice: biosynthesis, conjugation, and possible role. Plant Physiol 108:633–639

    CAS  PubMed  Google Scholar 

  • Subramaniam R, Desveaux D, Spickler C, Michnick SW, Brisson N (2001) Direct visualization of protein interactions in plant cells. Nat Biotechnol 19:769–772

    Article  CAS  PubMed  Google Scholar 

  • Takaichi M, Oeda K (2000) Transgenic carrots with enhanced resistance against two major pathogens, Erysiphe heraclei and Alternaria dauci. Plant Sci 153:135–144

    Article  CAS  PubMed  Google Scholar 

  • Thomma BP, Eggermont K, Penninckx IA, Mauch-Mani B, Vogelsang R, Cammue BPA, Broekaert WF (1998) Separate jasmonate-dependent and salicylate-dependent defense-response pathways in Arabidopsis are essential for resistance to distinct microbial pathogens. Proc Natl Acad Sci USA 95:15107–15111

    Article  CAS  PubMed  Google Scholar 

  • Tweddell RJ, Jabaji-Hare SH, Charest PM (1994) Production of chitinases and beta-1, 3-glucanases by Stachybotrys elegans, a mycoparasite of Rhizoctonia solani. Appl Environ Microbiol 60:489–495

    CAS  PubMed  Google Scholar 

  • Wally O, Jayaraj J, Punja ZK (2006) Carrot (Daucus carota. L). In: Wang K (ed) Agrobacterium protocols Volume 2. Humana Press Totowa, NJ.:,pp 3–12

  • Wally O, Jayaraj J, Punja ZK (2008) Comparative expression of beta-glucuronidase with five different promoters in transgenic carrot (Daucus carota L.) root and leaf tissues. Plant Cell Rep 27:279–287

    Article  CAS  PubMed  Google Scholar 

  • Wally O, Jayaraj J, Punja ZK (2009) Comparative resistance to foliar fungal pathogens in transgenic carrot plants expressing genes encoding for chitinase, beta-1, 3-glucanase and peroxidase. Eur J Plant Pathol 123:331–342

    Article  CAS  Google Scholar 

  • Yuan YX, Zhong SH, Li Q, Zhu ZR, Lou YG, Wang LY, Wang JJ, Wang MY, Li QL, Yang DL, He ZH (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

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

Funding for this research was provided by NSERC (Natural Sciences and Engineering Research Council of Canada), Discovery Grants program. We would like to thank Jasmine Norouzi, Lisa Leippi and Jutta Buchhop for assistance with tissue culture and nucleic acid analysis.

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Authors and Affiliations

  1. Department of Biological Sciences, Simon Fraser University, 8888 University Drive, Burnaby, BC, V5A 1S6, Canada

    Owen Wally, Jayaraman Jayaraj & Zamir K. Punja

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  1. Owen Wally
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  2. Jayaraman Jayaraj
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  3. Zamir K. Punja
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Correspondence to Zamir K. Punja.

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Wally, O., Jayaraj, J. & Punja, Z.K. Broad-spectrum disease resistance to necrotrophic and biotrophic pathogens in transgenic carrots (Daucus carota L.) expressing an Arabidopsis NPR1 gene. Planta 231, 131–141 (2009). https://doi.org/10.1007/s00425-009-1031-2

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