BcDR1, a putative gene, regulates the development and pathogenicity of Botrytis cinerea

Research Article
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Abstract

Botrytis cinerea is one of the important phytopathogenic fungi. Cloning of the genes related to their development and pathogenicity is fundamental to the pathogen control. A mutant (BCt160), which produces abnormal conidia and no sclerotia, was identified from Botrytis cinerea mutant library generated by Agrobacterium tumefaciens mediated transformation (ATMT). Southern blotting analysis showed that one T-DNA insertion occurred in the genome of the mutant. TAIL-PCR (thermal asymmetric interlaced PCR) and bioinformatic analysis indicated that the exogenous T-DNA insertion occurred in the second exon of a putative gene BC1G_12388.1, named as BcDR1 (B. cinerea development-related gene 1). The function analysis of BcDR1 gene showed that the BcDR1 was related to development, morphological differentiation, and pathogenicity of B. cinerea, suggesting that BcDR1 gene was required for the development and pathogenicity of B. cinerea.

Keywords

Botrytis cinerea T-DNA mutagenesis BcDR1 functional analysis 
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References

  1. Bahn Y S, Xue C, Idnurm A, Rutherford J C, Heitman J, Cardenas M E (2007). Sensing the environment: lessons from fungi. Nat Rev Microbiol, 5(1): 57–69PubMedCrossRefGoogle Scholar
  2. Balhadère P V, Foster A J, Talbot N J (1999). Identification of pathogenicity mutants of the rice blast fungus Magnaporthe grisea by insertional mutagenesis. Mol Plant Microbe Interact, 12(2): 129–142CrossRefGoogle Scholar
  3. Brito N, Espino J J, Gonzalez C (2006). The endo-beta-1,4-xylanase xyn11A is required for virulence in Botrytis cinerea. Mol Plant Microbe Interact, 19(1): 25–32PubMedCrossRefGoogle Scholar
  4. Choquer M, Fournier E, Kunz C, Levis C, Pradier J M, Simon A, Viaud M (2007). Botrytis cinerea virulence factors: new insights into a necrotrophic and polyphageous pathogen. FEMS Microbiol Lett, 277(1): 1–10PubMedCrossRefGoogle Scholar
  5. Cui Z, Ding Z, Yang X, Wang K, Zhu T (2009). Gene disruption and characterization of a class V chitin synthase in Botrytis cinerea. Can J Microbiol, 55(11): 1267–1274PubMedCrossRefGoogle Scholar
  6. de Groot M J, Bundock P, Hooykaas P J, Beijersbergen A G (1998). Agrobacterium tumefaciens-mediated transformation of filamentous fungi. Nat Biotechnol, 16(9): 839–842PubMedCrossRefGoogle Scholar
  7. Drenth A, Goodwin S B, Fry W E, Davidse L C (1993). Genotypic diversity of Phytophthora infestans in the Netherlands revealed by DNA polymorphisms. Phytopathology, 83(10): 1087–1092CrossRefGoogle Scholar
  8. Elad Y, Williamson B, Tudzynski P, Delen N (2004) Botrytis spp. And Diseases They Cause in Agricultural Systems—An Introduction. In: Elad Y, Williamson B, Tudzynski P, Delen N, eds. Botrytis: Biology, Pathology and Control. the Netherlands: Kluwer Academic PublishersGoogle Scholar
  9. Fillinger S, Chaveroche M K, Shimizu K, Keller N, D’Enfert C (2002). cAMP and ras signalling independently control spore germination in the filamentous fungus Aspergillus nidulans. Mol Microbiol, 44(4): 1001–1016PubMedCrossRefGoogle Scholar
  10. Jurick W N II, Rollins J A (2007). Deletion of the adenylate cyclase (sac1) gene affects multiple developmental pathways and pathogenicity in Sclerotinia sclerotiorum. Fungal Genet Biol, 44(6): 521–530PubMedCrossRefGoogle Scholar
  11. Kars I, McCalman M, Wagemakers L, van Kan J A (2005). Functional analysis of Botrytis cinerea pectin methylesterase genes by PCR-based targeted mutagenesis: Bcpme1 and Bcpme2 are dispensable for virulence of strain B05.10. Mol Plant Pathol, 6(6): 641–652PubMedCrossRefGoogle Scholar
  12. Lazo G R, Stein P A, Ludwig R A (1991). A DNA transformation-competent Arabidopsis genomic library in Agrobacterium. Nat Biotechnol, 9(10): 963–967CrossRefGoogle Scholar
  13. Liebmann B, Gattung S, Jahn B, Brakhage A A (2003). cAMP signaling in Aspergillus fumigatus is involved in the regulation of the virulence gene pksP and in defense against killing by macrophages. Mol Genet Genomics, 269(3): 420–435PubMedCrossRefGoogle Scholar
  14. Michielse C B, Hooykaas P J J, Hondel C A M J J, Ram A F J (2005). Agrobacterium-mediated transformation as a tool for functional genomics in fungi. Curr Genet, 48(1): 1–17PubMedCrossRefGoogle Scholar
  15. Mullins E D, Chen X, Romaine P, Raina R, Geiser D M, Kang S (2001). Agrobacterium-mediated transformation of Fusarium oxysporum: an efficient tool for insertional mutagenesis and gene transfer. Phytopathology, 91(2): 173–180PubMedCrossRefGoogle Scholar
  16. Nierman WC, Pain A, Anderson M J, Wortman J R, Kim H S, Arroyo J, Berriman M, Abe K, Archer D B, Bermejo C, Bennett J, Bowyer P, Chen D, Collins M, Coulsen R, Davies R, Dyer P S, Farman M, Fedorova N, Fedorova N, Feldblyum T V, Fischer R, Fosker N, Fraser A, García J L, García M J, Goble A, Goldman G H, Gomi K, Griffith-Jones S, Gwilliam R, Haas B, Haas H, Harris D, Horiuchi H, Huang J, Humphray S, Jiménez J, Keller N, Khouri H, Kitamoto K, Kobayashi T, Konzack S, Kulkarni R, Kumagai T, Lafton A, Latgé J P, Li W, Lord A, Lu C, Majoros W H, May G S, Miller B L, Mohamoud Y, Molina M, Monod M, Mouyna I, Mulligan S, Murphy L, O’Neil S, Paulsen I, Peñalva M A, Pertea M, Price C, Pritchard B L, Quail M A, Rabbinowitsch E, Rawlins N, Rajandream M A, Reichard U, Renauld H, Robson G D, de Córdoba S R, Rodríguez-Peña J M, Ronning C M, Rutter S, Salzberg S L, Sanchez M, Sánchez-Ferrero J C, Saunders D, Seeger K, Squares R, Squares S, Takeuchi M, Tekaia F, Turner G, de Aldana C R V, Weidman J, White O, Woodward J, Yu J H, Fraser C, Galagan J E, Asai K, Machida M, Hall N, Barrell B, Denning D W (2005). Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus. Nature, 438(7071): 1151–1156PubMedCrossRefGoogle Scholar
  17. Piers K L, Heath J D, Liang X, Stephens K M, Nester E W (1996). Agrobacterium tumefaciens-mediated transformation of yeast. Proc Natl Acad Sci USA, 93(4): 1613–1618PubMedCrossRefGoogle Scholar
  18. Rivera M C, Lopez M V, Lopez S E (2009). Mycobiota from Cyclamen persicum and its interaction with Botrytis cinerea. Mycologia, 101(2): 173–181PubMedCrossRefGoogle Scholar
  19. Rolland S, Jobic C, Fevre M, Bruel C (2003). Agrobacterium-mediated transformation of Botrytis cinerea, simple purification of monokaryotic transformants and rapid conidia-based identification of the transfer-DNA host genomic DNA flanking sequences. Curr Genet, 44(3): 164–171PubMedCrossRefGoogle Scholar
  20. Rui O, Hahn M (2007). The Slt2-type MAP kinase Bmp3 of Botrytis cinerea is required for normal saprotrophic growth, conidiation, plant surface sensing and host tissue colonization. Mol Plant Pathol, 8(2): 173–184PubMedCrossRefGoogle Scholar
  21. Segmuller N, Ellendorf U, Tudzynski B, Tudzynski P (2007). BcSAK1, a stress-activated mitogen-activated protein kinase, is involved in vegetative differentiation and pathogenicity in Botrytis cinerea. Eukaryot Cell, 6(2): 211–221PubMedCrossRefGoogle Scholar
  22. Takano Y, Komeda K, Kojima K, Okuno T (2001). Proper regulation of cyclic AMP-dependent protein kinase is required for growth, conidiation, and appressorium function in the anthracnose fungus Colletotrichum lagenarium. Mol Plant Microbe Interact, 14(10): 1149–1157PubMedCrossRefGoogle Scholar
  23. Tellier F, Fritz R, Kerhoas L, Ducrot P H, Einhorn J, Carlin-Sinclair A, Leroux P (2008). Characterization of metabolites of fungicidal cymoxanil in a sensitive strain of Botrytis cinerea. J Agric Food Chem, 56(17): 8050–8057PubMedCrossRefGoogle Scholar
  24. Thevelein J M, Gelade R, Holsbeeks I, Lagatie O, Popova Y, Rolland F, Stolz F, van de Velde S, van Dijck P, Vandormael P, van Nuland A, van Roey K, van Zeebroeck G, Yan B (2005). Nutrient sensing systems for rapid activation of the protein kinase A pathway in yeast. Biochem Soc Trans, 33(1): 253–256PubMedCrossRefGoogle Scholar
  25. Tudzynski P, Siewers V (2004). Approaches to Molecular Genetics and Genomics of Botrytis. In: Elad Y, Williamson B, Tudzynski P, Delen N, eds. Botrytis: Biology, Pathology and Control. The Netherlands: Kluwer Academic Press, 53–66Google Scholar
  26. van der Vlugt-Bergmans C J, Wagemakers C A, van Kan J A (1997). Cloning and expression of the cutinase A gene of Botrytis cinerea. Mol Plant Microbe Interact, 10(1): 21–29PubMedCrossRefGoogle Scholar
  27. van Kan J A (2006). Licensed to kill: the lifestyle of a necrotrophic plant pathogen. Trends Plant Sci, 11(5): 247–253PubMedCrossRefGoogle Scholar
  28. Vienken K, Fischer R (2006). The Zn(II)2Cys6 putative transcription factor NosA controls fruiting body formation in Aspergillus nidulans. Mol Microbiol, 61(2): 544–554PubMedCrossRefGoogle Scholar
  29. Williamson B, Tudzynski B, Tudzynski P, van Kan J A (2007). Botrytis cinerea: the cause of grey mould disease. Mol Plant Pathol, 8(5): 561–580PubMedCrossRefGoogle Scholar
  30. Yamauchi J, Takayanagi N, Komeda K, Takano Y, Okuno T (2004). cAMP-pKA signaling regulates multiple steps of fungal infection cooperatively with Cmk1 MAP kinase in Colletotrichum lagenarium. Mol Plant Microbe Interact, 17(12): 1355–1365PubMedCrossRefGoogle Scholar
  31. Zhao W, Panepinto J C, Fortwendel J R, Fox L, Oliver B G, Askew D S, Rhodes J C (2006). Deletion of the regulatory subunit of protein kinase A in Aspergillus fumigatus alters morphology, sensitivity to oxidative damage, and virulence. Infect Immun, 74(8): 4865–4874PubMedCrossRefGoogle Scholar
  32. Zheng L, Campbell M, Murphy J, Lam S, Xu J R (2000). The BMP1 gene is essential for pathogenicity in the gray mold fungus Botrytis cinerea. Mol Plant Microbe Interact, 13(7): 724–732PubMedCrossRefGoogle Scholar

Copyright information

© Higher Education Press and Springer-Verlag Berlin Heidelberg 2011

Authors and Affiliations

  1. 1.Molecular Plant Pathology LaboratoryAgricultural University of HebeiBaodingChina
  2. 2.Agriculture Bureau of LangfangLangfangChina
  3. 3.Millet InstituteHebei Academy of Agricultural and Forestry SciencesShijiazhuangChina

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