Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (15): 2971-2979.doi: 10.3864/j.issn.0578-1752.2014.15.007

• PLANT PROTECTION • Previous Articles     Next Articles

Function Analysis of BcKMO Gene in Growth, Development and Pathogenicity of Botrytis cinerea

 LI  Pei-Fen, ZHAO  Fu-Xin, DONG  Li-Ping, ZHENG  Hui-Xin, ZHAO  Bin, ZHANG  Jing, SI  He-Long, XING  Ji-Hong, HAN  Jian-Min, DONG  Jin-Gao   

  1. Laboratory of Mycotoxin and Molecular Plant Pathology, Agricultural University of Hebei, Baoding 071001, Hebei
  • Received:2014-01-27 Online:2014-08-01 Published:2014-04-23

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Abstract: 【Objective】The objective of this study is to investigate the function of BcKMO gene in growth, development and pathogenicity of Botrytis cinerea, to further clarify the molecular mechanism of the BcKMO gene in development and pathogenicity of the fungus and provide a theoretical basis for the gray mould control. 【Method】 Bioinformatics methods were used to analyze the BcKMO gene and its encoding product. The BcKMO gene coding region was amplified and cloned into pBARKS1-eGFP, and pBARKS1-BcKMO-eGFP was constructed. The protoplasts of mutant BCG183 were prepared and transformed with pBARKS1- BcKMO-eGFP by PEG-mediated method. PCR, Southern blotting and real-time PCR were used to identify the transformant. Phenotype including colony morphology, mycelium morphology, growth rate, conidial yield, and so on, and pathogenicity assays of wild-type strain BC22 (WT), mutant BCG183 and complemutant BCG183/BcKMO were performed to verify the function of the BcKMO gene in growth, development and pathogenicity. 【Result】The BcKMO gene encodes kynurenine 3-monooxygenase (KMO) having monooxygenase FAD conserved domain and four Aromatic-ring hydroxylase-like motifs. The BcKMO protein displayed high homology to rossmann-fold NAD(P)(+)-binding protein (gi156049701) of Sclerotinia sclerotiorum and FAD-dependent oxidoreductase (gi512192943) of Ophiostoma piceae. The expression vector of the BcKMO gene, pBARKS1-BcKMO-eGFP, was successfully constructed and transformed into the protoplasts of mutant BCG183. Results of PCR, Southern blotting and Real-time PCR showed that the BcKMO gene complementing mutant (BCG183/BcKMO) was successfully obtained. Phenotype assay showed that the mutant BCG183 growed slowly, did not produce conidia and sclerotia, and hyphae slender, but the phenotypes of the BCG183/BcKMO were all similar to that of WT. Compared with WT and the BCG183/BcKMO, the pathogenicity of the mutant BCG183 on kidney bean leaves and cucumber leaves were dramatically increased.【Conclusion】The BcKMO gene is a negative regulation in growth and development, and a positive regulation in pathogenicity of B. cinerea.

Key words: Botrytis cinerea , BcKMO , mutants , growth and development , pathogenicity

[1]Williamson B, Tudzynski B, Tudzynski P, van Kan J A L. Botrytis cinerea: The cause of grey mould disease. Molecular Plant Pathology, 2007, 8(5): 561-580.

[2]Elad Y, Williamson B, Tudzynski P, Delen N. Botrytis spp., and diseases they cause in agricultural systems-an introduction. In Botrytis: Biology, Pathology and Control, 2007: 1-8.

[3]Schumacher J, De Larrinoa I F, Tudzynski B. Calcineurin-responsive zinc finger transcription factor CRZ1 of Botrytis cinerea is required for growth, development, and full virulence on bean plants. Eukaryotic Cell, 2008, 7(4): 584-601.

[4]Schamber A, Leroch M, Diwo J, Mendgen K, Hahn M. The role of mitogen-activated protein (MAP) kinase signalling components and the Ste12 transcription factor in germination and pathogenicity of Botrytis cinerea. Molecular Plant Pathology, 2010, 11(1): 105-119.

[5]Rui O, Hahn M. The Slt2-type MAP kinase Bmp3 of Botrytis cinerea is required for normal saprotrophic growth, conidiation, plant surface sensing and host tissue colonization. Molecular Plant Pathology, 2007, 8(2): 173-184.

[6]Yan L, Yang Q, Sundin G W, Li H, Ma Z. The mitogen-activated protein kinase kinase BOS5 is involved in regulating vegetative differentiation and virulence in Botrytis cinerea. Fungal Genetics and Biology, 2010, 47(9): 753-760.

[7]Heller J, Ruhnke N, Espino J J, Massaroli M, Collado I G, Tudzynski  P. The mitogen-activated protein kinase BcSak1 of Botrytis cinerea is required for pathogenic development and has broad regulatory functions beyond stress response. Molecular Plant-Microbe Interactions, 2012, 25(6): 802-816.

[8]Segmuller N, Ellendorf U, Tudzynski B, Tudzynski P. BcSAK1, a stress-activated mitogen-activated protein kinase, is involved in vegetative differentiation and pathogenicity in Botrytis cinerea. Eukaryotic Cell, 2007, 6(2): 211-221.

[9]Choquer M, Fournier E, Kunz C, Levis C, Pradier J M, Simon A, Viaud M. Botrytis cinerea virulence factors: new insights into a necrotrophic and polyphageous pathogen. FEMS Microbiology Letters, 2007, 277(1): 1-10.

[10]张玉净, 郝志敏, 郑蒙, 张金林, 董金皋. 灰葡萄孢产孢缺陷菌株的遗传分析. 华北农学报, 2011, 26(3): 86-89.

Zhang Y J, Hao Z M, Zheng M, Zhang J L, Dong J G. Genetic analysis of sporulation defective in Botrytis cinerea. Acta Agriculturae Boreali-Sinica, 2011, 26(3): 86-89. (in Chinese)

[11]van Kan J A. Licensed to kill: The lifestyle of a necrotrophic plant pathogen. Trends in Plant Science, 2006, 11(5): 247-253.

[12]Schumacher J, Kokkelink L, Huesmann C, Jimenez-Teja D, Collado I G, Barakat R, Tudzynski P, Tudzynski B. The cAMP-dependent signaling pathway and its role in conidial germination, growth, and virulence of the gray mold Botrytis cinerea. Molecular Plant-Microbe Interactions, 2008, 21(11): 1443-1459.

[13]Zheng L, Campbell M, Murphy J, Lam S, Xu J R. The BMP1 gene is essential for pathogenicity in the gray mold fungus Botrytis cinerea. Molecular Plant-Microbe Interactions, 2000, 13(7): 724-732.

[14]王璇, 邢继红, 赵斌, 韩建民, 董金皋. 灰葡萄孢分生孢子产生相关基因的克隆及功能分析. 微生物学通报, 2013, 40(3): 533-543.

Wang X, Xing J H, Zhao B, Han J M, Dong J G. Cloning and functional analysis of a gene related to conidiospore formation in Botrytis cinerea. Microbiology China, 2013, 40(3): 533-543. (in Chinese)

[15]Choquer M, Fournier E, Kunz C, Levis C, Pradier J M, Simon A, Viaud M. Botrytis cinerea virulence factors: New insights into a necrotrophic and polyphageous pathogen. FEMS Microbiology Letters, 2007, 277(1): 1-10.

[16]Tudzynski P, Kokkelink L. Botrytis cinerea: Molecular aspects of a necrotrophic life style. Plant Relationships, 2009, 5: 29-50.

[17]Juge N. Plant protein inhibitors of cell wall degrading enzymes. Trends in Plant Science, 2006, 11(7): 359-367.

[18]Pinedo C, Wang C M, Pradier J M, Dalmais B, Choquer M, Le Pêcheur P, Morgant G, Collado I G, Cane D E, Viaud M. Sesquiterpene synthase from the botrydial biosynthetic gene cluster of the phytopathogen Botrytis cinerea. ACS Chemical Biology, 2008, 3(12): 791-801.

[19]Amaral M, Levy C, Heyes D J, Lafite P, Outeiro T F, Giorgini F, Leys D, Scrutton N S. Structural basis of kynurenine 3-monooxygenase inhibition. Nature, 2013, 496(7445): 382-385.

[20]Kurnasov O, Goral V, Colabroy K, Gerdes S, Anantha S, Osterman A, Begley T P. NAD biosynthesis: Identification of the tryptophan to quinolinate pathway in bacteria. Chemistry and Biology, 2003, 10: 1195-1204.

[21]Phillips R S. Structure, mechanism, and substrate specificity of kynureninase. Biochimica Biophysica Acta, 2011, 1814: 1481-1488.

[22]Kurnasov O, Jablonski L, Polanuyer B, Dorrestein P, Begley T, Osterman A. Aerobic tryptophan degradation pathway in bacteria: Novel kynurenine formamidase. FEMS Microbiology Letters, 2003, 227: 219-227.

[23]Matthijs S, Baysse C, Koedam N, Tehrani K A, Verheyden L, Budzikiewicz H, Schafer M, Hoorelbeke B, Meyer J M, De Greve H, Cornelis P. The Pseudomonas siderophore quinolobactin is synthesized from xanthurenic acid, an intermediate of the kynurenine pathway. Molecular Microbiology, 2004, 52: 371-384.

[24]Keller U, Lang M, Crnovcic I, Pfennig F, Schauwecker F. The actinomycin biosynthetic gene cluster of Streptomyces chrysomallus: A genetic hall of mirrors for synthesis of a molecule with mirror symmetry. Journal of Bacteriology, 2010, 192: 2583-2595.

[25]Hu Y, Phelan V, Ntai I, Farnet C M, Zazopoulos E, Bachmann B O. Benzodiazepine biosynthesis in Streptomyces refuineus. Chemistry and Biology, 2007, 14: 691-701.
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