European Journal of Plant Pathology

, Volume 131, Issue 2, pp 211–219 | Cite as

Cloning and characteristics of Brn1 gene in Curvularia lunata causing leaf spot in maize

  • Tong Liu
  • Shufa Xu
  • Lixing Liu
  • Feihong Zhou
  • Jumei Hou
  • Jie Chen
Article
First Online:
Accepted:

Abstract

The full length cDNA of the Brn1 was first cloned, and then expression of the Brn1 was analyzed and the function was identified by silencing technology. Results show that the full length cDNA of the C. lunata Brn1 gene contains 1001 base pairs and an 801 bp open reading frame encoding 267 amino acids. Semi-quantitative PCR analysis shows that the expression of Brn1 at 96 h is significantly higher than at 24 and 72 h (p < 0.05) in both the highly virulent isolate CX-3 and the weakly virulent isolate DD60. Brn1-silenced transformants were light brown in culture filtrate, and have significantly reduced toxin production relative to the wild-type. These results imply that Brn1 gene in C. lunata is not only involved in 1,8-dihydroxynaphthalene melanin synthesis, but is also relatively associated with toxin biosynthesis of the pathogen.

Keywords

Curvularia lunata Gene silencing 1,8-dihydroxynaphthalene Melanin Toxin 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgement

We thank Dr. He Ming, Shanghai Jiaotong University, P. R. China, for technical assistance in RT-PCR. This work was supported by grant of National Natural Science Foundation (30871610 and 30971949), China Agriculture Research System (CARS-02).

References

  1. Andersson, A., Jordan, D., Schneider, G., & Lindqvist, Y. (1996). Crystal structure of the ternary complex of 1, 3, 8-trihydroxynaphthalene reductase from Magnaporthe grisea with NADPH and an active-site inhibitor. Structure, 4, 1161–1170.PubMedCrossRefGoogle Scholar
  2. Ausubel, F. (2004). Short protocols in molecular biology. John Wiley and Sons Inc.Google Scholar
  3. Bell, A. A., & Wheeler, M. H. (1986). Biosynthesis and function of fungal melanins. Annual Review of Phytopathology, 24, 411–451.CrossRefGoogle Scholar
  4. Bell, A. A., Stipanovic, R. D., & Puhalla, J. E. (1976). Pentaketide metabolites of Verticillium dahliae: identification of (+)-scytalone as a natural precursor of melanin. Tetrahedron, 32, 1353–1356.CrossRefGoogle Scholar
  5. Butler, M. J., & Day, A. W. (1998). Fungal melanins: a review. Canadia Journal of Microbiology, 44, 1115–1136.CrossRefGoogle Scholar
  6. Chen, J., Yan, H. H., Gao, Z. G., Xie, C. S., & Zang, J. H. (2003). Identification techniques for physiological differentiation of Curvularia lunata in maize. ACTA Phytopathologica Sinica, 33, 121–125.Google Scholar
  7. Dai, F. C., Gao, W. D., Wu, R. J., & Jin, X. H. (1995). A noticeable corn disease: Curvularia leaf spot. ACTA Phytopathologica Sinica, 25, 330.Google Scholar
  8. Feng, B., Wang, X., Hauser, M., Kaufmann, S., Jentsch, S., Haase, G., et al. (2001). Molecular cloning and characterization of WdPKS1, a gene involved in dihydroxynaphthalene melanin biosynthesis and virulence in Wangiella (Exophiala) dermatitidis. Infection and Immunity, 69, 1781–1794.PubMedCrossRefGoogle Scholar
  9. Feng, J., Gao, Z. G., Xue, C. S., Zhuang, J. H., Chen, J., & Bai, S. Y. (2002). The pathogenesis of the cell-degrading enzymes produced by Curvularia lunata. Rain Fed Crops, 22, 164–166.Google Scholar
  10. Hamilton, A. J., & Gomez, B. L. (2002). Melanins in fungal pathogens. Journal of Medical Microbiology, 51, 189–191.PubMedGoogle Scholar
  11. Howard, R. J., & Ferrari, M. A. (1989). Role of melanin in appressorium function. Experimental Mycology, 13, 403–418.CrossRefGoogle Scholar
  12. Idnurm, A., & Howlett, B. J. (2001). Pathogenicity genes of phytopathogenic fungi. Molecular Plant Pathology, 2, 241–255.PubMedCrossRefGoogle Scholar
  13. Jensen, B. A., & Hahn, M. E. (2000). Molecular characterization and tissue-specific expression of the aryl hydrocarbon receptor in the Beluga, Delphinapterus leucas. Marine Environmental Research, 50, 61–81.CrossRefGoogle Scholar
  14. Kawamura, C., Tsujimoto, T., & Tsuge, T. (1999). Targeted disruption of a melanin biosynthesis gene affects conidial development and UV tolerance in the Japanese pear pathotype of Alternaria alternate. Molecular Plant-Microbe Interactions, 12, 59–63.PubMedCrossRefGoogle Scholar
  15. Kubo, Y., Takno, Y., Endo, N., Yasuda, N., Tajima, S., & Furusawa, I. (1996). Cloning and structural analysis of the melanin biosynthesis gene SCD1 encoding Scytalone Dehydratase in Collectotrichum lagenarium. Applied and Environmental Microbiology, 62, 4340–4344.PubMedGoogle Scholar
  16. Liu, T., Liu, L. X., Jiang, X., Huang, X. L., & Chen, J. (2009). A new furanoid toxin produced by Curvularia lunata, the causal agent of maize Curvularia leaf spot. Canadia Journal of Plant Pathology, 31, 22–27.CrossRefGoogle Scholar
  17. Liu, T., Liu, L. X., Jiang, X., Hou, J. M., Fu, K. H., Zhou, F. H., et al. (2010). Agrobacterium-Mediated transformation as a useful tool for molecular genetic study of phytopathogen Curvularia lunata. European Journal of Plant Pathology, 126, 363–371.CrossRefGoogle Scholar
  18. Meng, H., Li, H., & Wang, Y. (2004). Characterization of tissue expression of Peroxisome profilerator activated receptors in the chicken. Acta Genetica Sinica, 31, 682–687.PubMedGoogle Scholar
  19. Money, N. P. (1997). Mechanism linking cellular pigmentation and pathogenicity in rice blast disease. Fungal Genetics and Biology, 22, 151–152.PubMedCrossRefGoogle Scholar
  20. Nakayashiki, H., Hanada, S., Quoc, N. B., Kadotani, N., Tosa, Y., & Mayama, S. (2005). RNA silencing as a tool for exploring gene function in ascomycete fungi. Fungal Genetics and Biology, 42, 275–283.PubMedCrossRefGoogle Scholar
  21. Rizner, T. L., & Wheeler, M. H. (2003). Melanin biosynthesis in the fungus Curvularia lunata (teleomorph: Cochliobolus lunatus). Canadian Journal of Microbiology, 49, 110–119.CrossRefGoogle Scholar
  22. SAS/Stat User’s Guide. (1989). Version 6, Fourth Edition, Volume 1, Cry, NC: Sas Institute Inc., Cary, North Carolina.Google Scholar
  23. Schnitzler, N., Peltroche-Llacsahuanga, H., Bestier, N., Zundorf, J., Lutticken, R., & Haase, G. (1999). Effect of melanin and carotenoids of Exophiala (Wangiella) dermatitidis on phagocytosis, oxidative burst, and killing by human neutrophils. Infection and Immunity, 67, 94–101.PubMedGoogle Scholar
  24. Takano, Y., Kubo, Y., Kuroda, I., & Furosawa, I. (1997). Temporal transcriptional pattern of three melanin biosynthesis genes, PKS1, SCD1, and THR1, in apressorium-differentiating and nondifferentiating conidia of Colletotrichum lagenarium. Applied and Environmental Microbiology, 63, 351–354.PubMedGoogle Scholar
  25. Vidal-Cros, A., Viviani, F., Labesse, G., Boccara, M., & Gaudry, M. (1994). Polyhydroxynaphthalene reductase involved in melanin biosynthesis in Magnaporthe grisea. European Journal of Biochemistry, 219, 985–992.PubMedCrossRefGoogle Scholar
  26. Wheeler, M. H., Tolmsoff, W. J., Bell, A. A., & Mollenhauer, H. H. (1978). Ultrastructural and chemical distinction of melanins formed by Verticillium dahliae from (+)- scytalone, 1,8-dihydroxynaphthalene, catechol, and L- 3,4-dihydroxyphenylalanine. Canadia Journal of Microbiology, 24, 289–297.CrossRefGoogle Scholar
  27. Xu, S. F., Chen, J., Liu, L. X., Wang, X. F., Huang, X. L., & Zhai, Y. H. (2007). Proteomics associated with virulence differentiation of Curvularia lunata in maize (Zea maydis) in China. Journal of Integrative Plant Biology, 49, 1–10.Google Scholar
  28. Yan, H., Chen, J., Gao, Z., Xia, S., & Zhang, R. (2005). The heredity and variation of the interaction between Curvularia Lunata and Plant. Journal of Maize Sciences, 13, 119–120.Google Scholar

Copyright information

© KNPV 2011

Authors and Affiliations

  • Tong Liu
    • 1
    • 2
  • Shufa Xu
    • 1
  • Lixing Liu
    • 1
  • Feihong Zhou
    • 1
  • Jumei Hou
    • 2
  • Jie Chen
    • 1
  1. 1.School of Agriculture and Biology and Key laboratory of Urban Agriculture (South) of Ministry of AgricultureShanghai Jiaotong UniversityShanghaiChina
  2. 2.School of AgricultureHeilongjiang Bayi Agricultural UniversityDaqingPeople’s Republic of China

Personalised recommendations