Phoma stem canker disease on oilseed rape (Brassica napus) in China is caused by Leptosphaeria biglobosa ‘brassicae’
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Phoma stem canker of oilseed rape (Brassica napus) is a globally important disease that is caused by the sibling ascomycete species Leptosphaeria maculans and L. biglobosa. Sixty fungal isolates obtained from oilseed rape stems with phoma stem canker disease symptoms collected from four provinces in China in 1999, 2005 and 2006 were all identified as Leptosphaeria biglobosa, not L. maculans, by PCR diagnostics based on species-specific primers. There were no differences in cultural characteristics (e.g. pigmentation and in vitro growth) between these L. biglobosa isolates from China and those of 37 proven L. biglobosa isolates from Europe or Canada. In studies using amplified fragment length polymorphism (AFLP) markers, Chinese L. biglobosa populations were genetically more similar to European L. biglobosa populations than to the more diverse Canadian L. biglobosa populations. Sequencing of gene fragments of β-tubulin, actin and the internal transcribed spacer (ITS) region of rDNA from L. biglobosa isolates from China, Europe, Australia and Canada showed a closer taxonomic similarity of Chinese L. biglobosa to the European L. biglobosa ‘brassicae’ than to Canadian L. biglobosa ‘canadensis’ or to the Australian L. biglobosa ‘occiaustralensis’ or ‘australensis’ subclades. These results suggest that the Chinese L. biglobosa population in this study is in the same subclade as European L. biglobosa ‘brassicae’ populations.
KeywordsAFLP Blackleg Brassica napus Genetic structure ITS Leptosphaeria biglobosa subclades Phylogeny
This work was supported by the China Scholarship Council, Perry Foundation, Henry Lester Trust and Great Britain-China Education Trust, and the University of Hertfordshire. Rothamsted Research receives funding from the UK Biotechnology and Biological Sciences Research Council. The authors thank Aiming Qi for assistance with the analysis of the AFLP data, Jonathan West, Maria Eckert, Malgorzata Jedryczka, Hortense Brun, Marie-Hélène Balesdent, Randy Kutcher and Dilantha Fernando for providing isolates of L. biglobosa, Yongju Huang and John Hood for assistance with controlled environmental experiments, Georgia Mitrousia and Kevin King for assistance with the figures and molecular biological work, QiangSheng Li for Fig. 2a and b, and with Ziqin Li and many others for collecting diseased oilseed rape stems from China and other countries.
- Anderson, M. J. (2003). PCO: a FORTRAN computer program for principal coordinate analysis. Department of Statistics, University of Auckland, New Zealand. <http://www.stat.auckland.ac.nz/~mja/Programs.htm>.
- Brun, H., Levivier, S., Eber, F., Renard, M., and Chevre, A.M. (1997). Electrophoretic analysis of natural populations of Leptosphaeria maculans directly from leaf lesions. Plant Pathology, 46, 147–154.Google Scholar
- Chen, G. Y., Wu, C. P., Li, B., Su, H., Zhen, S. Z., & An, Y. L. (2010). Detection of Leptosphaeria maculans from imported Canola seeds. Journal of Plant Diseases and Plant Protection, 117, 173–176.Google Scholar
- Dilmaghani, A., Balesdent, M. H., Didier, J. P., Wu, C., Davey, J., Barbetti, M. J., Li, H., Moreno-Rico, O., Phillips, D., Despeghel, J. P., Vincenot, L., Gout, L., & Rouxel, T. (2009). The Leptosphaeria maculans-L. biglobosa species complex in the American continent. Plant Pathology, 58, 1044–1058.CrossRefGoogle Scholar
- Fitt, B. D. L., Evans, N., Howlett, B. J., & Cooke, B. M. (2006b) (Eds). Sustainable strategies for managing Brassica napus (oilseed rape) resistance to Leptosphaeria maculans (phoma stem canker). Springer, Dordrecht, the Netherlands. 126 pp.Google Scholar
- Hao, L., Song, P., Li, Z., Huangpu, H., & Li, Q. (2014). Genetic diversity of phoma stem canker pathogen Leptosphaeria biglobosa by ISSR. Chinese Journal of Oil Crop Sciences, 36, 98–105.Google Scholar
- Juska, A., Busch, L., & Tanaka, K. (1997). The blackleg epidemic in Canadian rapeseed as a “normal agricultural accident”. Ecological Applications, 7, 1350–1356.Google Scholar
- Li, C. X., Wratten, N., Salisbury, P. A., Burton, W. A., Potter, T. D., Walton, G., Li, H., Sivasithamparam, K., Banga, S. S., Banga, S., Singh, D., Liu, S. Y., Fu, T. D., & Barbetti, M. J. (2008). Response of Brassica napus and B. juncea germplasm from Australia, China and India to Australian populations of Leptosphaeria maculans. Australasian Plant Pathology, 37, 162–170.CrossRefGoogle Scholar
- Li, Q. S., Rong, S. B., Hu, B. C., Jiang, Y. F., Hou, S. M., Fei, W. X., Chen, F. X., Wu, X. J., Fan, Z. X., & Lei, W. X. (2013). Distribution of blackleg disease on oilseed rape in China and its pathogen identification. Chinese Journal of Oil Crop Sciences, 35, 415–423. in Chinese.Google Scholar
- Liu, S. Y., Liu, Z., Fitt, B. D. L., Evans, N., Foster, S. J., Huang, Y. J., Latunde-Dada, A. O., & Lucas, J. A. (2006). Resistance of Leptosphaeria maculans (phoma stem canker) in Brassica napus (oilseed rape) induced by L. biglobosa and chemical defence activators in field and controlled environments. Plant Pathology, 55, 401–412.CrossRefGoogle Scholar
- Nei, M. (1987). Molecular evolutionary genetics. New York: Columbia University Press.Google Scholar
- Payne, R. W., Harding, S. A., Murray, D. A., Soutar, D. M., Baird, D. B., Glaser, A. I., Welham, S. J., Gilmour, A. R., Thompson, R., & Webster, R. (2011). The Guide to GenStat Release 14, Part 2: Statistics (p. 997). Hemel Hempstead: VSN International Ltd.Google Scholar
- Purwantara, A., Barrins, J. M., Cozijnsen, A. J., Ades, P. K., & Howlett, B. J. (2000). Genetic diversity of isolates of the Leptosphaeria maculans species complex from Australia, Europe and North America using amplified fragment length polymorphism analysis. Mycological Research, 104, 772–781.CrossRefGoogle Scholar
- Toscano-Underwood, C., West, J. S., Fitt, B. D. L., Todd, A. D., & Jedryczka, M. (2001). Development of phoma lesions on oilseed rape leaves inoculated with ascospores of A‐group or B‐group Leptosphaeria maculans (stem canker) at different temperatures and wetness durations. Plant Pathology, 50, 28–41.CrossRefGoogle Scholar
- Van de Wouw, A. P., Thomas, V. L., Cozijnsen, A. J., Marcroft, S. J., Salisbury, P. A., & Howlett, B. J. (2008). Identification of Leptosphaeria biglobosa ‘canadensis’ on Brassica juncea stubble from northern New South Wales, Australia. Australasian Plant Disease Notes, 3, 124–128.CrossRefGoogle Scholar
- Voigt, K., Cozijnsen, A. J., Kroymann, J., Pöggeler, S., & Howlett, B. J. (2005). Phylogenetic relationships between members of the crucifer pathogenic Leptosphaeria maculans species complex as shown by mating type (MAT1-2), actin, and β-tubulin sequences. Molecular Phylogenetics and Evolution, 37, 541–557.PubMedCrossRefGoogle Scholar
- West, J. S., Balesdent, M. H., Rouxel, T., Nancy, J. P., Huang, Y. J., Roux, J., Steed, J. M., Fitt, B. D. L., & Schmit, J. (2002). Colonisation of winter oilseed rape tissues by A/Tox+ and B/Tox0 Leptosphaeria maculans (phoma stem canker) in France and England. Plant Pathology, 51, 311–321.CrossRefGoogle Scholar
- Yeh, F. C., Yang, R. C., Boyle, T., Ye, Z. H., & Mao, J. X. (1997). POPGENE: the user-friendly shareware for population genetic analysis. Molecular Biology and Biotechnology Centre, University of Alberta, Canada. (Available at http://www.ualberta.ca/~fyeh/).
- Zhang, X., White, R. P., Demir, E., Jedryczka, M., Lauge, R. M., Islam, M., Li, Z. Q., Huang, Y. J., Hall, A. M., Zhou, G., Wang, Z., Cai, X., Skelsey, P., & Fitt, B. D. L. (2014). Leptosphaeria spp., phoma stem canker and potential spread of L. maculans on oilseed rape crops in China. Plant Pathology, 63, 598–612.Google Scholar
- Zhou, Y., Fitt, B. D. L., Welham, S. J., Gladders, P., Sansford, C. E., & West, J. S. (1999). Effects of severity and timing of stem canker (Leptosphaeria maculans) symptoms on yield of winter oilseed rape (Brassica napus) in the UK. European Journal of Plant Pathology, 105, 715–728.CrossRefGoogle Scholar