Abstract
A method for simple and rapid detection of Ustilaginoidea virens in rice seeds was developed based on specific polymerase chain reaction (PCR). To design the specific primers for detection of U. virens, the comparison was made on 5.8S rDNA intra- and inter-specific variations in nucleotide sequences of U. virens and other pathogens: Fusarium verticillioides, Sclerotinia sclerotiorum, Tilletia barclayana, Fusarium graminearum and Magnaporthe oryzae. A 346-bp fragment could be amplified by using the specific primers uvr-F and uvr-R in U. virens, but not in other test fungi, indicating that the designed primers were specific for the detection of U. virens. U. virens was detected in nine of the 24 tested rice samples, indicating that 37.5% of the rice samples were contaminated with the false smut pathogen. The development of the simple and rapid detection technique using specific primers will be applicable for direct identification of U. virens in rice seeds to screen large numbers of samples.
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References
Altschul, S. F., Maddenn, T. L., Schaffer, A. A., Zhang, J., Zhang, Z., Miller, W., et al. (1997). Gapped BLAST and PSI-BLAST: a new generation of protein databases search programs. Nucleic Acids Research, 25, 3389–3402.
Ashizawa, T., Takahashi, M., Moriwaki, J., & Hirayae, K. (2010). Quantification of the rice false smut pathogen from soil in Japan using real-time PCR. European Journal of Plant Pathology, 128, 221–232.
Ashizawa, T., Takahashi, M., Moriwaki, J., & Hirayae, K. (2011). A refined inoculation method to evaluate false smut resistance in rice. Journal of General Plant Pathology, 77, 10–16.
Brooks, S. A., Anders, M. M., & Yeater, K. M. (2009). Effect of cultural management practices on the severity of false smut and kernel smut of rice. Plant Disease, 93, 1202–1208.
Brooks, S. A., Anders, M. M., & Yeater, K. M. (2010). Effect of furrow irrigation on the severity of false smut in susceptible rice varieties. Plant Disease, 94, 570–574.
Chen, Y., Zhang, Y., Yao, J., Li, Y. F., Yang, X., Wang, W. X., et al. (2013). Frequency distribution of sensitivity of Ustilaginoidea virens to four EBI fungicides, prochloraz, difenoconazole, propiconazole and tebuconazole, and their efficacy in controlling rice false smut in Anhui Province of China. Phytoparasitica, 41, 277–284.
Dodan, D. S., & Singh, S. R. (1996). False smut of rice: present status. Agricultural Review, 17, 227–240.
Hension, J. M., & French, R. C. (1993). The polymerase chain reaction and plant disease diagnosis. Annual Review of Phytopathology, 31, 81–109.
Ladhalakshmi, D., Laha, G. S., Singh, R., Karthikeyan, A., Mangrauthia, S. K., Sundaram, R. M., et al. (2012). Isolation and characterization of Ustilaginoidea virens and survey of false smut disease of rice in India. Phytoparasitica, 40, 171–176.
Lu, D., Mao, J., Wang, P., He, Z., & Chen, F. (2008). Resistant difference of indica rice hybrid varieties to rice false smut. Acta Phytophylacica Sinica, 35, 289–294.
Lu, D. H., Yang, X. Q., Mao, J. H., Ye, H. L., Wang, P., Chen, Y. P., et al. (2009). Characterising the pathogenicity diversity of Ustilaginoidea virens in hybrid rice in China. Journal of Plant Pathology, 91, 443–451.
Ou, S. H. (1972). Rice diseases. Kew, Surrey, UK: Commonwealth Mycological Institute.
Sambrook, J., Fritsch, E. F., & Maniatis, T. (1989). Molecular cloning (2nd ed., Vol. 1). New York, NY: Cold Spring Harbor Laboratory.
Tang, Y. X., Jin, J., Hu, D. W., Yong, M. L., Xu, Y., & He, L. P. (2013). Elucidation of the infection process of Ustilaginoidea virens (teleomorph: Villosiclava virens) in rice spikelets. Plant Pathology, 62, 1–8.
Tebeest, D. O., Jecmen, A., & Ditmore, M. (2010). Infection of rice by the false smut fungus, Ustilaginoidea virens. pp. 70–81. In: R. J. Norman & K. A. K. Moldenhauer, (Eds.) B. R. Wells Rice Research Studies.
Tsuda, M., Sasahara, M., Ohara, T., & Kato, S. (2006). Optimal application timing of simeconazole granules for control of rice kernel smut and false smut. Journal of General Plant Pathology, 72, 301–304.
Wang, S., Li, M., Dong, H., Liu, X. Z., Bai, Y. J., & Yang, H. (2008). Sporulation, inoculation methods and pathogenicity of Ustilaginoidea albicans, the cause of white rice false smut in China. Journal of Phytopathology, 156, 755–757.
Zhou, Y. L., Izumitsu, K., Sonoda, R., Nakazaki, T., Tanaka, E., Tsuda, M., et al. (2003). PCR-based specific detection of Ustilaginoidea virens and Ephelis japonica. Journal of Phytopathology, 151, 513–518.
Zhou, Y. L., Pan, Y. J., Xie, X. W., Zhu, L. H., Xu, J. L., Wang, S., et al. (2008). Genetic diversity of rice false smut fungus, Ustilaginoidea virens and its pronounced differentiation of populations in North China. Journal of Phytopathology, 156, 559–564.
Acknowledgments
This study was sponsored by the (i) Key Projects in the National Science & Technology Pillar Program during the Twelfth Five-Year Plan Period (No. 2011BAD16B06, 2012BAD04B09 & 2013BAD07B08); (ii) Special Fund for Agro-Scientific Research in the Public Interest from the Ministry of Agriculture, China (200903039 & 201303023); (iii) Special Fund for Quality Inspection Research in the Public Interest from the General Administration of Quality Supervision, Inspection, and Quarantine of the People’s Republic of China (201410076); (iv) National Natural Science Foundation of China (31201550); (v) Research Program from the General Administration of Quality Supervision, Inspection, and Quarantine of the People’s Republic of China (2012IK267 & 2010IK249); (vi) Anhui Provincial Scientific and Technological Project (1106c0805036); (vii) Innovation Team of Anhui Academy of Agricultural Sciences (12C1105 & 13C1113); (viii) 2012 Dean Youth Innovation Fund of Anhui Academy of Agricultural Sciences (12B1121); (ix) Seed Project of Anhui Academy of Agricultural Sciences (13D1107); (x) The Achievement Promotion Project of Anhui Academy of Agricultural Sciences for the Control of Rice False Smut (12E1107) and (xi) 2014 Science and Technological Fund of Anhui Province for Outstanding Youth.
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Yu Chen and Jian Yao share joint first authorship
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Chen, Y., Yao, J., Li, YF. et al. Simple and rapid detection of rice false smut pathogen Ustilaginoidea virens in rice seeds. Phytoparasitica 42, 371–375 (2014). https://doi.org/10.1007/s12600-013-0372-3
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DOI: https://doi.org/10.1007/s12600-013-0372-3