Plant Protect. Sci., 2015, 51(2):61-66 | DOI: 10.17221/43/2014-PPS

Real time PCR quantification of Sclerotium rolfsii in chilli tissue and soilOriginal Paper

Yu-Han GAO1, Wu MIAO2, Rong-Jun GUO1, Shi-Dong LI1
1 Key Laboratory of Pest Management in Crops of the Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, P.R. China
2 Xiangyan Seed Co., Changsha, Hunan, P.R. China

Root rot disease caused by Sclerotium rolfsii is one of the important factors limiting chilli production in southern China. Rapid methods of detection and identification of S. rolfsii are required for disease prediction and management. Herein, we adapted a four-step real time quantitative PCR method to detect S. rolfsii using new designed primer pair S301S/S301A. By using this assay, S. rolfsii was detected in ten soil samples and eight plant samples at concentrations as low as 10-7 g mycelium/g soil or plant, both with and without typical symptoms, while tissue-culturing method could only detect the fungus in two samples. This rapid and quantitative method is efficient for detection and determination of S. rolfsii population in soil and plants.

Keywords: chilli root rot; specific primer pair; pathogen identification; population detection; population determination

Published: June 30, 2015  Show citation

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GAO Y, MIAO W, GUO R, Shi-Dong L. Real time PCR quantification of Sclerotium rolfsii in chilli tissue and soil. Plant Protect. Sci. 2015;51(2):61-66. doi: 10.17221/43/2014-PPS.
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References

  1. Burgess L.W., Knight T.E., Tesorieo L., Phan H.T. (2008): Diagnostic manual for plant diseases in Vietnam. Canberra, Australian Centre for International Agricultural Research: 92-94.
  2. Cating R.A., Hoy M.A., Palmateer A.J. (2012): A comparison of standard and high-fidelity PCR: evaluating quantification and detection of pathogen DNA in the presence of orchid host tissue. Plant Disease, 96: 480-485. Go to original source... Go to PubMed...
  3. Jeeva M.L., Mishra A.K., Vidyadharan P., Misra R.S., Hegde V. (2010): A species-specific polymerase chain reaction assay for rapid and sensitive detection of Sclerotium rolfsii. Australasian Plant Pathology, 39: 517-523. Go to original source...
  4. Jenkins S.F., Averre C.W. (1986): Problems and progress in integrated control of southern blight of vegetables. Plant Disease, 70: 614-619. Go to original source...
  5. Jiménez-Fernández D., Montes-Borrego M., Navas-Cortés J.A., Jiménez-Díaz M., Landa B.B. (2010): Identification and quantification of Fusarium oxysporum in planta and soil by means of an improved specific and quantitative PCR assay. Applied Soil Ecology, 46: 372-382. Go to original source...
  6. Kalmesh M., Gurjar R.B.S. (2001): Sclerotium rolfsii- a new threat to chilli in Rajasthan. Journal of Mycology and Plant Pathology, 31: 261.
  7. Landa B.B., López-Díaz C., Jiménez-Fernández D., MontesBorrego M., Muñoz-Ledesma F.J., Ortiz-Urquiza A., Quesada-Moraga E. (2013): In-planta detection and monitorization of endophytic colonization by a Beauveria bassiana strain using a new-developed nested and quantitative PCR-based assay and confocal laser scanning microscopy. Journal of Invertebrate Pathology, 114: 128-138. Go to original source... Go to PubMed...
  8. Punja Z.K. (1985): The biology, ecology and control of Sclerotium rolfsii. Annual Review of Phytopathology, 23: 97-127. Go to original source...
  9. Punja Z.K. (1986): Progression of root rot on processing carrots due to Sclerotium rolfsii and the relationship of disease incidence to inoculum density. Canadian Journal of Plant Pathology, 8: 297-304. Go to original source...
  10. Qi R.D., Ding J.C., Gu J.T. Gao Z.M., Yue Y.D. (2001): Effects of media on pathogenicity of Phytophthora capsici. Journal of Anhui Agricultural Sciences, 29: 96-97.
  11. Shew B.B., Beute M.K., Campbell C.L. (1984): Spatial pattern of southern stem rot caused by Sclerotium rolfsii in six North Carolina peanut fields. Phytopathology, 74: 730-735. Go to original source...
  12. Xu Y.H. (2009): Occurrence and integrated management of chilli root rot caused by Sclerotium rolfsii. Plant Doctor, 22: 19-20.
  13. Yin J.L., Shackel N.A., Zekry A., McGuinness P.H., Richards C., Van der Putten K., McCaughan G.W., Eris J.M., Bishop G.A. (2001): Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for measurement of cytokine and growth factor mRNA expression with fluorogenic probes or SYBR Green I. Immunology and Cell Biology, 79: 213-221. Go to original source... Go to PubMed...
  14. Zhang C.Y., Zhang G.H., Yang M., Ben K. (2004a): Elimination of primer-dimer effect in SYBR green I real-time RT-PCR using 4-step program. Chinese Journal of Biochemistry and Molecular Biology, 20: 387-392.
  15. Zhang X.Y., Sun W.-Y., Gao Y.-N., Cheng S.-J. (2004b): To design PCR primers with Oligo 6 and Primer Premier 5. China Journal of Bioinformatics, 2: 15-18.

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