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Journal of Integrative Agriculture  2019, Vol. 18 Issue (6): 1415-1418    DOI: 10.1016/S2095-3119(19)62581-5
Agricultural Economics and Management Advanced Online Publication | Current Issue | Archive | Adv Search |
A rapid approach for isolating a single fungal spore from rice blast diseased leaves
FEI Li-wang1*, LU Wen-bo1*, XU Xiao-zhou1, YAN Feng-cheng1, ZHANG Li-wei1, LIU Jin-tao1, BAI Yuan-jun2, LI Zhen-yu3, ZHAO Wen-sheng1, YANG Jun1, PENG You-liang  
 
1 Key Laboratory of Pest Monitoring and Green Management, Ministry of Agriculture and Rural Affairs/College of Plant Protection, China Agricultural University, Beijing 100193, P.R.China
2 Institute of Plant Protection, Liaoning Academy of Agricultural Sciences, Shengyang 110161, P.R.China
3 Saline-Alkali Land Utilization and Research Institute, Liaoning Academy of Agricultural Sciences, Panjin 124010, P.R.China
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Abstract  
Single spore isolation is a fundamental approach in plant pathology and mycology to isolate and identify plant fungal pathogens from diseased samples.  However, routine single spore isolation procedure is time-consuming and has a high risk of contamination by other microorganisms.  In this study, we developed a rapid approach for isolating a single spore of the fungal pathogen, Pyricularia oryzae, from rice blast diseased leaves in the paddy field with low potential of contamination.  First, rice blast leaves with single lesions were selected in the paddy field, and a single lesion was cut out and pressed and dragged gently across the surface of water agar.  Next, a germinated single spore with a barely visible piece of agar was cut out of water agar with a dissecting needle under a stereomicroscope at approximately 120-fold magnification.  Last, the germinated single spore with water agar was transferred onto oatmeal tomato agar for growth and preservation.  Based on our experience, a skilled technician or student can successfully isolate single spore from over 150 independent diseased samples with nearly no contaminations in a single working day.  This approach is also suitable for isolating single spore from other fungal disease samples that produce abundant aerial conidia.
 
Keywords:  Pyricularia oryzae        single spore isolation        rice blast        field experiment  
Received: 26 September 2018   Accepted:
Fund: This work was supported by the earmarked fund for China Agricultural Research System (CARS-01-33) and the National Key Research and Development Program of China (2016YFD0300703).
Corresponding Authors:  Correspondence YANG Jun, Tel: +86-10-62732541, E-mail: yangj@cau.edu.cn    
About author:  FEI Li-wang, E-mail: Liwang_fei@cau.edu.cn; LU Wen-bo, E-mail: luwb@cau.edu.cn; * These authors contributed equally to this study.

Cite this article: 

FEI Li-wang, LU Wen-bo, XU Xiao-zhou, YAN Feng-cheng, ZHANG Li-wei, LIU Jin-tao, BAI Yuan-jun, LI Zhen-yu, ZHAO Wen-sheng, YANG Jun, PENG You-liang. 2019. A rapid approach for isolating a single fungal spore from rice blast diseased leaves. Journal of Integrative Agriculture, 18(6): 1415-1418.

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