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Research articles

ScienceAsia 48 (2022):ID 764-768 |doi: 10.2306/scienceasia1513-1874.2022.086

A novel, rapid and simple method for obtaining single-spore isolation of strongly parasitic fungi from diseased cherry leaves

Xiuming Ma, Jun Miao, Junfeng Wang, Wei Han*, Yang Sun*

ABSTRACT:     Spore isolation is an essential experimental approach in the fields of mycology and plant pathology, enabling researchers to isolate fungi from diseased plant samples for subsequent identification. Routine single-spore isolation strategies, however, are time-consuming and highly susceptible to contamination with other microorganisms. Herein, we developed a rapid approach to isolate individual spores of the fungal pathogen Passalora circumscissa from diseased cherry leaves in an orchard with a low risk for contamination. Diseased cherry leaves with individual lesions were first selected from the orchard, sterilized with 75% ethanol, and rinsed under deionized water. An acquisition tool was then used to isolate a single spore that was absorbed from the disease spot, and the spore was transferred onto the surface of water agar plate. Individual germinating spores were then excised from these agar plates using a cutting tool under a 10? objective magnification microscope. Lastly, the germinating spores were transferred from the water agar plates onto potato agar media for further growth and preservation. Based upon our findings, an experienced technician can utilize this strategy to isolate individual spores from over 100 independent diseased plant tissue samples in a single day with minimal risk of contamination. This strategy should also be well-suited for the isolation of individual fungal spores associated with other diseased sample types.

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a Institute of Vegetables and Flowers, Shandong Academy of Agricultural Sciences, Shandong Provincial Key Laboratory of Vegetable Biology, Shandong Branch of National Center for Vegetable Improvement, Jinan 250100 China

* Corresponding author, E-mail: sdscshw203@126.com, sunyang_729@163.com

Received 22 Jan 2021, Accepted 27 Feb 2022