Isolation of citrus epiphytic yeasts and identification of superior isolates in controlling green mold disease

Document Type : Research Paper

Authors

1 . M.Sc. Student, Department of Plant Protection, Faculty of Agricultural Sciences, University of Agricultural Sciences and Natural Resources, Sari, Mazandaran, Iran.

2 Professor, Department of Plant Protection, Faculty of Agricultural Sciences, University of Agricultural Sciences and Natural Resources, Sari, Mazandaran, Iran.

3 Assistant Professor, Iranian Research Institute of Plant Protection, Agricultural Research, Education and Extension Organization, Tehran, Iran.

4 Assistant Professor, Department of Plant Protection, Faculty of Agricultural Sciences, University of Agricultural Sciences and Natural Resources, Sari, Mazandaran, Iran.

10.22092/bcpp.2025.368126.382

Abstract

Green mold (Penicillium digitatum) is one of the most important diseases of citrus fruits during the postharvest and storage stages, which causes significant damage if not properly managed. In recent years, this pathogen has been controlled by the use of chemical fungicides. In order to reduce fungicide application and eliminate toxic residues, it is necessary to use other management methods. Use of microbial agents in biological control is one of these methods. Based on previous reports on the ability of yeasts to inhibit pathogens, the ability of native isolates to inhibit pathogen that cause green mold has been investigated. For this study, citrus fruit and leaf samples were gathered from various areas in Mazandaran province during autumn of 2023. Yeast colonies were then isolated and purified in the lab by using malt agar medium. Due to the large number of isolates, representatives of isolates were selected based on their phenotypic characteristics (shape, color and margin of the colony, physiological characteristics). The test was conducted on citrus fruits of the Thomson Novel variety and its Soluble solids concentration (SSC) content was determined to be 10 °Brix at the time of harvest. The biocontrol test was carried out on the citrus fruit with 44 yeast isolates. The fruit pieces were initially placed in a yeast suspension with a concentration of 10^8 cells per milliliter of sterile water, and after being stored for five hours at room temperature, 10 microliters of the green mold with a concentration of 10^4 cells per milliliter was added to the center of each cut piece. Fruits were stored under packinghouses conditions with over 90% humidity for six days until symptoms of disease appeared. The results of variance analysis based on the percentage of disease occurrence indicated that among the yeasts, there was a significant difference at a 1% level, regarding their ability to inhibit the green mold pathogen under storage conditions. Comparing the average percentage of disease incidence based on according to Duncan’s multiple range test at significance level 1% shows that the yeasts in the two statistical groups performed very well. Sequencing the D1/D2  domain within the Large subunit ribosomal ribonucleic acid (LSU rDNA) region was employed to detect these outstanding isolates. According to the results, the top–performing yeasts were Hanseniaspora uvarum, Hanseniaspora occidentalis, and Pichia kluyveri. So that after six days, they had complete inhibition of the occurrence and development of the pathogen compared to the control.
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References

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BioControl in Plant Protection
Volume 11, Issue 2
January 2025
Pages 53-65

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