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Volume 11
Issue 9
10.3390/jof11090640
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Article

Methods to Stimulate Sporulation and Freeze-Drying Strategies for the Conservation of Diplodia mutila, Diplodia seriata, Lasiodiplodia theobromae, and Neofusicoccum arbuti Isolated from Apple Trees with Canker and Dieback Symptoms

by
Adrián Valdez-Tenezaca
1,*,
Mauricio E. Gutiérrez
1,
Matías Guerra
2,
Jean Franco Castro
2,
Sergio A. Covarrubias
3 and
Gonzalo A. Díaz
1,*
1
Laboratorio de Patología Frutal, Departamento de Producción Agrícola, Facultad de Ciencias Agrarias, Universidad de Talca, Campus Talca, Av. Lircay s/n, Talca 3460000, Chile
2
Instituto de Investigaciones Agropecuarias (INIA) Quilamapu, Av. Vicente Méndez 515, Chillán 3800062, Chile
3
Academic Unit of Chemical Sciences, Campus Siglo XXI, University of Zacatecas, Carretera Zacatecas-Guadalajara km 6, La Escondida, Zacatecas 98160, Mexico
*
Authors to whom correspondence should be addressed.
J. Fungi 2025, 11(9), 640; https://doi.org/10.3390/jof11090640
Submission received: 25 July 2025 / Revised: 28 August 2025 / Accepted: 28 August 2025 / Published: 29 August 2025
(This article belongs to the Special Issue Control of Postharvest Fungal Diseases, 2nd Edition)
Review Reports Versions Notes

Abstract

Four Botryosphaeria spp. isolated from apple with dieback symptoms, Diplodia mutila, Diplodia seriata, Neofusicoccum arbuti, and Lasiodiplodia theobromae, were subjected to different conditions to induce sporulation, then freeze-dried and stored in glass vials and ampoules at a temperature of 4 °C using two protective media (skimmed milk powder in water at 20% and a mixture of skimmed milk powder at 20% plus 5% inositol). Viability was assessed after storage periods of 1, 90, 180, and 365 days. Low-nutrient growth conditions on 2% water agar supplemented with pine needles, incubated under UV light (λ = 350 nm) and at 24 °C, effectively stimulated sporulation of all four Botryosphaeriaceae species. The survival rate of the isolates was similar. Overall viability showed slight but significant differences depending on the type of protective medium and storage container used for the freeze-dried cultures (p < 0.001). Among the tested media, the highest viability was maintained in vacuum-sealed glass ampoules using either a medium containing 20% skimmed milk powder with 5% inositol or 20% skimmed milk powder alone.
Keywords: Botryosphaeria spp.; dieback; sporulation stimulation; freeze-drying; conidium; preservation Botryosphaeria spp.; dieback; sporulation stimulation; freeze-drying; conidium; preservation

Share and Cite

MDPI and ACS Style

Valdez-Tenezaca, A.; Gutiérrez, M.E.; Guerra, M.; Castro, J.F.; Covarrubias, S.A.; Díaz, G.A. Methods to Stimulate Sporulation and Freeze-Drying Strategies for the Conservation of Diplodia mutila, Diplodia seriata, Lasiodiplodia theobromae, and Neofusicoccum arbuti Isolated from Apple Trees with Canker and Dieback Symptoms. J. Fungi 2025, 11, 640. https://doi.org/10.3390/jof11090640

AMA Style

Valdez-Tenezaca A, Gutiérrez ME, Guerra M, Castro JF, Covarrubias SA, Díaz GA. Methods to Stimulate Sporulation and Freeze-Drying Strategies for the Conservation of Diplodia mutila, Diplodia seriata, Lasiodiplodia theobromae, and Neofusicoccum arbuti Isolated from Apple Trees with Canker and Dieback Symptoms. Journal of Fungi. 2025; 11(9):640. https://doi.org/10.3390/jof11090640

Chicago/Turabian Style

Valdez-Tenezaca, Adrián, Mauricio E. Gutiérrez, Matías Guerra, Jean Franco Castro, Sergio A. Covarrubias, and Gonzalo A. Díaz. 2025. "Methods to Stimulate Sporulation and Freeze-Drying Strategies for the Conservation of Diplodia mutila, Diplodia seriata, Lasiodiplodia theobromae, and Neofusicoccum arbuti Isolated from Apple Trees with Canker and Dieback Symptoms" Journal of Fungi 11, no. 9: 640. https://doi.org/10.3390/jof11090640

APA Style

Valdez-Tenezaca, A., Gutiérrez, M. E., Guerra, M., Castro, J. F., Covarrubias, S. A., & Díaz, G. A. (2025). Methods to Stimulate Sporulation and Freeze-Drying Strategies for the Conservation of Diplodia mutila, Diplodia seriata, Lasiodiplodia theobromae, and Neofusicoccum arbuti Isolated from Apple Trees with Canker and Dieback Symptoms. Journal of Fungi, 11(9), 640. https://doi.org/10.3390/jof11090640

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