Viability and pathogenicity of Rhynchosporium secalis after long-term storage

Arabi, M. I. E. and Alek, H. and Al-Shehadah, E. and Jawhar, M. (2019) Viability and pathogenicity of Rhynchosporium secalis after long-term storage. Acta Phytopathologica et Entomologica Hungarica, 54 (2). pp. 195-201. ISSN 0238-1249

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Abstract

Long-term storage of Rhynchosporium secalis cultures is a challenge for any lab managing a working collection of isolates. In this work, the viability and pathogenicity of R. secalis stock cultures were tested after four years of storage at −20 °C in different concentrations of glycerol. Germinability were measured after each storage by collecting spores by coverslips and placing them on water agar in closed Petri dishes at 20–22 °C in the dark and allowed to germinate for 24 h. Additionally, at the end of each storage treatment, conidia were collected by coverslips from sporulated leaf lesions of symptomatic barley leaves and placed under similar conditions as non-stored controls. Cultures of all stored isolates were viable with a spore germination rate of 72.28% (Rs22) after four years of storage at −20 °C in 60% glycerol. Low viability and contamination were observed when spores were stored in sterile distilled water and in Lima bean agar. All isolates continued to infect barley leaves after 4 years of storage. However, the pathogenicity was significantly (P <0.05) reduced in isolates stored in glycerol as compared with controls. This work helps to preserve R. secalis for a long term period at −20 °C without any contamination; therefore, due to the low costs our results could be applicable for laboratories that have limited resources.

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