Nachaat Sakr


Fusarium head blight (FHB), spot blotch (SB) and common root rot (CRR) are consistently of the most important cereal diseases globally, but few publications have appeared on preservation. We have stored seventy FHB, SB and CRR fungal isolates of five species (Fusarium culmorum, F. verticillioides, F. solani, F. equiseti and Cochliobolus sativus) in sterile distilled water and freezing in two independent experiments for 36 months. Fungi were tested for viability, purity and morphological stability. The storage treatments included fungal inoculum consisting of spores and hyphae suspended in sterile distilled water at 4°C and mycelial cultures on Petri-dishes with potato dextrose agar (PDA) by freezing at -16°C. The first experiment included 70 isolates was tested six times, after 6, 12, 18, 24, 30 and 36 months of storage. The second experiment with 16 isolates, out of the tested 70 isolates, was tested over periods from 1 to 36 months. The 70 three-year old water-stored and frozen cultures did grow when sub-cultured on PDA. Moreover, viable 16 cultures were recovered from the two storage methods having 100% revival at each time point for up to 3 years. All tested isolates were found to be pure, and maintained their original morphological features. Survival of cultures was not affected with preservation procedure, time in storage or taxonomic classification. The two techniques were found to be easy and reliable for preservation of important cereal fungi for up to 36 months. To our best knowledge, this is the first research reporting the possibility of preservation for FHB, SB and CRR causal agents by cold water and freezing for three years.


freezing; viability; water storage

Full Text:



Abd-Elsalam, K. A., Yassin, M. A., Moslem, M. A., Bahkali, A. H., Mckenzie, E. H. C., Stephenson, S. L., Cai, L. and K. D. Hyde. 2010. Culture collections, the new herbaria for fungal pathogens. Fungal Divers. 45: 21–32.

Akhtar, N., Hafeez, R. and A. Amna. 2016. Viability assessment of pure cultures of Mucorales preserved at 4°C. Pak. J. Phytopath. 28: 147-152.

Agrios, G. N. 2004. Plant pathology. Elsevier Academic, NJ. USA.

Arabi, M. I. E., Jawhar, M. and A. Al-Daoude. 2007. Viability of Cochliobolus sativus cultures after storage under different conditions. J. Plant Pathol. 89: 79-83.

Borman, A. M., Szekely, A., Campbell, C. K. and E. M. Johnson. 2006. Evaluation of the viability of pathogenic filamentous fungi after prolonged storage in sterile water and review of recent published studies on storage methods. Mycopathologia 161: 361–368.

Dhingra O. D. and J. B. Sinclair 1985. Basic Plant Pathology Methods. CRC Press, FL. USA

Elliott, M. L. 2005. Survival, growth and pathogenicity of Gaeumannomyces graminis var. graminis with different methods of long term storage. Mycologia 97: 901–907.

Ishikawa, M., Ide, H., Price, W. S., Arata, Y. and T. Kitashima. 2000. Freezing behaviours in plant tissues as visualized by NMR microscopy and their regulatory mechanisms. In: Engelmann, F., Takagi, H. (eds), Cryopreservation of Tropical Plant Germplasm: Current Research Progress and Application. Japan International Research Center for Agricultural Sciences, Owashi, pp. 22-56.

Jong, S. C. 1989. Microbial germplasm. In: Knutson, L., Stoner, A.K. (eds), Biotic diversity and germplasm preservation, global imperatives. Kluwer Academic Publication, Boston, pp. 241-273.

Kang, S., Blair, J. E., Geiser, D. M., Khang, C. H., Park, S. Y., Gahegan, M., O’Donnell, K, Luster, D. G., Kim, S. H., Ivors, K. L., Lee ,Y. H., Lee, Y. W., Grunwald, N. J., Martin, F. M., Coffey, M. D., Veeraraghavan, N. and I. Makalowska. 2006. Plant pathogen culture collections: It takes a village to preserve these resources vital to the advancement of agricultural security and plant pathology. Phytopathology 96: 920-925.

Karen, K. N., Stephen, W. P. and C. J. Shung. 2004. Preservation and distribution of fungal cultures. In: Muller, G.M. (eds), Biodiversity of Fungi. Elsevier Academic Press, San Diego, pp: 3-11.

Legard, D. E. and C. K. Chandler. 2000. Cryopreservation of strawberry pathogen in mechanical ultra-low temperature freezer. HortScience 35: 1357.

McGinnis, M. R., Padhye, A. A. and L. Ajello. 1974. Storage of stock cultures of filamentous fungi, yeasts, and some aerobic Actinomycetes in sterile distilled water. Appl. Microbiol. 28: 218–222.

Milosevic, M. B., Medic-Pap, S. S., Ignjatov, M. V. and D.N. Petrovic, 2007. Lyophilization as a method for pathogens long term preservation. Proc. Nat. Sci. Matica. Srpska. Novi. Sad. 113: 203-210.

Morris, G. J., Smith, D. and G. E. Coulson. 1988. A comparative study of the morphology of hyphae during freezing with the viability upon thawing of 20 species of fungi. J. Gen. Microbiol. 134: 2897-2906.

Roy, C. B., Srinivas, P. and C. K. Jacob. 2014. Relative efficacy of long-term storage methods on survival and virulence of Corynespora cassiicola and Phytophthora meadii pathogenic on rubber (Hevea brasiliensis). Rubber Sci. 27: 202-214.

Ryan, M. J., Smith, D. and P. Jeffries. 2000. A decision-based key to determine the most appropriate protocol for the preservation of fungi. World J. Microbiol. Biotechnol. 16: 183-186.

Sakr, N. 2018. Evaluation of two storage methods for fungal isolates of Fusarium sp. and Cochliobolus sativus. Acta Phytopathol. et Entomol. Hung. 53: 11-18.

Tariq, A., Naz, F., Chaudhary, A., Rauf, C. and G. Irshad 2015. Long term and least expensive preservation methods for various fungal cultures. Pak. J. Phytopath. 27: 147-151.

Windels, C. E., Burnes, P. M. and T. Kommedahll. 1993. Fusarium species stored on silica gel and soil for ten years. Mycologia 85: 21–23.


  • There are currently no refbacks.

Copyright (c) 2019 Nachaat Sakr

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Pakistan Journal of Phytopathological
ISSN: 1019-763X (Print), 2305-0284 (Online).
© 2013 Pak. J. Phytopathol. All rights reserved.