Stephan D, Da Silva AP, Bisutti IL. Optimization of a freeze-drying process for the biocontrol agent Pseudomonas spp. and its influence on viability, storability and efficacy. Biological control. 2016 Mar 1;94:74-81.

https://www.sciencedirect.com/science/article/abs/pii/S1049964415300621

i need to know how to lyophilisate savastanoi isolats obtained from olive knot for a long time;

dear prof; I have isolated bacteria from olive knot but some of them died because we have not freezer -80°C; i want to do lyophilisation but there more than protocol of lyophilisation ; you havr a great experience whith pseudomonas savastanoi from olive ; what did you think about this protocol in file attachement , because i want to stored them for a long time , think you ;

Sendos Sendosa Try more simple way:

Person, L. H. "Preservation and virulence of Xanthomonas vesicatoria isolates maintained in sterilized tap or distilled water." Plant disease reporter (1969). pp. 927-929

Iacobellis NS, DeVay JE. Long-term storage of plant-pathogenic bacteria in sterile distilled water. Applied and Environmental Microbiology. 1986 Aug;52(2):388-9. https://doi.org/10.1128/aem.52.2.388-389.1986

From Person (1969): "In 1956, Kelman (2) first showed that bacterial isolates could be maintained in a viable condition when stored in water. Isolates of Pseudomonas solanacearum were viable after 229 days' storage in tap water, distilled water or buffer, pH 7.0 at 21 C+3. Kelman and Person (3) found that stock cultures of P. solancearum isolates were still viable and pathogenic after storage in sterile distilled water at 22 C for a period of 18-24 months. DeVay and Schnathorst (1) successfully stored Agrobacterium tumefaciens, Corynebacterium insidiosum, and Pseudomonas spp. in steriled distilled water. one hundred singlecelled isolated of P. syringae originally obtained from soil and diseased plants were stored in this manner in March 1961. The pathogenecity of the isolates on peach seedlings and their production of a wide-spectrum antibiotic was not lessened after 20 months in storage. "

From Iacobellis & DeVay (1986): "After 20 or 24 years of storage in distilled water, a very high percentage (90 to 92%) of the isolates of Agrobacterium tumefaciens and Pseudomonas spp. were still alive. Moreover, 12 of 13 viable (after 24 years) isolates of P. syringae subsp. syringae maintained their ability to produce syringomycin and were pathogenic to bean seedlings."

In my collection, Xanthomonas isolates are stored since 1994, and still alive & pathogenic. For other species I have shorter period of stoorage, but successful as well. Temperature is very important - store them at 20-22oC. Elliminate any possible contamination and do not use isolates infected by bacteriophages.

thank you sir