Sugarolytic functions and molecular-biological diagnostics of pathogenic bacteria of white button mushroom Аgaricus bisporus

Abstract

Purpose. To carry out molecular ad biological diagnostics and to determine sugarolytic functions of pathogenic bacteria of Agaricus bisporus for accurate identification of their genus and species. Methods. Six bacterial cultures isolated from fruit bodies of white button mushroom produced by some farms in Ukraine were tested vs. reference strain CFBP 2068T Pseudomonas tolaasii as a positive control. There were performed such microbiological tests: for oxidase and catalase activity, for ability to hydrolyze gelatin and to coagulate or peptonize milk, for sugarolytic properties using method of color series. The tests were carried out with three replications. DNA were extracted from selected isolates of pathogenic bacteria was performed with the Marmur’s modified method. Results. All isolates are gram-negative bacteria that fluoresce on beef-extract agar and beef-extract broth + KNO3, do not hydrolyze gelatin, do not show oxidase activity (except for 4.1), but show catalase activity. The results on sugarolytic properties are as follows: isolate 1.1 uses maltose, lactose, mannitol, galactose, fructose and arabinose with acid formation as the only nutrition source; isolate 2.1 utilizes maltose, glucose (aerobically), xylose, lactose, mannitol, sucrose, fructose, galactose, arabinose; isolate 3.2 does maltose, glucose (aerobically), xylose, lactose, sucrose, fructose, galactose, arabinose; isolate 4.1 does maltose, glucose (aerobically), xylose, lactose, mannitol, sucrose, fructose, galactose, arabinose; isolate 5.1. does glucose, xylose, mannitol, fructose, galactose, arabinose; isolate 6.1 does lactose, mannitol, fructose, galactose, arabinose. There have been were chosen special primers for PCR analysis. Conclusions. The experiments suggest that the isolates studied are pathogenic and morphologically may be attributed to the genus Pseudomonas because they possessive the same morphological and biochemical properties as the reference strain. Identification of total and genomic DNA extracted is confirmed by PCR analysis. The results of the research would help to find effective ways of identifying the pathogens and to prevent spread of the disease during the early stages of the mushroom growth.