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Detection of Corynebacterium michiganense in tomato seed lots 1
Author(s) -
VAERENBERGH J. P. C.,
CHAUVEAU J. F.
Publication year - 1987
Publication title -
eppo bulletin
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.327
H-Index - 36
eISSN - 1365-2338
pISSN - 0250-8052
DOI - 10.1111/j.1365-2338.1987.tb00018.x
Subject(s) - inoculation , biology , isolation (microbiology) , horticulture , bioassay , bacteria , microbiology and biotechnology , genetics
Corynebacterium wilt of tomato causes severe losses in France and Belgium. The disease can be seed‐transmitted and sanitary selection of seedlots is a direct prophylactic control method. It must be selective, objective and sensitive and must provide information on the pathogenicity of the bacteria detected. A method is proposed that associates immunofluorescence (IF) staining with bioassay and isolation. In a preliminary comparison of inoculation on different host plants, tomato seedlings with two or four leaves were the most sensitive. Artificially and naturally contaminated seed extracts were IF‐screened and confirmed by isolation of the pathogen from inoculated seedlings. The detection method consists of maceration of tomato seeds in PBS‐cycloheximide buffer, filtration and centrifugation of the seed extract, and IF screening of the pellets with two antisera. Pellets with positive or suspect IF reading (threshold 5 × 10 3 cells in the pellet) are inoculated into tomato seedlings incubated at 25°C. Within 30 days the seedlings are inspected for typical wilt symptoms and isolation experiments are carried out from the vessels. The method has been tested on commercial seedlots.