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Partial Characterization of a Gene for a 31 kD Protein of Pseudomonas syringae pv. syringae Apparently Involved in Pathogenicity
Author(s) -
Niepold F.,
Nickel Antje,
Landsmann J.
Publication year - 1994
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/j.1439-0434.1994.tb04524.x
Subject(s) - pseudomonas syringae , biology , mutant , complementation , clone (java method) , plasmid , gene , microbiology and biotechnology , wild type , nicotiana tabacum , strain (injury) , hypersensitive response , genetics , plant disease resistance , anatomy
P. syringae pv. syringae strain R32 causes the bacterial brown spot disease on bush beans. A 31 kD protein was detected which is involved in the pathogenic response. Monospecific antibodies directed against this 31 kD‐protein were used to screen a protein expression gene bank made from the wild type strain R32. A 0.8 kb DNA insert of a clone which gave a positive reaction with the monospecific antibodies was used in hybrizations to clone a larger chromosomal fragment. A km‐cassette (kmresistance) was integrated into this chromosomal DNA‐fragment preventing the expression of the 31 kD‐protein. This construct was integrated into the chromosomal of the wild type strain R32 viahomologous recombination resulting in the 31 kD‐protein deficient mutant LMI. Biotests with the host plant (bean) and with tobacco leaves showed no symptoms or hypersensitive reaction (HR) when the mutant LMI was inoculated. However, atypical chlorotic and necrotic lesions compared to the wild type strain R32 were found on tobacco leaves when the mutant LMI was incubated for more than 2 weeks. Complementation of the LMI mutant with a plasmid harbouring the corresponding wild type R32 DNA fragment resulted in an isolate (LMIC) which showed a partial restoration of the HR on tobacco but no brown spot disease symptoms on bush beans. The 31 kD‐protein could be detected serologically in LMIC.