1887

Abstract

Plant-pathogenic bacteria may sense variations in environmental factors, such as temperature, to adapt to plant-associated habitats during pathogenesis or epiphytic growth. The bacterial blight pathogen of soybean, pv. PG4180, preferentially produces the phytotoxin coronatine at 18 °C and infects the host plant under conditions of low temperature and high humidity. A miniTn-based promoterless glucuronidase () reporter gene was used to identify genetic loci of PG4180 preferentially expressed at 18 or 28 °C. Out of 7500 transposon mutants, 61 showed thermoregulated expression as determined by a three-step screening procedure. Two-thirds of these mutants showed an increased reporter gene expression at 18 °C whilst the remainder exhibited higher expression at 28 °C. MiniTn- insertion loci from these mutants were subcloned and their nucleotide sequences were determined. Several of the mutants induced at 18 °C contained the miniTn- insertion within the 328 kb coronatine biosynthetic gene cluster. Among the other mutants with increased expression at 18 °C, insertions were found in genes encoding formaldehyde dehydrogenase, short-chain dehydrogenase and mannuronan C-5-epimerase, in a plasmid-borne replication protein, and in the locus, involved in pathogenicity of . Among the mutants induced at 28 °C, insertions disrupted loci with similarities to a repressor of conjugal plasmid transfer, UV resistance determinants, an isoflavanoid-degrading enzyme, a HU-like DNA-binding protein, two additional regulatory proteins, a homologue of bacterial adhesins, transport proteins, LPS synthesis enzymes and two proteases. Genetic loci from 13 mutants did not show significant similarities to any database entries. Results of plant inoculations showed that three of the mutants tested were inhibited in symptom development and multiplication rates. Temperature-shift experiments suggested that all of the identified loci showed a rather slow induction of expression upon change of temperature.

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2000-10-01
2019-10-17
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