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Cloning of genes required for hypersensitivity and pathogenicity inPseudomonas syringae pv.aptata

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Abstract

A genomic library ofPseudomonas syringae pv.aptata strain NCPPB 2664, which causes bacterial blight of sugar beet, lettuce and other plants, was constructed in the cosmid vector pCPP31. The 13.4 kbEcoRI fragment of the cosmid pHIR11, containing thehrp (hypersensitiveresponse andpathogenicity) gene cluster of the closely related bacteriumPseudomonas syringae pv.syringae strain 61, was used as a probe to identify a homologoushrp gene cluster inP. syringae pv.aptata. Thirty of 2500 cosmid clones, screened by colony hybridization, gave a strong hybridization signal with the probe, but none of these conferred to the non-pathogenic bacterium,Pseudomonas fluorescens, the ability to elicit the hypersensitive response (HR) in tobacco. Southern blot analysis ofEcoRI-digested genomic DNA ofP. syringae pv.aptata showed hybridizing bands of 12 kb and 4.4 kb. Only a 12 kb fragment hybridized in digests of the cosmids. Cosmid clone pCPP1069 was mutagenized with Tn10-minitet and marker-exchanged into the genome ofP. syringae pv.aptata. Three resulting prototrophic mutant strains failed to elicit the HR in tobacco and to cause disease in lettuce. The DNA flanking the Tn10-minitet insertions from mutated derivatives of pCPP1069 hybridized with the 10.6 kbBglII fragment of pHIR11. These results indicate thatP. syringae pv.aptata harbourshrp genes that are similar to, but arranged differently from, homologoushrp genes ofP. syringae pv.syringae.

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Abbreviations

HR:

hypersensitive response

Hrp:

mutant unable to induce HR and pathogenicity

Psa:

Pseudomonas syringae pv.aptata

Pss:

Pseudomonas syringae pv.syringae

Ea:

Erwinia amylovora

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Authors and Affiliations

  1. Department of Plant Pathology, Cornell University, 14853, Ithaca, NY, USA

    P. Minardi

  2. Institute of Plant Pathology, Bologna University, 40126, Bologna, Italy

    P. Minardi

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Minardi, P. Cloning of genes required for hypersensitivity and pathogenicity inPseudomonas syringae pv.aptata . Antonie van Leeuwenhoek 67, 201–210 (1995). https://doi.org/10.1007/BF00871214

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  • DOI: https://doi.org/10.1007/BF00871214

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