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Current Genetics

, Volume 25, Issue 1, pp 41–46 | Cite as

Restoration of pathogenicity of a penetration-deficient mutant ofCollectotrichum lagenarium by DNA complementation

  • Noel S. Perpetua
  • Yasuyuki Kubo
  • Tetsuro Okuno
  • Iwao Furusawa
Original Articles

Abstract

Infection byColletotrichum lagenarium requires formation of an appressorium and of a penetration peg. A mutant, 83 348, defective in morphogenesis of the penetration peg was unable to penetrate into cellulose membranes or infect cucumber leaves. DNA transformation using a wild-type genomic library constructed in pKVB resulted in two transformants, Ppr1 and Ppr2, with restored penetration peg formation, from 2,000 benomylresistant transformants. However, penetration into cellulose membranes by these transformats ranged from 30 to 40% compared to greater than 90% by wild-type. Southern-blot hybridization showed that a single copy of a cosmid clone had integrated into the genome of the transformants. A 12.0-kbp fragment of the cosmid vector with the flanking region of wild-type genomic DNA was recovered by plasmid rescue from Ppr1. Using the flanking DNA sequences as a probe for colony blot hybridization, a genomic clone was identified and designated pRP46. Transformants obtained following transformation with pRP46 were able to penetrate cellulose membranes. The penetration frequency of pRP46 transformants ranged from 25 to 65%. Transformants were also pathogenic on cucumber.

Key words

Fungal transformation Colletotrichum lagenarium Penetration peg formation DNA complementation 

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Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Noel S. Perpetua
    • 1
  • Yasuyuki Kubo
    • 1
  • Tetsuro Okuno
    • 1
  • Iwao Furusawa
    • 1
  1. 1.Laboratory of Plant Pathology, Faculty of AgricultureKyoto UniversityKyotoJapan

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