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Two pectin lyase genes, pnl-1 and pnl-2, from Colletotrichum gloeosporioides f. sp. malvae differ in a cellulose-binding domain and in their expression during infection of Malva pusilla^b

Department of Environmental Biology, University of Guelph, Guelph, Ontario, , Canada N1G 2W1¹

Author for correspondence: Paul H. Goodwin. Tel: +1 519 824 4120 ext. 2754. Fax: +1 519 837 0442. e-mail: pgoodwin{at}uoguelph.ca

Two pectin lyase genes, designated pnl-1 and pnl-2, were cloned from Colletotrichum gloeosporioides f. sp. malvae, a pathogen of round-leaved mallow (Malva pusilla). pnl-1 was isolated using cDNA from infected plant material; pnl-2 was isolated using cDNA from 3-day-old mycelia grown in mallow-cell-wall extract (MCWE) broth. pnl-1 is the first pectinase gene described thus far to encode a cellulose-binding domain (CBD), which is common in cellulases and xylanases, whereas pnl-2 encodes a pectin lyase that lacks a CBD. In pure culture, pnl-1 expression could be detected when purified pectin or glucose was the sole carbon source, but not when MCWE was the sole carbon source. The lack of pnl-1 expression appeared to be due to gene repression by some unknown factor(s) in the cell-wall extract. In contrast, expression of pnl-2 was detected in cultures when MCWE, but not when purified pectin or glucose, was the sole carbon source. In infected tissue, detection of pnl-1 expression by Northern-blot hybridization and by RT-PCR began with the onset of the necrotrophic phase of infection. Expression ofpnl-2 was not detectable by Northern-blot hybridization, but was observed byRT-PCR in both the biotrophic and necrotrophic phases of infection. The differences between pnl-1 and pnl-2 (i.e. pnl-1 encoding a CBD and differences in the expression patterns of both genes) may be related to the requirements of C. gloeosporioides f. sp. malvae to be able to grow in host tissue under the different conditions present during the biotrophic and necrotrophic phases of infection.

Keywords: anthracnose, biotrophy, hemibiotrophy, mallow, necrotrophy

Abbreviations: CBD, cellulose-binding domain; CWDE, cell-wall-degrading enzyme; Cgm, Colletotrichum gloeosporioides f. sp. malvae; LPH, large primary hyphae; MCWE, mallow-cell-wall extract; PDA, potato dextrose agar; PEL, pectate lyase; PNL, pectin lyase; RACE, rapid amplification of cDNA ends; TSH, thin secondary hyphae

^b The GenBank accession numbers for the sequences reported in this paper are AF158256 and AF156984.

^a These authors contributed equally to this work.

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