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Brassica napus possesses an expanded set of polygalacturonase inhibitor protein genes that are differentially regulated in response to Sclerotinia sclerotiorum infection, wounding and defense hormone treatment

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Abstract

Most plants encode a limited set of polygalacturonase inhibitor (PGIP) genes that may be involved in aspects of plant development, but more importantly in the inactivation of polygalacturonases (PG) secreted by pathogens. Previously, we characterized two Brassica napus PGIP genes, BnPgip1 and BnPgip2, which were differentially expressed in response to pathogen infection and wounding. Here we report that the B. napus genome encodes a set of at least 16 PGIP genes that are similar to BnPgip1 or BnPgip2. This is the largest Pgip gene family reported to date. Comparison of the BnPGIPs revealed several sites within the xxLxLxx region of leucine rich repeats that form β-sheets along the interacting face of the PGIP that are hypervariable and represent good candidates for generating PGIP diversity. Characterization of the regulatory regions and RT-PCR studies with gene-specific primers revealed that individual genes were differentially responsive to pathogen infection, mechanical wounding and signaling molecules. Many of the BnPgip genes responded to infection by the necrotic pathogen, Sclerotinia sclerotiorum; however, these genes were also induced either by jasmonic acid, wounding and salicylic acid or some combination thereof. The large number of PGIPs and the differential manner in which they are regulated likely ensures that B. napus can respond to attack from a broad spectrum of pathogens and pests.

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Abbreviations

BAC:

Bacterial artifical chromosome

cDNA:

Complementary deoxyribonucleic acid

DNA:

Deoxyribonucleic acid

DEPC:

Diethylpyrocarbonate

GUS:

β-Glucuronidase

JA:

Jasmonic acid

MS:

Minimal salts

NTP:

Nucleoside triphosphate

PCR:

Polymerase chain reaction

PDA:

Potato dextrose agar

PG:

Polygalacturonase

PGIP:

Polygalacturonase inhibitor protein

RT-PCR:

Reverse transcription polymerase chain reaction

SA:

Salicylic acid

SDS:

Sodium dodecyl sulphate

SSC:

Sodium chloride–sodium citrate

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Acknowledgments

We thank the Saskatchewan Agriculture Development Fund, the Saskatchewan Canola Development Commission and the Agriculture and Agri-Food Canada Matching Investments Initiative for providing funding to support this work.

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Authors and Affiliations

  1. Agriculture and Agri-Food Canada, 107 Science Place, Saskatoon, SK, S7N 0X2, Canada

    Dwayne D. Hegedus, Lone Buchwaldt, Isobel Parkin, Cathy Coutu, Diana Bekkaoui & S. Roger Rimmer

  2. Agriculture and Agri-Food Canada, Ottawa, K1A 0C6, Canada

    Rugang Li & Steve Whitwill

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  1. Dwayne D. Hegedus
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Correspondence to Dwayne D. Hegedus.

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Hegedus, D.D., Li, R., Buchwaldt, L. et al. Brassica napus possesses an expanded set of polygalacturonase inhibitor protein genes that are differentially regulated in response to Sclerotinia sclerotiorum infection, wounding and defense hormone treatment. Planta 228, 241–253 (2008). https://doi.org/10.1007/s00425-008-0733-1

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