Abstract

Cell walls are barriers that impair colonization of host tissues, but also are important reservoirs of energy-rich sugars. Growing hyphae of necrotrophic fungal pathogens, such as Botrytis cinerea (Botrytis, henceforth), secrete enzymes that disassemble cell wall polysaccharides. In this work we describe the annotation of 275 putative secreted Carbohydrate-Active enZymes (CAZymes) identified in the Botrytis B05.10 genome. Using RNAseq we determined which Botrytis CAZymes were expressed during infections of lettuce leaves, ripe tomato fruit, and grape berries. On the three hosts, Botrytis expressed a common group of 229 potentially secreted CAZymes, including 28 pectin backbone-modifying enzymes, 21 hemicellulose-modifying proteins, 18 enzymes that might target pectin and hemicellulose side-branches, and 16 enzymes predicted to degrade cellulose. The diversity of the Botrytis CAZymes may be partly responsible for its wide host range. Thirty-six candidate CAZymes with secretion signals were found exclusively when Botrytis interacted with ripe tomato fruit and grape berries. Pectin polysaccharides are notably abundant in grape and tomato cell walls, but lettuce leaf walls have less pectin and are richer in hemicelluloses and cellulose. The results of this study not only suggest that Botrytis targets similar wall polysaccharide networks on fruit and leaves, but also that it may selectively attack host wall polysaccharide substrates depending on the host tissue.

Highlights

  • The cell wall matrix is one of the first and largest plant structures that pathogens encounter when interacting with potential hosts

  • Twenty-three percent of the Carbohydrate-Active enZymes (CAZymes) genes encoded carbohydrate-binding proteins (CBMs), 16.48% coded for carbohydrate esterases (CEs) and 8.06 and 3.30% encoded glycosyltransferases (GTs) and www.frontiersin.org polysaccharide lyases (PLs), respectively

  • Eighty-eight of these CAZy proteins were previously detected in the secretomes of Botrytis-infected tomato fruit and Botrytis grown in host-free culture systems (Supplemental Table S2; Shah et al, 2009a,b; Espino et al, 2010; Fernández-Acero et al, 2010; Li et al, 2012; Shah et al, 2012.)

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Summary

Introduction

The cell wall matrix is one of the first and largest plant structures that pathogens encounter when interacting with potential hosts. Two co-extensive networks of polysaccharides comprise up to 80% of the mass of most plant cell walls. A network of cellulose microfibrils is cross-linked via hydrogen-bonded hemicelluloses and is embedded within a second network, a matrix of simple and branched pectin polysaccharides (Carpita and Gibeaut, 1993). The integrity of the hemicellulose-cellulose microfibril network provides much of the strength and rigidity of the cell wall (Harris and Stone, 2008; Scheller and Ulvskov, 2010). Structural glycoproteins, soluble proteins, ions and metabolites are located within the polysaccharide networks of most cell walls (Cassab, 1998; Keegstra, 2010)

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