Abstract

Sudden death syndrome (SDS) is caused by the fungal pathogen, Fusarium virguliforme, and is a major threat to soybean production in North America. There are two major components of this disease: (i) root necrosis and (ii) foliar SDS. Root symptoms consist of root necrosis with vascular discoloration. Foliar SDS is characterized by interveinal chlorosis and leaf necrosis, and in severe cases by flower and pod abscission. A major toxin involved in initiating foliar SDS has been identified. Nothing is known about how root necrosis develops. In order to unravel the mechanisms used by the pathogen to cause root necrosis, the transcriptome of the pathogen in infected soybean root tissues of a susceptible cultivar, ‘Essex’, was investigated. The transcriptomes of the germinating conidia and mycelia were also examined. Of the 14,845 predicted F. virguliforme genes, we observed that 12,017 (81%) were expressed in germinating conidia and 12,208 (82%) in mycelia and 10,626 (72%) in infected soybean roots. Of the 10,626 genes induced in infected roots, 224 were transcribed only following infection. Expression of several infection-induced genes encoding enzymes with oxidation-reduction properties suggests that degradation of antimicrobial compounds such as the phytoalexin, glyceollin, could be important in early stages of the root tissue infection. Enzymes with hydrolytic and catalytic activities could play an important role in establishing the necrotrophic phase. The expression of a large number of genes encoding enzymes with catalytic and hydrolytic activities during the late infection stages suggests that cell wall degradation could be involved in root necrosis and the establishment of the necrotrophic phase in this pathogen.

Highlights

  • Sudden death syndrome (SDS) is a serious, emerging soybean disease

  • Since only a small proportion of the RNA transcripts of infected soybean root tissues is encoded by F. virguliforme genes, deep sequencing of an individual RNA sample collected from three biological replications was conducted in a single lane to detect transcripts of most transcribed genes including those that are expressed at low levels

  • Increased expression of F. virguliforme gene g14667 encoding subtilase in the infected tissues especially during late infection suggests its possible involvement in nullifying host defense-related enzymes (S5 Table). In this comparative transcriptomic study, we were able to identify putative virulence factors by the following approaches: (i) investigation of the functions of infection-induced genes based on sequence homology using Blast2GO analyses; (ii) identification of secretory proteins and their GO annotation for possible functions; (iii) search for candidate virulence genes through sequence homology search with functionally characterized virulence genes; and (iv) studying the genes that are only induced in infected roots

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Summary

Introduction

Sudden death syndrome (SDS) is a serious, emerging soybean disease. It is caused by Fusarium virguliforme, which prefers cold and wet climates for developing the disease [1]. In the United States, the disease was first reported in Arkansas in 1971 [2]. The disease spread northward and it has been reported in soybean growing areas in the United States and Canada [2,3,4]. SDS can be divided into two components: (i) root necrosis and (ii) foliar SDS. Foliar SDS is characterized by interveinal chlorosis and necrosis of leaves, and in severe cases by flower and pod abscission [6]

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