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Transcriptome analyses suggest a disturbance of iron homeostasis in soybean leaves during white mould disease establishment

Calla, B., Blahut-Beatty, L., Koziol, L., Simmonds, D.H., Clough, S.J. (2014). Transcriptome analyses suggest a disturbance of iron homeostasis in soybean leaves during white mould disease establishment, 15(6), 576-588. http://dx.doi.org/10.1111/mpp.12113

Abstract

Sclerotinia sclerotiorum is a serious pathogen of numerous crops around the world. The major virulence factor of this pathogen is oxalic acid (OA). Mutants that cannot produce OA do not cause disease, and plants that express enzymes that degrade OA, such as oxalate oxidase (OxO), are very resistant to S.sclerotiorum. To examine the effect of OA on plants, we infiltrated soybean leaves with 5mm OA and examined the gene expression changes at 2h post-infiltration. By comparing the gene expression levels between leaves of a transgenic soybean carrying an OxO gene (OxO) and its parent AC Colibri (AC) infiltrated with OA (pH2.4) or water (pH2.4 or 5.5), we were able to compare the effects of OA dependent or independent of its pH. Gene expression by microarray analysis identified 2390 genes that showed changes in expression, as determined using an overall F-test P-value cut-off of 0.001. The additional requirement that at least one pairwise t-test false discovery rate (FDR)-corrected P value should be less than 0.001 reduced the list of the most highly significant differentially expressed genes to 1054. Independent of pH, OA altered the expression levels of 78 genes, with ferritin showing the strongest induction by OA. The combination of OA plus its low pH caused 1045 genes (99% of all significant genes) to be differentially expressed, with many of the up-regulated genes being related to basal defence, such as genes of the phenylpropanoid pathway and various cytochrome P450s. RNA-seq was also conducted on four samples: OxO and AC genotypes infiltrated with either OA pH2.4 or water pH2.4. The RNA-seq analysis also identified ferritin paralogues as being strongly induced by OA. As the expression of ferritin, a gene that encodes for an iron storage protein, is induced by free iron, these results suggest that S.sclerotiorum benefits from the ability of OA to free iron from plant proteins, as this induces host cell death, and also allows the uptake and assimilation of the iron for its own metabolic needs. © 2013 BSPP AND JOHN WILEY & SONS LTD.

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