Chlorogenic acid, a metabolite identified by untargeted metabolome analysis in resistant tomatoes, inhibits the colonization by Alternaria alternata by inhibiting alternariol biosynthesis

Abstract

Tomato fruits can be contaminated by saprophytic strains of Alternaria alternata which is the reason for the frequent occurrence of Alternaria toxins like alternariol, alternariol monomethylether or tenuazonic acid in these types of products. It was shown earlier that alternariol is a colonization factor for tomatoes. In the current analysis two different tomato genotypes were analysed by untargeted comprehensive two-dimensional gas chromatography mass spectrometry (GC×GC-MS). This analysis revealed clear differences in the metabolic profiles which were paralleled by differences in resistance towards Alternaria colonization. One of the genotypes was more resistant against A. alternata infection and contained high amounts of chlorogenic acid in contrast to the other genotype which was sensitive against infection. In in vitro analysis, chlorogenic acid reduced alternariol biosynthesis during the first days of growth of A. alternata. Expression analysis of the alternariol polyketide synthase gene, a key gene in the biosynthesis of alternariol, also revealed a temporal reduction in its expression in the first phases of growth. However by chromatographic analysis it could be demonstrated that chlorogenic acid was degraded over time. This degradation leads to a relief of inhibition resulting in an only temporal inhibition of alternariol biosynthesis. In vivo colonization experiments revealed that chlorogenic acid reduces colonization of tomatoes by A. alternata in a concentration dependent manner, which however is partly counteracted by the addition of alterariol.