Biocontrol treatments confer protection against Verticillium dahliae infection of potato by inducing antimicrobial metabolites.

Verticillium wilt, caused by Verticillium dahliae Kleb., is a serious potato (Solanum tuberosum L.) disease worldwide, and biocontrol represents a promising eco-friendly strategy to reduce its impact. We used extracts from Canada milk vetch (CMV) and a set of four V. dahliae-antagonistic bacterial strains to coat potato seeds at planting and examined the degree of protection provided against V. dahliae as well as accumulation of soluble phenolics as markers for induced resistance. All tested treatments were effective in reducing disease severity, and CMV showed the highest level of protection. In this treatment, flavonol-glycoside rutin was a highly abundant compound induced in potato tissues, with levels two to three times higher than those detected in noninoculated controls and V. dahliae-inoculated plants. We investigated dose-dependent effects of rutin on V. dahliae growth and sporulation in vitro and in planta. The effect of rutin on mycelial growth was inconsistent between disk assay and amended medium experiments. On the other hand, significant reduction of V. dahliae sporulation in vitro was consistently observed starting at 300 and 100 µM for isolates Vd-9 and Vd-21, respectively. We successfully detected 2-protocatechuoylphloroglucinolcarboxylic acid (2-PCPGCA) using ultra-performance liquid chromatography tandem mass spectrometry, indicating that V. dahliae dioxygenally oxidizes quercetin. Quercetin, as an aglycone, is freed from the sugar moiety by glucosidases and rhamnosidases produced by the fungus and is a substrate for quercetinases. The occurrence of quercetinases in V. dahliae provides a background to formulate a hypothesis about how by-product 2-PCPGCA may be interfering with potato defenses.

Treatment of chickpea with Rhizobium isolates enhances the expression of phenylpropanoid defense-related genes in response to infection by Fusarium oxysporum f. sp. ciceris.

Differential expression of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS) and isoflavone reductase (IFR) genes involved in phenylpropanoids metabolism was investigated using Northern blot analyses in chickpea seedlings bacterized with Rhizobium isolates (PchDMS and Pch43) and further challenged with Fusarium oxysporum f. sp. ciceris (Foc) race 0. Gene activation patterns in the moderately resistant accession INRAT87/1 were compared with those exhibited by the susceptible accession ILC482 at various time intervals after inoculation with Foc, to determine whether differences in levels or timing of transcript accumulation could be correlated with differences in the susceptibility of chickpea accessions to Foc. Gene activation was higher in the moderately resistant accession INRAT87/1 than in the susceptible ILC482. Pre-treatment of chickpea seedlings with Rhizobium isolates before inoculation with the pathogen enhanced the accumulation of the three genes' mRNA transcripts. In parallel, changes in the soluble phenolic pool produced through pathways involving these enzymes were analyzed in chickpea roots. A strong accumulation of these compounds was revealed at 72 hpi in both accessions. After that time, these high levels of phenolic compounds were maintained until the end of the experiment in the moderately resistant accession, while they have significantly declined in the susceptible accession. HPLC analyses revealed a very high accumulation of the constitutive isoflavones, formononetin and biochanin A and their glycoside conjugates in chickpea roots inoculated with Rhizobium isolates and/or challenged with Foc, as compared to the controls. Our results suggest that the increased accumulation of phenolic compounds, observed in chickpea seedlings inoculated with Foc, can be attributed to increased expression of genes in the phenylpropanoid pathway and that such gene expression is enhanced by pre-treatment with Rhizobium isolates.