Transcriptome analysis of Xanthomonas fragariae in strawberry leaves.

Xanthomonas fragariae is a quarantine bacterial pathogen that causes angular leaf spot on strawberry. The aim of our study was to analyse the mechanism of interaction of this bacterium with its host plant at the transcriptome level. For this purpose, mRNAs of X. fragariae growing in Wilbrink's medium and from infected strawberry cv. Elsanta plants were isolated and sequenced using the Illumina MiSeq platform. The expression profiles of the bacteria in Wilbrink's medium and in planta were very diverse. Of the 3939 CDSs recorded, 1995 had significantly different expression in planta (966 and 1029 genes were down- and upregulated, respectively). Among the genes showing increased expression in planta, those with eggNOG/COG (evolutionary genealogy of genes: Non-supervised Orthologous Groups/Cluster of Orthologous Groups) categories associated with bacterial cell motility, signal transduction, transport and metabolism of inorganic ions and carbohydrates and transcription were overrepresented. Among the genes with the most increased expression in planta, genes primarily associated with flagella synthesis and chemotaxis were found. It is also interesting to note that out of the 31 genes localized on a plasmid, 16 were expressed differently in planta, which may indicate their potential role in plant-pathogen interactions. Many genes with differentiated expression that were localized on chromosome and plasmid encode proteins of unknown function.

Mutual relations between jasmonic acid and acibenzolar-S-methyl in the induction of resistance to the two-spotted spider mite (Tetranychus urticae) in apple trees.

The possibility of inducing resistance to the two-spotted spider mite, Tetranychus urticae Koch, in 'Gala' apple trees growing under optimal fertilization or nitrogen-deficiency conditions was investigated. The effects of jasmonic acid (JA) at 1.5 and 2.5 mM, and acibenzolar-S-methyl (benzothiadiazole, BTH) at 0.5 and 1.5 mM, applied separately or together, on the fecundity of T. urticae females in a laboratory test as well as on the population growth of the pest in a greenhouse experiment were determined. The influence of both elicitors on the induction of LOX and PAL gene expression was assessed in a parallel experiment using real-time PCR. Jasmonic acid showed significantly higher effectiveness in inducing apple tree resistance to T. urticae, as compared to BTH. This was particularly evident in the reduction in pest numbers that was observed in the greenhouse experiment and was also confirmed by increased LOX gene expression after treatment with JA. BTH induced the expression of the PAL gene more strongly than jasmonic acid; however, this was not reflected in the performance of the two-spotted spider mite in the laboratory and greenhouse experiments. It was also found that the antagonistic effect of BTH on JA might lead to decreased effectiveness of the jasmonic acid used to induce apple tree resistance to the two-spotted spider mite. Although nitrogen fertilization stimulated the development of spider mite populations, the resistance induction mechanism was more effective in N-fertilized plants, which was especially evident at the higher jasmonic acid concentration.

Comparative transcriptome analysis of a lowly virulent strain of Erwinia amylovora in shoots of two apple cultivars - susceptible and resistant to fire blight.

BACKGROUND: Erwinia amylovora is generally considered to be a homogeneous species in terms of phenotypic and genetic features. However, strains show variation in their virulence, particularly on hosts with different susceptibility to fire blight. We applied the RNA-seq technique to elucidate transcriptome-level changes of the lowly virulent E. amylovora 650 strain during infection of shoots of susceptible (Idared) and resistant (Free Redstar) apple cultivars. RESULTS: The highest number of differentially expressed E. amylovora genes between the two apple genotypes was observed at 24 h after inoculation. Six days after inoculation, only a few bacterial genes were differentially expressed in the susceptible and resistant apple cultivars. The analysis of differentially expressed gene functions showed that generally, higher expression of genes related to stress response and defence against toxic compounds was observed in Free Redstar. Also in this cultivar, higher expression of flagellar genes (FlaI), which are recognized as PAMP (pathogen-associated molecular pattern) by the innate immune systems of plants, was noted. Additionally, several genes that have not yet been proven to play a role in the pathogenic abilities of E. amylovora were found to be differentially expressed in the two apple cultivars. CONCLUSIONS: This RNA-seq analysis generated a novel dataset describing the transcriptional response of the lowly virulent strain of E. amylovora in susceptible and resistant apple cultivar. Most genes were regulated in the same way in both apple cultivars, but there were also some cultivar-specific responses suggesting that the environment in Free Redstar is more stressful for bacteria what can be the reason of their inability to infect of this cultivar. Among genes with the highest fold change in expression between experimental combinations or with the highest transcript abundance, there are many genes without ascribed functions, which have never been tested for their role in pathogenicity. Overall, this study provides the first transcriptional profile by RNA-seq of E. amylovora during infection of a host plant and insights into the transcriptional response of this pathogen in the environments of susceptible and resistant apple plants.