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Background   Penicillium italicum  (blue mold) is one of citrus pathogens causing undesirable citrus fruit decay even at strictly-controlled low temperatures (< 10 °C) during shipping and storage.  P. italicum  isolates with considerably high resistance to sterol demethylation inhibitor (DMI) fungicides have emerged; however, mechanism(s) underlying such DMI-resistance remains unclear. In contrast to available elucidation on anti-DMI mechanism for  P. digitatum  (green mold), how  P. italicum  DMI-resistance develops has not yet been clarified.    Results  The present study prepared RNA-sequencing (RNA-seq) libraries for two  P. italicum  strains (highly resistant (Pi-R) versus highly sensitive (Pi-S) to DMI fungicides), with and without prochloraz treatment, to identify prochloraz-responsive genes facilitating DMI-resistance. After 6 h prochloraz-treatment, comparative transcriptome profiling showed more differentially expressed genes (DEGs) in Pi-R than Pi-S. Functional enrichments identified 15 DEGs in the prochloraz-induced Pi-R transcriptome, simultaneously up-regulated in  P. italicum  resistance. These included ATP-binding cassette (ABC) transporter-encoding genes, major facilitator superfamily (MFS) transporter-encoding genes, ergosterol (ERG) anabolism component genes  ERG2 ,  ERG6  and  EGR11  ( CYP51A ), mitogen-activated protein kinase (MAPK) signaling-inducer genes  Mkk1  and  Hog1 , and Ca 2+ /calmodulin-dependent kinase (CaMK) signaling-inducer genes  CaMK1  and  CaMK2 . Fragments Per Kilobase per Million mapped reads (FPKM) analysis of Pi-R transcrtiptome showed that prochloraz induced mRNA increase of additional 4 unigenes, including the other two  ERG11  isoforms  CYP51B  and  CYP51C  and the remaining kinase-encoding genes (i.e.,  Bck1  and  Slt2 ) required for Slt2-MAPK signaling. The expression patterns of all the 19 prochloraz-responsive genes, obtained in our RNA-seq data sets, have been validated by quantitative real-time PCR (qRT-PCR). These lines of evidence in together draw a general portrait of anti-DMI mechanisms for  P. italicum  species. Intriguingly, some strategies adopted by the present Pi-R were not observed in the previously documented prochloraz-resistant  P. digitatum  transcrtiptomes. These included simultaneous induction of all major  EGR11  isoforms ( CYP51A / B / C ), over-expression of  ERG2  and  ERG6  to modulate ergosterol anabolism, and concurrent mobilization of Slt2-MAPK and CaMK signaling processes to overcome fungicide-induced stresses.    Conclusions  The present findings provided transcriptomic evidence on  P. italicum  DMI-resistance mechanisms and revealed some diversity in anti-DMI strategies between  P. italicum  and  P. digitatum  species, contributing to our knowledge on  P. italicum  DMI-resistance mechanisms.

bmc genomics

Transcriptome analysis of fungicide-responsive gene expression profiles in two Penicillium italicum strains with different response to the sterol demethylation inhibitor (DMI) fungicide prochloraz