Phytophthora blight of pepper, which is caused by the
notorious
oomycete pathogen Phytophthora capsici, is a serious disease in global pepper production regions. Our previous
study had identified two WRKY transcription factors (TFs), CaWRKY01-10
and CaWRKY08-4, which are prominent modulators in the resistant pepper
line CM334 against P. capsici infection.
However, their functional mechanisms and underlying signaling networks
remain unknown. Herein, we determined that CaWRKY01-10 and CaWRKY08-4
are localized in plant nuclei. Transient overexpression assays indicated
that both CaWRKY01-10 and CaWRKY08-4 act as positive regulators in
pepper resistance to P. capsici. Besides,
the stable overexpression of CaWRKY01-10 and CaWRKY08-4 in transgenic Nicotiana benthamiana plants also significantly enhanced the resistance to P. capsici. Using comprehensive approaches including
RNA-seq, CUT&RUN-qPCR, and dual-luciferase reporter assays, we
revealed that overexpression of CaWRKY01-10 and CaWRKY08-4 can activate
the expressions of the same four Capsicum annuum defense-related genes (one PR1, two PR4, and one pathogen-related gene) by directly binding to their promoters.
However, we did not observe protein–protein interactions and
transcriptional amplification/inhibition effects of their shared target
genes when coexpressing these two WRKY TFs. In conclusion, these data
suggest that both of the resistant line specific upregulated WRKY
TFs (CaWRKY01-10 and CaWRKY08-4) can confer pepper’s resistance
to P. capsici infection by directly
activating a cluster of defense-related genes and are potentially
useful for genetic improvement against Phytophthora blight of pepper
and other crops.