Functional and Promoter Analysis of ChiIV3, a Chitinase of Pepper Plant, in Response to Phytophthora capsici Infection

Int J Mol Sci. 2017 Aug 1;18(8):1661. doi: 10.3390/ijms18081661.

Abstract

Despite the involvement of many members of the chitinase family in plant immunity, the precise functions of the majority of the members remain poorly understood. Herein, the gene ChiIV3 in Capsicum annuum encoding a chitinase protein containing a chitin binding domain and targeting to the plasma membrane was found to be induced by Phytophthora capsici inoculation (PCI) and applied chitin treatment. Besides its direct inhibitory effect on growth of Phytophthora capsici (P. capsici), ChiIV3 was also found by virus-induced gene silencing (VIGS) and transient overexpression (TOE) in pepper plants to act as a positive regulator of plant cell death and in triggering defense signaling and upregulation of PR (pathogenesis related) genes against PCI. A 5' deletion assay revealed that pChiIV3-712 to -459 bp was found to be sufficient for ChiIV3' response to PCI. Furthermore, a mutation assay indicated that W-box-466 to -461 bp in pChiIV3-712 to -459 bp was noted to be the PCI-responsible element. These results collectively suggest that ChiIV3 acts as a likely antifungal protein and as a receptor for unidentified chitin in planta to trigger cell death and defense signaling against PCI.

Keywords: Capsicum annuum; Phytophthora capsici; cell death; chitinase; promoter.

MeSH terms

  • Capsicum / enzymology*
  • Capsicum / genetics
  • Capsicum / immunology
  • Capsicum / microbiology*
  • Cell Death / drug effects
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism
  • Chitin / pharmacology
  • Chitinases / genetics*
  • Gene Expression Regulation, Plant / drug effects
  • Gene Silencing
  • Genes, Plant
  • Mutagenesis, Site-Directed
  • Mycelium / drug effects
  • Mycelium / growth & development
  • Nicotiana / metabolism
  • Phytophthora / drug effects
  • Phytophthora / growth & development
  • Phytophthora / physiology*
  • Plant Diseases / genetics
  • Plant Diseases / immunology
  • Plant Diseases / microbiology*
  • Plant Leaves / drug effects
  • Plant Leaves / metabolism
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Promoter Regions, Genetic / genetics*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sequence Deletion
  • Transcription, Genetic / drug effects

Substances

  • Plant Proteins
  • RNA, Messenger
  • Chitin
  • Chitinases