NbCZF1, a Novel C2H2-Type Zinc Finger Protein, as a New Regulator of SsCut-Induced Plant Immunity in Nicotiana benthamiana

Plant Cell Physiol. 2016 Dec;57(12):2472-2484. doi: 10.1093/pcp/pcw160. Epub 2016 Sep 20.

Abstract

SsCut, which functions as an elicitor, can induce plant immunity. In this study, we utilized Nicotiana benthamiana and virus-induced gene silencing to decrease the expression of > 2,500 genes individually. Using this forward genetics approach, several genes were identified that, when silenced, compromised SsCut-triggered cell death based on a cell death assay. A C2H2-type zinc finger gene was isolated from N. benthamiana Sequence analysis indicated that the gene encodes a 27 kDa protein with 253 amino acids containing two typical C2H2-type zinc finger domains; this gene was named NbCZF1 We found that SsCut-induced cell death could be inhibited by virus-induced gene silencing of NbCZF1 in N. benthamiana In addition, SsCut induces stomatal closure, accompanied by reactive oxygen species (ROS) production by NADPH oxidases and nitric oxide (NO) production. NbCZF1-silenced plants showed impaired SsCut-induced stomatal closure, decreased SsCut-induced production of ROS and NO in guard cells and reduced SsCut-induced resistance against Phytophthora nicotianae Taken together, these results demonstrate that the NbCZF1-ROS-NO pathway mediates multiple SsCut-triggered responses, including stomatal closure, hypersensitive responses and defense-related gene expression. This is the first report describing the function of a C2H2-type zinc finger protein in N. benthamiana.

Keywords: Elicitor; Hypersensitive response; Plant defense; SsCut; Stomatal closure.

MeSH terms

  • Amino Acid Sequence
  • CYS2-HIS2 Zinc Fingers
  • Cell Death
  • Gene Expression Regulation, Plant
  • Gene Silencing
  • Nicotiana / cytology
  • Nicotiana / genetics*
  • Nicotiana / immunology
  • Nicotiana / physiology
  • Nitric Oxide / metabolism
  • Phylogeny
  • Phytophthora / physiology*
  • Plant Diseases / immunology*
  • Plant Immunity*
  • Plant Leaves / cytology
  • Plant Leaves / genetics
  • Plant Leaves / immunology
  • Plant Leaves / physiology
  • Plant Proteins / genetics
  • Plant Proteins / metabolism*
  • Plant Stomata / cytology
  • Plant Stomata / genetics
  • Plant Stomata / immunology
  • Plant Stomata / physiology
  • Reactive Oxygen Species / metabolism
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Signal Transduction*

Substances

  • Plant Proteins
  • Reactive Oxygen Species
  • Nitric Oxide