K+ channels of Cf-9 transgenic tobacco guard cells as targets for Cladosporium fulvum Avr9 elicitor-dependent signal transduction

Plant J. 1999 Aug;19(4):453-62. doi: 10.1046/j.1365-313x.1999.00534.x.

Abstract

The Cf-9 gene encodes an extracytosolic leucine-rich repeat (LRR) protein that is membrane anchored near its C-terminus. The protein confers resistance in tomato to races of the fungus Cladosporium fulvum expressing the corresponding avirulence gene Avr9. In Nicotiana tabacum the Cf-9 transgene confers sensitivity to the Avr9 elicitor, and leads on elicitation to a subset of defence responses qualitatively similar to those normally seen in the tomato host. One of the earliest responses, both in the native and transgenic hosts, results in K+ salt loss from the infected tissues. However, the mechanism(s) underlying this solute flux and its control is poorly understood. We have explored the actions of Avr9 on Cf-9 transgenic Nicotiana using guard cells as a model. Much detail of guard cell ion channels and their regulation is already known. Measurements were carried out on intact guard cells in epidermal peels, and the currents carried by inward- (IK,in) and outward-rectifying (IK,out) K+ channels were characterized under voltage clamp. Exposures to Avr9-containing extracts resulted in a 2.5- to 3-fold stimulation of IK,out and almost complete suppression of IK,in within 3-5 min. The K+ channel responses were irreversible. They were specific for the Avr9 elicitor, were not observed in guard cells of Nicotiana lacking the Cf-9 transgene and, from kinetic analyses, could be ascribed to changes in channel gating. Both K+ channel responses were found to be saturable functions of Avr9 concentration and were completely blocked in the presence of 0.5 microM staurosporine and 100 microM H7, both broad-range protein kinase antagonists. These results demonstrate the ability of the Cf-9 transgene to couple Avr9 elicitation specifically to a concerted action on two discrete K+ channels and they indicate a role for protein phosphorylation in Avr9/Cf-9 signal transduction leading to transport control.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / pharmacology
  • Animals
  • Cladosporium / pathogenicity
  • Cladosporium / physiology*
  • Electric Conductivity
  • Fungal Proteins / antagonists & inhibitors
  • Fungal Proteins / pharmacology*
  • Ion Channel Gating / drug effects
  • Kinetics
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / physiology*
  • Membrane Potentials / drug effects
  • Nicotiana / cytology*
  • Nicotiana / drug effects
  • Nicotiana / metabolism
  • Nicotiana / microbiology
  • Patch-Clamp Techniques
  • Phosphorylation / drug effects
  • Plant Diseases / microbiology
  • Plant Proteins / genetics
  • Plant Proteins / physiology*
  • Plants, Genetically Modified
  • Plants, Toxic*
  • Potassium / metabolism
  • Potassium Channels / metabolism*
  • Potassium Channels, Inwardly Rectifying*
  • Potassium Channels, Tandem Pore Domain*
  • Protein Serine-Threonine Kinases / antagonists & inhibitors
  • Protein Serine-Threonine Kinases / metabolism
  • Signal Transduction / drug effects*
  • Staurosporine / pharmacology

Substances

  • Fungal Proteins
  • Membrane Glycoproteins
  • Plant Proteins
  • Potassium Channels
  • Potassium Channels, Inwardly Rectifying
  • Potassium Channels, Tandem Pore Domain
  • AVR9 protein, Cladosporium fulvum
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
  • Protein Serine-Threonine Kinases
  • Staurosporine
  • Potassium