Expression of green fluorescent protein in Aureobasidium pullulans and quantification of the fungus on leaf surfaces

Biotechniques. 1997 Oct;23(4):686-90. doi: 10.2144/97234st01.

Abstract

A red-shifted, mutated form of the jelly-fish green fluorescent protein (GFP) under control of a TEF promoter was expressed at high levels in the filamentous fungus Aureobasidium pullulans. In the three transformants studied, all morphotypes of the fungus, including pigmented chlamydospores, expressed GFP and fluoresced brightly. Confocal microscopy showed that the intra-cellular distribution of GFP was nonuniform. When applied to leaf surfaces, the transformants were readily visible and amenable to quantification by image analysis. Thus, GFP expression, together with quantitative image analysis, may provide a powerful method for ecological studies of plant-microbe relationships in nature.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Technical Report

MeSH terms

  • Blotting, Southern
  • Fluorescence
  • Gene Expression*
  • Green Fluorescent Proteins
  • Luminescent Proteins / analysis
  • Luminescent Proteins / genetics*
  • Microscopy, Confocal
  • Mitosporic Fungi / chemistry
  • Mitosporic Fungi / genetics*
  • Promoter Regions, Genetic
  • Transformation, Genetic

Substances

  • Luminescent Proteins
  • Green Fluorescent Proteins