Cotton metallothionein GhMT3a, a reactive oxygen species scavenger, increased tolerance against abiotic stress in transgenic tobacco and yeast

Abstract

A cDNA clone encoding a 64-amino acid type 3 metallothionein protein, designated GhMT3a, was isolated from cotton (Gossypium hirsutum) by cDNA library screening. Northern blot analysis indicated that mRNA accumulation of GhMT3a was up-regulated not only by high salinity, drought, and low temperature stresses, but also by heavy metal ions, abscisic acid (ABA), ethylene, and reactive oxygen species (ROS) in cotton seedlings. Transgenic tobacco (Nicotiana tabacum) plants overexpressing GhMT3a showed increased tolerance against abiotic stresses compared with wild-type plants. Interestingly, the induced expression of GhMT3a by salt, drought, and low-temperature stresses could be inhibited in the presence of antioxidants. H₂O₂ levels in transgenic tobacco plants were only half of that in wild-type (WT) plants under such stress conditions. According to in vitro assay, recombinant GhMT3a protein showed an ability to bind metal ions and scavenge ROS. Transgenic yeast overexpressing GhMT3a also showed higher tolerance against ROS stresses. Taken together, these results indicated that GhMT3a could function as an effective ROS scavenger and its expression could be regulated by abiotic stresses through ROS signalling.