Heterologous functional analysis and expression patterns of gibberellin 2-oxidase genes of barley (Hordeum vulgare L.)

The level of bioactive gibberellins (GAs) in plants is regulated partly by their inactivation, mainly by the action of GA 2-oxidases (GA2oxs). This study identified three new GA2ox genes in barley: HvGA2ox1, HvGA2ox3 and HvGA2ox6. Analysis of their nucleotide and putative amino acid sequences revealed that they share high sequence identity with other plant GA2oxs and their corresponding proteins. Phylogenetic analysis revealed the HvGA2ox1, HvGA2ox3 and HvGA2ox6 belong to GA2ox structural classes II, I, and III, respectively. Feeding the HvGA2ox1 and HvGA2ox3 recombinant proteins with the C19-GAs, GA1 and GA20, resulted in the production of GA8 and GA29, respectively, with no product detected when they were fed with the C20-GA, GA12. Whereas the HvGA2ox6 recombinant protein was able to convert GA12 to GA110, and no product was detected when it was fed with GA1 or GA20. HvGA2ox1 and HvGA2ox3 were highly expressed in internodes and the endosperm of maturing seeds while HvGA2ox6 was predominantly expressed in the embryos. Salinity stress upregulated the expression of all three genes in seedling tissues. Our results indicate that HvGA2ox1, HvGA2ox3 and HvGA2ox6 encode functional GA2oxs that can regulate GA levels, and therefore growth and development of a barley plant, and its interaction with environment.