4 partially characterized gibberellin 2-oxidases (GA2ox) (GA2ox1 to GA2ox4), 2 uncharacterized GA2oxs (GA2ox5 and GA2ox6), 4 other (GA2oxs GA2ox7 to GA2ox9, GA2ox10 is probably a pseudogene)
gibberellin 2-oxidase plays a key role in the GA catabolic pathway through 2beta-hydroxylation, activation of gibberellin 2-oxidase 6 decreases active gibberellin levels and creates a dominant semi-dwarf phenotype in rice, overview. GA biosynthesis inhibitor, paclobutrazol, positively regulates the OsGA2ox6 gene. The H032 mutant retains normal flowering and seed production, H032 mutant phenotype, overview
mutants overexpressing GA2ox9ACT, GA2ox5delta335-341ACT, and GA2ox6, motif III of the conserved motifs characteristic for C20 GA2oxs is necessary for C20 GA2ox activity, motif III is present in rice GA2ox5, GA2ox6, and GAsox9
isolation of a CaMV 35S-enhancer activation tagged mutant, H032, showing an increased level of OsGA2ox6 mRNA compared to the wild-type plants. RNAi and ectopic expression analysis of OsGA2ox6 show that the dwarf trait and the decreased levels of bioactive GAs in the H032 mutant are a result of the upregulation of the OsGA2ox6 gene. The H032 mutant retains normal flowering and seed production
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Escherichia coli with recombinant enzyme are centrifuged, resuspended in a BugBuster Protein Extraction Reagent, centrifuged, supernatants eluted through a GST-Bind resin with 50 mM Tris buffer, pH 8.0, containing 10 mM reduced glutathione, storage at -80°C
PCR-amplifiction of cDNA, inserted via pGEM-T Easy cloning vector and pAHC18 into pCAMBIA1301 and transferred into Agrobacterium tumefaciens (strain EHA105) and used to transform rice and tobacco (Nicotiana tabacum), or for activity assays subcloned into pGEX-5X expression vector to transform Escherichia coli BL21(DE3)
breeding plants with reduced height, increased root biomass, normal flowering and seed production by overexpression of C20 gibberellin 2-oxidases: first overexpression of GA2ox9ACT mutant generates semidwarf rice with only slightly reduced grain weight and fertility, increased tiller number (by 22%) compared to wild-type, second overexpression of C20 GA2oxs with defective motif III, such as GA2ox5delta335-341ACT mutant, generates a semidwarf rice variety with reduced grain weight (by 16%) and fertility (by 12%) and twofold increased tiller number, third overexpression of a selected C20 GA2ox gene, such as GA2ox6 with less effect on plant growth under the control of a weak promoter could be beneficial without sacrificing seed production
applications of the SBI(SV14) allele in rice breeding are an efficient strategy to develop elite rice varieties with improved lodging resistance and increased yield