150 mM NaCl represses the gibberellin 3-oxidase 1 gene in the wild-type and in the NTL8-deficient mutant (NTL8 acts downstream of the gibberellin biosynthesis), repression is present but less significant in aba3-1 mutant, deficient in the stress hormone abscisic acid (independent pathway dominates), expression of GA3ox1 is recovered after a longer incubation in NaCl and leads to germination
the enzyme is inhibited by Bacillus subtilis strain IJ-31 and hydrocinnamic acid, which is one of the allelochemicals produced by Bacillus subtilis strain IJ-31
wild type enzyme from soluble cell fraction, in 100 mM Tris-HCl pH 7.9 containing 4 mM ascorbate, 4 mM DTT, 4 mM 2-oxoglutarate, 0.5 mM FeSO4, 2 mg/ml bovine serum albumin and 1 mg/ml catalase
wild type enzyme after 12fold purification, in 100 mM Tris-HCl pH 7.9 containing 4 mM ascorbate, 4 mM DTT, 4 mM 2-oxoglutarate, 0.5 mM FeSO4, 2 mg/ml bovine serum albumin and 1 mg/ml catalase
treatment with either Bacillus subtilis strain IJ-31 culture extract or its allelochemicals, like hydrocinnamic acid, results in significant down-regulation of XTR9 gene expression, encoding xyloglucan endotransglucosylase/hydrolase protein 14, which is involved in plant cell elongation by cleavage of loadbearing cross-links between cellulose microfibrils, leading to wall-loosening and enabling elongation to occur, in Arabidopsis thaliana, as well as inhibition of gibberellin 3beta-hydroxxylase
transgenic plant lines with green fluorescence protein-anti gibberellin24 antibody fusion construct (GFP-scFv), called A24-1, 2, and 3, up-regulation in the transgenic plant line A24-2 enhances biosynthesis of gibberellin 24 which accumulates compared to wild-type. GFP-scFv reduces rosette leaf development, delays flower induction, reduces stem elongation of main culm, especially in the early stage of inflorescence growth, later stem elongation of the main culm and lateral shoot elongation is much less affected
transgenic plants overexpressing an active NTL8 form and T-DNA insertional ntl8-1 mutant, under standard conditions, germinate with the same rate, under high salinity stress (150 mM NaCl) the germination of the NTL8 overexpressing mutant is considerably reduced while the NTL8-deficient mutant is less sensitive, independent of absiasic acid (ABA), NTL8-deficient mutant is not impaired by gibberellin biosynthetic inhibitor paclabutrazol (PAC), gibberellin downregulates NTL8 lowering its threshold for germination
rosette leaves are frozen in liquid nitrogen and ground to a fine powder, protein extracted with 50 mM Tris-HCl, pH 8.0, containing 200 mM NaCl, 5 mM EDTA, 0.1% Tween 20 and a proteinase inhibitor mixture, SDS-PAGE
for the NTL8 complementation test, a sequence containing the NTL8 gene with its promoter is subcloned into pKGWFS7 Gateway vector, for histochemical staining an NTL8 promoter region is transcriptionally fused to the GUS-coding sequence, the fusion product transformed into Arabidopsis
the anti gibberellin 24 single monoclonal antibody gene (anti-GA24 scFv) is PCR-amplified with plasmid pH1L24B and introduced into the binary vector pBI-ER-D, transferred into Agrobacterium tumefaciens strain LBA4404 to transform Arabidospsis thaliana, the antibody is fused with the green fluorescence protein (GFP), a c-myc epitope tag and KDEL ER-retention signal