Involved in strigolactone (SL) signaling pathway. May function downstream of SL synthesis, as a component of hormone signaling or as an enzyme that participates in the conversion of SL to the bioactive form. Strigolactones are hormones that inhibit tillering and shoot branching through the MAX-dependent pathway, contribute to the regulation of shoot architectural response to phosphate-limiting conditions and function as rhizosphere signal that stimulates hyphal branching of arbuscular mycorrhizal fungi and trigger seed germination of root parasitic weeds. Strigolactone-dependent association of D14 with D3 and D53 (a repressor of SL signaling) triggers D53 ubiquitination and degradation . Hydrolyzes the butenolide ring of SLs . A reaction product D-OH is trapped in the cavity of D14, inducing the interaction with SLR1, and probably with other proteins such as D3 and D53 . Contributes to the negative regulation of gibberellin signaling .
Background References
1. Liu W.et.al.Identification and characterization of HTD2: a novel gene negatively regulating tiller bud outgrowth in rice.Planta 230:649-658(2009).
Immunofluorescence staining of paraffin- embedded rice tissue using anti-D14 rabbit polyclonal antibody.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.(sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes.) The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the antibody () at 1/100 dilution for 10 hours at 4℃ and detected using Alexa Fluor™ 488 conjugate-Goat anti-Rabbit IgG (H+L) Secondary Antibody at a dilution of 1:500 for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded rice tissue using anti-D14 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody () at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
Western blot analysis of D14 on Rice lysates with Rabbit anti-D14 antibody (ER1902-91) at 1/2,000 dilution.
Lysates/proteins at 15 µg/Lane. Exposure time: 3 minutes; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: ER1902-91, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 33 kDa Observed band size: 38 kDa
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