And fragments (1-6). Data shown as mean s.e.m. (n = 3; panels k-n). Various letters denote important variations (P 0.05, Duncan’s various variety test). o, OsGRF4 activates pOsAMT1.two, pOsGS2, pOsNADH-GOGAT2 and pOsFd-GOGAT promoter::Luciferase fusion constructs in transient transactivation assays. Information shown as mean s.e.m. (n = 3). P 0.05 as compared to control group by two-sided Student’s t-tests.Europe PMC Funders Author Manuscripts Europe PMC Funders Author ManuscriptsNature. Author manuscript; obtainable in PMC 2019 February 15.Li et al.Pagea, D-4-Hydroxyphenylglycine site relative abundance of NO3- uptake transporter-encoding OsNRT1.1B, OsNRT2.3a and OsNPF2.four mRNAs. Abundance shown relative to that in NJ6 (=1). Information shown as mean s.e.m. (n = three). Different letters denote substantial variations (P 0.05, Duncan’s multiple range test). b, Relative abundances of OsNIA1, OsNIA3 and OsNiR1 mRNAs encoding NO3- assimilation enzymes. Abundance shown relative to that in NJ6 (=1). Information shown as imply s.e.m. (n = 3). Distinctive letters denote considerable differences (P 0.05, Duncan’s a number of variety test). c-h, Flag-OsGRF4 mediated ChIP-PCR enrichment (relative to input) of GCGG-containing fragments (marked with ) from NO3- uptake transporter-encoding (c) OsNRT1.1B, (d) OsNRT2.3a and (e) OsNPF2.4 gene promoters; NO3- assimilation enzymeencoding (f) OsNIA1, (g) OsNIA3 and (h) OsNiR1 gene promoters. Information shown as mean s.e.m. (n = three). Distinctive letters denote substantial variations (P 0.05, Duncan’s multiple range test). i, OsGRF4 activates pOsNRT1.1B, pOsNRT2.3a, pOsNPF2.4, pOsNIA1, pOsNIA3 and pOsNiR1 promoter::Luciferase fusion constructs in transient transactivation assays. Data shown as imply s.e.m. (n = 3) in all panels. A two-sided Student’s t-test was applied to produce the P values.Extended Data Pyrroloquinoline quinone custom synthesis Figure three. OsGRF4 regulates expression of various NO3- metabolism genes.Europe PMC Funders Author Manuscripts Europe PMC Funders Author ManuscriptsExtended Data Figure four. GA promotes GS and NR activities.a, GS activities in roots of 2-week-old rice plants treated with 100 M GA (GA3) andor two M paclobutrazol (PAC), genotypes as indicated. Data shown as imply s.e.m. (n = three). A two-sided Student’s t-test was utilised to generate the P values. b, GS activities in shoots of plants treated with GA andor PAC, genotypes and treatment options as indicated within a. Information shown as imply s.e.m. (n = 3). A two-sided Student’s t-test was employed to produce the P values. c, NR activities in shoots of plants treated with GA andor PAC, genotypes and remedies as indicated inside a. Information shown as mean s.e.m. (n = 3). A two-sided Student’s t-test was applied to generate the P values.Nature. Author manuscript; available in PMC 2019 February 15.Li et al.PageEurope PMC Funders Author Manuscripts Europe PMC Funders Author ManuscriptsExtended Data Figure 5. BiFC visualisation of SLR1-OsGIF1-OsGRF4 interactions.a, Specifics of constructs expressing OsGRF4 and variants deleted for particular domains. OsGRF4 includes the QLQ (Gln, Leu, Gln) and WRC (Trp, Arg, Cys) domains, positions as indicated. b, BiFC assays. Constructs expressing OsGRF4 or deletion variants (shown as in a) tagged together with the N-terminus of YFP had been co-transformed into tobacco leaf epidermal cells, together with constructs expressing OsGIF1 or SLR1 tagged using the C-terminus of YFP, respectively. Scale bar, 60 m. c, BiFC assays. Constructs expressing OsGRF1 or connected OsGRFs and OsGIFs family members protein tagged using the N-terminus of YFP-tagged wereNature. Author ma.