And fragments (1-6). Data shown as mean s.e.m. (n = 3; panels k-n). Distinct letters denote considerable differences (P 0.05, Duncan’s various range 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 when compared with 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, 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). Various letters denote significant variations (P 0.05, Duncan’s various variety 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 mean s.e.m. (n = 3). Various letters denote substantial variations (P 0.05, Duncan’s numerous 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 imply s.e.m. (n = 3). Diverse letters denote significant variations (P 0.05, Duncan’s many range test). i, OsGRF4 activates pOsNRT1.1B, pOsNRT2.3a, pOsNPF2.4, pOsNIA1, pOsNIA3 and pOsNiR1 promoter::Luciferase fusion constructs in transient transactivation assays. Information shown as mean s.e.m. (n = three) in all panels. A two-sided Student’s t-test was made use of to produce the P 1-Methylpyrrolidine In stock values.Extended Data Figure three. OsGRF4 regulates expression of various NO3- metabolism genes.Europe PMC Funders Author Manuscripts Europe PMC Funders Author ManuscriptsExtended Information 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 2 M paclobutrazol (PAC), genotypes as indicated. Data shown as imply s.e.m. (n = three). A two-sided Student’s t-test was applied to create the P values. b, GS activities in shoots of plants treated with GA andor PAC, genotypes and therapies as indicated in a. Data 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 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 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 Information Figure five. BiFC visualisation of SLR1-OsGIF1-OsGRF4 interactions.a, Details of constructs expressing OsGRF4 and variants deleted for distinct 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 within a) tagged together with the N-terminus of YFP had been co-transformed into tobacco leaf epidermal cells, collectively 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 associated OsGRFs and OsGIFs household protein tagged with the N-terminus of YFP-tagged wereNature. Author ma.