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. 2020 Feb 18;183(1):289–303. doi: 10.1104/pp.19.01364

Figure 1.

Figure 1.

OsNAR2.1 interacts with OsNIT1 and OsNIT2 after nitrate supply to roots. A, Interaction test of OsNAR2.1 with OsNIT1 and OsNIT2 using the DUAL Membrane Pairwise Interaction Kit (Dualsystems Biotech). B, Interaction test of OsNAR2.1 with OsNIT1 and OsNIT2 using Co-IP. OE11 and OE16, two independent lines overexpressing OsNAR2.1 with 6×His tag; Negative, control line segregated from OsNAR2.1 with 6×His tag overexpression lines; pUbi::His, control line overexpressing 6×His tag only. Immunoblots were developed with anti-His antibody to detect OsNAR2.1 expression, and with anti-NIT2 to detect OsNIT2 expression. Anti-HSP used as positive control. C and D, Analysis of OsNAR2.1, OsNIT1, and OsNIT2 expression patterns in the roots of rice seedings (‘Nipponbare’) supplied with different concentrations of Ca(NO3)2. C, Expression patterns of OsNAR2.1, OsNIT1, and OsNIT2 revealed by RNA in situ hybridization in root tip and mature root area supplied with 2.5 mm of nitrate. All detection probes were based on the whole antisense strand. Negative control materials were incubated with water. D, Relative expression levels of OsNAR2.1, OsNIT1, and OsNIT2 in the roots treated with different nitrate concentrations as quantified by RT-qPCR. OsActin (LOC_Os03g50885) was used as an internal control. The values in (D) represent means ± se of three biological replicates.