Fig. 3: Validation of the MYB61 function.
From: MYB61 is regulated by GRF4 and promotes nitrogen utilization and biomass production in rice
a qPCR analysis of MYB61 in NP/WT and the NIL containing the 9311-MYB61 fragment, showing the relative expression to rice HNR. Transcription level in NP was set as 1. Error bars represent the mean ± SD of three biological replicates. b Schema of MYB61 gene structure and the mutation site in myb61. Boxes and lines in the diagram indicate exons and introns, respectively. c Schema of MYB61 protein. Green boxes indicate DNA binding domain (BD) and orange box indicates the transactivation domain (TD). Gray box indicates the truncated part in the mutant. d The cellulose content, µg in per mg of cell wall residues prepared from the internodes of the indicated plants. Error bars indicate the mean ± SD of four biological replicates. e Boxplot of the wall thickness of sclerenchyma fiber cells in the indicated plants. Box bounds represent the 25th and 75th percentile, center line represents the median, × indicates the mean, and whiskers represent the 25th percentile − 1.5 * the interquartile range and the 75th percentile + 1.5 * the interquartile range. n = 200 cells from three individual internodes of the indicated plants. f SEM graphs of sclerenchyma fiber cells from the internodes of the indicated plants. Bars = 2 μm. n = 3 biologically independent plants. g The leaf area of the indicated seedlings growing in the media with LN (0.2 mM NH4NO3) and HN (5 mM NH4NO3) supply. Error bars indicate the mean ± SD of at least 19 seedlings. h The expression level of MYB61 in the leaf sheaths of NP and the NIL seedlings treated in varied concentrations of NH4NO3 for 2 h. Rice HNR was used for normalization. Error bars represent the mean ± SD of four biological replicates. Statistical significance was calculated with two-tailed Welch’s unpaired t-test and p values are indicated in (a, d, e, g). Letters a–c in (d, e, g) indicate the different means according to Duncan’s multiple range test (p < 0.05). ComNP and Com9311 represent the plants expressing the complementary constructs shown in Supplementary Fig. 2e. The red vertical lines in (d, e, g) were used to separate the plants with different genetic background. NP, Nipponbare; NIL, near-isogenic line; ZH11, Zhonghua11. Source data underlying Fig. 3a, d, e, g, h are provided as a Source Data file.
