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. 2023 Nov;21(11):2169-2172.
doi: 10.1111/pbi.14167. Epub 2023 Aug 24.

CRISPR/Cas9-mediated elimination of OsHHO3, a transcriptional repressor of three AMMONIUM TRANSPORTER1 genes, improves nitrogen use efficiency in rice

Kexin Liu #  1 , Yasuhito Sakuraba #  1 , Namie Ohtsuki  1 , Mailun Yang  1 , Yoshiaki Ueda  1 , Shuichi Yanagisawa  1
Affiliations

CRISPR/Cas9-mediated elimination of OsHHO3, a transcriptional repressor of three AMMONIUM TRANSPORTER1 genes, improves nitrogen use efficiency in rice

Kexin Liu et al. Plant Biotechnol J. 2023 Nov.
No abstract available

Keywords: ammonium uptake; nitrogen acquisition; nitrogen deficiency; nitrogen use efficiency; rice; transcriptional repressor.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Association of OsHHO3 with growth and NH4+ uptake. (a–c) Shoot and root dry weight (a), chlorophyll content (b) and F v /F m ratio of the second leaf (c) of oshho3‐KO‐#1 (KO) and OsHHO3‐OX‐#1 (OX) rice. Seedlings were grown under control conditions (C, 1.42 mm NH4NO3) or N‐deficient conditions (low NH4+ [LA], 0.142 mm NH4+; low NO3− [LN], 0.142 mm NO3−) for 21 days. (d) Rice grown for 120 days in N‐deficient soil. (e, f) Tiller number (e) and per‐plant grain yield (f) of 150‐day‐old rice grown in regular or N‐deficient soil. (g) Filled and unfilled spikelets collected from one 150‐day‐old plant. (h) Venn diagram and scatterplot representing upregulated or downregulated genes (|log2FC| > 1, P < 0.05) in oshho3‐KO‐#1 seedlings. (i) AMT1;1, AMT1;2 and AMT1;3 expression levels in the roots of oshho3‐KO‐#1 and OsHHO3‐OX‐#1 seedlings grown under control conditions or in N‐free nutrient solution for 5 days. (j, k) NH4+ uptake activity (j) and content (k) of seedlings grown under control conditions. (l) Effects of 7 day N‐free treatment on NH4+ uptake activity. (m) OsHHO3 expression levels in the roots of WT seedlings subjected to N starvation for different periods. (n) OsHHO3 expression levels in roots of 13 rice cultivars. Seedlings were exposed to 7 days of control or low NH4+ conditions at the last stage of cultivation. (o, p) Correlation of OsHHO3 expression levels with AMT1;1, AMT1;2 and AMT1;3 expression levels under low NH4+ conditions (o), and with relative shoot and root dry weight (p). Relative dry weight is defined as the per‐seedling dry weight under low NH4+ conditions divided by the per‐seedling dry weight under control conditions. Data represent the mean ± SD of three to five biological replicates. In (a–c), (e), (f), (i–k), (m) and (n), different letters above bars indicate statistically significant differences (P < 0.05; Tukey's multiple comparison test). In (l), asterisks represent significant differences compared with the WT (**P < 0.01; Student's t‐test), and ‘N.D.’ indicates no significant difference. Gene expression levels were normalized first against the OsUBQ5 expression level and then against the value of a reference sample in each experiment.
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References

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