Blocking amino acid transporter OsAAP3 improves grain yield by promoting outgrowth buds and increasing tiller number in rice.

Amino acid transporters (AATs) play indispensable roles in nutrient allocation during plant development. In this study, we demonstrated that inhibiting expression of the rice amino acid transporter OsAAP3 increased grain yield due to a formation of larger numbers of tillers as a result of increased bud outgrowth. Elevated expression of OsAAP3 in transgenic plants resulted in significantly higher amino acid concentrations of Lys, Arg, His, Asp, Ala, Gln, Gly, Thr and Tyr, and inhibited bud outgrowth and rice tillering. However, RNAi of OsAAP3 decreased significantly Arg, Lys, Asp and Thr concentrations to a small extent, and thus promoted bud outgrowth, increased significantly tiller numbers and effective panicle numbers per plant, and further enhanced significantly grain yield and nitrogen use efficiency (NUE). The promoter sequences of OsAAP3 showed some divergence between Japonica and Indica rice, and expression of the gene was higher in Japonica, which produced fewer tillers than Indica. We generated knockout lines of OsAAP3 on Japonica ZH11 and KY131 using CRISPR technology and found that grain yield could be increased significantly. These results suggest that manipulation of OsAAP3 expression could be used to increase grain yield in rice.

Altered expression of OsNPF7.1 and OsNPF7.4 differentially regulates tillering and grain yield in rice.

The rice nitrate and di/tripeptide transporter (NPF) gene family plays an indispensable role in nitrogen transport and plant growth. In this study, 18 alternatively spliced OsNPF genes with 36 different forms of mRNAs were identified, and two of these, namely OsNPF7.1 and OsNPF7.4, showed opposite expression patterns in axillary buds under different nitrogen concentrations. Our results indicate that the expression levels of OsNPF7.1 and OsNPF7.4 determine the axillary bud outgrowth, especially for the second bud, and subsequently influence the tiller number in rice. The overexpression of either of the variants of OsNPF7.1 or the knockout of OsNPF7.4 increased the seedling biomass as well as the tiller number, filled grain number, and grain yield in rice. However, the RNAi-mediated silencing of OsNPF7.1 or the overexpression of either of the variants of OsNPF7.4 had an opposite effect. The overexpression of OsNPF7.1 or OsNPF7.4 could improve the uptake of nitrate, but the OsNPF7.4-overexpressing plants had lower biomass. It is possible that excessive nitrate in the OsNPF7.4-overexpressing plants led to the accumulation of amino acids in the leaf sheath, which inhibited seedling biomass. In addition, the reduced reutilization of nitrate in the seedlings also limited the plant biomass. However, the moderate increase in nitrate and amino acid concentrations in the OsNPF7.1-overexpressing plants could promote seedling biomass and enhance grain yield. In conclusion, our data suggest that different members in the NPF family have different roles in rice, and this study suggests an alternative way to modify rice architecture and enhance grain yield by regulating the expression of OsNPF7.1 and OsNPF7.4.

Rice nitrate transporter OsNPF7.2 positively regulates tiller number and grain yield.

BACKGROUND: Rice tiller number is one of the most important factors that determine grain yield, while nitrogen is essential for the crop growth and development, especially for tiller formation. Genes involved in nitrogen use efficiency processes have been identified in the previous studies, however, only a small number of these genes have been found to improve grain yield by promoting tillering. RESULTS: We constructed over-expression (OX) lines and RNA-interference (Ri) lines, and selected a mutant of OsNPF7.2, a low-affinity nitrate transporter. Our analyses showed that rice tiller number and grain yield were significantly increased in OX lines, whereas Ri lines and mutant osnpf7.2 had fewer tiller number and lower grain yield. Under different nitrate concentrations, tiller buds grew faster in OX lines than in WT, but they grew slower in Ri lines and mutant osnpf7.2. These results indicated that altered expression of OsNPF7.2 plays a significant role in the control of tiller bud growth and regulation of tillering. Elevated expression of OsNPF7.2 also improved root length, root number, fresh weight, and dry weight. However, reduced expression of OsNPF7.2 had the opposite result on these characters. OsNPF7.2 OX lines showed more significantly enhanced influx of nitrate and had a higher nitrate concentration than WT. The levels of gene transcripts related to cytokinin pathway and cell cycle in tiller bud, and cytokinins concentration in tiller basal portion were higher in OX lines than that in WT, suggesting that altered expression of OsNPF7.2 controlled tiller bud growth and root development by regulating cytokinins content and cell cycle in plant cells. Altered expression of OsNPF7.2 also was responsible for the change in expression of the genes involved in strigolactone pathway, such as D27, D17, D10, Os900, Os1400, D14, D3, and OsFC1. CONCLUSION: Our results suggested that OsNPF7.2 is a positive regulator of nitrate influx and concentration, and that it also regulates cell division in tiller bud and alters expression of genes involved in cytokinin and strigolactone pathways, resulting in the control over rice tiller number. Since elevated expression of OsNPF7.2 is capable of improving rice grain yield, this gene might be applied to high-yield rice breeding.