硝酸盐、脱落酸和赤霉素转运基因OsNPF3.1对水稻分蘖和氮利用效率至关重要

doi:10.1016/j.jia.2023.04.024

Journal of Integrative Agriculture ›› 2024, Vol. 23 ›› Issue (4): 1087-1104.DOI: 10.1016/j.jia.2023.04.024

所属专题：水稻遗传育种合辑Rice Genetics · Breeding · Germplasm Resources

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硝酸盐、脱落酸和赤霉素转运基因OsNPF3.1对水稻分蘖和氮利用效率至关重要

修回日期:2023-02-13 接受日期:2023-03-20 出版日期:2024-04-20 发布日期:2024-03-29

OsNPF3.1, a nitrate, abscisic acid and gibberellin transporter gene, is essential for rice tillering and nitrogen utilization efficiency

Junnan Hang^1*, Bowen Wu^1*, Diyang Qiu^{2, 3*}, Guo Yang¹, Zhongming Fang^1#, Mingyong Zhang^2#

¹Institute of Rice Industry Technology Research/Key Laboratory of Functional Agriculture, Guizhou Provincial Department of Education/Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region, Ministry of Education/Key Laboratory of Molecular Breeding for Grain and Oil Crops in Guizhou Province/College of Agricultural Sciences, Guizhou University, Guiyang 550025, China
²Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement/Guangdong Provincial Key Laboratory of Applied Botany/South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, China
³Institute of Fruit Tree Research, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China

Revised:2023-02-13 Accepted:2023-03-20 Online:2024-04-20 Published:2024-03-29
About author:#Correspondence Zhongming Fang, E-mail: zmfang@gzu.edu.cn; Mingyong Zhang, E-mail: zhangmy@scbg.ac.cn * These authors contributed equally to this study
Supported by:
We thank Chen Y and Miller A at John Innes Centre, UK, for assistance with the functional analysis of OsNPF3.1 in Xenopus laevis oocytes. This research was supported by the the Guizhou Provincial Excellent Young Talents Project of Science and Technology, China (YQK (2023) 002), the Guizhou Provincial Science and Technology Projects, China ((2022) Key 008), the Guizhou Provincial Science and Technology Support Plan, China ((2022) Key 026), the Key Laboratory of Molecular Breeding for Grain and Oil Crops in Guizhou Province, China ((2023) 008), and the Key Laboratory of Functional Agriculture of Guizhou Provincial Higher Education Institutions, China ((2023) 007).

摘要/Abstract

摘要：

低亲和硝酸盐转运基因成员已在水稻硝酸盐转运基因1/肽转运基因家族（NPF）的4-8亚家族中鉴定出来，但OsNPF3亚家族在硝酸盐和植物激素转运及水稻生长发育上还不清楚。本研究中，我们发现硝酸盐和植物激素转运基因OsNPF3.1在水稻分蘖和氮利用效率上起重要作用。OsNPF3.1的启动子序列在517个水稻品种中具有4种主要单倍型，其表达与分蘖数呈正相关。OsNPF3.1在水稻基部、茎和叶片中的表达量高于其他部位，且在水稻根部和地上部分被硝酸盐、脱落酸（ABA）和赤霉素3（GA3）强烈诱导表达。电生理实验表明，OsNPF3.1是一种pH依赖的低亲和硝酸盐转运基因，且水稻原生质体摄取实验表明它是ABA和GA3的转运基因。OsNPF3.1过表达后在高硝态氮浓度下显著促进了ABA在根系的积累和GA在基部的积累，进一步抑制了腋芽的伸长和水稻分蘖。在中低硝态氮浓度下OsNPF3.1的过表达植株氮利用效率增强，而在高硝态氮浓度下OsNPF3.1的突变体植株氮利用效率增加。以上结果表明，OsNPF3.1在不同硝态氮浓度下的不同水稻组织中转运硝酸盐和植物激素。OsNPF3.1表达改变的过表达植株或CRISPR植株分别在低硝酸盐和高硝酸盐浓度下提高氮利用效率。

Abstract:

Low-affinity nitrate transporter genes have been identified in subfamilies 4–8 of the rice nitrate transporter 1 (NRT1)/peptide transporter family (NPF), but the OsNPF3 subfamily responsible for nitrate and phytohormone transport and rice growth and development remains unknown. In this study, we described OsNPF3.1 as an essential nitrate and phytohormone transporter gene for rice tillering and nitrogen utilization efficiency (NUtE). OsNPF3.1 possesses four major haplotypes of its promoter sequence in 517 cultivars, and its expression is positively associated with tiller number. Its expression was higher in the basal part, culm, and leaf blade than in other parts of the plant, and was strongly induced by nitrate, abscisic acid (ABA) and gibberellin 3 (GA₃) in the root and shoot of rice. Electrophysiological experiments demonstrated that OsNPF3.1 is a pH-dependent low-affinity nitrate transporter, with rice protoplast uptake assays showing it to be an ABA and GA₃ transporter. OsNPF3.1 overexpression significantly promoted ABA accumulation in the roots and GA accumulation in the basal part of the plant which inhibited axillary bud outgrowth and rice tillering, especially at high nitrate concentrations. The NUtE of OsNPF3.1-overexpressing plants was enhanced under low and medium nitrate concentrations, whereas the NUtE of OsNPF3.1 clustered regularly interspaced short palindromic repeats (CRISPR) plants was increased under high nitrate concentrations. The results indicate that OsNPF3.1 transports nitrate and phytohormones in different rice tissues under different nitrate concentrations. The altered OsNPF3.1 expression improves NUtE in the OsNPF3.1-overexpressing and CRISPR lines at low and high nitrate concentrations, respectively.

Key words: rice tillering , grain yield , phytohormone , nitrate , transporter , nitrogen utilization efficiency

硝酸盐、脱落酸和赤霉素转运基因OsNPF3.1对水稻分蘖和氮利用效率至关重要 [J]. Journal of Integrative Agriculture, 2024, 23(4): 1087-1104.

Junnan Hang, Bowen Wu, Diyang Qiu, Guo Yang, Zhongming Fang, Mingyong Zhang.

OsNPF3.1, a nitrate, abscisic acid and gibberellin transporter gene, is essential for rice tillering and nitrogen utilization efficiency [J]. Journal of Integrative Agriculture, 2024, 23(4): 1087-1104.

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硝酸盐、脱落酸和赤霉素转运基因OsNPF3.1对水稻分蘖和氮利用效率至关重要

OsNPF3.1, a nitrate, abscisic acid and gibberellin transporter gene, is essential for rice tillering and nitrogen utilization efficiency

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