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华中农业大学李春龙团队揭示 CtrNAC019-CtrNPF2.1 模块调控枳耐盐性与氮素利用效率的双功能机制

  • 2026-06-04 04:00:25
华中农业大学李春龙团队揭示 CtrNAC019-CtrNPF2.1 模块调控枳耐盐性与氮素利用效率的双功能机制

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华中农业大学李春龙团队揭示 CtrNAC019-CtrNPF2.1 模块调控枳耐盐性与氮素利用效率的双功能机制

盐胁迫和氮肥过量施用是限制全球作物生产力的关键环境因子。氯离子(Cl-)是盐胁迫中毒性离子之一,在柑橘类作物中,氯离子而非钠离子是限制其耐盐性的主要因素。硝酸盐转运蛋白(NPF)家族成员参与硝酸盐和氯离子的跨膜运输,但NPF如何在液泡膜上协同调控氯离子区隔化和硝酸盐利用的分子机制尚不明确。NAC转录因子是植物逆境响应的核心调控因子,但其调控离子稳态和氮素利用的机制仍有待阐明。

华中农业大学李春龙团队以枳(Citrus trifoliata)为材料,通过NaCl/KCl处理联合转录组测序,从601个Cl-特异性响应差异基因中筛选到双响应基因CtrNPF2.1;经亚细胞定位、原位杂交证实其定位于根皮层细胞液泡膜。通过病毒诱导基因沉默(VIGS)、瞬时过表达、异源转化拟南芥发现,CtrNPF2.1 正调控耐盐性、促进根系生长并提升氮素同化效率。利用非洲爪蟾卵母细胞表达、双电极电压钳(TEVC)、¹⁵N 标记吸收与外排实验证实,CtrNPF2.1是双向Cl-转运蛋白、单向NO3-转运蛋白,介导液泡 Cl-区隔化与硝酸盐外排;通过双分子荧光互补(BiFC)荧光素酶互补成像(LCI)位点突变证明CtrNPF2.1可形成二聚体,TM3/TM6 界面负责Cl-转运。从转录组差异转录因子中,经酵母单杂交(Y1H)双荧光素酶报告实验(LUC)染色质免疫共沉淀- qPCR(ChIP-qPCR)凝胶迁移实验(EMSA)鉴定到CtrNAC019可直接结合 CtrNPF2.1 启动子 CGTG 基序并激活其转录;通过VIGS、过表达及回补实验证实,CtrNAC019 通过激活CtrNPF2.1增强耐盐性与氮素利用。最终通过盐硝酸盐互作处理验证,该模块介导盐增强氮素利用(SENU)与硝酸盐介导耐盐(NMST)的协同效应。

以上研究内容明确了 CtrNPF2.1 作为液泡膜双功能转运蛋白,通过区隔化 Cl⁻解毒、外排硝酸盐提升利用效率;解析了 CtrNAC019-CtrNPF2.1 模块的转录调控通路;阐明了盐胁迫与高氮条件下,该模块协同调控离子稳态与氮素代谢的分子机制,为培育耐盐、氮高效作物提供核心靶点。

Plant Biotechnology Journal., 18 May 2026

The Dual-Function of CtrNAC019-CtrNPF2.1 Module in Salt Tolerance and Nitrogen Use Efficiency Via Enhancing Vacuolar Chloride Sequestration and Nitrate Efflux in Citrus trifoliata

Author

Zeqi Zhao, Chengwei Yang, Xiangming Shang, Mengdi Li, Xiaoyang Su, Ji-Hong Liu, Chunlong Li*.

*:National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops,College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan, China.

Abstract

Salt stress and nitrogen utilisation efficiency (NUE) represent critical constraints affecting worldwide crop productivity. While nitrate transporter proteins (NPFs) have been implicated in saline chloride (Cl−) ion transport, the mechanistic linkage between chloride stress and NUE remains poorly understood. Through comparative transcriptomic profiling of Citrus trifoliata, we identified CtrNPF2.1 as a dual-responsive gene significantly upregulated in root tissues under both chloride salt and high nitrate conditions. The CtrNPF2.1 protein is localized to the vacuole membrane and highly expressed in root cortical cells. Transport functional characterisation combined with transgenic phenotypic analyses established that CtrNPF2.1 mediates dual transport mechanisms: facilitating vacuolar Cl− compartmentalisation while enabling nitrate efflux. This coordinated process enhances cellular ion detoxification and promotes nitrate redistribution for optimal root development under saline conditions. Molecular investigations further uncovered that CtrNAC019 transcriptionally regulates CtrNPF2.1 expression through direct binding to its promoter, forming a chloride/nitrate-responsive regulatory module. Notably, physiological validation reveals coordinated responses in which nitrate supplementation mitigates chloride toxicity while chloride availability enhances nitrogen utilisation in trifoliate orange, which may be explained by the CtrNAC019-CtrNPF2.1 expression regulation module. These results provide new insights into vacuolar chloride sequestration and nitrate efflux by CtrNAC019-CtrNPF2.1 under saline and high-nitrate conditions, offering a promising strategy for breeding salt-resilient and resource-efficient crops.

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  1. CONNECT:[ UseTime:0.000356s ] mysql:host=127.0.0.1;port=3306;dbname=h_mffb;charset=utf8mb4
  2. SHOW FULL COLUMNS FROM `fenlei` [ RunTime:0.000642s ]
  3. SELECT * FROM `fenlei` WHERE `fid` = 0 [ RunTime:0.001373s ]
  4. SELECT * FROM `fenlei` WHERE `fid` = 63 [ RunTime:0.000281s ]
  5. SHOW FULL COLUMNS FROM `set` [ RunTime:0.000482s ]
  6. SELECT * FROM `set` [ RunTime:0.000196s ]
  7. SHOW FULL COLUMNS FROM `article` [ RunTime:0.000498s ]
  8. SELECT * FROM `article` WHERE `id` = 536765 LIMIT 1 [ RunTime:0.000414s ]
  9. UPDATE `article` SET `lasttime` = 1781225542 WHERE `id` = 536765 [ RunTime:0.002195s ]
  10. SELECT * FROM `fenlei` WHERE `id` = 64 LIMIT 1 [ RunTime:0.000272s ]
  11. SELECT * FROM `article` WHERE `id` < 536765 ORDER BY `id` DESC LIMIT 1 [ RunTime:0.000406s ]
  12. SELECT * FROM `article` WHERE `id` > 536765 ORDER BY `id` ASC LIMIT 1 [ RunTime:0.000944s ]
  13. SELECT * FROM `article` WHERE `id` < 536765 ORDER BY `id` DESC LIMIT 10 [ RunTime:0.000754s ]
  14. SELECT * FROM `article` WHERE `id` < 536765 ORDER BY `id` DESC LIMIT 10,10 [ RunTime:0.003344s ]
  15. SELECT * FROM `article` WHERE `id` < 536765 ORDER BY `id` DESC LIMIT 20,10 [ RunTime:0.002099s ]
0.089005s