Cheng Rong, Xiao Qiang, Gong Jie, Sun Renwei, Du Yinke, Zhao Wei, Zheng Wei, Gao Shiqing
College of Grassland Science, Xinjiang Agricultural University, 830052 Urumqi, Xinjiang, China.
Institute of Hybrid Wheat, Beijing Academy of Agriculture and Forestry Sciences, 100097 Beijing, China.
Front Biosci (Landmark Ed). 2025 Jul 29;30(7):39679. doi: 10.31083/FBL39679.
Nitrate transporter NRT1/PTR family (NPF) proteins are crucial for plant nitrogen uptake and utilization. As an important hexaploid crop for grain and forage, oat ( L.) requires substantial levels of nitrogen. However, the oat nitrate transporter 1 (NRT1) family remains uncharacterized.
In this study, the oat subfamily members were identified through the Hmm and Pfam databases. Bioinformatics analysis was performed using the MEGA 11 and TBtools software to elucidate the physicochemical properties, evolutionary relationships, chromosomal localization, and gene structures. Furthermore, quantitative real-time polymerase chain reaction (qRT-PCR) analysis and the green fluorescent protein (GFP) fusion expression vector were utilized to investigate the candidate oat .
Phylogenetic classification categorized oat into eight subfamilies, with the most abundant being the NPF5 subfamily. Physicochemical property analysis revealed that the number of amino acids in the proteins encoded by these genes ranged from 235 to 673, with their molecular weights (MWs) ranging from 26 kDa to 74 kDa. Chromosomal localization revealed that these genes were unevenly distributed across all 12 oat chromosomes. Promoter analysis revealed that light-responsive elements appeared most frequently in the promoters of these genes (39.3%), followed by abscisic acid (ABA)-responsive elements (13.5%) and methyl jasmonate (MeJA)-responsive elements (9.4%). qRT-PCR analysis revealed that most of the genes exhibited tissue-specific expression patterns. Among them, was highly expressed in the leaves at 1 h post-low nitrogen (LN) treatment, while was highly expressed in the leaves at 12 h. Both these genes exhibited low expression levels in the roots. However, and were both highly expressed in the roots at 9 h post-LN treatment but exhibited low expression in the leaves. Subcellular localization revealed that all five proteins (AsNPF2.6, AsNPF4.5, AsNPF7.16, AsNPF6.8, and AsNPF7.19) were localized to the cytoplasm and cell membrane.
Our results demostrate the involvement of family members in nitrogen transport in oat, providing theoretical support for further investigation into the functions and molecular mechanisms of action of oat in nitrogen transport.
硝酸盐转运蛋白NRT1/PTR家族（NPF）蛋白对植物氮素吸收和利用至关重要。燕麦（ ）作为一种重要的粮饲兼用六倍体作物，需要大量的氮。然而，燕麦硝酸盐转运蛋白1（NRT1）家族仍未被鉴定。
在本研究中，通过Hmm和Pfam数据库鉴定燕麦 亚家族成员。使用MEGA 11和TBtools软件进行生物信息学分析，以阐明其理化性质、进化关系、染色体定位和基因结构。此外，利用实时定量聚合酶链反应（qRT-PCR）分析和绿色荧光蛋白（GFP）融合表达载体对候选燕麦 进行研究。
系统发育分类将燕麦 分为八个亚家族，其中NPF5亚家族最为丰富。理化性质分析表明，这些基因编码的蛋白质氨基酸数量在235至673之间，分子量（MW）在26 kDa至74 kDa之间。染色体定位表明，这些基因在燕麦的所有12条染色体上分布不均。启动子分析表明，光响应元件在这些基因的启动子中出现频率最高（39.3%），其次是脱落酸（ABA）响应元件（13.5%）和茉莉酸甲酯（MeJA）响应元件（9.4%）。qRT-PCR分析表明，大多数基因表现出组织特异性表达模式。其中， 在低氮（LN）处理后1小时在叶片中高表达，而 在12小时在叶片中高表达。这两个基因在根中均表现出低表达水平。然而， 和 在LN处理后9小时在根中均高表达，但在叶片中低表达。亚细胞定位表明，所有五种蛋白质（AsNPF2.6、AsNPF4.5、AsNPF7.16、AsNPF6.8和AsNPF7.19）均定位于细胞质和细胞膜。
我们的结果证明了 家族成员参与燕麦中的氮转运，为进一步研究燕麦 在氮转运中的功能和分子作用机制提供了理论支持。
