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作物杂志,2026, 第5期: 32–41 doi: 10.16035/j.issn.1001-7283.2026.05.004

• 植物激素与作物生长发育调控专题 • 上一篇    下一篇

转录组分析揭示小麦硝酸盐转运蛋白TaNPF6.2整合激素通路调控小麦发育

郭诗涵1,2(), 周佰慧2,3, 孔秀英2, 张立超2, 夏川2(), 倪志勇1()   

  1. 1 新疆农业大学生命科学学院极端环境生物生态适应与进化重点实验室， 830052， 新疆乌鲁木齐
    2 中国农业科学院作物科学研究所/作物基因资源与育种全国重点实验室， 100081， 北京
    3 安徽农业大学农学院， 230036， 安徽合肥
  • 收稿日期:2025-12-19 修回日期:2026-02-11 出版日期:2026-10-15 发布日期:2026-10-22
  • 通讯作者: 夏川，主要从事小麦重要农艺性状基因挖掘研究，E-mail：xiachuan@caas.cn；倪志勇为共同通信作者，主要从事生态适应与进化研究，E-mail：nzy@xjau.edu.cn
  • 作者简介:郭诗涵，主要从事小麦分蘖调控基因研究，E-mail：2175891933@qq.com
  • 基金资助:
    国家重点研发计划(2023YFF1000603);中国农业科学院创新工程项目

Transcriptome Analysis Reveals Wheat Nitrate Transporter TaNPF6.2 Integrates Hormone Pathways to Regulate Development

Guo Shihan1,2(), Zhou Baihui2,3, Kong Xiuying2, Zhang Lichao2, Xia Chuan2(), Ni Zhiyong1()   

  1. 1 Xinjiang Key Laboratory for Ecological Adaptation and Evolution of Extreme Environment Organisms, College of Life Sciences, Xinjiang Agricultural University, Urumqi 830052, Xinjiang, China
    2 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences / State Key Laboratory of Crop Gene Resources and Breeding, Beijing 100081, China
    3 School of Agronomy, Anhui Agricultural University, Hefei 230036, Anhui, China
  • Received:2025-12-19 Revised:2026-02-11 Online:2026-10-15 Published:2026-10-22

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摘要：

硝酸盐转运蛋白是介导植物氮素吸收的特异性膜蛋白，与氮素利用效率密切相关。本研究鉴定了小麦硝酸盐转运蛋白TaNPF6.2基因编辑（KO）株系的基因型。转录组比较发现，野生型与KO株系之间存在1981个差异表达基因（DEGs），GO富集分析显示上述基因显著富集于酰基转移酶活性及脂肪酸生物合成等过程。内源激素检测表明，KO株系中水杨酸和脱落酸的含量显著上升，且其合成与信号转导相关基因的表达也同步上调。结果表明，TaNPF6.2可能通过影响水杨酸和脱落酸介导的信号通路，参与小麦硝酸盐信号的感知与转导。其功能缺失激活了逆境相关激素信号，进而导致分蘖数、穗长和株高等农艺性状的变化。该研究为阐明小麦氮素响应机制和培育氮高效小麦品种提供了理论依据。

关键词: 小麦, 硝酸盐转运蛋白, 转录组, 激素

Abstract:

Nitrate transporters are specific membrane proteins that mediate nitrogen uptake in plants and are closely related to nitrogen use efficiency. In this study, the genotypes of gene-edited knockout (KO) lines of the wheat nitrate transporter TaNPF6.2 were identified. Transcriptome comparison revealed 1981 differentially expressed genes (DEGs) between the wild-type and KO lines, and GO enrichment analysis showed that these genes were significantly enriched in acyltransferase activity and fatty acid biosynthesis. Endogenous hormone detection indicated that the contents of salicylic acid and abscisic acid significantly increased in the KO lines, and the expression of genes related to their synthesis and signal transduction was also synchronously upregulated. These results suggested that TaNPF6.2 may participate in the perception and transduction of nitrate signaling in wheat by affecting salicylic acid- and abscisic acid-mediated signaling pathways. Loss of TaNPF6.2 function activated stress-related hormone signaling, thereby leading to changes in agronomic traits such as tiller number, spike length, and plant height. This study provided a theoretical basis for elucidating nitrogen response mechanisms and breeding nitrogen-efficient wheat varieties.

Key words: Wheat, Nitrate transporter (NPF), Transcriptome, Hormone

表1

TaNPF6.2基因特异性扩增引物

项目
Item		 正向特异性扩增引物
Forward specific amplification primer (5′-3′)		 反向特异性扩增引物
Reverse specific amplification primer (5′-3′)
A基因组A subgenome GGCGCGCTTGAAAATTCTCT CTGCTCGGTTTCCCTCCTTT
B基因组B subgenome TTCTCCTTTGCTTCCTCGGC CCAAGACTCCTGACTCGCTC
D基因组D subgenome CCCCCATCTCAGCTACCAAC CATGCCGATGATGGACAGGA

表2

qRT-PCR引物序列

基因ID Gene ID 基因名称Gene name 正向引物Forward primer (5′-3′) 反向引物Reverse primer (5′-3′)
TraesCS5B02G029300 NCED3 TCCGCGCAGCCCCTGGCCT ACACGTAGGCCGGCGTGCGT
TraesCS5B02G075800 SnRK2 ATTCGAAGTCATCGGTTCTTCACT CAACATCACGTAAAGGGTTACCC
TraesFLD5B01G508800 PAL GCCGAGGCCATTGATATAC ATCTGCATACGCGAACAC
TraesFLD7B01G073800 PR-10 CTGTTGAGGGAGAAGGCGG CGATGAGGGTCGATTTGCAC
TraesCS6D02G196300 TaGAPDH CGATTCAGAGCAGCGTATTGTTG AGTTGGTCGGGTCTCTTCTAAATG

图1

敲除材料KOTaNPF6.2的基因型鉴定

表3

测序数据质量统计

样本
Sample		 文库
Library		 原始读数
Raw reads		 原始碱基数
Raw bases (Gb)		 有效读数
Clean reads		 有效碱基数
Clean bases (Gb)		 错误率
Error rate (%)		 Q20值
Q20 value (%)		 Q30值
Q30 value (%)		 GC含量
GC content (%)
WT-1 FRAS250185109-1r 84631508 12.69 82192894 12.33 0.01 99.18 96.35 53.48
WT-2 FRAS250185110-1r 84659812 12.70 82036334 12.31 0.01 99.13 96.28 53.49
WT-3 FRAS250185111-1r 84415958 12.66 81516152 12.23 0.01 99.17 96.46 53.31
KO-1 FRAS250185112-1r 81627914 12.24 79531446 11.93 0.01 99.12 96.24 52.95
KO-2 FRAS250185113-1r 78440374 11.77 75364236 11.3 0.01 99.11 96.27 53.29
KO-3 FRAS250185114-1r 82075428 12.31 78843794 11.83 0.01 99.11 96.19 53.29

图2

差异表达分析和DEGs聚类分析

图3

DEGs的GO富集分析

表4

差异基因GO富集分析

类别Category GO编号GO code 描述Description Padj 数量Number
MF 0016746 转移酶活性（转移酰基） 0.0204451782464127 6
MF 0004867 丝氨酸型内肽酶抑制剂活性 0.0204451782464127 3
MF 0016747 转移酶活性（转移非氨酰基的酰基） 0.0204451782464127 5
BP 0006633 脂肪酸生物合成过程 0.1037956007872910 3
MF 0004866 内肽酶抑制剂活性 0.0226228547401966 3
MF 0030414 肽酶抑制剂活性 0.0226228547401966 3
MF 0061134 肽酶调节因子活性 0.0226228547401966 3
MF 0061135 内肽酶调节因子活性 0.0226228547401966 3

图4

DEGs的KEGG富集分析

表5

苗期内源激素含量测定

化合物
Compound		 分蘖节Crown node 茎Culm
WT KOTaNPF6.2 WT KOTaNPF6.2
SA 122.98±4.27 332.79±4.27 98.90±3.62 276.30±4.53
SA-2-O-β-葡萄糖苷SA-2-O-β-Glucoside 374.09±13.32 823.27±24.43 234.79±9.69 831.44±29.14
ABA 41.25±0.87 44.72±1.19 27.91±1.00 31.36±1.58
反式肉桂酸Trans-cinnamic acid 23.50±0.44 22.21±0.55 24.73±0.55 20.90±0.09
IAA-葡萄糖酯IAA-Glc 580.29±13.45 379.92±5.76 1138.49±28.71 1249.10±16.68
IAA-天冬氨酸IAA-Asp 1366.40±40.37 1534.95±34.73 785.02±22.54 1009.47±27.95
JA 1.31±0.03 1.08±0.03 5.82±0.34 7.45±0.22
顺(+)-12-氧代-植物二烯酸2-oxo-phytodienoic acid 69.38±3.51 86.77±2.37 304.57±2.35 355.81±10.36

图5

激素相关基因的qRT-PCR分析 “*”、“**”和“***”分别表示在P < 0.05、P < 0.01和P < 0.001水平差异显著。

[1] Fredes I, Moreno S, Díaz F P, et al. Nitrate signaling and the control of Arabidopsis growth and development. Current Opinion in Plant Biology, 2019, 47:112-118.
doi: 10.1016/j.pbi.2018.10.004
[2] 田纪春, 张忠义. 高蛋白和低蛋白小麦品种的氮素吸收和运转分配差异的研究. 作物学报, 1994, 20(1):76-83.
[3] 轩红梅, 王永华, 魏利婷, 等. 小麦幼苗叶片中硝酸盐转运蛋白NRT1和NRT2家族基因对氮饥饿响应的表达分析. 麦类作物学报, 2014, 34(8):1019-1028.
[4] 宋可欣, 王丽霞, 刘永霞, 等. 根际微生态环境中氮源转化及循环研究进展. 中国农学通报, 2024, 40(21):91-98.
doi: 10.11924/j.issn.1000-6850.casb2023-0716
[5] 白文钦, 胡明瑜, 王春萍, 等. 氮素在植物中的利用综述. 江苏农业科学, 2020, 48(4):1-11.
[6] 孙磊, 王弘, 李明月, 等. 氮磷钾肥施用量及施用时期对马铃薯干物质积累与分配的影响. 作物杂志, 2014(1):132-137.
[7] 于泳, 黄瑞冬, 赵尚文, 等. 不同施氮水平对高粱子粒淀粉积累规律的影响. 作物杂志, 2008(5):20-24.
[8] Li H, Hu B, Chu C C. Nitrogen use efficiency in crops: Lessons from Arabidopsis and rice. Journal of Experimental Botany, 2017, 68(10):2477-2488.
doi: 10.1093/jxb/erx101
[9] 金宏鑫, 裴占江, 李淑芹, 等. 污泥生物有机肥对大豆产量和氮磷吸收的影响. 作物杂志, 2012(1):92-95.
[10] 韦剑锋, 罗艺, 米超, 等. 应用15N标记对不同施氮方式甘蔗氮肥利用效率的研究. 作物杂志, 2012(1):76-79.
[11] Lamig L, Moreno S, Álvarez J M, et al. Molecular mechanisms underlying nitrate responses in plants. Current Biology, 2022, 32(9):433-439.
[12] 陈范骏, 米国华, 张福锁. 氮高效玉米新品种中农99的选育. 作物杂志, 2009(6):103-104.
[13] 陈颖, 周振翔, 周天阳, 等. 水稻氮高效吸收利用机制及栽培调控措施. 作物杂志, 2016(6):26-32.
[14] Schroeder J I, Delhaize E, Frommer W B, et al. Using membrane transporters to improve crops for sustainable food production. Nature, 2013, 497(7447):60-66.
doi: 10.1038/nature11909
[15] Xia X D, Fan X R, Wei J, et al. Rice nitrate transporter OsNPF2.4 functions in low-affinity acquisition and long-distance transport. Journal of Experimental Botany, 2015, 66(1):317-331.
doi: 10.1093/jxb/eru425 pmid: 25332358
[16] Li Y G, Ouyang J, Wang Y Y, et al. Disruption of the rice nitrate transporter OsNPF2.2 hinders root-to-shoot nitrate transport and vascular development. Scientific Reports, 2015, 5:9635.
doi: 10.1038/srep09635 pmid: 25923512
[17] Wei J, Zheng Y, Feng H M, et al. OsNRT2.4 encodes a dual- affinity nitrate transporter and functions in nitrate-regulated root growth and nitrate distribution in rice. Journal of Experimental Botany, 2018, 69(5):1095-1107.
doi: 10.1093/jxb/erx486
[18] Guan Y, Liu D F, Qiu J, et al. The nitrate transporter OsNPF7.9 mediates nitrate allocation and the divergent nitrate use efficiency between indica and japonica rice. Plant Physiology, 2022, 189(1):215-229.
doi: 10.1093/plphys/kiac044
[19] Hu B, Wang W, Ou S J, et al. Variation in NRT1.1B contributes to nitrate-use divergence between rice subspecies. Nature Genetics, 2015, 47(7):834-838.
doi: 10.1038/ng.3337 pmid: 26053497
[20] Ma X J, Wang W, Zhang J Y, et al. NRT1.1B acts as an abscisic acid receptor in integrating compound environmental cues for plants. Cell, 2025, 188(19):5231-5248.
doi: 10.1016/j.cell.2025.07.027 pmid: 40795855
[21] Jia L H, Hu D S, Wang J B, et al. Genome-wide identification and functional analysis of nitrate transporter genes (NPF, NRT2 and NRT3) in maize. International Journal of Molecular Sciences, 2023, 24(16):12941.
doi: 10.3390/ijms241612941
[22] Cao H R, Liu Z, Guo J, et al. ZmNRT1.1B (ZmNPF6.6) determines nitrogen use efficiency via regulation of nitrate transport and signalling in maize. Plant Biotechnology Journal, 2023, 22(2):316-329.
doi: 10.1111/pbi.v22.2
[23] Buchner P, Hawkesford M J. Complex phylogeny and gene expression patterns of members of the NITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER family (NPF) in wheat. Journal of Experimental Botany, 2014, 65(19):5697-5710.
doi: 10.1093/jxb/eru231 pmid: 24913625
[24] Kumar A, Sandhu N, Kumar P, et al. Genome-wide identification and in silico analysis of NPF, NRT2, CLC and SLAC1/SLAH nitrate transporters in hexaploid wheat (Triticum aestivum). Scientific Reports, 2022, 12(1):11227.
doi: 10.1038/s41598-022-15202-w pmid: 35781289
[25] Wang H H, Liu Y Y, Wu L F, et al. TaNPF6.2 improves agronomic traits via enhancing nitrogen uptake efficiency in wheat. Journal of Genetics and Genomics, 2025, 52(1):120-123.
doi: 10.1016/j.jgg.2024.11.004
[26] Krouk G, Lacombe B, Bielach A, et al. Nitrate-regulated auxin transport by NRT1.1 defines a mechanism for nutrient sensing in plants. Developmental Cell, 2010, 18(6):927-937.
doi: 10.1016/j.devcel.2010.05.008 pmid: 20627075
[27] Kanno Y, Kamiya Y, Seo M. Nitrate does not compete with abscisic acid as a substrate of AtNPF4.6/NRT1.2/AIT1 in Arabidopsis. Plant Signaling & Behavior, 2013, 8(12):e26624.
[28] Bouguyon E, Brun F, Meynard D, et al. Multiple mechanisms of nitrate sensing by Arabidopsis nitrate transceptor NRT1.1. Nature Plants, 2015, 1:15015.
doi: 10.1038/nplants.2015.15 pmid: 27246882
[29] Jayawardhane K N, Singer S D, Weselake R J, et al. Plant sn-glycerol-3-phosphate acyltransferases: biocatalysts involved in the biosynthesis of intracellular and extracellular lipids. Lipids, 2018, 53(5):469-480.
doi: 10.1002/lipd.12049 pmid: 29989678
[30] Wu C J, Yuan D Y, Liu Z Z, et al. Conserved and plant-specific histone acetyltransferase complexes cooperate to regulate gene transcription and plant development. Nature Plants, 2023, 9(3):442-459.
doi: 10.1038/s41477-023-01359-3
[31] Mugume Y, Ding G, Dueñas M E, et al. Complex changes in membrane lipids associated with the modification of autophagy in Arabidopsis. Metabolites, 2022, 12(2):190.
doi: 10.3390/metabo12020190
[32] Gaude N, Bréhélin C, Tischendorf G, et al. Nitrogen deficiency in Arabidopsis affects galactolipid composition and gene expression and results in accumulation of fatty acid phytyl esters. The Plant Journal, 2007, 49(4):729-739.
doi: 10.1111/tpj.2007.49.issue-4
[33] Sathee L, R S, Barman D, et al. Nitrogen at the crossroads of light: integration of light signalling and plant nitrogen metabolism. Journal of Experimental Botany, 2025, 76(3):803-818.
doi: 10.1093/jxb/erae437 pmid: 39540633
[34] Liu H, Gao X, Fan W, et al. Optimizing carbon and nitrogen metabolism in plants: from fundamental principles to practical applications. Journal of Integrative Plant Biology, 2025, 67(6):1447-1466.
doi: 10.1111/jipb.13919
[35] Persyn F, Smagghe W, Eeckhout D, et al. A nitrogen-specific interactome analysis sheds light on the role of the SnRK1 and TOR kinases in plant nitrogen signaling. Molecular & Cellular Proteomics, 2024, 23(10):100842.
doi: 10.1016/j.mcpro.2024.100842
[36] Zhu B, Zhang Y, Gao R, et al. Complete biosynthesis of salicylic acid from phenylalanine in plants. Nature, 2025, 645(8079):218-227.
doi: 10.1038/s41586-025-09175-9
[37] Hwang S H, Lee I A, Yie S W, et al. Identification of an OsPR10a promoter region responsive to salicylic acid. Planta, 2008, 227(5):1141-1150.
doi: 10.1007/s00425-007-0687-8
[38] Virlouvet L, Fromm M. Physiological and transcriptional memory in guard cells during repetitive dehydration stress. New Phytologist, 2015, 205(2):596-607.
doi: 10.1111/nph.13080 pmid: 25345749
[39] Tan W R, Zhang D W, Zhou H P, et al. Transcription factor HAT1 is a substrate of SnRK2.3 kinase and negatively regulates ABA synthesis and signaling in Arabidopsis responding to drought. PLoS Genetics, 2018, 14(4):e1007336.
doi: 10.1371/journal.pgen.1007336
[40] Hsu P K, Takahashi Y, Munemasa S, et al. Abscisic acid-independent stomatal CO2 signal transduction pathway and convergence of CO2 and ABA signaling downstream of OST1 kinase. Proceedings of the National Academy of Sciences of the United States of America, 2018, 115(42):9971-9980.
[41] Dixon L E, Greenwood J R, Bencivenga S, et al. TEOSINTE BRANCHED1 regulates inflorescence architecture and development in bread wheat (Triticum aestivum). The Plant Cell, 2018, 30(3):563-581.
doi: 10.1105/tpc.17.00961
[42] Dixon L E, Pasquariello M, Boden S A. TEOSINTE BRANCHED1 regulates height and stem internode length in bread wheat. Journal of Experimental Botany, 2020, 71(16):4742-4750.
doi: 10.1093/jxb/eraa252
[43] Lewis J M, Mackintosh C A, Shin S, et al. Overexpression of the maize Teosinte Branched1 gene in wheat suppresses tiller development. Plant Cell Reports, 2008, 27(7):1217-1225.
doi: 10.1007/s00299-008-0543-8
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