陆地棉硝酸盐转运体NRT基因家族鉴定及表达分析

doi:10.3724/SP.J.1006.2023.24159

Abstract

Abstract:

Nitrate transporters (NRTs) play an important role in plant nitrogen absorption, utilization, and storage. In this study, 106 GhNRT1/PTR (NPF) (Nitrate transporter 1 (NRT1)/Peptide Transporter (PTR) family (NPF)) and 14 GhNRT2 (Nitrate transporter 2 family) were identified from Gossypium hirsutum L. (TM-1) by HMM software and Blastp method. The conserved domains, phylogenetic relationships, physicochemical properties, subcellular localization, conserved motifs, gene structure, promoter cis-acting elements, and expression patterns of these GhNRTs were analyzed. The results showed that GhNPF had a typical PTR2 (Peptide Transporter 2 family) domain. Two PTR2 and/or other domains were found in individual proteins (GhNPF2.6bD, GhNPF4.1cA, and GhNPF2.14aD), indicating that GhNPF was less evolutionarily conserved. The GhNRT2 had a typical MFS_1 (Major Facilitator Superfamily) domain. Most proteins were located on the cytoplasmic membrane with hydrophobic properties. Phylogenetic analysis showed that these GhNRTs could be divided into 10 groups, and the same group had similar gene structure and motif distribution. The composition of cis-acting elements indicated that the relative expression levels of most GhNRTs could be related to plant hormones, abiotic stress, and light response. In addition, the relative expression patterns of GhNPF were different among the diverse subgroups, but the relative expression patterns of different members in the same subgroup were mostly conserved. GhNRT2 genes were mainly expressed in roots. Moreover, the transcriptome data with salt stress treatment revealed that the relative levels of nearly 1/5 GhNRTs were significantly up-regulated or down-regulated, indicating that they probably function in response to salt stress. Six GhNRTs were selected to detect the response of their expression in roots, young leaves, functional leaves, and old leaves to different NO₃^- supply levels. The results showed that GhNPF6.3dA and GhNPF7.3aA may have the ability to absorb NO₃^- with dual affinity, while GhNPF6.2bD may encode high-affinity NO₃^- transporter. The three may be involved in NO₃^- unloading in functional leaves and old leaves. These results were different from those reported in plants such as Arabidopsis. In conclusion, the results provide a reference for further functional characterization of nitrate transporters and provide a preliminary basis for the mechanism analysis of nitrogen absorption and utilization in cotton.

Key words: Gossypium hirsutum L., nitrate transporters, GhNPF, GhNRT2, expression pattern

MA Chun-Min, LI Wei-Xi, LI Fang-Jun, TIAN Xiao-Li, LI Zhao-Hu. Identification and expression analysis of nitrate transporter NRT gene family in upland cotton (Gossypium hirsutum L.)[J].Acta Agronomica Sinica, 2023, 49(6): 1496-1517.

Figures/Tables 12

Table 1

Fig. 1

Fig. 2

Table 2

Physicochemical properties and subcellular localization prediction of NRT proteins in Gossypium hirsutum L."

基因名称 Gene ID	分子质量 Molecular weight (kD)	等电点 pI	氨基酸数 Number of amino acids	亲水性 Grand average of hydropathicity	脂肪指数 Aliphatic index	不稳定系数Instability index	亚细胞定位 Subcellular localization
GhNPF1.1cD	63.711	9.13	579	0.352	101.93	43.69	plas
GhNPF1.2aA	66.070	8.65	603	0.342	100.15	46.12	plas
GhNPF2.6bA	107.252	8.60	982	0.521	114.58	35.36	plas
GhNPF2.6bD	122.915	9.21	1118	0.438	110.49	37.97	plas
GhNPF2.6aA	61.214	8.61	558	0.369	107.85	34.68	plas
GhNPF2.11aD	66.821	8.91	602	0.200	96.71	30.10	plas
GhNPF2.14aD	93.651	6.76	844	0.302	103.83	39.71	plas
GhNPF2.14bD	65.256	7.88	588	0.353	104.25	35.93	plas
GhNPF3.1dA	54.147	8.45	491	0.281	106.42	37.18	plas
GhNPF3.1aD	64.861	8.48	586	0.220	100.55	30.67	plas
GhNPF3.1cD	65.818	8.37	593	0.210	100.35	32.67	plas
GhNPF4.1aA	57.776	8.52	519	0.327	96.99	34.86	plas
GhNPF4.3bD	65.985	8.46	596	0.255	97.53	35.11	plas
GhNPF4.4aD	64.965	7.52	587	0.341	98.13	37.19	plas
GhNPF4.1cA	119.341	8.99	1078	0.362	102.18	29.89	plas
GhNPF4.6bD	64.247	9.16	582	0.339	102.94	43.31	plas
GhNPF4.6cD	63.787	8.54	580	0.356	101.43	42.33	plas
GhNPF4.6aA	56.049	9.33	511	0.448	102.90	39.84	plas
GhNPF4.7aA	57.238	8.89	519	0.385	97.17	42.30	plas
GhNPF5.2aD	66.315	9.16	594	0.243	98.15	28.95	plas
GhNPF5.4bD	63.908	9.06	578	0.274	102.04	30.58	plas
GhNPF5.7aD	66.067	8.60	594	0.276	106.72	35.40	plas
GhNPF5.9aA	60.359	7.86	542	0.367	101.62	37.62	plas
GhNPF5.10aA	62.338	8.76	571	0.411	104.31	34.02	plas
GhNPF5.10dA	64.148	9.22	579	0.149	91.16	36.18	plas
GhNPF6.1aA	70.503	8.91	634	0.290	99.54	39.06	plas
GhNPF6.2bD	63.343	9.33	582	0.375	98.04	29.66	plas
GhNPF6.3bA	64.680	8.91	586	0.317	105.00	35.83	plas
GhNPF6.4bA	65.734	8.95	595	0.277	106.00	24.31	plas
GhNPF7.1bA	65.353	5.96	598	0.315	99.60	26.46	plas
GhNPF7.2aD	66.711	6.47	600	0.216	94.93	27.77	plas
GhNPF7.3aA	66.570	6.24	599	0.207	95.73	29.23	plas
GhNPF8.1aA	63.838	8.60	569	0.200	96.98	23.26	plas
GhNPF8.2cA	64.192	7.89	576	0.152	93.18	21.88	plas
GhNPF8.3bA	64.679	5.97	584	0.248	93.89	30.46	plas
GhNRT2.4bD	57.456	9.31	530	0.345	87.28	41.81	plas
GhNRT2.4aA	57.526	8.82	532	0.331	88.05	41.58	plas
GhNRT2.3aD	57.938	9.21	536	0.392	86.51	32.60	plas
GhNRT2.2aA	56.878	8.59	526	0.319	87.60	35.91	plas
GhNRT2.5aD	54.632	9.15	506	0.457	94.68	38.68	plas

Table 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

Fig. 7

Fig. 8

Fig. 9

Table S1

Summary of identified substrates for NRT transporters in Arabidopsis thaliana and Oryza sativa"

物种 Species	名称 Name	先前名称 Old name	基因名称 Gene ID	底物 Substrates	参考文献 Reference(s)
拟南芥 Arabidopsis thaliana	AtNPF1.1	NRT1.12	At3g16180	NO₃^- (oocyte); ABA, GA, JA-Ile (yeast)	[1-2]
	AtNPF1.2	NRT1.11	At1g52190	NO₃^- (oocyte); GA, JA-Ile (yeast)	[1-2]
	AtNPF2.3		At3g45680	NO₃^- (liposome); GA (yeast)	[2-3]
	AtNPF2.4		At3g45700	GA, JA-Ile (yeast); Cl^- (oocyte)	[2,4]
	AtNPF2.5		At3g45710	ABA, GA (yeast); Cl^- (oocyte, yeast)	[2,5]
	AtNPF2.6		At3g45660	GA, JA-Ile (yeast)	[2]
	AtNPF2.7	NAXT1	At3g45650	NO₃^- (liposome); GA, JA-Ile (yeast)	[2,6]
	AtNPF2.9	NRT1.9	At1g18880	NO₃^-, 4 MTB (oocyte)	[7-8]
	AtNPF2.10	GTR1	At3g47960	NO₃^-, GA, JA-Ile, 4 MTB, 8 MTO (oocyte); GA, JA-Ile (yeast)	[2,7,9-11]
	AtNPF2.11	GTR2	At5g62680	4 MTB, 8 MTO, NO₃^-, GA (oocyte)	[7,9,11]
	AtNPF2.12	NRT1.6	At1g27080	NO₃^- (oocyte); GA (yeast)	[2,12]
	AtNPF2.13	NRT1.7	At1g69870	NO₃^-, 4 MTB (oocyte); GA, JA-Ile (yeast)	[2,7,13]
	AtNPF2.14		At1g69860	4 MTB (oocyte)	[7]
	AtNPF3.1	Nitr	At1g68570	NO₃^-, NO₂^-, GA (oocyte); GA, and JA-Ile (yeast)	[2,14-15]
	AtNPF4.1	AIT3	At3g25260	ABA, GA, JA-Ile (yeast); GA (oocyte)	[2,11,16]
	AtNPF4.2	AIT4	At3g25280	ABA, GA (yeast)	[2,16]
	AtNPF4.5	AIT2	At1g27040	ABA (yeast)	[16]
	AtNPF4.6	NRT1.2/AIT1	At1g69850	NO₃^- (oocyte); ABA (yeast, insect cell)	[2,16-18]
	AtNPF5.1		At2g40460	ABA, GA, JA-Ile (yeast)	[2]
	AtNPF5.2	PTR3	At5g46050	Dipeptides (yeast); ABA, GA (yeast)	[2,19-20]
	AtNPF5.3		At5g46040	ABA (yeast)	[2]
	AtNPF5.5		At2g38100	NO₃^-(oocyte)	[21]
	AtNPF5.6		At2g37900	GA (yeast)	[2]
	AtNPF5.7		At3g53960	ABA, GA, JA-Ile (yeast)	[2]
	AtNPF5.10		At1g22540	NO₃^- (oocyte)	[21]
	AtNPF5.11		At1g72130	NO₃^- (oocyte)	[22]
	AtNPF5.12		At1g72140	NO₃^- (oocyte)	[22]
	AtNPF5.16		At1g22550	NO₃^- (oocyte)	[22]
	AtNPF6.2	NRT1.4	At2g26690	NO₃^- (oocyte)	[23]
	AtNPF6.3	NRT1.1	At1g12110	NO₃^-,auxin (oocyte)	[24-26]
	AtNPF7.2	NRT1.8	At4g21680	NO₃^- (oocyte)	[27]
	AtNPF7.3	NRT1.5	At1g32450	NO₃^-,K⁺ (oocyte)	[28-29]
	AtNPF8.1	PTR1	At3g54140	Dipeptides (oocyte, yeast); JA-Ile (yeast)	[2,30-32]
	AtNPF8.2	PTR5	At5g01180	Dipeptides (oocyte, yeast); ABA, GA, JA-Ile (yeast)	[2,31-32]
	AtNPF8.3	PTR2	At2g02040	Dipeptides (yeast); Histidine	[33-34]
	AtNRT2.1		At1g08090	NO₃^- (oocyte)	[35]
	AtNRT2.2		At1g08100	NO₃^- (oocyte)	[35]
	AtNRT2.4		At5g60770	NO₃^- (oocyte)	[35]
	AtNRT2.5		At1g12940	NO₃^- (oocyte)	[35]
	AtNRT2.7		At5g14570	NO₃^- (oocyte)	[35]
水稻 Oryza sativa	OsNPF2.2	OsPTR2	Os12g44100	NO₃^- (oocyte)	[36]
	OsNPF2.4		Os03g48180	NO₃^- (oocyte)	[37]
	OsNPF7.2		Os02g47090	NO₃^- (oocyte)	[38]
	OsNPF7.3	OsPTR6	Os04g50950	Dipeptide (yeast)	[39]
	OsNPF8.9	OsNRT1	Os03g13274	NO₃^- (oocyte)	[40-41]
	OsNRT2.3b		Os01g50820	NO₃^- (oocyte)	[42]

Table S1

References 70

[1]	Iqbal A, Dong Q, Wang X, Gui H, Zhang H, Zhang X, Song M. Transcriptome analysis reveals differences in key genes and pathways regulating carbon and nitrogen metabolism in cotton genotypes under N starvation and resupply. Int J Mol Sci, 2020, 21: 1500. doi: 10.3390/ijms21041500
[2]	Xu G H, Fan X R, Miller A J. Plant nitrogen assimilation and use efficiency. Annu Rev Plant Biol, 2012, 63: 153-182. doi: 10.1146/annurev-arplant-042811-105532 pmid: 22224450
[3]	Wang Y Y, Cheng Y H, Chen K E, Tsay Y F. Nitrate transport, signaling, and use efficiency. Annu Rev Plant Biol, 2018, 69: 85-122. doi: 10.1146/arplant.2018.69.issue-1
[4]	Léran S, Varala K, Boyer J C, Chiurazzi M, Crawford N, Daniel-Vedele F, David L, Dickstein R, Fernandez E, Forde B, Gassmann W, Geiger D, Gojon A, Gong J M, Halkier B A, Harris J M, Hedrich R, Limami A M, Rentsch D, Seo M, Tsay Y F, Zhang M Y, Coruzzi G, Lacombe B. A unified nomenclature of NITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER family members in plants. Trends Plant Sci, 2014, 19: 5-9. doi: 10.1016/j.tplants.2013.08.008 pmid: 24055139
[5]	Wang H D, Wan Y F, Buchner P, King R, Ma H X, Hawkesford M J. Phylogeny and gene expression of the complete NITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER FAMILY in Triticum aestivum. J Exp Bot, 2020, 71: 4531-4546. doi: 10.1093/jxb/eraa210
[6]	Bai H, Euring D, Volmer K, Janz D, Polle A. The nitrate transporter (NRT) gene family in poplar. PLoS One, 2013, 8: e72126. doi: 10.1371/journal.pone.0072126
[7]	Tahir M M, Wang H, Ahmad B, Liu Y, Fan S, Li K, Lei C, Shah K, Li S H, Zhang D. Identification and characterization of NRT gene family reveals their critical response to nitrate regulation during adventitious root formation and development in apple rootstock. Sci Hortic, 2021, 275: 109642. doi: 10.1016/j.scienta.2020.109642
[8]	Wang X L, Cai X F, Xu C X, Wang Q H. Identification and characterization of the NPF, NRT2 and NRT3 in spinach. Plant Physiol Biochem, 2020, 158: 297-307. doi: 10.1016/j.plaphy.2020.11.017
[9]	Bouguyon E, Brun F, Meynard D, Kubes M, Pervent M, Léran S, Lacombe B, Krouk G, Guiderdoni E, Zazimalova E, Hoyerova K, Nacry P, Gojon A. Multiple mechanisms of nitrate sensing by Arabidopsis nitrate transceptor NRT1.1. Nat Plants, 2015, 1: 15015.
[10]	Buchner P, Hawkesford M J. Complex phylogeny and gene expression patterns of members of the NITRATE TRANSPORTER1/ PEPTIDE TRANSPORTER family (NPF) in wheat. J Exp Bot, 2014, 65: 5697-5710. doi: 10.1093/jxb/eru231 pmid: 24913625
[11]	Fan X R, Naz M, Fan X R, Xuan W, Miller A J, Xu G H. Plant nitrate transporters: from gene function to application. J Exp Bot, 2017, 68: 2463-2475. doi: 10.1093/jxb/erx011 pmid: 28158856
[12]	Hu B, Wang W, Ou S J, Tang J Y, Li H, Che R H, Zhang Z H, Chai X Y, Wang H R, Wang Y Q, Liang C Z, Liu L C, Piao Z Z, Deng Q Y, Deng K, Xu C, Liang Y, Zhang L H, Li L G, Chu C C. Variation in NRT1.1B contributes to nitrate-use divergence between rice subspecies. Nat Genet, 2015, 47: 834-838. doi: 10.1038/ng.3337
[13]	Huang N C, Liu K H, Lo H J, Tsay Y F. Cloning and functional characterization of an Arabidopsis nitrate transporter gene that encodes a constitutive component of low-affinity uptake. Plant Cell, 1999, 11: 1381-1392. doi: 10.1105/tpc.11.8.1381 pmid: 10449574
[14]	Segonzac C, Boyer J C, Ipotesi E, Szponarski W, Tillard P, Touraine B, Sommerer N, Rossignol M, Gibrat R. Nitrate efflux at the root plasma membrane: identification of an Arabidopsis excretion transporter. Plant Cell, 2007, 19: 3760-3777. doi: 10.1105/tpc.106.048173 pmid: 17993627
[15]	Filleur S, Dorbe M F, Cerezo M, Orsel M, Granier F, Gojon A, Daniel-Vedele F. An Arabidopsis T-DNA mutant affected in Nrt2 genes is impaired in nitrate uptake. FEBS Lett, 2001, 489: 220-224. doi: 10.1016/s0014-5793(01)02096-8 pmid: 11165253
[16]	Gu C S, Zhang X X, Jiang J F, Guan Z Y, Zhao S, Fang W M, Liao Y, Chen S M, Chen F D. Chrysanthemum CmNAR2 interacts with CmNRT2 in the control of nitrate uptake. Sci Rep, 2014, 4: e5833.
[17]	Barbier-Brygoo H, Angeli D A, Filleur S, Frachisse J M, Gambale F, Thomine S, Wege S. Anion channels/transporters in plants: from molecular bases to regulatory networks. Annu Rev Plant Biol, 2011, 62: 25-51. doi: 10.1146/annurev-arplant-042110-103741 pmid: 21275645
[18]	Lezhneva L, Kiba T, Feriabourrellier A B, Lafouge F, Boutet-Mercey S, Zoufan P, Sakakibara H, Daniel-Vedele F, Krapp A. The Arabidopsis nitrate transporter NRT2.5 plays a role in nitrate acquisition and remobilization in nitrogen-starved plants. Plant J, 2014, 80: 230-241. doi: 10.1111/tpj.12626
[19]	Du X Q, Wang F L, Li H, Jing S, Yu M, Li J G, Wu W H, Kudla J, Wang Y. The transcription factor MYB59 regulates K⁺/NO₃^- translocation in the Arabidopsis response to low K⁺ stress. Plant Cell, 2019, 31: 699-714. doi: 10.1105/tpc.18.00674
[20]	Wang Y Y, Tsay Y F. Arabidopsis nitrate transporter NRT1.9 is important in phloem nitrate transport. Plant Cell, 2011, 23: 1945-1957. doi: 10.1105/tpc.111.083618
[21]	Xia X D, Fan X R, Wei J, Feng H M, Qu H Y, Xie D, Miller A J, Xu G H. Rice nitrate transporter OsNPF2.4 functions in low- affinity acquisition and long-distance transport. J Exp Bot, 2015, 66: 317-331. doi: 10.1093/jxb/eru425
[22]	Tang Z, Fan X R, Li Q, Feng H M, Miller A J, Shen Q R, Xu G H. Knockdown of a rice stelar nitrate transporter alters long-distance translocation but not root influx. Plant Physiol, 2012, 160: 2052-2063. doi: 10.1104/pp.112.204461 pmid: 23093362
[23]	Hsu P K, Tsay Y F. Two phloem nitrate transporters, NRT1.11 and NRT1.12, are important for redistributing xylem-borne nitrate to enhance plant growth. Plant Physiol, 2013, 163: 844-856. doi: 10.1104/pp.113.226563
[24]	Chiu C C, Lin C S, Hsia A P, Su R C, Lin H L, Tsay Y F. Mutation of a nitrate transporter, AtNRT1:4, results in a reduced petiole nitrate content and altered leaf development. Plant Cell Physiol, 2004, 45: 1139-1148. pmid: 15509836
[25]	Fan S C, Lin C S, Hsu P K, Lin S H, Tsay Y F. The Arabidopsis nitrate transporter NRT1.7, expressed in phloem, is responsible for source-to-sink remobilization of nitrate. Plant Cell, 2009, 21: 2750-2761. doi: 10.1105/tpc.109.067603
[26]	Almagro A, Lin S H, Tsay Y F. Characterization of the Arabidopsis nitrate transporter NRT1.6 reveals a role of nitrate in early embryo development. Plant Cell, 2008, 20: 3289-3299. doi: 10.1105/tpc.107.056788 pmid: 19050168
[27]	Léran S, Garg B, Boursiac Y, Corratgé-Faillie C, Brachet C, Tillard P, Gojon A, Lacombe B. AtNPF5.5, a nitrate transporter affecting nitrogen accumulation in Arabidopsis embryo. Sci Rep, 2015, 5: 7962. doi: 10.1038/srep07962
[28]	Bouguyon E, Perrine-Walker F, Pervent M, Rochette J, Cuesta C, Benkova E, Martinière A, Bach L, Krouk G, Gojon A, Nacry P. Nitrate controls root development through posttranscriptional regulation of the NRT1.1/NPF6.3 transporter/sensor. Plant Physiol, 2016, 172: 1237-1248. pmid: 27543115
[29]	Chen C Z, Lyu X F, Li J Y, Yi H Y, Gong J M. Arabidopsis NRT1.5 is another essential component in the regulation of nitrate reallocation and stress tolerance. Plant Physiol, 2012, 159: 1582-1590. doi: 10.1104/pp.112.199257
[30]	Taochy C, Gaillard I, Ipotesi E, Oomen R, Leonhardt N, Zimmermann S, Peltier J B, Szponarski W, Simonneau T, Sentenac H, Gibrat R, Boyer J C. The Arabidopsis root stele transporter NPF2.3 contributes to nitrate translocation to shoots under salt stress. Plant J, 2015, 83: 466-479. doi: 10.1111/tpj.2015.83.issue-3
[31]	Li B, Byrt C, Qiu J, Baumann U, Hrmova M, Evrard A, Johnson A A, Birnbaum K D, Mayo G M, Jha D. Identification of a Stelar-localized transport protein that facilitates root-to-shoot transfer of chloride in Arabidopsis. Plant Physiol, 2016, 170: 1014-1029. doi: 10.1104/pp.15.01163
[32]	Li B, Qiu J, Jayakannan M, Xu B, Li Y, Mayo G M, Tester M, Gilliham M, Roy S J. AtNPF2.5 modulates chloride (Cl^-) efflux from roots of Arabidopsis thaliana. Front Plant Sci, 2017, 7: 2013.
[33]	Yang G, Tang H, Nie Y, Zhang X. Responses of cotton growth, yield, and biomass to nitrogen split application ratio. Eur J Agron, 2011, 35: 164-170. doi: 10.1016/j.eja.2011.06.001
[34]	储成才, 王毅, 王二涛. 植物氮磷钾养分高效利用研究现状与展望. 中国科学: 生命科学, 2021, 51: 1415-1423.
	Chu C C, Wang Y, Wang E T. Improving the utilization efficiency of nitrogen, phosphorus and potassium: current situation and future perspectives. Sci Sin (Vitae), 2021, 51: 1415-1423. (in Chinese with English abstract)
[35]	孙伊辰. 棉花吸收利用氮素的生理及分子机制研究. 中国农业大学硕士学位论文, 北京, 2019.
	Sun Y C. Investigation of Physiological and Molecular Mechanisms of Nitrogen Uptake and Utilization in Cotton Plant. MS Thesis of China Agricultural University, Beijing, China, 2019. (in Chinese with English abstract)
[36]	高美美. GhNRT2.5基因在棉花低氮胁迫反应中的功能分析. 西南大学硕士学位论文, 重庆, 2020.
	Gao M M. Function Analysis of GhNRT2.5 in Cotton Response to Low Nitrogen Stress. MS Thesis of Southwest University, Chongqing, China, 2020. (in Chinese with English abstract)
[37]	Subramanian B, Gao S, Lercher M J, Hu S, Chen W H. Evolview v3: a webserver for visualization, annotation, and management of phylogenetic trees. Nucleic Acids Res, 2019, 2: 270-275.
[38]	Wilkins M R, Gasteiger E, Bairoch A, Sanchez J C, Williams K L, Appel R D, Hochstrasser D F. Protein identification and analysis tools in the ExPASy server. Methods Mol Biol, 1999, 112: 531-552. pmid: 10027275
[39]	Horton P, Park K J, Obayashi T, Fujita N, Harada H, Adams- Collier C J, Nakai K. WoLF PSORT: protein localization predictor. Nucleic Acids Res, 2007, 35: 585-587. pmid: 17517783
[40]	Voorrips R E. MapChart: software for the graphical presentation of linkage maps and QTLs. J Hered, 2002, 93: 77-78. doi: 10.1093/jhered/93.1.77 pmid: 12011185
[41]	Bailey T L, Williams N, Misleh C, Li W W. MEME: discovering and analyzing DNA and protein sequence motifs. Nucleic Acids Res, 2006, 34: 369-373. pmid: 16845028
[42]	Chen C J, Chen H, Zhang Y, Thomas H R, Frank M H, He Y H, Xia R. TBtools: an integrative toolkit developed forinteractive analyses of big biological data. Mol Plant, 2020, 13: 1194-1202. doi: 10.1016/j.molp.2020.06.009
[43]	An J, Hu P G, Li F J, Wu H H, Shen Y, White J C, Tian X L, Li Z H, Giraldo J P. Emerging investigator series: molecular mechanisms of plant salinity stress tolerance improvement by seed priming with cerium oxide nanoparticles. Environ Sci Nano, 2020, 7: 2214-2228. doi: 10.1039/D0EN00387E
[44]	Yang D D, Li F J, Yi F, Eneji A E, Tian X L, Li Z H. Transcriptome analysis unravels key factors involved in response to potassium deficiency and feedback regulation of K⁺ uptake in cotton roots. Int J Mol Sci, 2021, 22: 3133. doi: 10.3390/ijms22063133
[45]	Li F J, Wu Q, Liao B P, Yu K K, Huo Y N, Meng L, Wang S M, Wang B M, Du M W, Tian X L, Li Z H. Thidiazuron promotes leaf abscission by regulating the crosstalk complexities between ethylene, auxin, and cytokinin in cotton. Int J Mol Sci, 2022, 23: 2696. doi: 10.3390/ijms23052696
[46]	Hoagland D R, Arnon D I. The water-culture method for growing plants without soil. Cali Agric Exp Sta, 1950, 357: 1-39.
[47]	Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCTmethod. Met, 2001, 25: 402-408.
[48]	袁丁. 5种木本植物 NRT2 基因家族的生物信息学分析与分子进化研究. 西北农林科技大学硕士学位论文, 陕西杨凌, 2012.
	Yuan D. Bioinformatical Analysis and Molecular Evolutionary Investigation of NRT2 Gene Family in 5 Woody Plants. MS Thesis of Northwest A&F University, Yangling, Shaanxi, China, 2012. (in Chinese with English abstract)
[49]	Kiba T, Feria-Bourrellier A B, Lafouge F, Lezhneva L, Boutet-Mercey S, Orsel M, Bréhaut V, Miller A, Daniel-Vedele F, Sakakibara H, Krapp A. The Arabidopsis nitrate transporter NRT2.4 plays a double role in roots and shoots of nitrogen-starved plants. Plant Cell, 2012, 24: 245-258. doi: 10.1105/tpc.111.092221
[50]	Chopin F, Orsel M, Dorbe M F, Chardon F, Truong H N, Miller A J, Krapp A, Daniel-Vedele F. The Arabidopsis ATNRT2.7 nitrate transporter controls nitrate content in seeds. Plant Cell, 2007, 19: 1590-1602. doi: 10.1105/tpc.107.050542
[51]	Wang W, Hu B, Yuan D Y, Liu Y Q, Che R H, Hu Y C, Ou S J, Liu Y X, Zhang Z H, Wang H R, Li H, Jiang Z M, Zhang Z L, Gao X K, Qiu Y H, Meng X B, Liu Y X, Bai Y, Liang Y, Wang Y Q, Zhang L H, Li L G, Sodmergen, Jing H C, Li J Y, Chu C C. Expression of the nitrate transporter gene OsNRT1.1A/OsNPF6.3 confers high yield and early maturation in rice. Plant Cell, 2018, 30: 638-651. doi: 10.1105/tpc.17.00809
[52]	Zhao F J, McGrath S P, Meharg A A. Arsenic as a food chain contaminant: mechanisms of plant uptake and metabolism and mitigation strategies. Annu Rev Plant Biol, 2010, 61: 535-559. doi: 10.1146/arplant.2010.61.issue-1
[53]	Wen J, Li P F, Ran F, Guo P C, Zhu J T, Yang J, Zhang L L, Chen P, Li J N, Du H. Genome-wide characterization, expression analyses, and functional prediction of the NPF family in Brassica napus. BMC Genomics, 2020, 21: 871. doi: 10.1186/s12864-020-07274-7
[54]	Wang Q, Liu C H, Dong Q L, Huang D, Li C Y, Li P M, Ma F W. Genome-wide identification and analysis of apple NITRATE TRANSPORTER 1/PEPTIDETRANSPORTER family (NPF) genes reveals MdNPF6.5 confers high capacity for nitrogen uptake under low-nitrogen conditions. Int J Mol Sci, 2018, 19: 2761. doi: 10.3390/ijms19092761
[55]	Steiner H Y, Naider F, Becker J M. The PTR family: a new group of peptide transporters. Mol Microbiol, 1995, 16: 825-834. pmid: 7476181
[56]	Zhang G B, Yi H Y, Gong J M. The Arabidopsis ethylene/ jasmonic acid-NRT signaling module coordinates nitrate reallocation and the trade-off between growth and environmental adaptation. Plant Cell, 2014, 26: 3984-3998. doi: 10.1105/tpc.114.129296
[57]	Kanno Y, Hanada A, Chiba Y, Ichikawa T, Nakazawa M, Matsui M, Koshiba T, Kamiya Y, Seo M. Identification of an abscisic acid transporter by functional screening using the receptor complex as a sensor. Proc Natl Acad Sci USA, 2012, 109: 9653-9658. doi: 10.1073/pnas.1203567109 pmid: 22645333
[58]	Chiba Y, Shimizu T, Miyakawa S, Kanno Y, Koshiba T, Kamiya Y, Seo M. Identification of Arabidopsis NRT1/PTR FAMILY (NPF) proteins capable of transporting plant hormones. J Plant Res, 2015, 128: 679-686. doi: 10.1007/s10265-015-0710-2
[59]	Wen Z, Tyerman S D, Dechorgnat J, Ovchinnikova E, Dhugga K, Kaiser B N. Maize NPF6 proteins are homologs of Arabidopsis CHL1 that are selective for both nitrate and chloride. Plant Cell, 2017, 29: 2581-2596. doi: 10.1105/tpc.16.00724
[60]	Krouk G, Lacombe B, Bielach A. Nitrate-regulated auxin transport by NRT1.1 defines a mechanism for nutrient sensing in plants. Dev Cell, 2010, 18: 927-937. doi: 10.1016/j.devcel.2010.05.008 pmid: 20627075
[61]	Zhang L, Ma H J, Chen T T, Pen J, Yu S X, Zhao X H. Morphological and physiological responses of cotton (Gossypium hirsutum L.) plants to salinity. PLoS One, 2014, 9: e112807.
[62]	汪芳珍. 荒漠植物沙芥NAXT家族基因的克隆及其成员PcNPF2.7的功能分析. 兰州大学硕士学位论文, 甘肃兰州, 2021.
	Wang F Z. Cloning of NAXT Homologous Genes and Functional Analysis of One Member PcNPF2.7 in the Xerophyte Pugionium cornutum. MS Thesis of Lanzhou University, Lanzhou, Gansu, China, 2021. (in Chinese with English abstract)
[63]	Goel P, Singh A K. Abiotic stresses downregulate key genes involved in nitrogen uptake and assimilation in Brassica juncea L. PLoS One, 2015, 10: e0143645.
[64]	Liu K H, Huang C Y, Tsay Y F. CHL1 is a dual-affinity nitrate transporter of Arabidopsis involved in multiple phases of nitrate uptake. Plant Cell, 1999, 11: 865-874. doi: 10.1105/tpc.11.5.865 pmid: 10330471
[65]	Léran S, Muños S, Brachet C, Tillard P, Gojon A, Lacombe B. Arabidopsis NRT1.1 is a bidirectional transporter involved in root-to-shoot nitrate translocation. Mol Plant, 2013, 6: 1984-1987. doi: 10.1093/mp/sst068
[66]	Okamoto M, Vidmar J J, Glass A D M. Regulation of NRT1 and NRT2 gene families of Arabidopsis thaliana: responses to nitrate provision. Plant Cell Physiol, 2003, 44: 304-317 doi: 10.1093/pcp/pcg036 pmid: 12668777
[67]	Lin S H, Kuo H F, Canivenc G, Lin C S, Lepetit M, Hsu P K, Tillard P, Lin H L, Wang Y Y, Tsai C B, Gojon A, Tsay Y F. Mutation of the Arabidopsis NRT1.5 nitrate transporter causes defective root-to-shoot nitrate transport. Plant Cell, 2008, 20: 2514-2528. doi: 10.1105/tpc.108.060244
[68]	Sakuraba Y, Chaganzhana, Mabuchi A, Iba K, Yanagisawa S. Enhanced NRT1.1/NPF6.3 expression in shoots improves growth under nitrogen deficiency stress in Arabidopsis. Commun Biol, 2021, 4: 256. doi: 10.1038/s42003-021-01775-1
[69]	Tang W, Ye J, Yao X M, Zhao P Z, Xuan W, Tian Y L, Zhang Y Y, Xu S, An H Z, Chen G M, Yu J, Wu W, Ge Y W, Liu X L, Li J, Zhang H Z, Zhao Y Q, Yang B, Jiang X Z, Peng C, Zhou C, Terzaghi W, Wang C M, Wan J M. Genome-wide associated study identifies NAC42-activated nitrate transporter conferring high nitrogen use efficiency in rice. Nat Commun, 2019, 10: 5279. doi: 10.1038/s41467-019-13187-1 pmid: 31754193
[70]	Wittkopp P J, Kalay G. Cis-regulatory elements: molecular mechanisms and evolutionary processes underlying divergence. Nat Rev Genet, 2012, 13: 59-69.

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基因名称 Gene name	引物序列 Primer sequence (5'-3')
GhActin9	F: GCCTTGGACTATGAGCAGGA	R: AAGAGATGGCTGGAAGAGGA
GhNPF4.6cA	F: CCAATCTTCTTCTTTGGCTCTGG	R: AGAAGGGCTTGCAGAGAGGTTC
GhNPF6.1aA	F: GCTGCCTTCAAGAAACGACA	R: GAGTGCTGCCTTATCCAAGC
GhNPF6.2bD	F: CTCTTGTGTTTGCTTGGTGGC	R: AATGTGCCAGTCCCAAGTGT
GhNPF6.3dA	F: CCCAAATCGCTGCAGTGTTT	R: CGAAACTGCTTGCTGTGAGGC
GhNPF7.3aA	F: GTTGAAGAGCAGAAGAAGGGAAC	R: ACCTTGCTCAACGAATAGGGATG
GhNRT2.4aA	F: CCAGTGGGGTAGCATGTTTC	R: GCTTTCCACGCTCAGACCG

Identification and expression analysis of nitrate transporter NRT gene family in upland cotton (Gossypium hirsutum L.)

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