Identification and Expression Analysis of NRT2 Gene Family of Chenopodium quinoa

doi:10.7668/hbnxb.20193363

Abstract

Abstract:

Nitrate nitrogen is one of the two main form of nitrogen uptaken by plants.Nitrate transporter 2(NRT2)plays a leading role in nitrogen uptake under nitrate deficiency conditions.In order to explore characteristics and expression pattern of NRT2 gene in Chenopodium quinoa,in the present study,fifteen members of C.quinoa NRT2s were identified based on bioinformatics analysis,and their encoding protein,chromosome location,gene structure,systematic evolution,promoter elements,tissue expression specificity and expression profile under different nitrogen levels were analyzed.The results showed that 15 NRT2s of C.quinoa were distributed on 7 chromosomes,each member contained 1-9 introns and 2-10 exons,respectively.These C.quinoa NRT2 proteins were predicted to be located on plasma membrane and belong to hydrophobic proteins.The evolutionary analysis found that the NRT2 protein family of quinoa belonged to two subfamilies,and they showed closer relationship to Arabidopsis thaliana.The cis-acting element analysis exhibited that the promoter regions of NRT2 had elements related to growth and stress responses.NRT2s showed varied expression profile in different C.quinoa tissues where more CqNRT2 members were higher expressed in roots,stems and leaves than those in other organs.The expression of CqNRT2.4,CqNRT2.7 and CqNRT2.15 was up-regulated under low nitrogen conditions where the expression levels of CqNRT2.7 and CqNRT2.15 were up-regulated sharply with 0 and 25% nitrogen treatment.The study might provide a reference basis for subsequent cloning,functional and nitrogen stress analysis of CqNRT2s gene.

Key words: Chenopodium quinoa, Nitrate, Nitrate transporter 2, Gene family, Nitrogen stress

YANG Liyan, GAO Yuan, ZHU Manxi, DENG Yan, WANG Chuangyun. Identification and Expression Analysis of NRT2 Gene Family of Chenopodium quinoa[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(S1): 8-18. doi: 10.7668/hbnxb.20193363.

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Figures/Tables 11

References 37

[1]	Crawford N M. Molecular and physiological aspects of nitrate uptake in plants[J]. Trends in Plant Science, 1998, 3(10): 389-395.doi :10.1016/S1360-1385(98)01311-9. doi: 10.1016/S1360-1385(98)01311-9 URL
[2]	Mueller N D, West P C, Gerber J S, MacDonald G K, Polasky S, Foley J A. A tradeoff frontier for global nitrogen use and cereal production[J]. Environmental Research Letters, 2014, 9(5): 054002. doi:10.1088/1748-9326/9/5/054002. doi: 10.1088/1748-9326/9/5/054002
[3]	袁丁, 李红, 郝文芳, 罗志斌. 毛果杨全基因组硝酸根转运蛋白家族(NRT2s)序列分析[J]. 西北林学院学报, 2012, 27(5): 80-86. doi:10.3969/j.issn.1001-7461.2012.05.15. doi: 10.3969/j.issn.1001-7461.2012.05.15
	Yuan D, Li H, Hao W F, Luo Z B. Genome-wide sequence analysis of high affinity nitrate transporter(NRT2s)gene family in Populus trichocarpa[J]. Journal of Northwest Forestry University, 2012, 27(5): 80-86.
[4]	叶玲, 张洁, 郭永正, 赵雄伟, 王兴春. 谷子NRT2基因家族的鉴定及生物信息学分析[J]. 山西农业科学, 2020, 48(3): 283-290. doi:10.3969/j.issn.1002-2481.2020.03.02. doi: 10.3969/j.issn.1002-2481.2020.03.02
	Ye L, Zhang J, Guo Y Z, Zhao X W, Wang X C. Identification and bioinformatics analysis of NRT2 gene family in millet[J]. Journal of Shanxi Agricultural Sciences, 2020, 48(3): 283-290.
[5]	张合琼, 张汉马, 梁永书, 南文斌. 植物硝酸盐转运蛋白研究进展[J]. 植物生理学报, 2016, 52(2): 141-149.doi: 10.13592/j.cnki.ppj.2015.0621. doi: 10.13592/j.cnki.ppj.2015.0621
	Zhang H Q, Zhang H M, Liang Y S, Nan W B. Research progress of nitrate in plant transport mechanism[J]. Plant Physiology Journal, 2016, 52(2): 141-149. doi: 10.1111/j.1399-3054.1981.tb06047.x URL
[6]	Li W B, Wang Y, Okamoto M, Crawford N M, Siddiqi M Y, Glass A D M. Dissection of the AtNRT2.1: AtNRT2.2 inducible high-affinity nitrate transporter gene cluster[J]. Plant Physiology, 2006, 143(1): 425-433. doi:10.1104/pp.106.091223. doi: 10.1104/pp.106.091223 URL
[7]	Remans T, Nacry P, Pervent M, Filleur S, Diatloff E, Mounier E, Tillard P, Forde B G, Gojon A. The Arabidopsis NRT1.1 transporter participates in the signaling pathway triggering root colonization of nitrate-rich patches[J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(50): 19206-19211. doi:10.1073/pnas.0605275103. doi: 10.1073/pnas.0605275103 pmid: 17148611
[8]	Remans T, Nacry P, Pervent M, Girin T, Tillard P, Lepetit M, Gojon A. A central role for the nitrate transporter NRT2.1 in the integrated morphological and physiological responses of the root system to nitrogen limitation in Arabidopsis[J]. Plant Physiology, 2006, 140(3): 909-921. doi:10.1104/pp.105.075721. doi: 10.1104/pp.105.075721 pmid: 16415211
[9]	Feng H M, Yan M, Fan X R, Li B Z, Shen Q R, Miller A J, Xu G H. Spatial expression and regulation of rice high-affinity nitrate transporters by nitrogen and carbon status[J]. Journal of Experimental Botany, 2011, 62(7): 2319-2332. doi:10.1093/jxb/erq403. doi: 10.1093/jxb/erq403 pmid: 21220781
[10]	宋田丽, 周建建, 徐晨曦, 蔡晓锋, 戴绍军, 王全华, 王小丽. 植物硝酸盐转运蛋白功能及表达调控研究进展[J]. 上海师范大学学报(自然科学版), 2017, 46(5): 740-750.doi: 10.3969/J.ISSN.1000-5137.2017.05.019. doi: 10.3969/J.ISSN.1000-5137.2017.05.019
	Song T L, Zhou J J, Xu C X, Cai X F, Dai S J, Wang Q H, Wang X L. Progress in function and regulation of nitrate transporters in plants[J]. Journal of Shanghai Normal University (Natural Sciences), 2017, 46(5): 740-750.
[11]	商海军, 蒋丽君, 於春, 江本利, 闫晓明. 藜麦的营养功能及其蛋白和皂苷提取的研究进展[J]. 中国食物与营养, 2021, 27(4): 43-48.doi: 10.19870/j.cnki.11-3716/ts.2021.04.003. doi: 10.19870/j.cnki.11-3716/ts.2021.04.003
	Shang H J, Jiang L J, Yu C, Jiang B L, Yan X M. Nutritional function of quinoa and extraction status of quinoa protein and saponin[J]. Food and Nutrition in China, 2021, 27(4): 43-48.
[12]	郭慧敏, 耿艳楼, 吕玮, 杨修仕. 藜麦开发利用研究进展[J]. 粮食与油脂, 2021, 34(3): 9-11.doi: 10.3969/j.issn.1008-9578.2021.03.003. doi: 10.3969/j.issn.1008-9578.2021.03.003
	Guo H M, Geng Y L, Lü W, Yang X S. Research progress on development and utilization of quinoa[J]. Cereals & Oils, 2021, 34(3): 9-11.
[13]	侯丽媛, 董艳辉, 李亚莉, 王育川, 赵佳, 刘江, 秦永军, 吴慎杰. 藜麦抗旱性研究进展与展望[J]. 江苏农业科学, 2021, 49(11): 22-28.doi: 10.15889/j.issn.1002-1302.2021.11.004. doi: 10.15889/j.issn.1002-1302.2021.11.004
	Hou L Y, Dong Y H, Li Y L, Wang Y C, Zhao J, Liu J, Qin Y J, Wu S J. Research progress and prospect of drought tolerance of quinoa[J]. Jiangsu Agricultural Sciences, 2021, 49(11): 22-28.
[14]	黎雅楠. 藜麦在盐碱地改良中的应用前景[J]. 绿色科技, 2018(21): 104-105, 108.doi: 10.16663/j.cnki.lskj.2018.21.041. doi: 10.16663/j.cnki.lskj.2018.21.041
	Li Y N. Application Prospect of Chenopodium quinoa Willd in the improvement of saline-alkali land[J]. Journal of Green Science and Technology, 2018(21): 104-105,108.
[15]	Jacobsen S E. The situation for quinoa and its production in southern bolivia: from economic success to environmental disaster[J]. Journal of Agronomy and Crop Science, 2011, 197(5): 390-399.doi: 10.1111/j.1439-037x.2011.00475.x. doi: 10.1111/j.1439-037x.2011.00475.x URL
[16]	朱满喜, 张玉荣, 杨雅舒, 杨小兰, 王创云, 邓妍, 赵丽, 张丽光, 秦丽霞, 杨利艳. 藜麦NLP转录因子家族的鉴定及表达分析[J]. 华北农学报, 2021, 36(4): 37-46.doi: 10.7668/hbnxb.20191913. doi: 10.7668/hbnxb.20191913
	Zhu M X, Zhang Y R, Yang Y S, Yang X L, Wang C Y, Deng Y, Zhao L, Zhang L G, Qin L X, Yang L Y. Identification and expression analysis of NLP transcription factor family of Chenopodium quinoa willd[J]. Acta Agriculturae Boreali-Sinica, 2021, 36(4): 37-46.
[17]	von Wittgenstein N V, Le C H, Hawkins B J, Ehlting J. Evolutionary classification of ammonium,nitrate,and peptide transporters in land plants[J]. BMC Evolutionary Biology, 2014, 14(1): 11.doi: 10.1186/1471-2148-14-11. doi: 10.1186/1471-2148-14-11 URL
[18]	钱瑜, 察倩倩, 孔敏, 刘照坤, 李英, 侯喜林, 刘同坤. 植物NRT2家族的分子进化[J]. 江苏农业学报, 2015, 31(1): 45-54.doi: 10.3969/j.issn.1000-4440.2015.01.007. doi: 10.3969/j.issn.1000-4440.2015.01.007
	Qian Y, Cha Q Q, Kong M, Liu Z K, Li Y, Hou X L, Liu T K. Molecular evolution of NRT2 gene family in plant[J]. Jiangsu Journal of Agricultural Sciences, 2015, 31(1): 45-54.
[19]	Forde B G. Nitrate transporters in plants: structure,function and regulation[J]. Biochimica et Biophysica Acta, 2000, 1465(1/2): 219-235.doi: 10.1016/s0005-2736(00)00140-1. doi: 10.1016/s0005-2736(00)00140-1
[20]	Boudet N, Aubourg S, Toffano-Nioche C, Kreis M, Lecharny A. Evolution of intron/exon structure of DEAD helicase family genes in Arabidopsis,Caenorhabditis,and Drosophila[J]. Genome Research, 2001, 11(12): 2101-2114. doi:10.1101/gr.200801. doi: 10.1101/gr.200801 pmid: 11731501
[21]	Babenko V N, Rogozin I B, Mekhedov S L, Koonin E V. Prevalence of intron gain over intron loss in the evolution of paralogous gene families[J]. Nucleic Acids Research, 2004, 32(12): 3724-3733. doi:10.1093/nar/gkh686. doi: 10.1093/nar/gkh686 pmid: 15254274
[22]	Quesada A, Galván A, Fernández E. Identification of nitrate transporter genes in Chlamydomonas reinhardtii[J]. Heliyon, 1994, 5(3): 407-419. doi:10.1111/j.1365-313x.1994.00407.x. doi: 10.1111/j.1365-313x.1994.00407.x
[23]	Orsel M, Krapp A, Daniel-Vedele F. Analysis of the NRT2 nitrate transporter family in Arabidopsis structure and gene expression[J]. Plant Physiology, 2002, 129(2): 886-896. doi:10.1104/pp.005280. doi: 10.1104/pp.005280 URL
[24]	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[J]. The Plant Cell, 2007, 19(5): 1590-1602. doi:10.1105/tpc.107.050542. doi: 10.1105/tpc.107.050542 URL
[25]	Krapp A, David L C, Chardin C, Girin T, Marmagne A, Leprince A S, Chaillou S, Ferrario-Méry S, Meyer C, Daniel-Vedele F. Nitrate transport and signalling in Arabidopsis[J]. Journal of Experimental Botany, 2014, 65(3): 789-798. doi:10.1093/jxb/eru001. doi: 10.1093/jxb/eru001 pmid: 24532451
[26]	李婧, 左欣欣, 赵培伶, 张陆玉, 徐文鸾, 张徐杨, 倪德江, 王明乐. 茶树高亲和硝酸盐转运蛋白家族基因NRT2的鉴定与表达[J]. 应用与环境生物学报, 2022, 28(1): 50-56.doi: 10.19675/j.cnki.1006-687x.2021.02056. doi: 10.19675/j.cnki.1006-687x.2021.02056
	Li J, Zuo X X, Zhao P L, Zhang L Y, Xu W L, Zhang X Y, Ni D J, Wang M L. Identification and expression analysis of high-affinity nitrate transporter family genes NRT2 in Camellia sinensis[J]. Chinese Journal of Applied and Environmental Biology, 2022, 28(1): 50-56.
[27]	Lawton-Rauh A. Evolutionary dynamics of duplicated genes in plants[J]. Mol Phylogenet Evol, 2003, 29(3):396-409. doi:10.1016/j.ympev.2003.07.004. doi: 10.1016/j.ympev.2003.07.004 pmid: 14615182
[28]	Barbier-Brygoo H, De Angeli A, Filleur S, Frachisse J M, Gambale F, Thomine S, Wege S. Anion channels/transporters in plants: From molecular bases to regulatory networks[J]. Annual Review of Plant Biology, 2011, 62: 25-51. doi:10.1146/annurev-arplant-042110-103741. doi: 10.1146/annurev-arplant-042110-103741 pmid: 21275645
[29]	Lezhneva L, Kiba T, Feria-Bourrellier 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[J]. The Plant Journal, 2014, 80(2): 230-241. doi:10.1111/tpj.12626. doi: 10.1111/tpj.12626 pmid: 25065551
[30]	Cerezo M, Tillard P, Filleur S, Muños S, Daniel-Vedele F, Gojon A. Major alterations of the regulation of root NO₃ -uptake are associated with the mutation of Nrt2.1 and Nrt2.2 genes in Arabidopsis[J]. Plant Physiology, 2001, 127(1): 262-271. doi:10.1104/pp.127.1.262. doi: 10.1104/pp.127.1.262 pmid: 11553754
[31]	Asim M, Ullah Z, Oluwaseun A, Wang Q, Liu H. Signalling overlaps between nitrate and auxin in regulation of the root system architecture: Insights from the Arabidopsis thaliana[J]. International Journal of Molecular Sciences, 2020, 21(8): E2880. doi:10.3390/ijms21082880. doi: 10.3390/ijms21082880
[32]	李志康, 严冬, 薛张逸, 顾逸彪, 李思嘉, 刘立军, 张耗, 王志琴, 杨建昌, 顾骏飞. 细胞分裂素对植物生长发育的调控机理研究进展及其在水稻生产中的应用探讨[J]. 中国水稻科学, 2018, 32(4): 311-324.doi: 10.16819/j.1001-7216.2018.8027. doi: 10.16819/j.1001-7216.2018.8027
	Li Z K, Yan D, Xue Z Y, Gu Y B, Li S J, Liu L J, Zhang H, Wang Z Q, Yang J C, Gu J F. Regulations of plant growth and development by cytokinins and their applications in rice production[J]. Chinese Journal of Rice Science, 2018, 32(4): 311-324. doi: 10.16819/j.1001-7216.2018.8027
[33]	唐贤礼, 张月, 张盾, 夏新莉, 尹伟伦. 毛果杨基因PtNRT2.7的功能初步鉴定与分析[J]. 北京林业大学学报, 2016, 38(8): 18-27.doi: 10.13332/j.1000-1522.20160041. doi: 10.13332/j.1000-1522.20160041
	Tang X L, Zhang Y, Zhang D, Xia X L, Yin W L. Functional identification and analysis of PtNRT2.7 gene from Populus trichocarpa[J]. Journal of Beijing Forestry University, 2016, 38(8): 18-27.
[34]	Bassett C L, Baldo A M, Moore J T, Jenkins R M, Soffe D S, Wisniewski M E, Norelli J L, Farrell R E. Genes responding to water deficit in apple(Malus×domestica Borkh.)roots[J]. BMC Plant Biology, 2014, 14: 182. doi:10.1186/1471-2229-14-182. doi: 10.1186/1471-2229-14-182 pmid: 25004790
[35]	Garnett T, Conn V, Plett D, Conn S, Zanghellini J, Mackenzie N, Enju A, Francis K, Holtham L, Roessner U, Boughton B, Bacic A, Shirley N, Rafalski A, Dhugga K, Tester M, Kaiser B N. The response of the maize nitrate transport system to nitrogen demand and supply across the lifecycle[J]. The New Phytologist, 2013, 198(1): 82-94. doi:10.1111/nph.12166. doi: 10.1111/nph.12166 URL
[36]	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[J]. Frontiers in Plant Science, 2016, 7: 2013. doi:10.3389/fpls.2016.02013. doi: 10.3389/fpls.2016.02013
[37]	Little D Y, Rao H, Oliva S, Daniel-Vedele F, Krapp A, Malamy J E. The putative high-affinity nitrate transporter NRT2.1 represses lateral root initiation in response to nutritional cues[J]. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102(38): 13693-13698. doi:10.1073/pnas.0504219102. doi: 10.1073/pnas.0504219102 pmid: 16157886

基因 Gene	引物(5'-3') Primer(5'-3')	引物(5'-3') Primer(5'-3')	产物长度/bp Product size
18SrRNA	CAACCATAAACGATGCCGA	AGCCTTGCGACCATACTCC	112
CqNRT2.2	GGGTGTTACAGAAGAAGAC	CTCTCATTCCTACTGCCT	108
CqNRT2.4	AATTCGCCGAAGGTTGTAGG	AACATCAATCGCAGGACGG	89
CqNRT2.7	AGTACGATGACGACGAGAA	TTGTGTAGGTGGAGTAGCA	116
CqNRT2.15	GTCTTACCTACATGGGAGTCAT	AGCCTCCTCATTAGCATTCTTA	116

基因 Gene	引物(5'-3') Primer(5'-3')	引物(5'-3') Primer(5'-3')	产物长度/bp Product size
18SrRNA	CAACCATAAACGATGCCGA	AGCCTTGCGACCATACTCC	112
CqNRT2.2	GGGTGTTACAGAAGAAGAC	CTCTCATTCCTACTGCCT	108
CqNRT2.4	AATTCGCCGAAGGTTGTAGG	AACATCAATCGCAGGACGG	89
CqNRT2.7	AGTACGATGACGACGAGAA	TTGTGTAGGTGGAGTAGCA	116
CqNRT2.15	GTCTTACCTACATGGGAGTCAT	AGCCTCCTCATTAGCATTCTTA	116

基因ID Gene ID	基因名称 Gene name	编码序列长度/bp CDS length	等电点 pI	分子质量/ku MW	氨基酸数目/个 Amino acid number	不稳定系数 Instability index	总平均亲水性 Grand average of hydropathicity	跨膜结构域 Transmembrane domain
AUR62006663	CqNRT2.1	2 082	9.12	75.93	693	39.96	0.28	15
AUR62006662	CqNRT2.2	1 401	8.51	50.99	466	38.30	0.45	11
AUR62014088	CqNRT2.3	2 829	8.66	101.59	942	37.46	0.45	21
AUR62006661	CqNRT2.4	1 452	8.77	52.60	483	37.34	0.51	12
AUR62001493	CqNRT2.5	1 500	8.51	53.57	499	45.32	0.48	9
AUR62001495	CqNRT2.6	1 578	9.07	56.76	525	40.13	0.36	11
AUR62001496	CqNRT2.7	1 581	8.91	56.92	526	41.40	0.37	10
AUR62003509	CqNRT2.8	1 527	8.68	54.98	508	36.46	0.41	11
AUR62000316	CqNRT2.9	1 449	8.81	52.69	482	36.67	0.45	12
AUR62000317	CqNRT2.10	1 479	8.78	53.94	492	41.08	0.40	12
AUR62000318	CqNRT2.11	1 794	9.26	64.67	597	39.74	0.31	13
AUR62005427	CqNRT2.12	1 428	9.05	51.63	475	36.50	0.35	9
AUR62009877	CqNRT2.13	1 563	8.20	56.05	520	39.99	0.45	11
AUR62009875	CqNRT2.14	1 539	8.90	55.76	512	40.25	0.37	10
AUR62009874	CqNRT2.15	1 581	8.71	56.94	526	39.83	0.37	10

基因ID Gene ID	基因名称 Gene name	编码序列长度/bp CDS length	等电点 pI	分子质量/ku MW	氨基酸数目/个 Amino acid number	不稳定系数 Instability index	总平均亲水性 Grand average of hydropathicity	跨膜结构域 Transmembrane domain
AUR62006663	CqNRT2.1	2 082	9.12	75.93	693	39.96	0.28	15
AUR62006662	CqNRT2.2	1 401	8.51	50.99	466	38.30	0.45	11
AUR62014088	CqNRT2.3	2 829	8.66	101.59	942	37.46	0.45	21
AUR62006661	CqNRT2.4	1 452	8.77	52.60	483	37.34	0.51	12
AUR62001493	CqNRT2.5	1 500	8.51	53.57	499	45.32	0.48	9
AUR62001495	CqNRT2.6	1 578	9.07	56.76	525	40.13	0.36	11
AUR62001496	CqNRT2.7	1 581	8.91	56.92	526	41.40	0.37	10
AUR62003509	CqNRT2.8	1 527	8.68	54.98	508	36.46	0.41	11
AUR62000316	CqNRT2.9	1 449	8.81	52.69	482	36.67	0.45	12
AUR62000317	CqNRT2.10	1 479	8.78	53.94	492	41.08	0.40	12
AUR62000318	CqNRT2.11	1 794	9.26	64.67	597	39.74	0.31	13
AUR62005427	CqNRT2.12	1 428	9.05	51.63	475	36.50	0.35	9
AUR62009877	CqNRT2.13	1 563	8.20	56.05	520	39.99	0.45	11
AUR62009875	CqNRT2.14	1 539	8.90	55.76	512	40.25	0.37	10
AUR62009874	CqNRT2.15	1 581	8.71	56.94	526	39.83	0.37	10

蛋白名称 Protein name	细胞质膜 Plasma membrane	内质网 Endoplasmic reticulum	液泡膜 Vacuolar membrane	叶绿体 Chloroplast	高尔基体 Golgi	细胞质 Cytoplasm	细胞核 Nucleus	线粒体 Mitochondrion
CqNRT2.1	9	2	-	2	-	-	-	1
CqNRT2.2	9	1	2	-	-	1	-	-
CqNRT2.3	11	1	-	-	2	-	-	-
CqNRT2.4	10	3	1	-	-	-	-	-
CqNRT2.5	11	-	1	-	1	1	-	-
CqNRT2.6	8	1	3	-	-	1	-	-
CqNRT2.7	8	-	4	-	-	1	-	-
CqNRT2.8	7	1	3	-	3	-	-	-
CqNRT2.9	9	4	1	-	-	-	-	-
CqNRT2.10	8	3	2	-	-	-	-	-
CqNRT2.11	9	3	-	1	-	-	-	1
CqNRT2.12	10	1	1	-	1	-	1	-
CqNRT2.13	9	-	2	-	2	2	-	-
CqNRT2.14	8	2	3	-	1	-	-	-
CqNRT2.15	7	1	4	-	-	1	-	-

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