Cloning and Expression Analysis of NRT2.4 Gene in Kentucky Bluegrass

doi:10.7668/hbnxb.20191685

Abstract

Abstract: Nitrate transporter (NRT) in plants can effectively regulate and transport NO₃^- and improve nitrogen utilization efficiency. In order to investigate the relationship between Poa pratensis L. NRT2.4 and nitrogen stress, Poa pratensis L. was selected as the material for the cloning and bioinformatics analysis of NRT2.4 gene, and the regulation of this gene expression was observed under treatment by nitrogen nutrient solution. The cloned NRT2.4 gene of Poa pratensis L.belonged to the Nitrate/nitrite transporter NarK superfamily, and was highly homologous to that of Brachypodium distachyon (L.) Beauv. and Aegilops tauschii Coss. Between the 6th and 7th transmembrane domain (TMD), there was a sequence with signature of G-X3-D-X2-G-X-R unique to the MFS family. The 4th TMD had a typical NO₃^-/NO₂^- transporter signature sequence of A-G-W-G/A-N-M-G. There was a conservative protein kinase C motif (S/T-X-R/K):SKR after the 8th TMD.The expression level of Poa pratensis L.NRT2.4 gene was tissue-specific, with the most expressed in leaves. Nitrate nitrogen was conducive to the expression of NRT2.4 gene, both nitrogen starvation and the treatment by nitrogen nutrient solution with a high NaNO₃ were beneficial to the expression of NRT2.4 gene. The results above have provided a theoretical bad for the application of Poa pratensis L.NRT2.4 in stress-tolerance gene engineering in turfgrass.

Key words: Turf grass, Kentucky bluegrass, NRT2.4 gene, Nitrogen nutrition deficiency, Clone, Expression

CLC Number:

CHEN Yang, JIN Yifeng, JIN Zhongmin, WANG Yushu, CAI Rongjian, SHAO Zhulin, WANG Jie. Cloning and Expression Analysis of NRT2.4 Gene in Kentucky Bluegrass[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(2): 1-8. doi: 10.7668/hbnxb.20191685.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.20191685

http://www.hbnxb.net/EN/Y2021/V36/I2/1

References

[1] Guo T C, Xuan H M, Yang Y Y, Wang L, Wei L T, Wang Y H, Kang G Z. Transcription analysis of genes encoding the wheat root transporter NRT1 and NRT2 families during nitrogen starvation[J]. Journal of Plant Growth Regulation, 2014, 33(4):837-848.doi:10.1007/s00344-014-9435-z.
[2] Tsay Y F, Chiu C C, Tsai C B, Ho C H, Hsu P K. Nitrate transporters and peptide transporters[J]. FEBS Lett, 2007,581(12):2290-2300. doi:10.1016/j.febslet.2007.04.047.
[3] Rexach J, Llamas A, Fern ndez E, Galván A. The activity of the high-affinity nitrate transport system I (NRT2;1, NAR2) is responsible for the efficient signalling of nitrate assimilation genes in Chlamydomonas reinhardtii[J]. Planta, 2002, 215(4):606-611. doi:10.1007/s00425-002-0778-5.
[4] Yan M, Fan X R, Feng H M, Miller A J, Shen Q R, Xu G H. Rice OsNAR2.1 interacts with OsNRT2.1, OsNRT2.2 and OsNRT2.3a nitrate transporters to provide uptake over high and low concentration ranges[J]. Plant Cell Environ, 2011,34(8):1360-1372. doi:10.1111/j.1365-3040.2011.02335.x.
[5] Kotur Z, Mackenzie N, Ramesh S, Tyerman S D, Kaiser B N, Glass A D. Nitrate transport capacity of the Arabidopsis thaliana NRT2 family members and their interactions with AtNAR2.1[J]. The New Phytol, 2012,194(3):724-731. doi:10.1111/j.1469-8137.2012.04094.x.
[6] 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[J]. The Plant Cell, 2012, 24(1):245-258. doi:10.1105/tpc.111.092221.
[7] Wang J, Hüner N, Tian L. Identification and molecular characterization of the Brachypodium distachyon NRT2 family, with a major role of BdNRT2.1[J]. Physiol Plant, 2019,165(3):498-510. doi:10.1111/ppl.12716.
[8] Zou X, Liu M Y, Wu W H, Wang Y. Phosphorylation at Ser28 stabilizes the Arabidopsis nitrate transporter NRT2.1 in response to nitrate limitation[J]. Journal of Integrative Plant Biology, 2020, 62(6):865-876. doi:10.1111/jipb.12858.
[9] Zou L P,Qi D F, Sun J B, Zheng X, Peng M.Expression of the cassava nitrate transporter NRT2.1 enables Arabidopsis low nitrate tolerance[J]. Journal of Genetics, 2019, 98(3):74.doi:10.1007/s12041-019-1127-9.
[10] Santino A, Taurino M, De Domenico S, Bonsegna S, Poltronieri P, Pastor V, Flors V. Jasmonate signaling in plant development and defense response to multiple (a) biotic stresses[J]. Plant Cell Rep, 2013,32(7):1085-1098. doi:10.1007/s00299-013-1441-2.
[11] Hu Y B, Fernández V, Ma L. Nitrate transporters in leaves and their potential roles in foliar uptake of nitrogen dioxide[J]. Front Plant Sci, 2014,5:360.doi:10.3389/fpls.2014.00360.
[12] Okamoto M, Vidmar J J, Glass A D M. Regulation of NRT1 and NRT2 gene families of Arabidopsis thaliana:responses to nitrate provision[J]. Plant Cell Physiol, 2003,44(3):304-317. doi:10.1093/pcp/pcg036.
[13] Tong J F, Walk T C, Han P P, Chen L Y, Shen X J, Li Y S, Gu C M, Xie L H, Hu X J, Liao X, Qin L. Genome-wide identification and analysis of high-affinity nitrate transporter 2(NRT2) family genes in rapeseed (Brassica napus L.) and their responses to various stresses[J]. BMC Plant Biol,2020, 20(1):464. doi:10.1186/s12870-020-02648-1.
[14] 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 Physiol, 2007,143(1):425-433. doi:10.1104/pp.106.091223.
[15] 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]. Plant Journal, 2014, 80(2):230-241.doi:10.1111/tpj.12626.
[16] 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[J]. Plant Physiol, 2012,160(4):2052-2063. doi:10.1104/pp.112.204461.
[17] Chen J G, Liu X Q, Liu S H, Fan X R, Zhao L M, Song M Q, Fan X R, Xu G H.Co-overexpression of OsNAR2.1 and OsNRT2.3a increased agronomic nitrogen use efficiency in transgenic rice plants[J]. Frontiers in Plant Science, 2020, 11:1245. doi:10.3389/fpls.2020.01245.
[18] Huang S J, Chen S, Liang Z H, Zhang C M,Yan M,Chen J G,Xu G H,Fan X R,Zhang Y L. Knockdown of the partner protein OsNAR2.1 for high-affinity nitrate transport represses lateral root formation in a nitrate-dependent manner[J]. Scientific Reports, 2015, 5(1):18192. doi:10.1038/srep18192.
[19] Wei J, Zheng Y, Feng H M, Qu H Y, Fan X R, Yamaji N, Ma J F, Xu G H. OsNRT2.4 encodes a dual-affinity nitrate transporter and functions in nitrate-regulated root growth and nitrate distribution in rice[J]. Journal of Experimental Botany,2018,69(5):1095-1107. doi:10.1093/jxb/erx486.
[20] 黄化刚, 申燕, 王卫峰,连文力,陈雪,翟欣,喻奇伟,杨振智,贾宏昉.烟草硝酸盐转运蛋白基因 NtNRT2.4 的克隆及表达分析[J].中国烟草学报,2016,22(1):84-91.
Huang H G, Shen Y, Wang W F,Lian W L,Chen X, Zhai X,Yu W Q,Yang Z Z,Jia H F. Cloning and characterization of NtNRT2.4 gene from Nicotiana tabacum L.[J]. Acta Tabacaria Sinica, 2016,22(1):84-91.

Please choose a citation manager

Content to export