Cloning and Expressing Analysis of a Nitrogen Transporter 2.5 Gene from Tea Plant[Camellia sinensis (L.)]

FENG Suhua; WANG Liyuan; CHEN Changsong; LIN Zhenghe; CHENG Hao; WEI Kang; ZHANG Chengcai

doi:10.13305/j.cnki.jts.2014.04.007

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Journal of Tea Science ›› 2014, Vol. 34 ›› Issue (4) : 364-370. DOI: 10.13305/j.cnki.jts.2014.04.007

Cloning and Expressing Analysis of a Nitrogen Transporter 2.5 Gene from Tea Plant[Camellia sinensis (L.)]

FENG Suhua¹, WANG Liyuan^1,*, CHEN Changsong², LIN Zhenghe², CHENG Hao^1,*, WEI Kang¹, ZHANG Chengcai¹

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Abstract

The full cDNA length of NRT2.5 gene was obtained via rapid amplification of cDNA ends (RACE) from tea plant [Camellia sinessis (L.)] leaves. The cDNA sequence of this gene was 2 457 bp and opening reading frame (ORF) was 1 362 bp, encoding for 454 amino acids. The putative protein of this gene had an isoelectric point of 9.63 and a calculated molecular weight of 48.7 kD. Bioinformatics analysis showed that NRT2.5 of tea plant was highly homologous to the genes such as TcNRT2.5 and AtNRT2.5, and its encoded protein had the common structural characteristics of NRT family. Fluorescence quantitative real-time PCR analysis showed that NRT2.5 gene were detected in all the tested tissues of tea plant but was mainly detected in the mature leaves and roots. The expression level in different nitrogen concentration showed that the expression of NRT2.5 gene in low N concentration was significant higher than that in high concentration.

Key words

Camellia sinessis / NRT2.5 gene / clone / expression analysis

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FENG Suhua, WANG Liyuan, CHEN Changsong, LIN Zhenghe, CHENG Hao, WEI Kang, ZHANG Chengcai. Cloning and Expressing Analysis of a Nitrogen Transporter 2.5 Gene from Tea Plant[Camellia sinensis (L.)][J]. Journal of Tea Science. 2014, 34(4): 364-370 https://doi.org/10.13305/j.cnki.jts.2014.04.007

References

[1] Lee R B, Clarkson D T.Nitrogen-13 studies of nitrate fluxes in barley roots. Compartmental analysis from measurements of ¹³N efflux[J]. The EMBO Journal, 1986, 5: 1753-1767.
[2] Tsay Y F, Chu C C, Tsai C B, et al. Nitrate transporters and peptide transporters[J]. Febs Letters, 2001, 581: 2290-2300.
[3] Pao SS, Paulsen IT, Saier MH.Major facilitator super-family[J]. Molecular Biology Reviews, 1998, 62(1): 1-34.
[4] Galvan A, Fernandez E.Eukaryotic nitrate and nitrite transporters[J]. Cellular and Molecular Life Science, 2001, 58: 225-233.
[5] Okamoto M, Jone Vidmar J, Glass A D M. Regulation of NRT1 and NRT2 gene families of Arabidosis thaliana: Responses to nitrate provision[J]. Plant Cell Physoil, 2003, 44(3): 304-317.
[6] Brian G.Nitrate transporters in plants: Structure, function and regulation[J]. Forde Biochimica er Biophysica Acta, 2000, 1465(1/2): 219-235.
[7] Orsel M, Krapp A, Daniel-Vedele F.Analysis of the NRT2 nitrate transporter family in Arabidopsis. Structure and gene expression.[J]. Plant Physiol, 2002, 129: 886-896.
[8] Ryoichi A, Hiroshi H.Expression of rice gene involved in high-affinity nitrate transport during the period of nitrate induction[J]. Breeding Sci, 2006, 56: 295-402.
[9] Trueman L J, Richardson A, Forde B G.Molecular cloning of higher plant homologues of the high-affinity nitrate transporters of Chlamydomonas reinhardtii and Aspergillus nidulans[J]. Gene, 1996, 175(1/2): 223-231.
[10] Vidmar J J, Zhuo D, Siddiqi M Y, et al. Isolation and characterization of HvNRT2.3 and HvNRT2.4, cDNAs encoding high-affinity nitrate transporters from roots of barley[J]. Plant Physiology, 2000, 122(3): 783-792.
[11] Amarasinghe B H, de Bruxelles G L, Braddon M, et al. Regulation of Gmnrt2 expression and nitrate transport activity in roots of soybean (Glycine max)[J]. Planta, 1998, 206(1): 44-52.
[12] Quaggiotti S, Ruperti B, Borsa P, et al. Expression of a putative high-affinity NO₃- transporter and of an H+-ATPase in relation to whole plant nitrate transport physiology in two maize genotypes differently responsive to low nitrogen availability[J]. Journal of Experiment Botany, 2003, 54: 1023-1031.
[13] 赵学强, 李玉京, 刘建中, 等. 小麦NO₃-转运蛋白基因TaNRT2.3的克隆和表达分析[J]. 植物学报, 2004, 46(3): 347-354.
[14] Forde B G.Nitrate transporters in plants: structure, function and regulation[J]. Biochim Biophys Acta, 2000, 1465: 219-235.
[15] 赖灯妮. 高亲和性硝酸盐转运基因型水稻品种初筛及该基因的诱导表达研究[D]. 长沙: 湖南农业大学, 2009.
[16] 蔡超. 禾谷类作物高亲和力NO₃-吸收系统基因表达调控研究[D]. 北京: 中国科学院生态环境研究中心, 2007.
[17] 王新亮. 果树根系硝态氮信号响应关键基因的克隆与功能分析[D]. 泰安: 山东农业大学, 2012.
[18] 石乐松, 刘进平. 文心兰RNA不同提取方法比较[J]. 生物技术, 2012, 22(6): 42-45.
[19] Kivak KJ, Schmittgen TD.Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCT method[J]. Methods, 2001, 25(4): 402-408.
[20] 许金兰, 谢小东, 冯爽, 等. 烟草硝酸盐转运蛋白基因的克隆及表达分析[J]. 烟草农学, 2013, 312(7): 68-71.
[21] 孔敏, 杨学东, 候喜林, 等. 白菜NRT2基因的克隆及表达模式分析[J]. 园艺学报, 2011, 38(12): 2309-2316.
[22] 徐海荣, 谷俊涛, 路文静, 等. 水稻新型硝酸盐转运蛋白基因OsTNrt1的编码蛋白特征和表达[J]. 作物学报, 2007, 33(5): 723-730.
[23] 钟丽华, 宋世威, 陈日远. 菜薹硝酸盐转运蛋白基因NRT2的克隆与表达分析[J]. 园艺学报, 2013, 40: 2650.