Identification of NtNRAMP2 Gene and Its Function Analysis in Cadmium Absorption in Nicotiana tabacum
YANG Yuan-Yuan1, CHENG Lan1, XU Jia-Di1, LI Yan-Yan1, DENG Zhao-Long1, HAO Hao-Hao2, WANG Ping-Ping3, JIN Wei-Huan1,*, GUO Hong-Xiang1,*
1 College of Life Sciences, Henan Agricultural University, Zhengzhou 450002, China; 2 Zhumadian Branch of Henan Province Tobacco Company, Zhumadian 463000, China; 3 Shaanxi Branch of China National Tobacco Corporation, Xi'an 710061, China
Abstract：Natural resistance associated macrophage proteins (NRAMPs) play an important role in metal transport in the transmembrane. To investigate the role of tobacco (Nicotiana tobacum) NRAMP genes in heavy metal Cd uptake and transport, a NRAMP subfamily gene, named NtNRAMP2 (GenBank No. OP972862), was cloned from cultivated tobacco 'K326' in this study. Bioinformatics analysis and subcellular localization were carried out to clarify the biological function of the NtNRAMP2, and its expression level in tissue specificity and under exogenous Cd, ABA and ethylene treatment were analyzed by qPCR. Sequence analysis showed that the full-length CDS of tobacco NtNRAMP2 gene was 1 629 bp, encoding 542 amino acids, containing 4 exons and 10 transmembrane structural domains. Evolutionary analyses revealed that NtNRAMP2 was most closely related to tomato (Lycopersicon esculentum) as well as potato (Solanum tuberosum), subcellularly localised in the endoplasmic reticulum and vesicles. qPCR results showed that NtNRAMP2 gene was expressed in different tissues of tobacco, but the expression was highest in the roots. NtNRAMP2 expression was significantly increased under different concentrations and times of Cd stress, and NtNRAMP2 gene expression was also triggered in tobacco roots by both exogenous ABA and ethylene. Meanwhile, NtNRAMP2 overexpression vector was constructed and 18 NtNRAMP2 overexpressing transgenic tobacco plants were obtained by the Agrobacterium tumefaciens mediated transformation method. This study provides a reference for further exploration to study the function of NtNRAMP2.
杨园园, 程岚, 许佳迪, 李燕燕, 邓兆龙, 郝浩浩, 王平平, 金维环, 郭红祥. 烟草NtNRAMP2基因的鉴定及其在镉吸收中功能分析[J]. 农业生物技术学报, 2023, 31(10): 2035-2047.
YANG Yuan-Yuan, CHENG Lan, XU Jia-Di, LI Yan-Yan, DENG Zhao-Long, HAO Hao-Hao, WANG Ping-Ping, JIN Wei-Huan, GUO Hong-Xiang. Identification of NtNRAMP2 Gene and Its Function Analysis in Cadmium Absorption in Nicotiana tabacum. 农业生物技术学报, 2023, 31(10): 2035-2047.
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