Cloning and expression pattern analysis of tobacco high affinity nitrate transporter gene NtNRT2.5
[Background and Objective]High-affinity nitrate transporter NRT2 family genes play an important role in nitrate absorption,transport and reuse in plants.This study aims to explore the role of tobacco NRT2 family genes NtNRT2.5 in regulating nitrogen in tobacco.[Methods]The high affinity nitrate transporter gene NtNRT2.5 was cloned from tobacco variety K326,and its bioinformation and expression pattern were analyzed.K326 transgenic tobacco material with GUS reporter gene fusion of NtNRT2.5 promoter was created,and GUS tissue chemical staining was performed.[Results]The full-length ORF of the NtNRT2.5 gene is 1509 bp,encoding 502 amino acids,and is a stable hydrophobic protein.Its transmembrane protein domain has the structural characteristics of the NRT2 family.Tobacco NtNRT2.5 is closely related to Arabidopsis AtNRT2.5 and shares similar function.The gene promoter contains a variety of hormone and stress response elements.The NtNRT2.5 protein is localized to the plasma membrane of the cell.The NtNRT2.5 gene was expressed in all tissues of tobacco,with the strongest expression in the lower leaves,and its expression was enhanced under nitrogen-deficient conditions.GUS tissue staining results showed that,under both regular and low nitrogen conditions,the GUS reporter gene can be detected in the roots(axis)and lower leaves(veins)of transgenic tobacco,displaying a blue hue,with a deeper blue and stronger expression under low nitrogen conditions[Conclusion]Tobacco NtNRT2.5 encodes a high affinity nitrate transporter,and its expression is enhanced under nitrogen-deficient conditions,suggesting its potential involvement in tobacco nitrate transport and reuse.
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