Cloning and Functional Analysis of Gene NtNRAMP3b in Nicotiana tabacum

doi:10.13560/j.cnki.biotech.bull.1985.2022-0382

Abstract

Abstract:

Iron（Fe）affects plant growth and development，and NRAMP（natural resistance-associated macrophage protein）in animals and plants is mainly responsible for the absorption and transport of metals ions such as Fe，Mn and Cd. Studying the transport mechanism of NtNRAMP3b to Fe is to provide a theoretical basis for clarifying the mechanism of NtNRAMP3b in tobacco（Nicotiana tabacum）transport iron. Taking the cDNA of tobacco cultivar K326 as a template，the tobacco’s NtNRAMP3b was cloned and its sequence characteristics were analyzed，the instantaneous expression of NtNRAMP3b was determined by RT-qPCR. Using transgenic technology to obtain NtNRAMP3b-OE（NtNRAMP3b overexpression），and its iron concentration was determined. NtNRAMP3b was 1 530 bp long and encoded 509 amino acid sequences. NtNRAMP3b was most closely related to AtNRAMP3，with 74.85% homogeneity. RT-qPCR results showed that NtNRAMP3b was expressed the most in leaves，and the relative expression after iron deficiency treatment increased. Compared with the wild type，NtNRAMP3b-OE biomass increased，chlorophyll concentration increased，and iron content decreased after iron deficiency treatment. NtNRAMP3b is mainly involved in the transport of iron from vacuoles to organelles，improving the absorption and reuse of iron in leaves.

Key words: Nicotiana tabacum, NtNRAMP3b, iron deficiency, iron content

YIN Zhuo-ran, XUAN Dong-dong, LI Chen-yi, LI Chang, CHAI Zhe, WANG Kun-yao, ZHAO Meng-qi, PENG Jing-yuan, DONG Jie, JIA Hong-fang. Cloning and Functional Analysis of Gene NtNRAMP3b in Nicotiana tabacum[J]. Biotechnology Bulletin, 2022, 38(12): 175-183.

Figures/Tables 7

Fig. 1 Cross-membrane domain analysis of NtNRAMP3b

Fig. 2 Homology analysis

Fig. 3 Phylogenetic tree and motif analysis

Fig. 4 Promoter cis-element analysis

Fig. 5 NtNRAMP3b relative expression Different letters indicate significant differences in the P< 0.01 level

Fig. 6 Biomass under treatment with different cadmium concentrations In the same treatment, *and ** indicate OE1 and OE2 has a significant difference in the P<0.05 or P< 0.01 compared to WT. The same below

Fig. 7 Transient expressions of genes under iron deficiency

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