Preliminary Study on Transformation of Nicotiana benthamiana with AtHKT1 Promoter

doi:10.7668/hbnxb.2018.S1.002

Abstract

Abstract: The HKT protein family is mainly involved in controlling the absorption of K⁺ and the selective transport of K⁺/Na⁺, which plays an important role in improving the stress resistance of plants.In order to study the relationship between the tissue expression level of AtHKT1 and salt tolerance of plants,the homology of HKT proteins,the expression of AtHKT1 gene and the cis-acting elements of AtHKT1 promoter were analyzed and predicted using bioinformatics techniques. On the basis,the AtHKT1 promoter was introduced into Nicotiana benthamiana. GUS staining results were used to analyze the tissue expression level of AtHKT1 gene in Nicotiana benthamiana. The results of bioinformatics analysis showed that Arabidopsis thaliana and wheat were located in different evolutionary branches and their genetic relationship was far, suggesting that the function of AtHKT1 might be different from the corresponding protein in wheat. AtHKT1 gene was abundantly expressed in many organs and tissues of Arabidopsis thaliana, especially in leaves, roots and flowers, which confirmed that AtHKT1 gene was likely to have important physiological functions. The AtHKT1 promoter might be a stress-responsive promoter containing a number of important elements that responded to environmental stress. Therefore, the expression of AtHKT1 gene was likely to be regulated by environmental stress. Results of GUS staining showed that the coloration of leaves,vascular system,roots and flowers of the seedlings with pHKT1-gus was deeper,further confirming that the expression level of AtHKT1 in these regions was high. The above results indicated that the regulation of AtHKT1 gene expression was conducive to the translocation of Na⁺,thereby regulating the salt tolerance of plants.In addition, there might be other unknown features that further increase the complexity of AtHKT1 functionality. At present, the mechanism of K⁺ and Na⁺ transport of HKT proteins is not clear. The expression level of AtHKT1 gene in Nicotiana benthamiana was analyzed to provide reference for further understanding the action mechanism of AtHKT1 gene.

Key words: AtHKT1 gene, Nicotiana benthamiana, Promoter, Expression level, GUS staining

CLC Number:

Q78
S572.03

LU Yujian, LI Zhen, ZHANG Hongyang, JIANG Cuifeng, HAO Shulei, LIN Jianhui, ZHANG Yonglei, WU Tao. Preliminary Study on Transformation of Nicotiana benthamiana with AtHKT1 Promoter[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(S1): 10-16. doi: 10.7668/hbnxb.2018.S1.002.

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