Promoter cloning and analysis of low temperature-induced expression of sikFBA4 from Saussurea involucrata

doi:10.11686/cyxb2017527

Abstract

Abstract: Saussurea involucrata tolerates severe abiotic stresses, especially cold conditions. Therefore, it is a useful plant for studies on cold adaptability. Previous studies have shown that the cold tolerance of Lycopersicon esculentum was significantly improved by over-expressing sikFBA4. To further analyze sikFBA4, changes in its transcript levels under a 4 ℃ treatment were determined by qRT-PCR. Hi-Tail PCR was used to clone the promoter sequence of sikFBA4 from genomic DNA, and PlantCARE software was used to predict and analyze promoter elements in PsikFBA4. To evaluate the activity and study the expression characteristics of PsikFBA4, we constructed GUS gene reporter constructs for transient expression in Nicotiana tabacum leaves. The results showed that sikFBA4 was strongly induced by low temperature. Its transcript levels peaked at 1 hour of cold treatment before declining slowly. The bioinformatics analysis indicated that a low temperature response element was located at -1648 bp in the gene promoter. The result that GUS activity was regulated by PsikFBA4 in response to a cold treatment confirmed that PsikFBA4 is a cold-inducible promoter. These results provide a foundation for further research on the expression and regulation mechanism of sikFBA genes.

Key words: Saussurea involucrata, sikFBA4, Hi-Tail PCR, promoter, GUS

ZHANG Yao, LI Zhong-qing, WANG Ai-ying, ZHU Jian-bo. Promoter cloning and analysis of low temperature-induced expression of sikFBA4 from Saussurea involucrata[J]. Acta Prataculturae Sinica, 2018, 27(12): 199-207.

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