A novel cold-inducible promoter, PThCAP from Tamarix hispida, confers cold tolerance in transgenic Arabidopsis thaliana

Abstract

Our previous studies have revealed that the ThCAP gene plays a vital role in transgenic Populus (P. davidiana × P. bolleana) in response to cold stress. However, the regulatory mechanism of ThCAP gene expression has been unclear. In this study, the 5′ flanking region of the ThCAP promoter (PThCAP) was cloned using a genome-walking method. By analyzing cis-acting regulatory elements of PThCAP, a DRE motif and MYC and MYB elements were found to be located in the promoter. To identify the regulatory elements that control the expression of the ThCAP gene promoter, a series of deletion derivatives of PThCAP, P1–P5, from the translation start code (−1538, −1190, −900, −718 and −375 bp), were fused to the GUS reporter gene, and then each deletion was stably introduced into Arabidopsis thaliana plants. Deletion analysis of the promoter suggested that only the P2 fragment had strong GUS expression in leaves and roots of A. thaliana exposed to low temperature stress. These results suggest that this 290-bp region (−1190 to −900 bp), as an important part in PThCAP, was associated with cold tolerance of A. thaliana. Our results provide evidence for the regulatory mechanism of ThCAP gene involved in the response to cold stress, and that the gene is promising candidate gene for genetic improvement of crops.

Keywords

Arabidopsis thaliana / Cis-acting elements / GUS expression / PThCAP / ThCAP gene

References

[1] Basehoar AD, Zanton SJ, Pugh BF. Identification and distinct regulation of yeast TATA box-containing genes. Cell, 2004, 116: 699-709.

[2] Company N, Nadal A, Ruiz C, Pla M. Production of phytotoxic cationic α-helical antimicrobial peptides in plant cells using inducible promoters. PLoS One, 2014, 9: e109990.

[3] Doyle JJ, Doyle JL. Isolation of plant DNA from fresh tissue. Focus, 1990, 12: 13-15.

[4] Fang ZW, Xu XY, Gao JF, Wang PK, Liu ZX, Feng BL. Characterization of FeDREB1 promoter involved in cold- and drought-inducible expression from common buckwheat (Fagopyrum esculentum). Genet Mol Res, 2015, 14: 7990-8000.

[5] Fei J, Wang YS, Jiang ZY, Cheng H, Zhang JD. Identification of cold tolerance genes from leaves of mangrove plant Kandelia obovata by suppression subtractive hybridization. Ecotoxicology, 2015, 24: 1686-1696.

[6] Guo XH, Jiang J, Lin SJ, Wang BC, Wang YC, Liu GF, Yang CP. A ThCAP gene from Tamarix hispida confers cold tolerance in transgenic Populus (P. davidiana × P. bolleana). Biotechnol Lett, 2009, 31: 1079-1087.

[7] Jefferson RA. The GUS reporter gene system. Nature, 1989, 342: 837-838.

[8] Kovalchuk N, Jia W, Eini O, Morran S, Pyvovarenko T, Fletcher S, Bazanova N, Harris J, Beck-Oldach K, Shavrukov Y, Langridge P, Lopato S. Optimization of TaDREB3 gene expression in transgenic barley using cold-inducible promoters. Plant Biotechnol J, 2013, 11: 659-670.

[9] Li H, Wang Y, Jiang J, Liu G, Gao C, Yang C. Identification of genes responsive to salt stress on Tamarix hispida roots. Gene, 2009, 433: 65-71.

[10] Mayer BF, Ali-Benali MA, Demone J, Bertrand A, Charron JB. Cold acclimation induces distinctive changes in the chromatin state and transcript levels of COR genes in Cannabis sativa varieties with contrasting cold acclimation capacities. Physiol Plant, 2015, 155: 281-295.

[11] Nataliya KC, Wei J, Omid E, Sarah M, Tatiana P, Stephen F, Natalia B, John H, Kontanze BO, Yuri S, Peter L, Sergiy L. Optimization of TaDREB3 gene expression in transgenic barley using cold-inducible promoters. Plant Biotechnol J, 2013, 11: 659-670.

[12] Shinozaki K, Yamaguchi-Shinozaki K. Gene expression and signal transduction in water-stress response. Plant Physiol, 1997, 115: 327-334.

[13] Trivedi DK, Gill SS, Tuteja N (2016) Abscisic acid (ABA): biosynthesis, regulation, and role in abiotic stress tolerance. In: Tuteja N, Gill SS (eds) Abiotic stress response in plants, Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. doi:10.1002/9783527694570.ch15

[14] Yamaguchi-Shinozaki K, Shinozaki K. Organization of cis-acting regulatory elements in osmotic-and cold-stress-responsive promoters. Trends Plant Sci, 2005, 10: 88-94.

[15] Yu X, Liu Y, Wang S, Tao Y, Wang Z, Shu Y, Ma H. CarNAC4, a NAC-type chickpea transcription factor conferring enhanced drought and salt stress tolerances in Arabidopsis. Plant Cell Rep, 2016, 35: 613-627.

[16] Zhang JY, Huang SN, Wang G, Xuan JP, Guo ZR. Overexpression of Actinidia deliciosa pyruvate decarboxylase1 gene enhances waterlogging stress in transgenic Arabidopsis thaliana. Plant Physiol Biochem, 2016, 106: 244-252.

[17] Zong JM, Li XW, Zhou YH, Wang FW, Wang N, Dong YY, Yuan YX, Chen H, Liu XM, Yao N, Li HY. The AaDREB1 transcription factor from the cold-tolerant plant adonis amurensisenhances abiotic stress tolerance in transgenic plant. Int J Mol Sci, 2016