title: Construction of efficient, tuber-specific, and cold-inducible promoters in potato.
abstract: Promoter activity is crucial for precise gene expression. Previously, a synthetic tuber-specific and cold-inducible promoter, pCL, containing a C-repeat/dehydration-responsive element (CRT/DRE) cassette and a tuber-specific fragment, was constructed in order to regulate cold-induced sweetening (CIS) in potatoes. However, the utility of pCL is limited due to its low activity. To improve its inducibility in response to low temperatures, we modified the CRT/DRE and flanking sequences. In particular, promoter activity was significantly improved by site-specific mutation of flanking sequences next to the core element (CCGAC) of CRT/DRE. We also inserted a modified CRT/DRE cassette into pCL; although this enhanced activity, it was not more effective than mutation of the flanking sequences. Indeed, up to 20-fold enhanced pCL activity could be achieved by replacing the CRT/DRE cassette in pCL with tandem repeats of two mutated CRT/DRE cassettes. This improvement was due to an enhanced affinity between the CRT/DRE cassette(s) and the StCBF1 transcription factor. Together, these data suggest that altering the structure of CRT/DRE can enhance CBF-related transcription complex formation and thus improve the activity of this cold-inducible promoter.
authors.affiliation: National Center for Vegetable Improvement (Central China)/Key Laboratory of Horticulture Plant Biology (Huazhong Agricultural University), Ministry of Education/Huazhong Agricultural University, Wuhan 430070, People's Republic of China; Central South University of Forestry and Technology, Changsha, People's Republic of China. Electronic address: limeng0422@foxmail.com.
authors.forename: Meng
authors.initials: M
authors.lastname: Li
authors.affiliation: National Center for Vegetable Improvement (Central China)/Key Laboratory of Horticulture Plant Biology (Huazhong Agricultural University), Ministry of Education/Huazhong Agricultural University, Wuhan 430070, People's Republic of China.
authors.forename: Conghua
authors.initials: C
authors.lastname: Xie
authors.affiliation: National Center for Vegetable Improvement (Central China)/Key Laboratory of Horticulture Plant Biology (Huazhong Agricultural University), Ministry of Education/Huazhong Agricultural University, Wuhan 430070, People's Republic of China.
authors.forename: Botao
authors.initials: B
authors.lastname: Song
authors.affiliation: National Center for Vegetable Improvement (Central China)/Key Laboratory of Horticulture Plant Biology (Huazhong Agricultural University), Ministry of Education/Huazhong Agricultural University, Wuhan 430070, People's Republic of China.
authors.forename: Yongbin
authors.initials: Y
authors.lastname: Ou
authors.affiliation: National Center for Vegetable Improvement (Central China)/Key Laboratory of Horticulture Plant Biology (Huazhong Agricultural University), Ministry of Education/Huazhong Agricultural University, Wuhan 430070, People's Republic of China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lin
authors.affiliation: National Center for Vegetable Improvement (Central China)/Key Laboratory of Horticulture Plant Biology (Huazhong Agricultural University), Ministry of Education/Huazhong Agricultural University, Wuhan 430070, People's Republic of China.
authors.forename: Xun
authors.initials: X
authors.lastname: Liu
authors.affiliation: National Center for Vegetable Improvement (Central China)/Key Laboratory of Horticulture Plant Biology (Huazhong Agricultural University), Ministry of Education/Huazhong Agricultural University, Wuhan 430070, People's Republic of China.
authors.forename: Huiling
authors.initials: H
authors.lastname: Zhang
authors.affiliation: National Center for Vegetable Improvement (Central China)/Key Laboratory of Horticulture Plant Biology (Huazhong Agricultural University), Ministry of Education/Huazhong Agricultural University, Wuhan 430070, People's Republic of China. Electronic address: liujun@mail.hzau.edu.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
mesh_terms: D017360:Arabidopsis / Q000235:genetics; D050260:Carbohydrate Metabolism / Q000235:genetics*; D003080:Cold Temperature; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D005818:Genetic Engineering / Q000379:methods; D009154:Mutation; D010940:Plant Proteins / Q000235:genetics; D035281:Plant Tubers / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D020218:Response Elements; D011198:Solanum tuberosum / Q000235:genetics* / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic; D015533:Transcriptional Activation
chemical_list: D010940:Plant Proteins; D015534:Trans-Activators; D014157:Transcription Factors
keywords: Cold-inducible C-repeat/dehydration-responsive element; Flanking sequence; Potato (Solanum tuberosum L.); Promoter