Abstract
Main conclusion
The structure of the cotton uceA1.7 promoter and its modules was analyzed; the potential of their key sequences has been confirmed in different tissues, proving to be a good candidate for the development of new biotechnological tools.
Abstract
Transcriptional promoters are among the primary genetic engineering elements used to control genes of interest (GOIs) associated with agronomic traits. Cotton uceA1.7 was previously characterized as a constitutive promoter with activity higher than that of the constitutive promoter from the Cauliflower mosaic virus (CaMV) 35S gene in various plant tissues. In this study, we generated Arabidopsis thaliana homozygous events stably overexpressing the gfp reporter gene driven by different modules of the uceA1.7 promoter. The expression level of the reporter gene in different plant tissues and the transcriptional stability of these modules was determined compared to its full-length promoter and the 35S promoter. The full-length uceA1.7 promoter exhibited higher activity in different plant tissues compared to the 35S promoter. Two modules of the promoter produced a low and unstable transcription level compared to the other promoters. The other two modules rich in cis-regulatory elements showed similar activity levels to full-length uceA1.7 and 35S promoters but were less stable. This result suggests the location of a minimal portion of the promoter that is required to initiate transcription properly (the core promoter). Additionally, the full-length uceA1.7 promoter containing the 5′-untranslated region (UTR) is essential for higher transcriptional stability in various plant tissues. These findings confirm the potential use of the full-length uceA1.7 promoter for the development of new biotechnological tools (NBTs) to achieve higher expression levels of GOIs in, for example, the root or flower bud for the efficient control of phytonematodes and pest-insects, respectively, in important crops.
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Abbreviations
- GOIs:
-
Genes of interest
- NBTs:
-
New biotechnological tools
- CaMV:
-
Cauliflower mosaic virus
- UTR:
-
Untranslated region
- TSS:
-
Transcriptional start site
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Acknowledgements
We are grateful to EMBRAPA, CAPES, CNPq, INCT PlantStress Biotech, and FAP-DF for providing financial support for this scientific research.
Funding
MFB is grateful to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for a post-doctoral research fellowship (PDJ 150936/2018-4). This work was supported by grants from CAPES, CNPq, FAP-DF, INCT, and EMBRAPA.
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425_2020_3348_MOESM1_ESM.tif
Supplementary file1 (TIF 6714 kb) Overview of the four uceA1.7 modules (highlighted in bold background) aligned with the full-length uceA1.7 promoter. a Module 1 containing 681 bp. b Module 2 containing 792 bp. c Module 3 containing 581 bp. d Module 4 containing 251 bp were planned around the core promoter elements predicted in this study or by Viana et al. (2011). ORF, open reading frame; SC, translation start codon; TSS, transcription start site; 5’ UTR, 5’ untranslated region
425_2020_3348_MOESM2_ESM.tif
Supplementary file2 (TIF 9023 kb) Genetic transformation of Arabidopsis thaliana mediated by Agrobacterium tumefaciens GV3101 strain. a Six independent events from each full-length promoter (uceA1.7 and CaMV 35S) or four uceA1.7 modules (Mod 1 to 4) were evaluated. b PCR detection of the transgene in A. thaliana T1 events using specific primers for the gfp (green fluorescent protein) reporter gene (Suppl. Table 1). Marker: 1.0-kb DNA ladder (Invitrogen® Cat. #10787018); WT, wild-type A. thaliana line used as a negative control for PCR and RT-qPCR assays; C+, DNA plasmid used as a positive control for the PCR assay
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Basso, M.F., Lourenço-Tessutti, I.T., Busanello, C. et al. Insights obtained using different modules of the cotton uceA1.7 promoter. Planta 251, 56 (2020). https://doi.org/10.1007/s00425-020-03348-8
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DOI: https://doi.org/10.1007/s00425-020-03348-8