Marcos Fernando Basso1, Isabela Tristan Lourenço-Tessutti2, Carlos Busanello3, Clidia Eduarda Moreira Pinto2,4, Elínea de Oliveira Freitas2,4, Thuanne Pires Ribeiro2, Janice de Almeida Engler5, Antonio Costa de Oliveira3, Carolina Vianna Morgante2,6, Marcio Alves-Ferreira7, Maria Fatima Grossi-de-Sa8,9. 1. Embrapa Genetic Resources and Biotechnology, PqEB Final, W5 Norte, PO Box 02372, Brasília, DF, 70770-901, Brazil. marcosbiotec@gmail.com. 2. Embrapa Genetic Resources and Biotechnology, PqEB Final, W5 Norte, PO Box 02372, Brasília, DF, 70770-901, Brazil. 3. Federal University of Pelotas, Capão Do Leão, RS, 96160-000, Brazil. 4. Federal University of Brasília, Brasília, DF, 70910-900, Brazil. 5. UMR Institut Sophia Agrobiotech INRA/CNRS/UNS, Sophia Antipolis, France. 6. Embrapa Semi Arid, Petrolina, PE, 56302-970, Brazil. 7. Federal University of Rio de Janeiro, Rio de Janeiro, RJ, 21941-901, Brazil. 8. Embrapa Genetic Resources and Biotechnology, PqEB Final, W5 Norte, PO Box 02372, Brasília, DF, 70770-901, Brazil. fatima.grossi@embrapa.br. 9. Catholic University of Brasília, Brasília, DF, 71966-700, Brazil. fatima.grossi@embrapa.br.
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. 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.
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. 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.
Authors: Wen Zhi Wang; Ben Peng Yang; Xiao Yan Feng; Zheng Ying Cao; Cui Lian Feng; Jun Gang Wang; Guo Ru Xiong; Lin Bo Shen; Jun Zeng; Ting Ting Zhao; Shu Zhen Zhang Journal: Front Plant Sci Date: 2017-09-28 Impact factor: 5.753