Development of sugarcane synthetic promoters responsive to water deficit stress

• Grant number: 24/01328-4
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: May 01, 2024
• End date: February 29, 2028
• Field of knowledge: Biological Sciences - Genetics - Plant Genetics
• Principal Investigator: Marcelo Menossi Teixeira
• Grantee: Bruno Spinassé Floreste
• Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Company: Ministério da Agricultura, Pecuária e Abastecimento (Brasil). Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA). Embrapa Sede
• Associated research grant: 22/04006-2 - Center for Plant Molecular Breeding (CeM²P), AP.PCPE

Abstract

Sugarcane is a crop of great economical importance due to the production of sugar and ethanol. Water deficit caused by decreases in precipitation and periods of drought result in significant loss of productivity. Several biotechnological tools have been applied to improve plant tolerance to water deficit, and in most cases constitutive promoters have been used, which can lead to waste of energy and deleterious effects. Synthetic promoters have been developed to fine tune transgene expression, avoiding these kinds of issues. In this work we will use transcription factor binding sites motifs from sugarcane to develop synthetic promoters responsive to water deficit stress. The DNA motifs will be separated by random generated spacers and associated with a minimal 35S promoter. The promoters will be assembled by Golden Gate cloning in vectors containing a GFP cassette and a CaMV35S double promoter expressing a RFP as control. The fluorescence will be analyzed after protoplast transfection using PEG as a water deficit agent and the optimal architecture selected. Then, fusions of the synthetic promoter with the GUS gene will be validated in planta, by transformation of the cultivar SP80-3280. Full grown plants will be subjected to a water withholding treatment and samples from multiple tissues will be analyzed for GUS enzymatic activity for quantification of promoter expression. (AU)