| Grant number: | 16/20486-3 |
| Support type: | Regular Research Grants |
| Duration: | March 01, 2017 - August 31, 2019 |
| Field of knowledge: | Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms |
| Principal researcher: | Maria Helena de Souza Goldman |
| Grantee: | Maria Helena de Souza Goldman |
| Home Institution: | Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil |
Abstract
This project aims to study the protein SCI1(Stigma / style Cell Cycle Inhibitor-1) of Nicotiana tabacum and the mechanisms by which it regulates the development and influence the final size of the female reproductive organ of plants (pistil). Notably, this project aims to advance the understanding of how SCI1 regulates cell proliferation. The objectives of this project are: 1) To investigate the hypothesis of SCI1 receiving post-translational modifications. This investigation will be done primarily by mass spectrometry of protein extracted from stigmas / styles. 2) To confirm and expand knowledge of the interactome of SCI1 and NtCDKG 2. The analysis of the clones resulting from the yeast two-hybrid screening (Y2H), using NtCDKG; 2 as bait, will be continued. Additionally, some interaction partners identified with SCI1 will be tested with NtCDKG, 2 and vice-versa, by Y2H and Bimolecular Fluorescence complementation (BiFC). 3) To investigate the hypothesis of SCI1 playing a role in processes related to RNA splicing. The interactions of SCI1 with NtU2a' (spliceosome), NtRS2Z33 (splicing factor), and some candidates of goal 2, will be tested. 4) To investigate the hypothesis of SCI1 playing a role in processes related to RNA transcription. The results of the RNASeq of stigmas/styles of wild-type and SCI1 RNAi plants will be analyzed, the differentially expressed genes will be identified and RT-qPCR experiments will be performed to confirm the results. 5) To investigate the hypothesis SCI1 being expressed in the floral meristem and having a role in controlling cell proliferation in this meristem. In situ hybridization of SCI1 in floral meristems will be done, as well as the production and analysis of transgenic plants SCI1prom:: SCI-GFP. The production of SCI1 knockout plants by CRISPR/Cas 9 will be initiated. (AU)