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Study of Nicotiana tabacum pistil development and function: characterization of the stigma/style expressed genes and protein-protein interactions


In a previous project we identified a gene specifically expressed in stigma/style, encoding a CDK inhibitor (SCI1 - stigma/style cell cycle inhibitor 1), which is responsible for controlling cell proliferation/differentiation, probably as a component of the auxin signaling pathway. To fulfill its Junction, SCI1 interacts with cyclins A and probably with several other proteins. Therefore, the objectives of the present project are: 1) To study the protein interactions established by SCI1: a) Construction and screening of a tobacco stigma/style (S/S) cDNA library, in yeast two-hybrid vector; b) to obtain the SCI1 recombinant protein, produce polyclonal antibody and carry out co-immunoprecipitation; c) Perform pulldown with SCI1 and S/S proteins; d) Perform in vitro and/or in planta experiments to confirm the interactions with the putative partners identified. 2) To study the role of auxin in the pistil development of N. tabacum and Arabidopsis. a) To obtain transgenic plants expressing the chimeric gene STIG1pro::iaaM; b) To analyze the phenotype of the transgenic plants; c) To analyze the expression of SCI1 and auxin-related genes in wild-type and tobacco transgenic plants; d) To analyze the phenotype of Arabidopsis mutants for SCI1 and auxin biosynthesis genes. 3) To identify and characterize genes preferentially/specifically expressed in the pistil: a) To analyze their expression by real time RT-PCR in the vegetative and reproductive organs; b) Elect 3 genes for detailed expression analysis; c) To analyze the subcellular localization of the corresponding proteins by GFP fusion; d) To obtain transgenic plants silenced and overexpressing the elected genes. (AU)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
BRITO, MICHAEL S.; DEPAOLI, HENRIQUE C.; COSSALTER, VIVIANI; AVANCI, NILTON C.; FERREIRA, PEDRO B.; AZEVEDO, MARIANA S.; STRINI, EDWARD J.; QUIAPIM, ANDREA C.; GOLDMAN, GUSTAVO H.; PERES, LAZAR E. P.; GOLDMAN, MARIA HELENA S. A novel cysteine-rich peptide regulates cell expansion in the tobacco pistil and influences its final size. Plant Science, v. 277, p. 55-67, DEC 2018. Web of Science Citations: 0.

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