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| Author(s): |
Mariana Crotti Franco Grandi
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Campinas, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia |
| Defense date: | 2021-12-17 |
| Examining board members: |
Paulo Mazzafera;
José Francisco de Carvalho Gonçalves;
Ricardo Alfredo Kluge;
Sara Adrián López de Andrade;
Pedro Araújo
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| Advisor: | Paulo Mazzafera |
| Abstract | |
Several Pilocarpus Vahl species are native to Brazilian territory. Popularly known as jaborandi, they have among other secondary metabolites, the alkaloid pilocarpine. Pilocarpine has pharmacological interest and is extract and commercialized in Brazil from cultivated areas and/or sustainable harvest programs. Until now there is no definition of the genes that are involved in natural pilocarpine production, being important to uncover the biosynthetic pathway to optimize pilocarpine concentrations in jaborandi leaves. The present study aims to score genes that may be present at pilocarpine biosynthetic pathway through assays of modulation of alkaloids concentration in leaves and through proteomics and transcriptomics data. Two jaborandi species (P. spicatus and P. microphyllus) were chosen to this study by their contrasting pilocarpine concentrations. All generated data were used to search enzymes from amino acids catabolism, in special from L-threonine and L-histidine that could be involved as jaborandi alkaloids precursors or CYP450 enzymes that can be present along the pathway. Proteomics data provided a total of 4,168 proteins that were analyzed and classified at their respective biological process. From transcriptomic data, the sequenced reads were de novo assembled and analyzed to obtain a list of different expressed genes between the two studied species. From 13,497 different expressed genes, 7,777 were mapped and used to search genes presented in pilocarpine biosynthetic pathway. Threonine ammonia-lyase and phenylalanine ammonia-lyase were found at the sequenced data. Those enzymes can act in the amino acids catabolism in jaborandi alkaloids pathway. CYP450s were mapped through 45 genes with different expression levels between the two jaborandi studied species. Most of the CYP450 genes found in this work were described in other species to be involved in alkaloids biosynthetic pathways. Therefore, the present work brings a database of genes and proteins from two jaborandi species with indication for enzymes that could be acting in the pilocarpine biosynthetic pathway and that can be used in future works (AU) | |
| FAPESP's process: | 14/16488-5 - Identification of the pilocarpine biosynthesis route in jaborandi using genetic variability and biochemistry and molecular techniques |
| Grantee: | Mariana Crotti Franco |
| Support Opportunities: | Scholarships in Brazil - Doctorate |