Construction of plant models for functional study of sugarcane Thi1 gene variants

• Grant number: 15/15419-2
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)
• Start date: August 30, 2015
• End date: August 29, 2016
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Marie-Anne Van Sluys
• Grantee: Andréia Prata Vieira
• Supervisor: Michael Andrew Lawton
• Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: Rutgers The State University of New Jersey, New Brunswick, United States
• Associated to the scholarship: 15/05058-2 - At-thi1 homologs in sugarcane: a molecular and functional study, BP.DD

Abstract

Sugarcane (Saccharum spp.) is a C4 photosynthesis monocot and is an important tropical crop grown for sucrose production. The thi1 gene was the first reported to be involved in plant thiamine biosynthesis. THI1 protein is known to be involved in the synthesis of the thiazole ring, a thiamine (vitamin B1) component, which is an essential co-factor in several carbohydrate and amino acid metabolic pathways. While in A. thaliana and other plants thi1 is single copy gene, sugarcane has two copies, like other C4 grasses with the genome sequenced. Previously results showed that each copy presents differences in expression profile. Taken together, combined with the fact that previous studies support that A. thalina thi1 mutant accumulated sucrose in tissues leads to the importance of studying the sugarcane homologues. A previously complementation assay using a yeast auxotrophic to thiamine was not conclusive about the function of the sugarcane thi1 probably because the assay used a heterologous organism. Therefore, the aim of this project is to generate two plant models with modified thi1 genes, which will enable functional studies of sugarcane thi1 genes (sc-thi1.1 and sc-thi1.2). Physcomitrella patens and Brachypodium distachyon will be used. P. patens is a moss considered as a plant model organism as it shares fundamental genetic and physiological processes with vascular plants and has advantages in cultivation based on its small size and life cycle. B. distachyon is a grass related to the major cereal grains and used as model as it has a relatively small genome, short life cycle and small size. To produce the thi1 modified plant lineages, two technologies will be applied - Homologous recombination for P. patens, and CRISPR/Cas9 technology for P. patens and B. distachyon. (AU)