Eucalyptus urograndis growth under CO2-enriched atmosphere: changes in the chloroplast proteome

Author(s)

Bruna Marques dos Santos

• Document type: Master's Dissertation
• Press: Jaboticabal. 2016-05-24.
• Institution: Universidade Estadual Paulista (Unesp). Faculdade de Ciências Agrárias e Veterinárias. Jaboticabal
• Defense date: 2016-03-30
• Advisor: Tiago Santana Balbuena

Abstract

Carbon dioxide (CO2) emissions from human activities have increased since the industrial revolution. Global projections indicate that there will be a significant increase in the atmospheric concentration of this gas in the coming years. This fact can result in metabolic changes in plants and, consequently, affect the Brazilian forest sector. Chloroplasts are key organelles in carbon fixation and early carbon partitioning in plants. Changes in the availability of CO2 may affect the metabolism of these organelles. The goal of the present study was to assess whether the cultivation of seedlings of Eucalyptus urograndis under a CO2 enriched environment could result in changes in the chloroplast proteome. For this purpose, different chloroplast isolation methods were evaluated to the following parameters: chloroplast morphology observed in bright-field microscopy (1); protein yield after plastid isolation (2); degree of contamination by non-plastidic proteins (3); and abundance in the number of identified proteins described as plastidic (4). After determining the best methodology for the isolation of the chloroplast proteome, E. urograndis seedlings about three months old were grown under CO2 controlled atmospheric concentrations (400 and 1000 ppm) for ten weeks. Evaluation of the plastid proteome, using stringent search against a protein sequence database from Eucalyptus grandis, resulted in the identification of 816 proteins in E. urograndis, from which 80% were already described as plastidic. In silico metabolic pathway mapping resulted in the identification of all proteins involved in the Calvin-Benson cycle and detection of a slight but significant increase in the abundance of key enzymes: PGK, GAPDH, FBA, FBPase, SBPase, and RPI. Although the assessment of the quantum efficiency of photosystem II suggested the absence of changes in the photosynthesis rate, plants treated with 1000 ppm of CO2 presented stomatal closure in response to the imposed environmental condition. A decreased area of the leaf vascular tissue was also detected in young leaves. This is the first characterization of chloroplast proteome of the genus Eucalyptus. Our results indicate that the CO2 enriched atmosphere stimulated metabolic responses, including an increase in the abundance of proteins involved in carbon fixation. Results showed here will assist on the understanding of the biochemical responses stimulated by an increase in the atmospheric CO2 concentration in C3-type plants, and contribute to breeding programs that aim to obtain plants adapted to future climate conditions. (AU)

• FAPESP's process: 14/07454-0 - Characterization of the chloroplast proteome of young plants of Eucalyptus urograndis grown in an atmosphere enriched with carbon dioxide
• Grantee: Bruna Marques dos Santos
• Support Opportunities: Scholarships in Brazil - Master