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Analysis of expression and phenotypic evaluation of plants of sugar cane genetically transformed with the gene-C4 PEPC

Grant number: 12/12268-5
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): November 01, 2012
Effective date (End): June 30, 2013
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Helaine Carrer
Grantee:Flávio dos Santos
Home Institution: Escola Superior de Agricultura Luiz de Queiroz (ESALQ). Universidade de São Paulo (USP). Piracicaba , SP, Brazil
Associated research grant:08/52066-7 - Functional genomics of photosynthetic genes in sugarcane, AP.BIOEN.TEM

Abstract

To analyze the phenotypic plant cane sugar overexpressing C4-PEPC gene in normal conditions and under water stress. Total RNA extraction perform for analysis of expression of PEPC in C4-transformed plants. Plants that are experiencing the highest levels of expression in real-time quantitative PCR will be selected for the experiments. Will be judged on condition of daily watering and dry, the following parameters:- Physiological: CO2 assimilation (A), transpiration (E), stomatal conductance (gS), intercellular CO2 concentration (Ci), apparent carboxylation efficiency (A / Ci), water use efficiency (WUE = A / E ) intrinsic efficiency of water use (WUEI = a / gS) and relative amount of water on the leaves (RWC) (Barrs and Weatherley, 1962).- Biometric: shoot growth and root, stem diameter, leaf number, tiller number, fresh weight and dry weight.- Biochemical parameters: total soluble sugars (Dubois et al., 1956), reducing sugars (Nelson, 1944), total soluble protein (Bradford, 1976), proline content (Bates et al., 1973).The physiological parameters are measured with the IRGA unit Infra Red Gas Analyzer.Expression analysis will be conducted in quantitative PCR in a thermocycler One Step Plus (Applied Biosystems).All data will be subjected to statistical analysis (ANOVA), the averages are compared by Tukey test at 5% probability.

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