O metaboloma da cana-de-açúcar (Saccharum sp.) na resposta à herbivoria

Abstract

The growing world demand for renewable energy production in substitution of fossil fuels has given great prominence in the culture of sugarcane (Saccharum sp.). This has been considered the most efficient crop for energy production, as Brazil being the world's largest producer, with a production of 655.6 million tons in 2015/2016 harvest in an area of 8.5 million hectares. One of the major obstacles to the production of sugarcane is still the attack by pest and estimates that about 10% of the crop losses are caused by insects, being a sugarcane borer (Diatraea Saccharalis) the most importante pest. The plants, during its evolution, to reduce the damages caused by the attack of pests, they have developed a series of defense mechanisms, among them, physical barriers, proteins and toxic metabolites and volatile metabolic flags. Many works have been developed to decipher the mechanisms of defense of sugarcane to the attack of herbivorous insects, an attempt of genetic breeding programs and information biotechnology for the development of more resistant plants, however, many of the mechanisms still remain to be clarified. These works are mostly restricted to the study of genes and proteins and global studies of transcript and proteome analyzes of the plant. The current work proposes to perform an analysis of the metabolism of two varieties of sugarcane (RB92579 and SP791011) through direct and indirect extraction methods in response to herbivory by Diatraea saccharalis using NMR spectroscopy and multivariate statistical analysis from data by principal component analysis (PCA) and least squares analysis of discrimination (OPLS-DA). In the metabolomic analysis of sugarcane of the variety RB92579 was studied using the direct extraction method (leaves) in the 4 herbivory times (24, 48, 72 and 96 hours), only after 48, 72 and 96 hours of stress by herbivory, it was possible to get differences between the control groups and herbivore groups, which the greater timing of response was obtained in 72 hours, that indicated the increase of the primary metabolites asparagine, aspartic acid, dimethylamine, glutamic acid , isoleucine, leucine, malic acid, tyrosine and phenylalanine. The variety of sugarcane SP791011, the method of indirect extraction of the leaves of the plant was applied in times 24, 48 and 72 hours by stress induced by caterpillars of Diatraea saccharalis and, coincidentally, showed in the time 72 hours a greater number of herbivory response metabolites, which caused a depletion of aconitic acid, formic acid, asparagine, alanine and elevation of acetic acid and chlorogenic acid. Despite their different metabolic profiles in response to herbivory, the elucidated metabolites suggest the metabolic pathway of shikimic acid to produce phenylpropanoids due to the increase of tyrosine, phenylalanine in the leaves of the sugarcane variety RB92579, and increase of chlorogenic acid in the leaves of the cane. In addition to the herbivory test, it was carried out a biological assay with chlorogenic acid inserted in the artificial diet of Diatraea saccharalis caterpillars, which demonstrated a decrease in the development time of the pupae comparing with pupae of Caterpillars Control, which caused an outbreak of moths with deformations at all concentrations of chlorogenic acid used in the bioassay. These results may help in the development of sugarcane varieties more resistant to the attack by Diatraea Saccharalis.