Quitosana na indução de resistência e controle in vitro de mofo cinzento, podridão parda e podridão amarga

Abstract

With the rapid population growth and the reduction of areas for cultivation, the increased productivity of foods of all kinds is essential to meet the world's population demand. However post-harvest losses from the time of harvest until the arrival to the consumer, limits the supply of fruits to the consumer. The losses in the post-harvest of apples, strawberries and peaches, caused by the incidence of rot led by phytopathogenic fungi, is responsible for most of the losses. These rots are caused by various fungi, among these are the Colletotrichum sp., cause of bitter rot on apples, Botrytis cinerea causative agent of gray mold on strawberries and M. fructicola on peaches which causes brown rot. These phytopathogens are fungi with high ability to spread infection and therefore cause serious damage to fruits, generating losses in post-harvest. Usually the fungi control is conducted with the use of fungicides. However, the use of chemicals in the process concerns consumers, since there may be residues in fruits and the environment. Therefore, alternative methods like the resistance induction can be used to control the disease-causing microorganisms in the postharvest fruits. The induction consists on stimulating the plants defenses through (inducing) elicitor molecules, specifically the synthesis of compounds that act directly on the pathogen as phenols, protein-RPs, or producing structural reinforcement of tissues adjacent to the site of infection of the fungus. Currently, chitosan extracted from crustacean shells is an alternative elicitor molecule of low cost and no risk to the consumer that has been used in the induction of the resistance in postharvest fruits. This biopolymer has the ability to trigger the defensive responses of the fruits; apple, peach, and strawberry against fungi Colletotrichum sp., Botrytis cinerea and Monilinia fructicola, respectively. The inductor was applied at concentrations of 0.25, 0.5; 1.0 and 2.0% and on the control (distilled water). The treatments were arranged in a completely randomized design with four replications of 20 apple fruits, 20 strawberry fruits and 15 peach fruits. The fruits were selected and standardized, and subsequently the apples and peaches were subjected to treatment by immersion in solutions of chitosan, and strawberries were treated by spraying the chitosan-containing solution directly onto the fruits. After 24 hours, the fruits were inoculated with a solution containing conidia of the phytopathogen Colletotrichum sp. on apples, B. cinerea on strawberries and M. fructicola on peaches, in concentrations of [5.10]^(-3) conidia/ml directly on the fruit, with the help of a spray bottle or with a micropipette in the case of apples, directly inoculating the solution in an wound to the 2 mm bark. After completion of the treatments, the fruits were placed in BOD at 26 ± 1 ° C for apples and peaches, and 10 ± 1 ° C for strawberries, and evaluated after 24 hours to determine the following parameters; weight loss, physical and chemical analysis (solids and total soluble, titratable acidity, firmness, decay incidence) and biochemical (protein, reducing sugars, total sugars, anthocyanins, flavonoids, FAL, peroxidase, chitinase and β-glucanase). An initial sample of the fruits was taken to carry out the initial analyzes, using this data as comparative parameters. In a second experiment, the fungus (Colletotrichum sp, Botrytis cinerea and M. fructicola) were grown in the middle of culture containing the different concentrations of chitosan, to verify the existence of fungitoxic or fungistatic effect of the biomolecule in vitro. The fungi were previously cultivated in clean plates, and subsequently transferred to plates containing PDA medium with the chitosan concentrations (0, 0.25, 0.5, 1 and 2), and after 24 and 48 hours, were performed perpendicular measurements of the diameter of the colony to verify its mycelial growth. Data from experiments were submitted to analysis of normality and variance, and measures were compared by Tukey test and regression test (p = 0.05), with the assistance of Assistat software. The results demonstrated the interference of chitosan on the induction of resistance to control the incidence of bitter rot in postharvest Gala apple, activating the PRPs B-1-3 glucanase and control of Colletotrichum sp in vitro with fungistatic action. On strawberries, the inductor controlled the gray mold by activating the peroxidase, chitinase and β-1-3-glucanase, directly under the fungus B. cinerea in vitro. On peaches, the action was on the maintenance of fruit quality, on the induction, activating genes of chitinase, β 1-3 glucanase, and on the same way, on the in vitro of M. fructicola. Therefere, it has been concluded that chitosan has great potential in the induction of fruit resistance in post-harvest, activating the defense against pathogenic fungi, and directly over the latter with fungistatic activity and in vitro fungitoxic activity.