Uso do ácido salicílico e do ácido jasmônico na produção de metabólitos secundários no cultivo de calos de Bacupari (Garcinia brasiliensis Mart.)

Abstract

Garcinia brasiliensis, known as Bacupari, is a species that stands out for presenting secondary metabolites with different biological activities. However, the nature of the production thereof is very small. To increase production of secondary metabolites, economic and medical interest, biotechnological strategies have been studied as production of these compounds in vitro. The addition of elicitors, such that salicylic acid (SA) and jasmonic acid (AJ), in the culture medium for cell culture may be an alternative to maximize the production of these compounds. In this context, the work was to evaluate the effect of elicitor AS and AJ in the production of phenolics and total flavonoids in callus of zygotic embryos of G. brasiliensis. For callus induction were tested different combinations of 2-iP (0; 9; 18; 36; 72 µM) and 2,4-D (0; 2.05; 10.26; 18.47; 28.73 µM) in the medium of culture Wood Plant Medium (WPM). Callus with 20 days were stained with toluidine blue and lugol for histological evaluation. The areas covered by callus on explants were measured through ImageJ® program. For induction of phenolic compounds and total flavonoid callus were subcultured on WPM containing AS (0.1, 1, 10, 100 µM) in the absence of 2-iP and 2,4-D and AS (0.1, 1, 10, 100 µM) with presence of 72 uM 2-iP and 28.73 mM of 2,4-D and AJ in concentrations 0.1, 1, 10;. 100 uM. The callus were incubated for 150 days at 25 ° C and absence of photoperiod. Every 30 days, callus were subjected to phenolic content analysis and total flavonoid. To determine the kinetics of calluses, these were weighed every 30 days (for treatments with AS) and 15 days (for treatments AJ) to the growth curve. Callus with 30 and 90 days of incubation were analyzed for content of fukugetin and 7-epiclusianone. By making the histological evaluations can be noted that the cells of calluses exposed to the conditions that gave greater callus formation (72 µM of 2-iP with 28.73 µM of 2,4-D) have proved very varied sizes and intercellular space, featuring a disorganized tissue. The area of ​​callus, it was found that the treatment gave better callus induction was 72 uM 2-iP with 28.73 uM 2,4-D. In elicitation, high concentrations of AS (10 and 100 µM) and longer exposure time (120 and 150 days) did not entail greater accumulation of phenolic compounds and total flavonoids. The concentration of 0.1 µM AS was efficient in producing fukugetina and 7-epiclusianone in callus when they are exposed 30 days in culture medium. But when the AS was added to the medium with 2-iP and 2,4-D, the concentration that elicited the production of more fukugetina and 7-epiclusianone was 10 uM AS with 90 days of exposure. The concentration of 0.1 µM AJ promoted the increase of the mass of calluses and G. brasiliensis, increasing the exposure time was favorable production of phenols and flavonoids. The highest concentrations tested AJ (10 and 100 µM) were more efficient in stimulating the production of fukugetina and 7-epiclusianone when the calli 90 days remained exposed to the elicitor. For the production of fukugetin, the most favorable condition for the exposure was 30 days in WPM containing 10 µM of AJ. Anyway, it can conclude that for better callus induction in Bacupari embryos is necessary high concentrations of 2-iP and 2,4-D. The AS and AJ shown to be effective abiotic elicitors on the optimization of secondary metabolites in callus of G. brasiliensis. Taking into account the growth curve of bacupari callus, it can be concluded that the best period for the extraction of secondary metabolites of G. brasiliensis callus was the early stationary phase.