620 Testing Neem Leaf Extract to Control Contamination in Guava Tissue Culture

Abstract

Shoot tip explants from field-grown guava (Psidium guajava L.) trees, which frequently show a high rate of contamination, were cultured on the MS medium with neem leaf (Azadirechta indica L.) extract in H2O. Ten grams of neem powder prepared from crushed dried leaves was made to 200 mL aqueous solution and was left on a laboratory bench for 24 h. The amber-color neem extract was filtered to separate and discard solid. The liquid was refrigerated until needed. To 950 mL basic medium in deionized H2O containing 25 mL macronutrients, 5 mL micronutrients, 5 mL vitamins, 5 mL EDTA + Fe, 2 mL BA, 30 g sucrose, and 7 g agar, 50 mL neem extract was added either before or after autoclaving. No neem extract was added to the control. Medium was adjusted to 5.7 pH. Unused medium was refrigerated. Shoot tips from actively growing young twigs of field-grown L-49 guava trees that showed variable contamination in previous studies were harvested during midmorning hours. Samples were washed in running water and disinfected with 15% Clorox solution with few drops of Tween-20 for 15 min. There were 20 explants in each treatment. Explants were subcultured a week later. None of the four test studies showed contamination-free cultures from field trees. There was no consistency due to autoclaving neem extract. The neem leaf extract did not show complete elimination of contamination, although it delayed it for few days. It was concluded that neem extract was not effective at concentrations used in getting rid of contamination. Therefore, further investigate effect of neem for this purpose higher concentrations need to be examined.