Indução de calos em espécies amazônicas do gênero Theobroma

Abstract

Many works have been done on cocoa (Theobroma cacao) in vitro culture, with few studies being published for other species of the same genus, as cupuassu (T. grandiflorum), whose planted area is increasing expressively, and others that could be used as a source of genes for those with recognized economical importance. Protocols to obtain in vitro somatic embryos from T. cacao,T. grandiflorum,T. speciosum and the hybrid T. grandiflorum x T. obovatum from two sources of explants, staminodes and petals (formed by ligues and cogules) were evaluated, using a primary callus growth medium made of DKW salts, supplemented with 20 g l-1 of sucrose, 250 mg l-1 of glutamine, 200 mg l-1 of myo-inositol, 0.2 mg l-1 of thyamine-HCl; 0.1 mg l-1 of nicotinic acid; 0.2 mg l-1 of glycine; 2 mg l-1 of 2,4-D; 2.2 g l-1 of Gelrite® and the pH adjusted to 5.8. To this media was added different concentrations of thidiazuron (0; 5 and 10 µg l-1). Cultures were maintained at dark for 14 days, at a temperature of 25 ± 2 ºC, and so transferred for the secondary callus growth, made with WPM salts, Gamborg vitamins, 20 g l-1 of sucrose, 2 mg l-1 of 2,4 D; 0.3 mg l-1 of cinetin, 50 ml l-1 of coconut milk, 2.2 g l-1 of Gelrite® and the pH adjusted to 5.8. Callus formation occurred in all species. Somatic embryos were obtained only for T. cacao. Callus formation was influenced by genotype and was higher on staminodes.