In vitro culture of zygotic embryos of Ilex species

Plants of Ilex argentina L., I. brasiliensis (S.) L., I. brevicuspis R., I. dumosa R., I. integerrima (V.C.) L., I. microdonta R., I. pseudoboxus R., and I. theezans C.M. were obtained by immature embryo culture. Heart-stage zygotic embryos were removed from immature fruits and cultured aseptically on quarter-strength Murashige and Skoog medium,vith 3% sucrose, 0.65% agar, and 0.1 mg.L-1 zeatin. Cultures were incubated at 27 +/- 2 degreesC for 4 weeks, in darkness and subsequently transferred to a culture roam with a 14-hour photoperiod (116 mu mol.m(-2).s(-1)) for another 4 weeks. Seedlings with two leaves, derived from germinated embryos, were successfully transplanted to pots containing 1 peat: 1 perlite : 1 sand (v/v) and were maintained in greenhouse conditions. From 95% to 100% of transplanted seedlings survived. Chemical name used: 6-(4-hydroxy-3-methylbut-2-enylamino) purine (zeatin).