Plant regeneration from Ilex spp. (Aquifoliaceae) in vitro

In vitro plant regeneration from nodal segments (containing one axillary bud) of seven species of the genus Ilex (I. argentina, I. brevicuspis, I. dumosa, I. microdonta, I. pseudoboxus, I. taubertiana and I. theezans) were readily achieved through three steps: 1) shoot regeneration by in vitro culture of nodal segments in MS medium at 1/4 strength, plus 3% sucrose and 0.65% agar (1/4MS) and 0.5 microM BA (45 days of culture); 2) Induction of rooting from regenerated shoots with 1/4MS (solidified with 2.5 g.L-1 "Phytagel") with 7.3 microM IBA (7 days) and, 3) subculture of shoot on a fresh medium (1/4MS lacking plant growth regulators) during 21 days. Shoot regeneration of other three species (I. aquifolium, I. brasiliensis and I. integerrima) were also obtained by in vitro culture of nodal segments. Shoot regeneration of I. aquifolium, I. brasiliensis, I. integerrima, I. microdonta, I. pseudoboxus, and I. taubertiana were also obtained by culture shoot tips on 1/4MS and 0.5 microM BA. Shoot regeneration from meristems of I. argentina, I. brevicuspis, I. dumosa, and I. theezans were readily achieved by in vitro culture on the same medium.