Micropropagation of Ilex glabra (L.) Gray

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American Society for Horticultural Science

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Nodal segments containing one axillary bud (1 to 1.5 cm) were disinfected using 10% bleach and were established on a Murashige and Skoog (MS) medium without hormones at 27 °C and with a 16-h photoperiod. The sprouted shoots (≈1.0 cm) were cultured on a MS medium supplemented with 6-benzylaminopurine (BAP), kinetin (KIN), or zeatin (ZT) at 2.3, 4.5, 9.1, or 18.2 μM. After 38 d, ZT and BAP significantly induced multiple shoot formation with multiplication rates of 4 to 6, whereas the multiplication rate of KIN was less than 2. Shoots cultured on ZT grew significantly taller than those on BAP and KIN. The height of the longest shoots treated with ZT was 4.6 cm, which was 1.6 to 2.2 times greater than those treated with BAP or KIN. To induce rooting, shoots (≈2 cm) were subcultured on one-fourth strength MS (1/4 MS) medium containing either 3-indolebutyric acid (IBA) or 1-naphthylacetic acid (NAA) at 2.6, 5.1, or 10.3 μM. Adventitious roots formed in vitro after 2 to 4 weeks. IBA at 10.3 μM produced the best rooting (100%) compared with other treatments after 38 d of culture. The average number of roots per shoot for IBA was ≈15, which was 1.6 to 3.1 times as many as that of other treatments. All rooted plantlets were then transplanted into a mix of peatmoss and perlite (1:1 v/v) and acclimatized in a mist system. Average plantlet survival was 73.6% after 35 d. After acclimatization, they were grown in a pot with Metro-mix under greenhouse conditions for 10 weeks where 95% of plants survived and grew up to 6.8 cm high. The micropropagation procedure, i.e., nodal segments containing one axillary bud proliferated on MS with 4.5 μM ZT followed by in vitro rooting on 1/4 MS plus 10.3 μM IBA, could be used for commercial mass production of new inkberry cultivars.

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