Transformation of <Emphasis Type="Italic">Liquidambar formosana</Emphasis> L. via <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> using a mannose selection system and recovery of salt tolerant lines |
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Authors: | Guirong Qiao Jing Zhou Jing Jiang Yuehua Sun Luanyin Pan Honggai Song Jingmin Jiang Renying Zhuo Xiaojuan Wang Zongxiu Sun |
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Institution: | (1) Postgraduate and Research Department of Botany, St. Thomas College, Pala, Arunapuram (P.O), Kottayam (Dt.), Kerala, 686 574, India;(2) Field Science Centre for Northern Biosphere, Hokkaido University, Kita 11, Nishi 10, Kita-ku, Sapporo 060-0811, Japan; |
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Abstract: | Malaxis acuminata is a terrestrial orchid that grows in shady areas of semi-evergreen to shrubby forests. It is highly valued for its medicinal
properties as dried pseudo-bulbs are important ingredients of several Ayurvedic preparations. In this study, adventitious
shoot buds were induced from internodal explants of M. acuminata grown on Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kn),
and thidiazuron (TDZ). Of the three cytokinins used, TDZ at 3 mg l−1 induced the highest frequency (82%) of organogenic explants. However, all responding explants produced only a single adventitious
shoot irrespective of the type and concentration of the cytokinin. Adding 0.5 mg l−1 α naphthaleneacetic acid (NAA) to the medium enhanced adventitious shoot formation. In the presence of 3 mg l−1 TDZ and 0.5 mg l−1 NAA, frequency of organogenesis was 96% with a mean number of 6.1 shoots per explant. Prolonged culture or subculture on
the same medium did not promote further shoot production. However, transfer of these cultures to MS medium supplemented with
3 mg l−1 TDZ and 0.5 mg l−1 NAA and various concentrations of different polyamines (PAs), including spermine, spermidine, and putrescine, significantly
increased mean shoot number per explant. The highest frequency of shoot induction (100%) and mean shoot number per explant
(14.6) was observed on MS medium with 3 mg l−1 TDZ, 0.5 mg l−1 NAA, and 0.4 mM spermidine. Regenerated shoots were excised and subcultured on an elongation medium consisting of MS medium
with 3 mg l−1 BA. Moreover, the highest frequency of rooting (96%) and mean number of roots per shoot (3.3) was observed on MS medium with
4 mg l−1 indole-3-butyric acid (IBA) and 1.5 mg l−1 activated charcoal (AC). Almost 90% of rooted shoots were successfully acclimatized and established ex vitro. |
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