In vitro regeneration of loblolly pine and random amplified polymorphic DNA analyses of regenerated plantlets |
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Authors: | W Tang |
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Institution: | (1) State Key Laboratory of Biochemical Engineering, Institute of Chemical Metallurgy, Chinese Academy of Sciences, Beijing 100080, P. R. China e-mail: wtang@unity.ncsu.edu Fax: +1-919-5157801, CN |
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Abstract: | Adventitious buds were induced from organogenic callus derived from mature zygotic embryos of three lines (E-311, E-440,
and E-822) of loblolly pine (Pinus taeda L.) within 27 weeks of culture. The influence of cytokinins, silver nitrate, and low-temperature treatment on the differentiation
of adventitious buds was analyzed. Elongation of adventitious buds was achieved on TE medium supplemented with 0.5 mg/l indole-3-butyric
acid (IBA) and 1 mg/l 6-benzyladenine (BA). After adventitious shoots had rooted on TE medium supplemented with 0.5 mg/l IBA,
2 mg/l BA, and 0.5 mg/l gibberellic acid, 498 regenerated plantlets were transferred to a perlite:peatmoss:vermiculite (1 :>: 1)
soil mixture; 351 of these survived in the field. Total DNA was extracted from 21 regenerated plantlets randomly chosen from
the 151 regenerated plantlets of line E-822. Random amplified polymorphic DNA (RAPD) analysis using 80 arbitrary oligonucleotide
10-mers showed that 21 primers gave 107 clear reproducible bands, with the amplification products being monomorphic for all
of the plantlets of line E-822 tested. A total of 2,247 bands obtained from these studies exhibited no aberration in RAPD
banding patterns among the tested plantlets. These results suggest that somatic organogenesis can be used for clonal micropropagation
of some lines of loblolly pine without the fear of the appearance of unwanted somaclonal variants.
Received: 5 August 2000 / Revision received: 5 September 2000 / Accepted: 10 October 2000 |
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Keywords: | Pinus taeda L Organogenesis Plant regeneration Random amplified polymorphic DNA |
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