Plant regeneration from leaf mesophyll protoplasts of the tropical woody plant,passionfruit (Passiflora edulis fv flavicarpa Degener.): the importance of the antibiotic cefotaxime in the culture medium |
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Authors: | F B d'Utra Vaz A V P dos Santos G Manders E C Cocking M R Davey J B Power |
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Institution: | (1) Plant Genetic Manipulation Group, Department of Life Science, University of Nottingham, NG7 2RD Nottingham, UK;(2) Universidade Federal de Alagoas, Campus Rio Largo, Rio Largo, Alagoas, Brazil;(3) Department of Forest Genetics, Swedish University of Agriculture, P.O. Box 7027, S-75007 Uppsala, Sweden |
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Abstract: | Enzymatic digestion of newly expanded leaves of glasshouse-grown seedlings of passionfruit released protoplasts which exhibited highest division frequency (38.6%) when plated at a density of 1.5×105 ppts ml–1 in agarose-solidified droplets of KM8P medium containing the antibiotic cefotaxime (250 g ml–1). Cefotaxime was essential for sustained cell division. Protoplast-derived calli were cultured on agarsolidified MS medium with 5.0 mg H NAA, 0.25 mg l–1 BAP and additional vitamins. These calli regenerated shoots on transfer to MS medium with 1.0 mg l–1 BAP. Regenerated shoots were rooted in half-strength MS medium with 3.0 mg l–1 IBA and 0.5 mg l–1 NAA (7 d), followed by sub-culture to MS medium lacking growth regulators. The ability to regenerate plants from protoplasts of passionfruit is discussed in relation to the application of somatic cell techniques for the genetic improvement of this economically important tropical woody plant.Abbreviations B5
medium after Gamborg et al. (1968)
- BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- d
day
- FDA
fluorescein diacetate
- FPE
final plating efficiency
- f. wt
fresh weight
- h
hour
- 1BA
4-indole-3yl-butyric acid
- IPE
initial plating efficiency
- MES
2-N-morpholinoethane sulphonic acid
- MS
medium after Murashige and Skoog (1962)
- NAA
-naphthaleneacetic acid
- PVP-10
polyvinylpyrrolidone (M. Wt. 10,000)
- rpm
rotations per minute |
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