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High efficiency protoplast isolation from in vitro cultures and hairy roots of Maesa lanceolata

Ellen Lambert (UGent) and Danny Geelen (UGent)
(2010) AFRICAN JOURNAL OF BIOTECHNOLOGY. 9(42). p.7071-7078
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Abstract
In vitro cultures of the medicinal plant Maesa lanceolata were established to enable the cultivation of plant material for the production of protoplasts. Callus cultures were initiated using leaves collected from shoot cultures and the root tips from hairy root cultures obtained upon Agrobacterium rhizogenes transformation. For the isolation of protoplast, the different explant material of M. lanceolata was exposed to an enzyme mixture consisting of 1.5% cellulase, 0.5% macerozyme R-10 and 0.5 M mannitol. About 6 x 106 protoplasts g-1 fresh weight were obtained from leaf material and 5 x 105 protoplasts g-1 fresh weight from callus. To obtain high amounts of hairy root protoplasts, the cultures were pretreated with the auxin indole-3-butyric acid (IBA) that stimulated the formation of novel root tips. Using the dissected root tips as starting material, 8 x 105 protoplasts g-1 fresh weight were obtained per preparation. The protoplast isolation method will enable further studies on the transformation and fusion of protoplasts from M. lanceolata.
Keywords
protoplast isolation., Maesa lanceolata, tissue culture, in vitro conservation, TRITERPENOID SAPONINS, BIOLOGICAL-ACTIVITIES, HYOSCYAMUS MUTICUS, PLANT-REGENERATION, TRANSGENIC PLANTS, TISSUE CULTURES, SOMATIC HYBRIDS, TRANSFORMATION, CALLUS, GENE

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Chicago
Lambert, Ellen, and Danny Geelen. 2010. “High Efficiency Protoplast Isolation from in Vitro Cultures and Hairy Roots of Maesa Lanceolata.” African Journal of Biotechnology 9 (42): 7071–7078.
APA
Lambert, Ellen, & Geelen, D. (2010). High efficiency protoplast isolation from in vitro cultures and hairy roots of Maesa lanceolata. AFRICAN JOURNAL OF BIOTECHNOLOGY, 9(42), 7071–7078.
Vancouver
1.
Lambert E, Geelen D. High efficiency protoplast isolation from in vitro cultures and hairy roots of Maesa lanceolata. AFRICAN JOURNAL OF BIOTECHNOLOGY. 2010;9(42):7071–8.
MLA
Lambert, Ellen, and Danny Geelen. “High Efficiency Protoplast Isolation from in Vitro Cultures and Hairy Roots of Maesa Lanceolata.” AFRICAN JOURNAL OF BIOTECHNOLOGY 9.42 (2010): 7071–7078. Print.
@article{1057768,
  abstract     = {In vitro cultures of the medicinal plant Maesa lanceolata were established to enable the cultivation of plant material for the production of protoplasts. Callus cultures were initiated using leaves collected from shoot cultures and the root tips from hairy root cultures obtained upon Agrobacterium rhizogenes transformation. For the isolation of protoplast, the different explant material of M. lanceolata was exposed to an enzyme mixture consisting of 1.5\% cellulase, 0.5\% macerozyme R-10 and 0.5 M mannitol.
About 6 x 106 protoplasts g-1 fresh weight were obtained from leaf material and 5 x 105 protoplasts g-1 fresh weight from callus. To obtain high amounts of hairy root protoplasts, the cultures were pretreated with the auxin indole-3-butyric acid (IBA) that stimulated the formation of novel root tips. Using the dissected root tips as starting material, 8 x 105 protoplasts g-1 fresh weight were obtained per preparation. The protoplast isolation method will enable further studies on the transformation and fusion of protoplasts from M. lanceolata.},
  author       = {Lambert, Ellen and Geelen, Danny},
  issn         = {1684-5315},
  journal      = {AFRICAN JOURNAL OF BIOTECHNOLOGY},
  keyword      = {protoplast isolation.,Maesa lanceolata,tissue culture,in vitro conservation,TRITERPENOID SAPONINS,BIOLOGICAL-ACTIVITIES,HYOSCYAMUS MUTICUS,PLANT-REGENERATION,TRANSGENIC PLANTS,TISSUE CULTURES,SOMATIC HYBRIDS,TRANSFORMATION,CALLUS,GENE},
  language     = {eng},
  number       = {42},
  pages        = {7071--7078},
  title        = {High efficiency protoplast isolation from in vitro cultures and hairy roots of Maesa lanceolata},
  volume       = {9},
  year         = {2010},
}

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