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Effect of enzyme concentrations on protoplast isolation and protoplast culture of Spathiphyllum and Anthurium

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Abstract
Vital protoplasts from Spathiphyllum wallisii ‘Alain’ and Anthurium scherzerianum ‘238’ were isolated from both somatic embryos and leaves. The highest yields were obtained when 1.5% cellulase, 0.5% macerase and 0.5% driselase were used for Spathiphyllum wallisii leaves and 0.5% cellulase, 0.3% macerase and 0.5% driselase for Anthurium scherzerianum embryos. About 1 × 106 protoplasts g−1 and 1 × 105 protoplasts g−1 could be isolated from leaves and embryos, respectively. For protoplast fusion Spathiphyllum wallisii ‘Alain’ and Anthurium scherzerianum ‘238’ were mixed in a 1:1 ratio in a fusion solution containing 1 mM CaCl2·2H2O, 1 mM MES and 0.5 M mannitol. Fusion was performed by protoplast alignment under 500 V cm−1 alternating current for 60 s and subsequent generation of two pulses of 4500 V cm−1 direct current during 50 μs. Development until colony stage was achieved using agarose beads for protoplast culture.
Keywords
FERTILE PLANTS, BRASSICA-NAPUS, Cellulase, Araceae, Driselase, SOLANUM-MELONGENA, MOLECULAR ANALYSIS, PLANT-REGENERATION, ARABIDOPSIS-THALIANA, MICROPROTOPLAST FUSION, ASYMMETRIC SOMATIC HYBRIDS, Somatic embryos, Agarose beads, Macerase, Electrofusion, HYBRIDIZATION, S-TORVUM

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Chicago
Duquenne, Barbara, Tom Eeckhaut, Stefaan Werbrouck, and Johan Van Huylenbroeck. 2007. “Effect of Enzyme Concentrations on Protoplast Isolation and Protoplast Culture of Spathiphyllum and Anthurium.” Plant Cell Tissue and Organ Culture 91 (2): 165–173.
APA
Duquenne, B., Eeckhaut, T., Werbrouck, S., & Van Huylenbroeck, J. (2007). Effect of enzyme concentrations on protoplast isolation and protoplast culture of Spathiphyllum and Anthurium. PLANT CELL TISSUE AND ORGAN CULTURE, 91(2), 165–173. Presented at the 20th Conference on Tissue Culture Facing the Future.
Vancouver
1.
Duquenne B, Eeckhaut T, Werbrouck S, Van Huylenbroeck J. Effect of enzyme concentrations on protoplast isolation and protoplast culture of Spathiphyllum and Anthurium. PLANT CELL TISSUE AND ORGAN CULTURE. 2007;91(2):165–73.
MLA
Duquenne, Barbara, Tom Eeckhaut, Stefaan Werbrouck, et al. “Effect of Enzyme Concentrations on Protoplast Isolation and Protoplast Culture of Spathiphyllum and Anthurium.” PLANT CELL TISSUE AND ORGAN CULTURE 91.2 (2007): 165–173. Print.
@article{682797,
  abstract     = {Vital protoplasts from Spathiphyllum wallisii {\textquoteleft}Alain{\textquoteright} and Anthurium scherzerianum {\textquoteleft}238{\textquoteright} were isolated from both somatic embryos and leaves. The highest yields were obtained when 1.5\% cellulase, 0.5\% macerase and 0.5\% driselase were used for Spathiphyllum wallisii leaves and 0.5\% cellulase, 0.3\% macerase and 0.5\% driselase for Anthurium scherzerianum embryos. About 1 {\texttimes} 106 protoplasts g\ensuremath{-}1 and 1 {\texttimes} 105 protoplasts g\ensuremath{-}1 could be isolated from leaves and embryos, respectively. For protoplast fusion Spathiphyllum wallisii {\textquoteleft}Alain{\textquoteright} and Anthurium scherzerianum {\textquoteleft}238{\textquoteright} were mixed in a 1:1 ratio in a fusion solution containing 1 mM CaCl2{\textperiodcentered}2H2O, 1 mM MES and 0.5 M mannitol. Fusion was performed by protoplast alignment under 500 V cm\ensuremath{-}1 alternating current for 60 s and subsequent generation of two pulses of 4500 V cm\ensuremath{-}1 direct current during 50 \ensuremath{\mu}s. Development until colony stage was achieved using agarose beads for protoplast culture.},
  author       = {Duquenne, Barbara and Eeckhaut, Tom and Werbrouck, Stefaan and Van Huylenbroeck, Johan},
  issn         = {0167-6857},
  journal      = {PLANT CELL TISSUE AND ORGAN CULTURE},
  keyword      = {FERTILE PLANTS,BRASSICA-NAPUS,Cellulase,Araceae,Driselase,SOLANUM-MELONGENA,MOLECULAR ANALYSIS,PLANT-REGENERATION,ARABIDOPSIS-THALIANA,MICROPROTOPLAST FUSION,ASYMMETRIC SOMATIC HYBRIDS,Somatic embryos,Agarose beads,Macerase,Electrofusion,HYBRIDIZATION,S-TORVUM},
  language     = {eng},
  location     = {Gembloux, Belgium},
  number       = {2},
  pages        = {165--173},
  title        = {Effect of enzyme concentrations on protoplast isolation and protoplast culture of Spathiphyllum and Anthurium},
  url          = {http://dx.doi.org/10.1007/s11240-007-9226-3},
  volume       = {91},
  year         = {2007},
}

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