Isolation of protoplasts from callus of Maesa species

Maesa species are used in traditional medicine in many African and Asian countries. Maesa plants could, however, also play a role in modern medicine, for example in the cure of cancer or Leishmania. Combination of the genome of different Maesa species may result in a change in the secondary metabolism. Protoplast fusion is one possible strategy for making Maesa hybrids. The aim of this study was to isolate protoplasts from callus material of Maesa species. We found that the optimal enzyme mixture consisted of 1% Cellulase R-10, 0.5% Macerozyme R-10 and 0.6 M mannitol. Using this enzyme mixture we were able to isolate 7.7 x 10(5) protoplasts g(-1) fresh material for Maesa perlarius, 5.0 x 10(5) protoplasts g(-1) fresh material for Maesa balansae and 4.2 x 10(5) protoplasts g(-1) fresh material for Maesa lanceolata. In addition, a new method for automatically counting protoplasts was introduced.Maesa species are used in traditional medicine in many African and Asian countries. Maesa plants could, however, also play a role in modern medicine, for example in the cure of cancer or Leishmania. Combination of the genome of different Maesa species may result in a change in the secondary metabolism. Protoplast fusion is one possible strategy for making Maesa hybrids. The aim of this study was to isolate protoplasts from callus material of Maesa species. We found that the optimal enzyme mixture consisted of 1% Cellulase R-10, 0.5% Macerozyme R-10 and 0.6 M mannitol. Using this enzyme mixture we were able to isolate 7.7 x 10(5) protoplasts g(-1) fresh material for Maesa perlarius, 5.0 x 10(5) protoplasts g(-1) fresh material for Maesa balansae and 4.2 x 10(5) protoplasts g(-1) fresh material for Maesa lanceolata. In addition, a new method for automatically counting protoplasts was introduced.P