A procedure for producing pineapple [Ananas comosus (L.) Merr.] transgenic plants was developed that involved selection by micropropagation in temporary immersion bioreactors (TIBs). Pineapple calluses ranging in size from 1.5 mm to 2.0 mm that were co-cultivated with Agrobacterium tumefaciens strains AT2260 (pIG121Hm) and LBA4404 (pTOK233) for 24 h produced the highest percentage (40%) of GUS + calluses. Phosphinothricin and hygromycin, but not kanamycin, were effective selection markers in TIBs. Large-scale transformation experiments with AT2260 (pHCA58) and AT2260 (pHCG59) resulted in up to a 6.6% efficiency of transgenic plant recovery. TIB technology was found to be more efficient for transgenic plant selection than conventional micropropagation. Polymerase chain reaction and genomic Southern blot analyses confirmed the nonchimeric nature of the transgenic plants recovered from TIBs.
Morinda royoc L. (Rubiaceae) root cultures were established for the production of anthraquinones (AQs). Three independent experiments were performed to evaluate the effects of different levels of indole-3-acetic acid (0-22.8 lM), culture duration (15-75 days) and subculture number (0-4). The following indicators were recorded: root fresh weight per Erlenmeyer and intracellular and extracellular AQ production. The experiments performed in this study allowed an increase of intracellular AQ content up to a maximum of 4.5 mg g -1 of fresh mass, after 30 days of culture in a medium 5.7 lM of IAA. In addition, isolation and identification of seven AQs from M. royoc L. roots is described, one of them being reported for the first time for this species. The structures of isolated compounds were determined from 1 H-NMR data. To the best of our knowledge, this is the first report on AQ production from root culture of this plant.
The infusion rate of a slug of tracer into an anchor agitated 100-1iter batch mixer was characterized by a decay rate constant. This constant was then used to define a dimensionless mixing-rate number which was related to the stirrer Reynolds number. This correlationship allows the calculation of time or rotational speed needed to achieve any desired degree of uniformity of the mixture. 99mTc was used as radiotracer and the mixing process was followed by a scintillation NaI(TI) counter situated on the reactor wall near the injection point.
The dichloromethane extract and seven anthraquinones isolated from in vitro cultured roots of Morinda royoc L. were tested for their antimicrobial activity against seven yeast and seven bacterial strains. The extract showed a minimum inhibitory concentration (MIC) value of 15.6 μg/m against all species of Candida tested; except C. glabrata (MIC 1.95 μg/mL), and it inhibited the growth of oxacillin-resistant Staphylococcus aureus (MIC 31.2 μg/mL). Only morindone showed activity against all yeast strains (MIC 1.9 μg/mL), and against oxacillin-resistant Staphylococcus aureus (MIC 15 μg/mL).
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