Growth and Plating of Cell Suspension Cultures of Datura innoxia

Abstract

AbstractSuspension cultures of Datura innoxia Mill, were successfully grown on a modified Murashige and Skoog medium with 2,4–D, NAA or BAP as growth substances, provided the micronutrient levels were reduced to 1/10. Normal amounts of micronutrients were toxic. Attempts to identify the toxic elements did not succeed.Cultures grew exponentially on a shaker at 27°C in the light. Their doubling times varied from 1.1 days on 2,4–D (10–6M) or NAA (10−5M)+ 1 g/1 casein hydrolysate to 2.7 days on BAP (3 × 10−7M) and 5.1 days on supraoptimal levels of 2,4‐D (10−5M). Cultures grew on NH4+‐N alone (from ammonium malate) or on NO3−‐N alone. Dry weight yield was proportional to the amount of nitrate‐N added (47 mg/mg N).Filtered suspension cultures containing single cells (plating cultures) could be grown in agar in petri dishes when NAA or 2,4‐D were used as growth substances. Cells grew at densities above 500 units/ml in the agar. Most colonies grew from cell aggregates but division in single cells was observed. The highest plating efficiency was about 50% on 10−6 M 2,4‐D + 1 g/1 casein hydrolysate.