Effect of aggregate size in cell cultures of Tagetes patula on thiophene production and cell growth

The effect of the size of Tagetes patula (marigolds) cell aggregates on growth and thiophene production in MS-medium was studied. A heterogeneous aggregate suspension was aseptically divided into 7 fractions, each with a defined aggregate diameter range, with subsequent inoculation of the fractions into MS growth medium. Growth occurred in all aggregate fractions and thiophene production increased with increasing aggregate diameter starting at about 3 mm, an effect possibly due to an increasing lack of oxygen in the aggregate centre. Calculations of oxygen concentration profiles in the aggregates showed namely, that the critical aggregate diameter where the oxygen concentration in the aggregate centre becomes very low, is about 3 mm. Aggregates with a diameter exceeding 1.2 cm showed a decreased thiophene production, however, these aggregates were hollow. The thiophenes produced mainly consisted of 5-(4-hydroxy-1-butenyl)1-2,2′-bithienyl, which was excreted into the medium.