Cellular Growth Dynamics Affects Allelopathic Activity in Coffee Cell Culture

Cellular growth dynamics and allelopathic activity in coffee cell cultures were examined as follows: First, we compared allelopathic activity of seven woody plant calli, Coffea canephora, Derris indica, Ficus carica L., Juniperus confer-ta, Prunus persica, Punica granatum, and Sonneratia ovata, using a modified “sandwich method bioassay” and found that coffee callus showed the strongest growth inhibition to lettuce seedling nearly 90% of hypocotyl and 96% of root. This coffee callus actively proliferated, with a 21-fold increase during five weeks of subculture, with a growth curve comprising two typical phases: a lag phase of 0 - 2 weeks of culture and an exponential phase of 3 - 5 weeks of culture. Allelopathic activity varied depending on the growth phase of the coffee callus. The strongest allelopathic activity was detected in 1 - 2-week-old callus showing nearly 100% inhibitory effect on lettuce seedling growth. As the allelopathic activity of coffee calli is extremely high, beyond the natural level in coffee leaves and green beans, we focused on analyzing the allelopathic activity of its aqueous extracts using high-performance liquid chromatography. Several prominent peaks, including two reference alkaloids, theobromine and caffeine, which are known allelochemicals in coffee plants, and three distinct unknown peaks were identified at 270 nm in coffee calli during the lag phase (1 - 2 weeks of culture). The higher value of the total phenolic content in the lag phase also suggested a key biosynthetic pathway in relation to the allelopathic activity of coffee callus will be activated in the lag phase.