Analysis of cytokinin activity in commercial aqueous seaweed extract

Abstract

TTC viability staining method as a bioassay for cytokinin activity was refined using the soybean (Glycine max L.) suspension cultures by detecting cell viability and plotting a standard curve (R-2 = 0.98) and using different concentrations of kinetin in soybean medium as control. When kinetin concentrations are between 0.02 and 2 mg/l, it can improve cell viability. Soybean cell cultures are sensitive to the presence of kinetin. When cytex dilutions are between 5x and 5000x, it can improve cell viability as well. Thus it is concluded that cytex contains the biological activity similar to cytokinin. Establishment of a standard curve (R-2 = 0.91) using enzyme-linked immunosorbent assay (ELISA) for a natural cytokinin (DHZR), we found that cytex contains about 6.5 pmole/0.1ml DHZR which shows cytokinin like biological activity in soybean cell culture. The results are consistent with TTC method. We conclude that TTC can be one of the bioassays for cytokinin in suspension cell culture.