Changes in morphology and biochemical indices in browning callus derived from Jatropha curcas hypocotyls

Callus browning is a typical feature of callus cultures derived from the hypocotyl of Jatropha curcas. Brown callus results in decreased regenerative ability, poor growth and even death. In this study, we investigated the effect of browning on callus morphology and biochemical indices. Light microscopy and scanning electron microscopy showed striking differences in callus morphology. During browning, chlorophylls and carotenoids concentrations decreased steadily. Polyphenol oxidase (PPO) and peroxidase (POD) enzymatic activities patterns were similar during callus culture with a higher activity level at week 3 compared to week 2 or later weeks. Grey relation degree analysis indicated that PPO played a more important role than POD in enzymatic callus browning. Polyacrylamide gel electrophoresis results showed differences between browning and non-browning callus. Gas chromatography-mass spectrometry results showed that saturated and unsaturated fatty acid quantities differed significantly but there was little difference in fatty acid composition between non-browning and browning callus. Differences in 17, 18.4 and 25 kDa protein concentrations were also observed in browning and non-browning callus using sodium dodecyl sulfate-polyacrylamide gel electrophoresis.