A method to evaluate the bioactive function of fruit extracts of Chinese wild Citrus with microtubular activity

Abstract

China is one of the most important centers of origin for Citrus genetic resources. Due to the high content of secondary metabolites, mining wild Chinese Citrus for novel medical applications is promising. In this study, extracts of Chinese wild species from different taxonomical groups were screened for potential effects on microtubules (MTs) in vitro. MT density as a readout for nucleation, and frequency distribution over MT lengths as a readout for elongation and decay were determined by quantitative image analysis via a standardized coverslip assay using fluorescently labelled neurotubulin. Extract from peels of Citrus ichangensis Swing. strongly increased the density of MTs; whereas, extract from peels of Citurs limon (L.) Burm.f. exerted the opposite effect. Extract from pulp of Citrus limonia Osbeck promoted MT elongation, and in addition induced a small population of very long MTs. These data suggest that wild Chinese Citrus harbour compounds that act specifically on different aspects of MT nucleation, elongation, and decay.

Keywords

• fruit extract
• natural product-based pharmaceuticals
• Chinese wild Citrus
• microtubules secondary plant metabolites