Abstract
Marine organisms especially sessile invertebrates, such as soft corals, gorgonians and sponges, can survive in the competitive environment mainly relying on their second metabolites with chemoecological effects including allelopathy and algal growth inhibition. It is well known that the microscale models are urgently needed in marine chemoecology assessment to evaluate the algal growth inhibition activity of trace quantity natural products. In this work, a microalgal growth inhibition model was established for microalgal inhibition evaluation of marine natural products with 96-well microplate by automatic fluorescence observation using microplate reader. Subsequently, this model was applied to bioassay-guided isolation and preliminary bioactivity screening of the secondary metabolites from soft corals, gorgonians, sponges and their symbiotic microbes collected from the South China Sea. As a result, fifteen compounds (1‒15) were found to exhibit microalgal growth inhibition activities against at least one of marine microalgae, Karenia mikimotoi, Isochrysis galbana, and Heterosigma akashiwo. Specifically, altersolanol C (13) demonstrated potent activity against K. mikimotoi with the 96h-EC50 value of 1.16 µg/mL, more than four times stronger than that of the positive control K2Cr2O7. It was suggested that the microalgal growth inhibition microscale model is suitable for bioassay-guided isolation and preliminary bioactivity screening of marine natural products.
Highlights
A convenient and operable microalgal growth inhibition microscale model was established by modifying the microalgal microscale model with optimized conditions involving in target marine microalgae, microalgal tested period, cosolvent, excitation wavelength for fluorescence observation, and calculation method
The microaglal growth inhibition activity was tested by using marine microalgae, 96-well microplate, together with fluorescence measurement to quantify microaglal biomass
The practicality and feasibility of the established model were validated by application of this model to the evaluation of microalgal growth inhibition activity of marine natural products derived from marine organisms
Summary
Qing Zhao 1,2, An-Na Chen[1,2], Shun-Xin Hu3, Qian Liu[3], Min Chen[1,4], Lu Liu[1,2], Chang-Lun Shao[1,2], Xue-Xi Tang3 & Chang-Yun Wang[1,2,5]. A microalgal growth inhibition model was established for microalgal inhibition evaluation of marine natural products with 96-well microplate by automatic fluorescence observation using microplate reader This model was applied to bioassay-guided isolation and preliminary bioactivity screening of the secondary metabolites from soft corals, gorgonians, sponges and their symbiotic microbes collected from the South China Sea. As a result, fifteen compounds (1‒15) were found to exhibit microalgal growth inhibition activities against at least one of marine microalgae, Karenia mikimotoi, Isochrysis galbana, and Heterosigma akashiwo. Based on the above consideration, in the present study, we modified a microalgal growth inhibition microscale model by using 96-well microplates with three microalgae and measuring the chlorophyll fluorescence based on microalgal biomass This improved model would be suitable for the assessment of the microalgal inhibition activities, for micro- or trace marine natural products. We report how to evaluate the microalgal inhibition activity of the micro- or trace marine natural products in a convenient and efficient way
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