Alternative strategies to by-pass the plant-based Azadirachtin-A production

Abstract

All parts of Neem (Azadirachta indica A. Juss) show a broad spectrum efficacy against insect pests including insecticidal, anti-feedant or insect repellent activities. Several studies have shown that plant cell cultures can produce azadirachtins. We induced more than 40 novel Neem cell lines in modified Murashige and Skoog (MS) media containing different concentrations of auxins and cytokinins. To enhance the Azadirachtin production from 1 mg/l, it was necessary to optimize the media composition separately for biomass and secondary metabolite production. In light of this complex challenge we used our novel fully automated high-throughput microbioreactor system that allows us a fast and controlled batch and fedbatch screening in 48-well microtiter plates. There is increasing evidence that plants like Azadirachta indica contain endophytes which are able to colonize internal plant tissue without causing visible disease symptoms. The estimated high species diversity of endophytes suggests a rich and almost untapped source of new secondary metabolites. We isolated more than 340 endophytes from various plant tissues and tested if they were able to produce Azadirachtin-A. Here, we present data on isolation of endophytes and induction of callus as well as first results of our microbioreactor system.