Larvicidal activity and microencapsulation of tobacco(Nicotiana tabacum)extract on Malacosoma neustria testacea larvae

To understand and improve the stability of the insecticidal activity of tobacco extract,the 3 rd instar larvae of Malacosoma neustria testacea was determined by the leaf film method.Spectrophotometry identified extract effects on activities of several enzymes.In addition,to improve the stability of the extract,microcapsules were prepared by complex coacervation and phase separation with the extract as core material,and gelatin and gum arabic as wall material.With the embedding rate as the evaluation index,the response surface method was used to optimize the preparation process of the microcapsules.The results show that the extract had a strong insecticidal activity on the larvae,with inhibitory effects on several enzymes examined of carboxylesterase,acetylcholinesterase,glutathione-S transferase,catalase,and superoxide dismutase.The inhibition rate increased with time.The best preparation process of tobacco extract microcapsules was 25% mass fraction of emulsifier,2.05% mass fraction of gelatin,3% mass fraction of gum arabic,1.34 wall core ratio,36 min of complex coacervation time.The embedding rate was 58.4% which is approximately the theoretical embedding rate(58.9%).The microcapsules prepared by this method have a smooth surface,good combining form and particle size distribution,and a median diameter of 8.6 μm.Infrared characteristic peaks of the extracts were preserved at 877.55 cm^(-1) and2922.13 cm^(-1).Microencapsulation can improve the thermal stability of the tobacco extract.Indoor toxicity tests showed that LC_(50) of extract microcapsules was 20.2 mg·mL^(-1),equivalent to the toxicity level of the tobacco extract itself,indicating that microencapsulation did not reduce extract insecticidal effects.This research may provide a reference for the optimization of the tobacco extract microcapsule preparation process.

Biosynthesis of Callose and Cellulose by Detergent Extracts of Tobacco Cell Membranes and Quantification of the Polymers Synthesized in vitro

The conditions that favor the in vitro synthesis of cellulose from tobacco BY-2 cell extracts were determined. The procedure leading to the highest yield of cellulose consisted of incubating digitonin extracts of membranes from 11-day-old tobacco BY-2 cells in the presence of 1 mM UDP-glucose, 8 mM Ca^2＋ and 8 mM Mg^2＋. Under these conditions, up to nearly 40% of the polysaccharides synthesized in vitro corresponded to cellulose, the other polymer synthesized being callose. Transmission electron microscopy analysis revealed the occurrence of two types of structures in the synthetic reactions. The first type consisted of small aggregates with a diameter between 3 and 5 nm that associated to form fibrillar strings of a maximum length of 400 nm. These structures were sensitive to the acetic/nitric acid treatment of Updegraff and corresponded to callose. The second type of structures was resistant to the Updegraff reagent and corresponded to straight cellulose microfibrils of 2-3 nm in diameter and 200 nm to up to 5 μm in length. In vitro reactions performed on electron microscopy grids indicated that the minimal rate of microfibril elongation in vitro is 120 nm/min. Measurements of retardance by liquid crystal polarization microscopy as a function of time showed that small groups of microfibrils increased in retardance by up to 0.047 nm/min per pixel, confirming the formation of organized structures.