Study On The Absorption Mechanism And Visualization Of Nono Gold Conjugated Rotenonein By Tobacco Cells

The visual study of guided nano pesticides' absorption mechanism and distribution law in plant cells is an important direction to study guided pesticides. In the previous study of this group, different monosaccharide and the conjugates of gold nanoparticles were synthesized, and the sudy shows that the recognizability of monosaccharid transport proteins to D-glucose is stronger than that of D-galactose and D-mannose, then it synthesized D-glucose and Au nanoparticles(Au NPs–Glc–FITC?Au NPs–R–FITC and Au NPs–R–Glc–FITC) protected by rotenone biligands, D-glucose labled by FITC and Au nanoparticles(Au NPs–Glc–FITC ? Au NPs–R–FITC and Au NPs–R–Glc–FITC) protected by rotenone biligands,and L-cysteine and Au nanoclusters(Au NCs–Cys and Au NCs–Cys–R) protected by rotenone biligands. This experiment aims to investigate or study monosaccharide and amino acids' effect on absorption,distribution and visualization of rotenone nano pesticides in plant cells.This experiment uses tobacco BY–2 cells as biological materials and membrane potential sensitive probes Di BAC4(3) as indication to test the effect of 50 ?g/m L(Au concentration) of Au NPs–Glc, Au NPs–R, Au NPs–R–Glc, Au NCs–Cys and Au NCs–Cys–R on protoplasts of tobacco BY–2 cells. The result shows the above-mentioned guided nano pesticides have no obvious depolarization effect on membrane potential of tobacco BY–2 cells' protoplasts, and it concludes that guided nano pesticides of the test concentration have no potential toxicity to tobacco BY–2 cells and this plant cell can be used as experimental materials.In order to investigate the effect of D-glucose on tobacco BY–2 cells' s absorption of nano pesticides, we use ICP-OES to have a quantitative detection of cells per gram(dryweight) respectively to Au NPs–Glc, Au NPs–R and Au NPs–R–Glc, and the results are as follows : Au NPs–Glc(33.23 ?g/g), Au NPs–R–Glc(21.34 ?g/g), Au NPs–R(12.16?g/g) and Au NPs(11.49 ?g/g),which indicates that the introduction of glucose groups can obviously improve the cell absorbtivity to gold nano pesticides. To have a visal observation of distribution of D-glucose labled by FITC and gold nanoparticles protected by rotenone biligands in tobacco BY–2 cell wth CLSM, it can be found that nano pesticides distribute both in cytoplasm and vacuoles, but the strongest fluorescence intensity of cells treated by Au NPs–Glc–FITC, followed by Au NPs–R–Glc–FITC and Au NPs–R–FITC, and the result shows glucose as coupling groups can promote nano pesticide's accumulation in tobacco BY–2 cells. The inhibitor test further explores tobacco BY–2 cells' s absorption mechanism to glycosyl nano pesticides, and the study found both substrate glucose and energy inhibitor CCCP can inhabit the absorption of nano pesticides Au NPs–R–Glc.In order to study gold nanoclusters Au NCs–Cys protected by L-Cysteine can be used as nano fluorescent probs to label pesticide molecules, we bulit Au NCs–Cys and Au NCs–Cys–R,whose average size is respectively 2.33 nm and 5.81 nm. We used TEM,FTIR, UV–vis, fluorescence spectrometer, Zeta potential measuring apparatus, etc to have characterization and spectral attribution to its surface structure. Analysing the 6h' culture of Au NCs–Cys and Au NCs–Cys–R in tobacco BY–2 cells by ICP-OES, whose absorbtivity is respectively 32.58 ?g/g and 20.88 ?g/g. The visual study finds that fluorescence intensity in tobacco BY–2 cells treated by Au NCs–Cys is stronger and the bule-fluorescence distributes in the cytoplasm, and the test shows Au NCs–Cys can be used as nano fluorescent probe to label rotenone.The study indicates thatf gold nanoparticles can be applied to test and fluorescence label of pesticide molecules, provides scientific evidence for clearing action mechanism of pesticides and developing guided pesticide theory, and make a good basis of experiment for the visual method of guided pesticides.