| Phellodendron amurense Rupr. is commonly used Chinese medicine of cortex, which is a rare and endangered species exist in the wild state barriers to self-renewal, resulting in the decline of wild populations. Preliminary study by our group confirmed that soil seed bank of wild species in different depth of soil all exist activity seeds of Phellodendron amurense Rupr. Original seed germination simulation experiments confirmed the existence of peel and deciduous were an important factor in seed germination. Further experiments confirmed that allelochemicals present in the low-temperature aqueous extracts, water extract and ethanol extract about peel can inhibit the seed germination and seeding growth, which can inhibit seed germination, including the seeds of Phellodendron amurense Rupr. and others.The experiments on water-soluble substances of peel, separation of which on seed germination and growth inhibitory compounds, determine their structure, and to study the compounds on wheat, rape, seed of Phellodendron amurense Rupr. germination and growth. Finally, compounds content of peel in different regions were determined.Experiment first peel aqueous extracts of protein, polysaccharides and other macromolecules to study activity by activity analysis, the two sub stances on seed germination and growth inhibition was not obvious, but the small molecular from water extract substances were significantly inhibited. Small molecular subatances on silica gel column chromatography, chloroform, methanol gradient elution for further isolation and biological activity tracking, inhibit the active component in the isolated compounds by 13C-NMR, FAB-MS and HPLC-ELSD to determine the compounds:D-fructose and D-glucose.The content of D-fructose and D-glucose were measured by HPLC-ELSD, respectively 25.58% and 21.81%. Use of D-fructose and D-glucose reference standard configuration and content of aqueous extracts of the same solution:D-fructose processing solution, D-glucose solution, D-fructose and D-glucose solution combined treatment group, study wheat, rape and seed of Phellodendron amurense Rupr. germination and growth, and preliminary analysis of the reasons for suppression. The results showed that the activity of seed, the three seed germination rate an germination index, the seed development including height, root length, shoot weight, root weight were significantly inhibited by D-fructose and D-glucose combined group of treatment solution containing the equivalent of peel 50mg/mL or more, and inhibition increased with the concentration increased. In the concentration range of 75to125mg/mL, D-fructose and D-glucose combined group can complete inhibit seed germination and seed growth. After the culture medium used, D-fructose and D-glucose combined group on the D-glucose utilization rate is higher, while the treating solution of pH reached 2.0to3.5. Microscopic observation of changes in root by culture on D-fructose and D-glucose combined group, aqueous extracts, with increasing time, tip of the light transmission decreases, looks like cell wall thickening, and appears shrunk, fluorescence intensity gradually decreased.Prepare the treating solutions with D-fructose and D-glucose respectively, which has the same osmotic pressure with the D-fructose and D-glucose combined group of treatment solution, to investigate the osmotic pressure on seed germination and seed growth. Seed germination and growth inhibition and no significant relationship between osmotic, osmotic pressure effects can be ruled out.After the experiment, seed germination and seed growth were inhibited when there were D-fructose and D-glucose in the peel, and the content of D-fructose and D-glucose were higher in the peel. To determine two kinds of sugar content in 4 main producing areas, both contents are generally higer. D-fructose content of 14.44%～27.01%, D-glucose content of 10.14%～22.78%. |
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