Study On Genetoxic And Mutagenicity Of The Secondary Metabolites Of Rhizotonia Leguminicola

Subject: Prepare the secondary metabolites of Rhizoctonia Leguminicola, use the reducing Karber's method to determine its LD50, This work can establish the foundation for the continued toxicity test. Study genetoxic of the secondary metabolites of Rhizoctonia Leguminicola from three angles: chromosome, cell granule and DNA. Research its mutagenicity through bone marrow micronucleus test and Ames test. According to these results, we can evaluate the safety of the secondary metabolites of Rhizoctonia Leguminicola, All these can provide laboratory data for it's used in the clinic.Method:①C ultivate the stable strain had been kept in our lab, which named 94-2A, the strain was cultured by improved Czapek's culture medium, and the secondary metabolites of Rhizoctonia Leguminicola had been got.②Using the acute toxicity test (the reducing Karber's method) to determine LD50 of the secondary metabolites of Rhizoctonia Leguminicola. We can obtain the organ of body in the experiment and make paraffin section, HE stain, observe the pathology changes by optics microscope.③On the basis of government standard, we can research the genetoxic of the secondary metabolites of Rhizoctonia Leguminicola through the tests as follows: chromosome aberration test of bone marrow cell and didymus, sperm shape abnormality test in mice and single cell gel electrophoresis assay (SCGE, comet assay)and at same time we can measure the blood routine of blood which from vena caudalis.④The mutagenicity of the secondary metabolites was studied by experiments including bone marrow micronucleus assay (stained by giemsa and feuglen) and ames assay.Result:①G ot the strains of Rhizoctonia Leguminicola named 94-2A. The secondary metabolite is deep yellow, clear and got less suspended material.②Using the reducing Karber's method to determine LD50 is 226.46mg/kg, the 95% confidengce interval is 158.49mg/kg～323.59mg/kg. The LD50 is about 14～84 times than the dose of anti-tumor. So we can say that the secondary metabolite is a kind of safety medicine. It was found that the histopathological lesions were characterized by the vacuolar degeneration in the liver, kidney cell. The histopathological changes were similar to that of the toxicity caused by swainsonine.③T he indicate of the data analysis of chromosome aberration test and sperm shape...