| The biological characteristics and toxicity of Russula subnigricans Hongo was studied for the first time from ecology and morphologic characteristics and histology, the orthogonal experiment of the optimum culture condition, the analysis of components, apoptosis of the cells from little white rat liver and kidney induced by extract of Russula Subnigricans Hongo,to the histopathologic changes observation of little white rat liver and kidney through ecological observation, light microscopy and scanning electron microscopy, reversed-phase high performance liquid chromatography, agarose gel electrophoresis, transmission electron micioscopy.The result showed as below:Based on ecological observation of Russula Subnigricans Hongo,its ecological environment was investigated in order to simulate its ecological environment when they are cultivated. Russula Subnigricans Hongo was studied with light microscopy and scanning electron microscopy techniques.The result showed that Russula Subnigricans Hongo is a kind of ecotomycorrhizal fungi, Russula Subnigricans Hongo emerged and developed in the relatively steady ecological environment where average temperature,humidity and soil water content were 24 ~ 32 , 50 ~ 80% and 30% respectively.Among the three environmental factors,soil water content was the leadig factor. Russula Subnigricans Hongo grows fast after shower in the later summer and early autumn when it was hot.They emerged in the forest stand in which crown density was 0.9 or so.Soil pH was 5.75,and Fagaceae species dominated.The optimum culture condition of R.Subnigricans was investigatedby the orthogonal experiment in four factors and three levels of nutrient concentration.The basic culture conditions were pH5.6;culture temparature,26.2 ;culture duration 45d.The optimum culture condition was selected by testing and analyzing diameter of the R.subnigricans mycelium.It was (NH4)2HPO4,1g/1000mL;mannose45g/1000mL; KH2 -PO4,0.5g/1000mL;vitaminB1,10 g/1000mL. The nitrogen compo- und was the most important tropho-factor for the growth of Russula Subnigricans Hongo.The analysis of amino acids in Russula subnigricans Hongo was made with reversed-phase high performance liquid chromatography,using phenylisothiocyanate as a precolum derivatizing agent(i.e.pico-Tag method).Components of Russula Subnigricans Hongo were studied with Anthrone colorimetric analysis and reversed-phase high performance liquid chromatography.The result showed that sixteen kinds of amino acid(included kinds of essential amino acids)existed in Russulusa subnigricans Hongo,except that tryptophane destroyed by hydrolysis is not checked out.The sum of amino acids was 179.652 g/mg.Te percentage of essential aminoacids was 35.37%.Soluble sugar content of Russula Subnigricans Hongo is 2.93%,and the coarse fibre was 7.55%.Vitamin C 0.1484 mg/100g ,Nicotinic acid 0.1096 mg/100g,Vitamin B1 0.0586 mg/100g,Vitamin B2 0.0096 mg/100g. The extract of Russula Subnigricans Hongo contains -amanitin.The apoptosis induced by extract of Russula subnigricans Hongo was investigated in little white rat liver and kidney cells by agarose gel electrophoresis.The result showed that agarose gel electrophoresis of DNA extracted from poisoned little white rat liver and kidney cellsrevealed typical 180 ~ 200bp integer-fold "ladder" 'bands.Apoptosis induced by extract of Russula subnigricans Hongo was dose-and time-dependentThe result indicated that extract of Russula subnigricans Hongo could induce apoptosis in little white rat liver and kidney cells.Toxicity tests in white rats with the crud extracts of Russula subnigricans Hongo were carried out.The LD50 of the crud extracts of Russula subnigricans Hongo was determined.Transmission electron microscopy was used to observe the histopathologic changes in the poisoned rats.The result showed that the LD50 of the crud extracts of Russula subnigricans Hongo was 2852.1mg/kg.Electron microscopy analysis showed significant changes in kidney cells of the poisoned rats,including cell nuclear membrance swelling,mitoc...
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