The Study On Biology Effects Of Secondary Metabolites Of Rhizoctonia Leguminicola

Subject: Pick up strains of Rhizoctonia leguminicola that can product large amounts of swainsonine (SW) and grow stably. This work can establish the foundation for zymology study on the strains of Rhizoctonia leguminicola. We can also acquire the systemic identification datum of the vegetal and modal character, and study the antitumor effects of pathology and immunology in the body of animal by the secondary metabolites of Rhizoctonia Leguminicola. Thereby we could know the reason of antitumor by SW. At last, we could explore damage effect and cytobiology relationship of animal and plant cell by Rhizoctonia leguminicola and its secondary metabolites. All these can provide new thoughtfulness for the study on tumor pathology and toxicology. Method: ① By way of strains separation, purification, revival of this epiphyte in field and the Rhizoctonia leguminicola had been kept in our lab since 1997 respectively, we could get some strains of growth stably, then modal character identification had been processed by optics microscope and SEM, and the experimentation of hypha amalgamation had been done to distinguish different amalgamation groups. ② The stable strains was cultured by improved Czapek's culture medium, and the secondary metabolites of Rhizoctonia Leguminicola had been got, then gas chromatography mensuration was used in the secondary metabolites to show the content of SW. ③ We built cancer mold by transplanting Hepatoma 22 cells to rats, and poured the secondary metabolites of Rhizoctonia Leguminicola which had swainsonine into the stomach of the rats, and the doses of swainsonine were 2.7mg/kg/d and 16.2mg/kg/d separately. MTT method was used to observe the T lymphocytes proliferation after 0d, 4d, 8d, 12d and 16d. The swallow percentages of peritoneal macrophages were determined after 4d and 8d. We also determined the ratio of tumor inhibition and observed the pathology changes by optics microscope and TEM. ④ We interfused the Rhizoctonia Leguminicola and the secondary metabolites of Rhizoctonia Leguminicola into feedingstuff of the rabbit, and we fed these rabbit for 17 days, then put the rabbit to death and the pathology obserbvation of the brain, liver, kidney by optics microscope and TEM had been done. Result and brief summary: ① We got 6 strains of Rhizoctonia Leguminicola that growth stably, and these strains belonged to different amalgamation groups in the same genus (Rhizoctonia Leguminicola). They had modal character of Rhizoctonia Leguminicola. The secondary metabolites of Rhizoctonia Leguminicola can product large amount of SW. Among those strains, metabolism products of 02-6B and 02-3A had the largest amount of SW, and the amounts were 1093.17mg/L and 1199.64mg/ml respectively. ② In the experiment of antitumor effects, the result showed that the secondary metabolites can enhance immune response and inhibit tumor growth and metastasis, especially the dose of swainsonine that is 16.2mg/kg/d had obvious result. The SI was 1.670±0.527 in group Ⅲ within 8d,and this data was higher than 1.133±0.053〔P＜0.05〕in group Ⅰ. The swallow percentages of peritoneal macrophages was 33.99±1.56% higher than group Ⅰ,and the ratio of tumor inhibition were 26.44％and33.91％in 8d, 46.98％and47.24％in 16d. In the experiment of pathology observation, group Ⅱand group Ⅲ had badly fatty degeneration and large area putrescence in tumor tissue, and the putrescence area was meshwork shape ,and cut the tumor cell into some grids .Above all, the secondary metabolites of Rhizoctonia Leguminicola had similar function with swainsonine. ③ In the experiment of the damage effects of cell from animal and plant body by the secondary metabolites of Rhizoctonia Leguminicola, the rabbit brain and lymphocyte had put up vacuolar degeneration, and the granule of ribosome had had broken off in neurocyte, and also expressed dilatation of RER.; And the plant root tissue had expressed the structure damage, and the cell had showed vacuolar degeneration and breach of the inner membrane. These results show that the poisoni...