Identification And Studies On The Bioactive Compounds Of Five Psychoactive Mushrooms

Psychoactive mushrooms, also called magic mushrooms, are a cohort of macro-fungi that contain active tryptamines in their fruiting bodies or mycelium, which will cause psychedelic intoxication for human and other vertebrates. Acter tryptamines have an indole group and a dimethylamine group in structure. Including psilocybin, psilocin found in nature. The hallucinogenic mushrooms are widely distributed throughout the world, including Psilocybe, Panaeolus, Gymnopillus, Stropharia, Inocybe, Conocybe, Amanita, Hypboloma, Pluteus, Agrocybe etc.The symptoms of intoxication of the psychoactive mushrooms and their toxins,cause nervous excitement,can speed up heart rate,respiratory rate and body temperature increases, systolic blood pressure increases, face flushing, cause numbness of the lips or mouth, tongue, shivering or sweating, anxiety, altered sensory perception hallucinations and alterations of body imagetic. Because of the abused of the mushrooms and toxins by some young people, causing a certain degree of jeopardy to the society, the use and proliferation of the toxins or mushrooms were restricted in some western countries. However, the potentials of the active component and mushrooms in the study and treatment of some psychosis, neurological disorder, fear,in treatment of giving up smoking and drinking and cluster headache, in study of human brain, the study and application of the toxins and mushrooms have never been interrupted. The habitat investigations identification, extraction,detection,purification, structure identification metabolism and secretion synthesis of active tryptamines were studied. These study in China are relatively backward except for some investigations of habitats of some species. The synthesis and application of the toxins are not carried out.The identification and active component analysis of the five mushrooms collected from different place were studied by means of light microscopy, molecular biology, reversed-phase high performance liquid chromatography, MS and NMR. The study included the morphological characteristics of the five mushrooms, the ITS sequence analysis, the strains isolation and the optimization of liquid medium of mycelium, identifying of toxins and the toxicity to rats, isolating and identifying of active component, and the results were as followings:1.The 18S rDNA(partial)-ITS1-5.8S rDNA-ITS2-28SrDNA(partial) sequence of the five mushrooms were assayed. The comparison and analysis of the sequences with some related fungus in gene library suggested that analysis of ITS DNA sequence is very useful in mushroom identification to genus level. Strains-443, strains-518 and strains-ningxiang were identified to be Psilocybe samuiensis by their ITS sequence and morphological characteristics. Strains-581 and Strains- miluo are identified to be Galerina but not to be species unless combined with other means.2.The morphological characteristics of the five mushrooms were studied by means of light microcopy and fluorescence microcopy techniques. The basidia is 4-spores, spore is ellipse, brown, germination aperture is obvious, Cheilocysticlia are abundant in gill.Clamp connection of the mycelium is often found and the mycelium is dual-core. The species of 443,518 and ningxiang were identified to be Psilocybe samuiensis by their ITS sequence and morphological characteristics.3. The strains were got by means of the isolation of tissue. The best liquid culture medium cow dung juice medium was selected from the basic medium, wort medium and the cow dung juice medium by calculating the yield of mycelium.4. Purple-brown spot (Rf=0.16) was separated from crude venom by TLC and qualitative paper method, which indicated the psilocybin was a component of crude venom and the five mushrooms were Psychoactive. The toxicity of crude extract of the Psilocybe samuiensis fruitings body was assayed by the rats intraperitoneal injection.The results suggested that the crude extract can cause the rats exciting, and the degree of intoxication is proportional to the amounts of crude venom, which showed that the Psilocybe samuiensis is a kind of hallucinogenic mushroom. In the toxicity assays, no rat died,which predicted the Psilocybe samuiensis are less toxic.5. Four components were separated from Psilocybe samuiensis mycelium by HPLC method. MS and NMR were performed after the collection. The first component named Uridine is the one of the important Components in Psilocybe samuiensis. The other three components are being assayed.