The Inonotus Phylogenetic Relationships And Artificial Domestication And Cultivation Of,

Inonotus obliquus is a very valuable and rare officinal epiphyte. Sclerotium hasthe properties of medicine. It belongs to Basidiomycotina, Hymenomycetes,Aphyllophorales, Hymenochaetaceae. The mycelium of Inonotus obliquus canextremely suffer chillness. Inonotus obliquus mainly distribute in the area of theNorthern Hemisp-herenorth latitude 45°～50°. Foreign countries do much researchand development work on Inonotus obliquus, however, our research is little andlimited to the morphology. The reserch of domesticated cultivation and molecularbiology is almost nothing. At present domesticated cultivation is a essential workbecause the wild resource is confronted with drying up increasingly. Studies onconsanguineous relationship of Inonotus obliquus through antagonistic effect,isczymes and PAPD molecule marker, on pure mycelium which is isolated from wildsclerotium in different areas from 2004 to 2006. The main research results asfollowing:1,This article analyses antagonism of mycelium of Inonotusobliquus(Fr.)Pilat.The results showed that the difference among each strain is distributeon. Theconsang-uinity of CX01 and CX02 is the closest, JL03, JL01 and JL02is much closer.2,By using different extracting agent to extract peroxidase isozyme of Inonotuscbliquus under the same condition, applying polyacrylamide gel electrophoresis toconduct comparative study of peroxidase isozyme bands of Inonotus obliquus myeelia.The results showed that the sample which applied the extracting agent with Tris-HC1(pH6.9) had the best extracting effect of electrophoresis. It was affluent in bands andthe bands were also clear; the worst extracting effect was Tris-glyeine(pH 8.3).3,With polyacrylamide gel electrophoresis, we analysed peroxidase isozymes ofInonotus obliquus(Fr.) Pilat species from 6 different areas. The results showed thatthere were 2～8 enzyme bands in all strains, the characteristic enzyme bands isRf=0.75 and Rf=0.98. The Fuzzy cluster method was used for studying and themembership grade is between 0.250 and 1.000. The writer take taxonomy on strains at0.250 (classification set). The consanguinityof CX and JL was close. Theconsanguinity was far between them and HLJ. The difference among isozyme bandsof each atrains is distribute(latitude). The consanguinityof some strains is close.4,An improved CTAB method is applied to obtaining genomic DNA. And it wasused for template and then to optimized the reaction system of Amygdalus. The result indicated that: optimal 25uL total reaction volume of RAPD-PCR contained:ternplate DNA 40ng/μL, primer 10pmol, Mg²⁺ 2mmol/L, Taq DNA polymerase lunitdoraestic, dNTP 100μmol/L, others complement with ddH₂O. The reaction programwas: initial denaturation at 94℃for 5 minutes; denaturation at 94℃for 1 minutes,annealing at 40℃for 1 minutes, elongation at 72℃for 1.5minutes, 45 cycles; finalelongation at 72℃for 5 minutes.5,The relationship of Inonotus obliquus strains isolated from sclerotia collectedfrom different areas also was studied by use of random amplified polymorphic DNA(RAPD) markers. The DNA fingerprints of different strains showed that there wasgenesic diversity among the strains tested. The results indicated that 12 randomprimers generated 167 bands in total, of which 101 bands showed polymorphism andthe average polymorphism rate was 60.5％. There are a little intraspecific differencearnong fingerprints of each strains under the same condition. The consanguinity ofsome strains are close. At the level of 0.508 (genetic distance), CX01 and CX02(latitude 39.01°-40.80°) were clustered in the same group and the second groupincluded JL01, JL02, JL03, JL04 and JL05 (latitude 42.93°-43.35°). HLJ01 (latitude47.20°-50.13°) is independent. The consanguinityof CX and JL was close. Theconsanguinity was far between them and HLJ. The clusters results of strains is samewith geographic distribution (latitude) difference. Indicate that there is intraspeciesand geographic distribution difference among each strains, the consanguinity of partstrains is close.6,Choosing wild sclerotium oflnonotus obliguus as material, by applying tissueisolate method to obtain pure strain, studied on all levels spawn and domesticatedcultivation, the results showed that The optimum mother culture medium was glucose20g,soybean powder 10g,KH₂PO₄lg,MgSO₄0.5g,water 1000ml, pH nature;The optimum composition of pathogen culture medium was corn, the optimumformulae of compost of sclerotia artificial cultivation was birch sawdust 52％,corncob26％, bran20％, sucrosel％, gypsuml％and moisture 65％and thebiolegical efficiency reached to 30.8％. The formation temperature of sclerotium isamong 22～28℃. Other preferable formation condition is no placket on cultivationbag, no light and moisture is 60％.