Studies On The Relationships Of Phellinus Linteus Strains

Phellinus igniarius is a kind of rare medicinal fungi. It is placed in the Basidiomycota, Hymenomycetes, Aphyllophorales, Hymenochaetaceae and Phellinus. It was reported that there were 106 species from Hymenochaetaceae at present in China, 40 species are identified as Phellinus. P. igniarius is recognized as the highest efficiency of medicinal fungus in the field of internationally anti-cancer biology.This article Summarized the research status of P. igniarius, including growth character, artificial cultivation, liquid fermentation, phamacodynamic function and so forth. The antagonistic reaction, peroxidase isozyme and esterase isozyme, P. igniarius genomic DNA extraction and PCR products amplified polymorphism and optimization of P. igniarius strains which collected by edible fungi lab of agricultural college of Yanbian university were tested in this paper. The results of antagonistic reaction showed that S₃, S₆, S₇ strains had affinity; and peroxidase isozyme showed the zymogram among S₃, S₆, S₇ strains are very same indicated that they had nearer affinity; S₆ and S₇ had much nearer affinity, but the others had much farther affinity or had no affinity, And 1 band in the esterase isozyme zymogram at Rf=0.805 was all the same for P. igniarius strains tested; The method of genomie DNA extraction and the optimization of RAPD analytic conditions were studied in P. igniarius, The result showed that the high-quality genomie DNA was obtained by the revised CTAB method; The optimal PCR system for RAPD analysis was as follows: 14.67μl ddH₂O, 100μmol. L^-1dNTP, 2.5mmol. L^-1 Mg²⁺, 10 pmol random primer, 30 ng template, 1 U Taq polymerase, 2.5μl Buffer in 25μl reaction system. The program of amplifying reaction was as follows: After pre-denaturing at 94℃for 5 min, under the condition of denaturing at 94℃for 1 min, annealing at 40℃for 1 min, extension at 72℃for 1.5 min, amplifying for 40 cycles, and extension at 72℃for 5 min at last. The random amplified polymorphic DNA(RAPD) analysis was conducted with 20 random primers in various strains of P. igniarius which collected from different localities. The result showed that 17 of the 20 random primers were polymorphic ones. The DNA fragments derived from each primer amplifying in all of the tested strains were found to range from 10 to 33. The size of the amplified DNA fragments ranged from 250 to 2000 base pairs. Of each primer tested, there was observed a wide variation in handing profiles among the strains of P. igniarius. A total of 377 band positions were scored for all of the tested strains, which differed significantly among bands from different primers. UPGMA cluster analysis subdivided the tested strains into two groups which was helpful to find cut the difference among the tested strains and distinguish them directly.Isozyme and RAPD analysis of the cluster results showed P. igniarius strains can be divided into two broad categories: S₃, S₆, S₇ have close genetic relationship belong to a class for their express positions and the number of bands are much similar; S₁, S₂, S₄, S₅ for a single category, showed a close genetic relationship between their own. Antagonistic test, esterase isozyme, peroxidase isozyme and RAPD analysis result have the basic agreement. Therefore, the method of antagonistic effect was effective in the preliminary identification of strains of P. igniarius; Isozyme, RAPD technique may be ascribed as one of the effective methods to distinguish strains.