Study On The Mating System Of Wolfporia Cocos

Wolfporia cocos (Schw.) Ryv.&Gilbn., a high basidiomycete, which belongs toHymenomycetes, Aphyllophorales, Polyporaceae, Wolfporia. It is also a type of famoustraditional Chinese medicine with enormously officinal value. This study primarylyincluded three facets: mating test, isozyme electrophoresis and random amplifiedpolymorphic DNA. Moreover, the protoplast preparing conditions had been optimized.The concrete results as followed:The suitable protoplast preparing conditions were preliminarily studied for 4 strains ofWolfporia cocos. Taking production and regeneration rate into account, the most suitableenzyme dissolving temperature was 30℃, the most suitable enzyme dissolving time was4h and the concentrations of Lywallzyme was 2%.The mating test between protoplast isolates, spore isolates and the original strain showedthat the most of mating between spore isolates whatever they from the same strain or thedifferent generated antagonistic lines. It is just opposite that all mating between protoplastisolates whatever they from the same strain or the different could grow harmoniously,only one mating combination generated antagonistic lines. All mating between protoplastisolates and spore isolates generated antagonistic lines. The protoplast isolates and itsoriginal strain could grow harmoniously in mating test.The protoplasts of strain 876 of Wolfporia cocos were prepared. Sixteen protoplastisolates were chosen from 40 protoplast regenerative colonies, there were remarkabledifference in mycelia growth rate and/or CMC enzyme relative activity between whichand strain 876. Analysis of isozyme and RAPD indicated that the pattern of POD isozymeof protoplast isolates could be divided into two types compared to 876, namely type Awith secondary belts but without primary belts and type B with primary belts but withoutsecondary belts. Simultaneously, the pattern of EST isozyme in some isolates and thepattern of RAPD in the other isolates markedly differed from that in 876, whichcorresponded to type A and type B respectively. Five monokaryons protoplast isolates,type A P12 and type B P38,P29,P34 and P3, were obtained ultimately. Analysis of POD isozyme of mating products indicated that the original pattern was maintained by themating pairs between identical type isolates, while the pattern of hybrids of inter-type wassimilar to that of 876.Base on the results above, we deduce that the two nuclei in a basiaispore of Wolfporiacocos are heterogeneous and complementary. In addition, a single spore could completelife cycle. Consequently, Wolfporia cocos is likely to be a secondary homothallismfungus.