Study On Technique Of Molecular Identification Of Edible Fungi A.auricular、G.frondosa、P.nameko、P.adiposa、P.linteus、A.blazei、H.erinaceus、Cordyceps And A.polytricha

In this study,the ITS sequences and ITS-RFLP analysis were utilized for studying the molecular identified technology among the Grifola frondosa、Pholiota nameko、Pholiota adiposa、Phellinus linteus、Agaricus blazei、Hericium erinaceus、Cordyceps、Auricularia polytricha、Auricularia auricula strains,which were collected by Fujian Edible Fungi Germplasm Resource Collection Center,and the strains which had the same name in the different species were weeded out.The main results were as follows:1.By combining the restriction fragment length polymorphism(ITS-RFLP) obtained from digestions with 3 enzymes Hhal、HaeⅢand Dral,the 32 Grifola frondosa strains were efficiently classified.And it was found that Restriction Fragments of Gr0004 strain were different from the others.By sequencing and aligning with Pleurotus eryngii which belongs to Pleurotus,Gr0004 was judged to be a Pleurotus eryngii strain.2.By combining the restriction fragment length polymorphism(ITS-RFLP) obtained from digestions with 2 enzymes NruⅠand NcoⅠamong the Pholiota nameko and Pholiota adiposa strains,it was found that restriction Fragments were in consistent within species,but were different within Genera.The both were distinguished by the two enzymes.In addition,the results from ITS-RFLP were corresponded with Pholiota nameko morphological characters.3.Applying antagonistic test and ITS sequence analysis in this study,six strains of Phellinus linteus were classified and identified.The result of antagonistic test showed that Ph.l 0001、Ph.l 0003、Ph.l 0003+1 and Ph.l 0005 were corresponded with Phellinus linteus colony morphology;and antagonistic lines were obvious in strains Ph.l 0002 and Ph.l 0004 with other strains,and antagonistic lines were relatively weak among the others.And the result of ITS sequence analysis showed that there was 99.86%sequence identity among strains Ph.l 0001、Ph.l 0003、Ph.l 0003+1 and Ph.l 0005,and obvious differences existed among the others.Through antagonistic test and alignments on Online,strains Ph.l 0001、Ph.l 0003、Ph.l 0003+1 and Ph.l 0005 were considered to be Phellinus linteus;strain Ph.l 0004 was Phellinus pini;and strain Ph.l 0002 had a near relationship with Phellinus badius and its species property can not be determined.4.The length of ITS region exhibited polymorphism among the Agaricus blazei strains tested.Based on the ITS sequences analysis,all strains were divided into three kinds,which were Agaricus blazei、Aspergillus、Hypsizygus marmoreus.By alignment,and combining the restriction fragment length polymorphism obtained from digestions with 2 enzymes HaeⅢand XbaⅠamong the fourteen Agaricus blazei strains,it was found that restriction Fragments were in consistent within species,and showed that Species Property was consistent among the fourteen strains.5.Based on the genetic diversity study of Hericium erinaceus,the ITS sequences of H0003 and H0029 were sequenced.By combining the restriction fragment length polymorphism(ITS-RFLP) obtained from digestions with BamHI enzyme among the Hericium erinaceus strains,it was found that the experiment result was different with the analysis result;and the result was proved to be the same after different time of enzymes digestion,and different enzyme amount.By cloning PCR products and then sequencing further,it was proved that there could be two kinds of ITS sequences at least.6.By combining the restriction fragment length polymorphism(ITS-RFLP) obtained from digestions with 3 enzymes ApaⅠ、HaeⅢand AluⅠamong the Cordyceps strains,it was found that all strains were divided into three kinds,which were Cordyceps、Tolypocladium、Gibberella.7.Internal transcribed spacers(ITS) was amplified via the PCR from 102 Auricularia auricula strains and subjected to restriction fragment length polymorphism analysis;and all strains were identified and classified with HaeⅢ、MspⅠ、XbaⅠand HpaⅠenzymes.The ITS-RFLP results showed that three restriction enzymes HaeⅢ、XbaⅠand HpaⅠcould distinguish Auricularia auricular from the other species,and the MspⅠcould not.These results were in consistency with the results of cultivation experiments.Through sequencing further,it was found that Au.a 0004、Au.a 0005、Au.a 0050、Au.a 0051、Au.a 0052、Au.a 0077 strains are Auricularia polytricha strains.Whereafter,identifying 60 Auricularia polytricha strains with HaeⅢ、XbaⅠand HpaⅠenzymes,it was found that Au.p 0038 strain is Auricularia auricula strain.In addition,through analysing the fragments by the enzyme HpaⅠ,the result showed that there could be two different gene sequences,and showed that the different ITS regions of Auricularia have been homogenized at different extent.By cloning PCR products and then sequencing further,it was proved that there are two kinds of ITS sequences at least.