Identification Of Two Wild Species From Amanita

Amanita in the fungi community belongs to the genus Agaricales,Basidiomycotina,Amanitaceae,Amanita.At present,there are about 600 species described in the world,of which about 120 species of Amanita have been reported in China.Amanita has edible amanita,such as Amanita caesarea,Amanita ovoidea.There are also poisonous amanitas,such as Amanita phalloides and Amanita pantherina.Amanita has attracted the attention of mycologists since the very beginning of scientific mycology.Moreover,a large majority of the species in this genus form ectomycorrhizal(ME)relationships with vascular plants and play important roles in ecosystems.The taxonomy of Amanita has shifted from traditional morphological identification to molecular biology identification.The gene fragments commonly used in fungal identification have also started from ribosome deoxyribonucleic acid(rDNA)to novel protein-labeled genes(RPB2,?-tubulin,EFI-?).The continuous development of technology has led to a rapid increase in the classification and awareness of Amanita,and significant progress has been made in recent years about the diversity,phylogeography and population genetics of Amanita.At the same time,Amanita toxin has applications in different research areas of new drug development,biological therapy and transcription mechanisms.In this paper,two strains of wild Amanita in Sichuan Yajiang and Shanxi Zuoquan were used as research materials.Genomic DNA was extracted by CTAB method.The DNA fragments of ITS,LSU,RPB2,?-tubulin and EFI-? were amplified by PCR and then cloned to T vector and then sequenced.These sequences were used to BLAST in Gen Bank databases.These sequences together with referece sequences obtained from the Gen Bank database,were used to construct a phylogenetic tree.Based on the sequences of ITS and LSU,it was regarded as Amanita cinnamomescens in Sichuan Yajiang.However,it was considered asAmanita hemibapha var.ochracea on account of the results of RPB2,?-tubulin and EFI-?.Combinded dataset with ITS,LSU,RPB2,?-tubulin,and EFI-?,the species of Amanita were determined to be Amanita hemibapha var.ochracea.Similarly through ITS,LSU,RPB2 and EF?-?the Amanita of Shanxi Zuoquan was identified as Amanita caesareoides.Finally,the toxicity of two amanita species was detected by the bud-inhibited assay.The results showed that both amanitas were free of peptide toxoids and were non-toxic amanita,which was edible.