Phylogenetic And Population Genetic Analysis Of The Cantharellus Cibarius Species Complex Across Yunnan And Five Countries In Europe

Chanterelle is currently one of the four famous mushroom（matsutake,truffle,chanterelle,boletus）in the word.They are valuable wild edible fungi with important edible,medicinal,economic,and research values.They are mainly distributed in Southwestern China.However,the resource status and genetic diversity of the Cantharellus cibarius species complex in this region are not well understood.Most of the classifications and nomenclature are based on existing European and American names,which is extremely unreasonable.In this study,we used multi-gene fragments as molecular markers,based on their molecular data and microstructural characteristics,Systematic classification and population genetics of the Cantharellus cibarius species complex in Yunnan Province and five European countries,in order to understand the richness of the species diversity of the Cantharellus cibarius species complex in Eurasia,explore whether the chanterelles in the Eurasian region belong to the same species or different species,and the patterns of genetic variation in different geographical regions,the aim is to provide valuable clues for the classification of chanterelles,the pattern of population reproduction,genetics of mitochondrial genes and sustainable use of resources.The main results are as follows:1.232 Mushroom fruiting bodies with morphological characteristics similar to Cantharellus cibarius were collected in 25 counties and cities of Yunnan Province,144fruiting bodies were obtained from 6 regions of Europe.A total of 376 chanterelle fruiting bodies were collected.Then we amplified five gene loci of 376 samples:ITS,TEF1,RPB1,ATP6 and CHS1.Finally,376 ITS sequences,376 TEF1 sequences,265RPB1 sequences,244 CHS1 sequences and 245 ATP6 sequences were obtained for subsequent phylogenetic and population genetic analysis.2.A total of 49 TEF1 genotypes were identified from 376 TEF1 sequences.The highest and lowest values of genotype diversity appeared in Yunnan;the genotype diversity in Europe is in the middle,between 0.14-0.53.Phylogenetic analysis shows that our samples were distributed among six clades,most Yunnan samples clustered in the clade1,close to C.tuberculosporus,most Europe samples clustered in the clade3,close to C.cibarius.A total of 9 new phylogenetic species were identified,combining the shape and microstructure characteristics of Cantharellus fungi,two samples with fresh fruiting bodies were morphologically described,and the species composition diversity and phylogenetic relationship of Cantharellus were discussed.3.Analysis of the haplotype sequences of four gene fragments from Yunnan and Europe samples revealed that samples were not clustered phylogenetically together according to geographical origin,and gene flow existed among different geographic groups.Population genetic analysis was performed on the samples of the largest clade in Yunnan and Europe.Analysis of molecular variation(F_ST)and population structure showed that the molecular variation of the C.tuberculosporus in Yunnan was mainly distributed within local population,and there were obvious gene flow among the populations,and there is a positive correlation between genetic differentiation and geographical distance.Population genetics analysis of Europe samples shows that there is no positive correlation between the degree of genetic differentiation and geographic distance.Population structure analysis showt here is obvious genetic flow between the groups.Hardy-Weinberg equilibrium detection and linkage equilibrium analysis based on polygenic haplotype data proved that sexual spores play an important role in the transmission of chanterelles among Yunnan and Europe.4.Two hybrid samples DL7-6 and LF-13 were found in the mitochondrial ATP6fragment,Basic Local Alignment Search Tool（BLAST）and absolute quantitative experiments were indicate the presence of two mitochondrial genomes coexisting within individuals instead of numts,this stable and interspecific mitochondrial reorganization was first discovered in the natural population of Basidiomycetes.The results of ancestral state reconstruction show that the Mitochondrial ATP6 CA genotype is the earliest and the most frequent,so it is most likely the ancestor of all four genotypes,but other genotypes also have corresponding probability.It was also found that the levels of TEF1 haplotype diversity and ATP6 heterogeneity locus genetic diversity were different in the same geographic population.Differentiation patterns of nuclear gene diversity and mitochondrial recombinant genotype diversity of Cantharellus follow different evolutionary patterns.