| After a long period of coevolution,most of the bark beetles and Ophiostoma fungi have formed mutually beneficial concomitant relationships.It covers many aspects,such as the recognition of fungi by insects,the special alienation of insects and fungi,the carrying and dissemination of fungi,and so on,which is a dynamic and comprehensive complex process.The mutualistic symbiotic relationship between the two not only promoted the better reproduction and development of bark beetle,occupied new habitats,but also promoted the spread of fungi.The main purpose of this project is to combine traditional microbiology with modern molecular biology to study the five regions of Dali,Chuxiong and Shangri-La in Kunming,Lijiang,Dali,Chuxiong and Shangri-La areas of Yunnan Province.An indepth study on the associated Ophiostoma fungi of Tomicus minor Hartig was carried out in the same place.It mainly involves three directions of research,including species diversity of associated fungi,morphological observation and molecular identification of associated fungi,screening and evaluation of DNA bar codes of Ophiostoma fungi.The project collected epidemic wood and insect samples in Yunnan Province in August 2017 and transferred them to the laboratory for isolation and purification from the collection bag.The original isolation plate reached 254,and 400 pure strains were obtained after purification.Extraction of DNA,from400 strains of bacteria and amplification of ITS gene sequence fragment.After sequencing,it was confirmed that there were 50 genera involved,and 6 genera were obtained.a total of112 strains of fungi were obtained: Ceratocystiopsis,Graphilbum,Leptographium,Ophiostoma,Raffaelea,Sporothrix,accounted for 28% of all species.Through the indepth molecular study of Opiostoma,based on the sequencing and phylogenetic analysis of polygenes: ITS、BT2、TEF,The phylogenetic tree constructed by maximum likelihood method was used to identify five suspected new species of the Ophiostoma fungi :YML185、 YML208、YML209、YML210 and YML212.Morphological observation and image acquisition are used to describe and report it.Sporulation structure was observed in 5strains of bacteria.Sexual morphological structure was observed by YML208 and YML210.YML185、YML209 and YML212 observed the asexual structure.Based on the results of molecular phylogeny and morphological observation,five new species of strains were described and reported.Based on the study of the associated fungi of the Tomicus minor Hartig in Yunnan Province,the species diversity and dominant species of the associated fungi were preliminarily identified.It provides a theoretical basis for the control of the Tomicus minor Hartig,the understanding of the fungi of the Ophiostoma and the concomitant relationship between the insects and the beetles in Yunnan Province.It also points out the direction of exploration and control ideas for the quarantine and monitoring of diseases and insect pests in this area. |
