Phylogenetic Analysis Of Wild Morchella In Southern Yan’an And Selection Of Fine Strains

Recently,the scale of artificial cultivation of Morchella sp.has been expanding rapidly,with the rapid development of the edible mushroom industry.Problems hinder the development of the morel industry,such as lack of cultivable varieties and unstable cultivation traits.Therefore,The development of Morchella sp.quality resources,the breeding of fine varieties and the development of cultivation techniques have become the hotspots of research.The northern of Shaanxi has a typical temperate continental monsoon climate,which provides excellent natural conditions for the occurrence of wild Morchella.In this study,the resources of wild Morchella were systematically studied,in southern Yan’an.Based on these studies which include morphological,phylogenetic analysis,biological characteristics research and artificial cultivation experiments.The strains M52 and M53 show fast mycelium growth,late pigment production,more sclerotia,black ascoma,high cap,short stipe,and thick flesh,and their yields can reach342.13g/m~2 and 164.33g/m~2.In the meanwhile,the factory-based cultivation mode of Morchella was initially explored.The box-type cultivation successfully obtained ascomas with a yield of 138g/m2,and the bag-type cultivation mode did’t produce fruit.The main research contents and results of this paper follows as:(1)Collection and morphological study of wild Morchella:This study investigates the distribution of wild Morchella in southern Yan’an,such as Huanglong,Luochuan,Fuxian,Yichuan,Baota,etc.A total of 12 specimens were collected,The macroscopic and microscopic morphological characteristics,M55 has high morphological similarity with Morchella elata,M32,M37,M38,M39,M40,M41,M42 are similar to the Esculenta Clade,M51,M52,M53,M54,M55 are similar to the Elata Clade.(2)The molecular phylogenetic analysis of wild Morchella:Molecular identification of wild samples were carried out by ITS sequence identification.The results showed that M32 and Mes-6 sequences had more than 99%identies,and M37,M38,M39,M40,M41,M42 were the same species as Mes-26.M51,M52,M53,M54 are the same species as Mel-13.M55 is the same species as Morchella elata.The comparison of ITS,LSU,RPB1and RPB2 showed that M32,M37,M38,M39,M40,M41,M42 had 100%identies with Mes-21,and M51,M52,M53,M54,M55 had 99.18%identities with Mel-13.Which combined morphological and molecular phylogenetic analysis showed that M32,M37,M38,M39,M40,M41 and M42 belonged to the Esculenta Clade and the same species as Mes-21,while M51,M52,M53,M54,M55 belonged to the Elata Clade and the same species as Mel-13.(3)Breeding of excellent strains of Morchella:The mycelium growth rate,microscopic morphological characteristics of mycelium,morphological characteristics of sclerotia,and morphological characteristics of colony were taken as evaluation criteria,The result showed that M32,M52,M53,M55 were good.Their mycelium growth rate is fast;Their pigment production time is late,and they can produce sclerotia and neat colonies;The angle are mainly acute angles,between the main mycelium and the secondary mycelium.The artificially cultivated strains M.sextelata M56,M57 as domesticated and cultivated strains.The trial planting showed that the M57 strain produced the fastest fruiting,the yield reached 695.48g/m~2,and the wild varieties M52and M53 could form ascomas with yields of 342.13 g/m~2 and 164.33 g/m~2,respectively,with excellent characters.It can be used as a material for the breeding of fine strains.(4)Study on the factory cultivation mode of Morchella:Two modes of box cultivation and bag cultivation were used to carry out the factory cultivation experiment of Morchella.After a cultivation period of 85-92 days,the box-type cultivation successfully formed ascomas,and the yield was as high as 138g/m~2,and a set of box-type cultivation process parameters was finally determined.