Phylogenetic Analysis Of Morchella Spp.(True Morel) In The North Qinling Mt.and The Promotion Of Morchella Crassipes For Maize And Winter Wheat Growth

In recent years,true morel?Morchella spp?species diversity was reported from worldwide.China is a mainland country rich of natural morel recourses,half of the 61phylospecies were discovered in China,many cryptic species are still undiscovered in complex and remote region,e.g.Qinling Mt.,a boundary of south China and north China,where habituated plenty of plants of Quaternary Glacier Relics.In additional,M.crassipes can promote plant growth by improving host nutrition absortion and defence.Given the diversity of plant microbes it is not surprising that new plant growth-promoting microbes continue to be discovered.Species of morel fungi?genus Morchella?have only recently been shown to be beneficial symbionts of grasses.In this study,31 collections of ascocarps from north Qinling Mt.,4 collections of commercial cultivars from south Qinling Mt.and 3 Morchella mycelium clones of commercial cultivars were investigated by the genealogical concordance phylogenetic species recognition?GCPSR?method.The maximum-likelihood method was employed for the construction of phytogenic trees with highly homologous sequences downloaded from the NCBI and MLST databases.The GCPSR study resolved 4 phylogenetic species of Dist.Esculenta among the 38collections in north Qinling Mt.,namely,Morchella sp.Mes-9,Morchella sp.Mes-13,Morchella sp.Mes-25,and Morchella chensiensis,in addition to a false morel?Verpa bohemica?.The cultivated Morchella sp.Mel-6,Mel-10,and Mel-12 coincided with their commercial names in farms.As many as 75%?3/4?were new records in the Qinling region except Mes-19,and a novel monophyletic linkage,Morchella chensiensis,was distinct from the previously reported phylospecies by single gene and combined genes analysis and was ultimately resolved as a new phylospecies in this study.Among all phylogenetic trees,the false morel collection displayed a closer relationship with Elata Morchella than Esculenta Morchella and formed the base of these two clades.All collections in this study showed continental endemism,and all Qinling Mt.collections were grouped together in all phylogenic trees and represented a basal clade of the Esculenta clade.More specifically,M.crassipes from Shaanxi,Northwest in China has been shown to increase growth and defense of sweet corn,Zeamays var.saccharata.Here,we examined the effects of M.crassipes inoculations on maize growth?dent corn,Zeamays var.indentata cv.‘Shaandan 985'?.This time we included as plant response variables the following:plant height and stem diameter,leaf area,root development,N,P,and K of roots,stems and leaves,chlorophyll and photosynthetic activity,four defense enzyme activities?SOD,CAT,PAL and POD?.We included six soil variables as sustainability metrics:soil moisture,mineral NO₃,NH₄,P,K,pH of soil.We employed three levels of inoculum of M.crassipes:50,100,and150 mL of mycelial suspension?1 petrish per 100 mL H₂O?.Inoculation with M.crassipes significantly increased measures of maize growth?e.g.,fresh and dried biomass?,and it also affected other variables in tissue-and dosage-dependent ways.In particular,soil moisture and available K were increased.Thus,our findings overall suggest that inoculation with M.crassipes might affect not only yield of maize but also the sustainability of the cropping system.Perhaps most significantly the promotion of growth by M.crassipes was not accompanied by increases in defense enzyme activities,unlike the urea control which increased the latter.To identify the role of biochar-morel agent for winter wheat growing in the salt stressed soil,winter wheat seeds were arranged in pot array after biochar-morel agents were input to the soil,and growth index was dated and evaluated in green house.High performance liquid chromatography?HPLC?and histological dying after artificial inoculation were employed to detect the phytohormone secreted by morel in the liquid suspending and symbiosis structure,respectively,biochar was prepared in an optimized program from trimmed branch of Cedrus deodara?cedar?in winter.Results showed seeds germinated highest at concentration 1.5%of this mixture.Without salt stress?namely,the positive control PC2?,biochar-morel could improve seedlings root/shoot ratio significantly?P<0.05?at concentration 2.0%,2.5%,with ratio of 0.469,0.549,respectively.However,under 0.5%salt stress,the biochar-morel agent could improve root/shoot ration of seedlings significantly at tested concentration from 1.0%to 2.5%?P<0.05?,with ratio of 0.515,0.653,0.573,0.580,respectively,which are even higher than those results from positive control PC2.No significant difference was found on seedlings height,fresh weight,and biomass,but most of them are higher than the control except 0.5%tested concentrations whether under salt stress or not.Higher concentration showed higher improvement for wheat growth index.Averagely,salt stress can contribute 21%inhibition ratio for seeds germination,21.8%for seedlings height,36.3%for seedlings fresh mass,12%for biomass,and 23.9%for root/shoot,respectively.However,biochar-morel can relieve salt stress at ratio 33.3%for seeds germination,3%for height,21%for fresh mass,36.4%for biomass,and 471.4%for root/shoot ratio,respectively.Winter wheat improve itself salt resistance by enhancing root/shoot ratio,Morel in this study can secret phytohormons,IAA,SA and ABA,and so further stimulate the root system developing.Biochar in this agent can help reserving the activity of morel by concentrating nutrition and humidity around the morel mycelium,the interaction between morel and biochar gets a win-win result for wheat growing.This is a novel soil improving agent with potential application in practise.This study had provided further informative classification of Morchella spp.species diversity in north Qinling Mt.and also suggested the guidnaces,technique and basical theory of M.crassipes application.