Response surface (RSM) methodology based on a three-level three-factor Box-Behnkendesign of experiment was used to optimize the exopolysaccharide content (EPC) and themycelium biomass in submerged cultivation by Clito...Response surface (RSM) methodology based on a three-level three-factor Box-Behnkendesign of experiment was used to optimize the exopolysaccharide content (EPC) and themycelium biomass in submerged cultivation by Clitocybe sp. AS 5.112. The criticalfactors selected for the investigation were cultivation temperature, time and volume ofmedium, based on the results of previous Plackett-Burman design. By analyzing theresponse surface plots, the optimum ranges of cultivation temperature, time and mediumvolume for obtaining over 1 253.00gmL-1 of EPC lie in 24.325.8℃, 9.710.2d and 76.090.0mL, respectively. While for obtaining over 8.32 mg mL-1 of dry cell weight (DCW), theabove variables would be in the range of 23.824.8℃, 9.610.3d and 71.098.0mL,respectively. By solving the inverse matrix from the quadratic regression equations, theoptimal conditions to gain 1 265.45gmL-1 of EPC were 25.0℃, 9.9d and 83.4mL, to gain8.50mg mL-1 of DCW were 24.4℃, 9.9d and 87.1mL. In order to obtain the maximum yield ofEPC and DCW at the same time, the above conditions would be 24.5℃, 9.9d and 84.7mL,respectively, in this situation, the maximum predicted EPC and DCW were 1 261.60gmL-1and 8.47mgmL-1, respectively. The experimental data under various conditions have validatedthe theoretical values.展开更多
In vitro Kirby-Bauer disc-diffusion assays coupled with bio-imaging software techniques were used to assess native forest dwelling “fairy ring” forming fungi (Clitocybe nebularis) and co-habitant forest tree-root co...In vitro Kirby-Bauer disc-diffusion assays coupled with bio-imaging software techniques were used to assess native forest dwelling “fairy ring” forming fungi (Clitocybe nebularis) and co-habitant forest tree-root colonising non pathogenic, antibiotic producing bacteria (Bacillus licheniformis, Bacillus pumilis) for their antagonism towards Japanese larch dieback oomycetes phytopathogens which also affects ornamental alternative hosts. The aqueous extracts of C. nebularis exhibited the highest clearance (inhibitory) zone of 21.4 × 105 pixels = 573%) against Phytophthora ramorum than growth/clear zone Cartesian integrates recorded in untreated (control) disc (3.7 × 105 pixels = 100%) over 3-day incubation. The fairy ring fungal extracts also exhibited substantive antagonism against P. kernoviae (147%), P. lateralis (347%) and a solanaceous crop infecting P. infestans (86%). Quite encouragingly, the soil oomycete phytopathogen P. ramorum was inhibited strongly (mean ~ 177%) by both forest bacilli. Aqueous extracts of non-forest antifungal herbaceous plants (garlic and elderberry) expressed similar inhibitory effects (mean ~ 70%). A seaweed fungal elicitor component fucoidan showed moderate levels (mean ~ 85%) of antagonism against P. ramorum, P. kernoviae, P. lateralis and P. infestans. The results in this in vitro study highlight the intensity and vigor of antagonistic forest microflora and non-forest herbaceous antifungal agents such as garlic and other plant extracts as serious candidates for suppression of the oomycete Phytophthora pathogenic fungi in forest soils. This study calls for urgent scoping and impact assessment studies in pot experiments and mini-plot forest trials to gauge the fitness of these natural resources for field level potential biotechnological applications to combat the devastating dieback disease in the native woodlands and horticulture.展开更多
The Clitocybaceae is a recently established family.Currently,the infrafamilial divisions and relationships within the family are vague due to limited sampling and genes employed for phylogenetic analysis.Some mushroom...The Clitocybaceae is a recently established family.Currently,the infrafamilial divisions and relationships within the family are vague due to limited sampling and genes employed for phylogenetic analysis.Some mushrooms of the family contain the neurotoxic muscarine,which has caused many severe and even deadly poisonings worldwide.However,the taxonomic distribution and evolution of the toxin within the family is largely unknown.In this study,phylogenetic analyses based on nucleotide sequences of ITS and of six molecular loci(ITS,LSU,TEF1,RPB1,RPB2 and ATP6),plus a phylogenomic analysis based on 485 single-copy orthologous genes,were performed to reconstruct the framework of Clitocybaceae.BEAST analysis was used to estimate the divergence times within the family.Additionally,biochemical analysis for muscarine was conducted of 32 representative species.Based on these analyses,an updated classification of Clitocybaceae into six genera(Clitocybe,Collybia,Dendrocollybia,Lepista,Pseudolyophyllum,and Singerocybe)is proposed.The genus Collybia is emended to accommodate four subgenera(Collybia,Crassicybe,Leucocalocybe,and Macrosporocybe).Seventeen new Chinese species and 15 new combinations are proposed.Keys to the genera of Clitocybaceae and the subgenera of Collybia,as well as to the known species of Clitocybe and Collybia subgen.Collybia in China,are presented.In addition,muscarine was detected in 18 species,and these muscarine-containing species formed a major monophyletic clade within Collybia subgen.Collybia.Finally,our phylogenetic,phylogenomic,chemotaxonomic and molecular dating results indicate that the Clitocybaceae is a natural group estimated to have arisen some 60 million years ago,and in this family,muscarine has evolved only once circa 20 million years ago without later losses.展开更多
A new classification of several clitocyboid taxa is here proposed to accommodate results from multigenic phylogenetic inference.The analysis of ITS rDNA as well as a combined dataset including 18S and 28S rDNA,tef1 an...A new classification of several clitocyboid taxa is here proposed to accommodate results from multigenic phylogenetic inference.The analysis of ITS rDNA as well as a combined dataset including 18S and 28S rDNA,tef1 and rpb2 data,support significantly a shared monophyletic origin of the genera Pseudoclitocybe,Musumecia and Pogonoloma,and the species Clitocybe alexandri and C.harperi.The new family Pseudoclitocybaceae is here proposed to name this clade,characterized by the presence of loop-like clamp connections in most species,absence of cystidia,and parallel hymenophoral trama with broad,cylindrical hyphae with intracellular granulations.The new genera Clitopaxillus and Harmajaea are proposed to accommodate the type species C.alexandri and H.harperi,as well as the combination H.wellsiae.In addition,two new species are described:C.fibulatus has a differential distribution of clamp connections in the basidiome,while H.guldeniae is,by now,an exclusively European taxon with brownish pileus,somewhat decurrent gills,ovoid spores and basidia longer than those of H.harperi.Finally,the species concept within Pseudoclitocybe and Pogonoloma is discussed and descriptions of the most representative species are provided.展开更多
文摘Response surface (RSM) methodology based on a three-level three-factor Box-Behnkendesign of experiment was used to optimize the exopolysaccharide content (EPC) and themycelium biomass in submerged cultivation by Clitocybe sp. AS 5.112. The criticalfactors selected for the investigation were cultivation temperature, time and volume ofmedium, based on the results of previous Plackett-Burman design. By analyzing theresponse surface plots, the optimum ranges of cultivation temperature, time and mediumvolume for obtaining over 1 253.00gmL-1 of EPC lie in 24.325.8℃, 9.710.2d and 76.090.0mL, respectively. While for obtaining over 8.32 mg mL-1 of dry cell weight (DCW), theabove variables would be in the range of 23.824.8℃, 9.610.3d and 71.098.0mL,respectively. By solving the inverse matrix from the quadratic regression equations, theoptimal conditions to gain 1 265.45gmL-1 of EPC were 25.0℃, 9.9d and 83.4mL, to gain8.50mg mL-1 of DCW were 24.4℃, 9.9d and 87.1mL. In order to obtain the maximum yield ofEPC and DCW at the same time, the above conditions would be 24.5℃, 9.9d and 84.7mL,respectively, in this situation, the maximum predicted EPC and DCW were 1 261.60gmL-1and 8.47mgmL-1, respectively. The experimental data under various conditions have validatedthe theoretical values.
文摘In vitro Kirby-Bauer disc-diffusion assays coupled with bio-imaging software techniques were used to assess native forest dwelling “fairy ring” forming fungi (Clitocybe nebularis) and co-habitant forest tree-root colonising non pathogenic, antibiotic producing bacteria (Bacillus licheniformis, Bacillus pumilis) for their antagonism towards Japanese larch dieback oomycetes phytopathogens which also affects ornamental alternative hosts. The aqueous extracts of C. nebularis exhibited the highest clearance (inhibitory) zone of 21.4 × 105 pixels = 573%) against Phytophthora ramorum than growth/clear zone Cartesian integrates recorded in untreated (control) disc (3.7 × 105 pixels = 100%) over 3-day incubation. The fairy ring fungal extracts also exhibited substantive antagonism against P. kernoviae (147%), P. lateralis (347%) and a solanaceous crop infecting P. infestans (86%). Quite encouragingly, the soil oomycete phytopathogen P. ramorum was inhibited strongly (mean ~ 177%) by both forest bacilli. Aqueous extracts of non-forest antifungal herbaceous plants (garlic and elderberry) expressed similar inhibitory effects (mean ~ 70%). A seaweed fungal elicitor component fucoidan showed moderate levels (mean ~ 85%) of antagonism against P. ramorum, P. kernoviae, P. lateralis and P. infestans. The results in this in vitro study highlight the intensity and vigor of antagonistic forest microflora and non-forest herbaceous antifungal agents such as garlic and other plant extracts as serious candidates for suppression of the oomycete Phytophthora pathogenic fungi in forest soils. This study calls for urgent scoping and impact assessment studies in pot experiments and mini-plot forest trials to gauge the fitness of these natural resources for field level potential biotechnological applications to combat the devastating dieback disease in the native woodlands and horticulture.
基金This study was supported by Yunnan Xingdian Talents Support Plan-Science and Technology Leading Talents Program(202305AB350004)Yunnan Ten-Thousand-Talents Plan-Yunling Scholar Project(YNWR-YLXZ-2018-011)Postdoctoral Directional Training Foundation of Yunnan Province.
文摘The Clitocybaceae is a recently established family.Currently,the infrafamilial divisions and relationships within the family are vague due to limited sampling and genes employed for phylogenetic analysis.Some mushrooms of the family contain the neurotoxic muscarine,which has caused many severe and even deadly poisonings worldwide.However,the taxonomic distribution and evolution of the toxin within the family is largely unknown.In this study,phylogenetic analyses based on nucleotide sequences of ITS and of six molecular loci(ITS,LSU,TEF1,RPB1,RPB2 and ATP6),plus a phylogenomic analysis based on 485 single-copy orthologous genes,were performed to reconstruct the framework of Clitocybaceae.BEAST analysis was used to estimate the divergence times within the family.Additionally,biochemical analysis for muscarine was conducted of 32 representative species.Based on these analyses,an updated classification of Clitocybaceae into six genera(Clitocybe,Collybia,Dendrocollybia,Lepista,Pseudolyophyllum,and Singerocybe)is proposed.The genus Collybia is emended to accommodate four subgenera(Collybia,Crassicybe,Leucocalocybe,and Macrosporocybe).Seventeen new Chinese species and 15 new combinations are proposed.Keys to the genera of Clitocybaceae and the subgenera of Collybia,as well as to the known species of Clitocybe and Collybia subgen.Collybia in China,are presented.In addition,muscarine was detected in 18 species,and these muscarine-containing species formed a major monophyletic clade within Collybia subgen.Collybia.Finally,our phylogenetic,phylogenomic,chemotaxonomic and molecular dating results indicate that the Clitocybaceae is a natural group estimated to have arisen some 60 million years ago,and in this family,muscarine has evolved only once circa 20 million years ago without later losses.
文摘A new classification of several clitocyboid taxa is here proposed to accommodate results from multigenic phylogenetic inference.The analysis of ITS rDNA as well as a combined dataset including 18S and 28S rDNA,tef1 and rpb2 data,support significantly a shared monophyletic origin of the genera Pseudoclitocybe,Musumecia and Pogonoloma,and the species Clitocybe alexandri and C.harperi.The new family Pseudoclitocybaceae is here proposed to name this clade,characterized by the presence of loop-like clamp connections in most species,absence of cystidia,and parallel hymenophoral trama with broad,cylindrical hyphae with intracellular granulations.The new genera Clitopaxillus and Harmajaea are proposed to accommodate the type species C.alexandri and H.harperi,as well as the combination H.wellsiae.In addition,two new species are described:C.fibulatus has a differential distribution of clamp connections in the basidiome,while H.guldeniae is,by now,an exclusively European taxon with brownish pileus,somewhat decurrent gills,ovoid spores and basidia longer than those of H.harperi.Finally,the species concept within Pseudoclitocybe and Pogonoloma is discussed and descriptions of the most representative species are provided.
