A Preliminary Screening of Optimum Medium and Symbiotic Fungi for Paphiopedilum dianthum

[Objectives]This study aimed to screen the suitable medium and symbiotic fungi for tissue culture of Paphiopedilum dianthum and to establish a rapid propagation system of Paphiopedilum,so as to promote the development of planting industry and commercial application of Paphiopedilum.[Methods]Using P.dianthum as an experimental materials,the optimal culture medium for P.dianthum was selected from DE medium,Harvais modified medium and(bark+DE)liquid medium.Then,six fungal species isolated from the original habitat soil of Paphiopedilum were co-cultured with P.dianthum on the selected optimal culture medium.By comparing the morphology,the growth amount and the physiological indexes,such as chlorophyll content,soluble sugar content and root vitality of tissue culture plantlets of P.dianthum co-cultured with the six fungal species,the suitable fungal strains for symbiosis with P.dianthum were screened out.[Results]In three kinds of culture media,P.dianthum grew normally.The growth rate was higher than that in the mediums containing DE,and there were no significant differences in the content of soluble sugar from the leaves of tissue culture plantlets,which were symbiotic with the six different fungi species.Among the six fungi species growing on the optimal medium,the growing rate of XH-A was the fastest,and that of YY001 was the slowest.The growth and physiological indexes of P.dianthum co-cultured with DY1 in DE medium were higher than those in the control group.[Conclusions]DE medium was the optimal tissue culture medium for P.dianthum,and the optimal symbiotic fungus was DY1.

Ectomycorrhizal fungi and dark septate endophyte inoculation improve growth and tolerance of Pinus tabulaeformis under cadmium stress

Forest trees can establish symbiotic associations with dark septate endophytes(DSEs)and ectomycorrhizal fungi(ECMF)simultaneously.However,the combined effects of these two fungi on the growth and cadmium(Cd)tolerance of host plants remain largely unexplored.To address this knowledge gap,a pot experiment was conducted to examine the effects of the interaction between an ECMF strain(Suillus granulatus)and a DSE strain(Pseudopyrenochaeta sp.)on Pinus tabulaeformis under Cd stress,by assessing plant growth and physiological parameters,nutrient uptake,and soil properties.Notably,the colonization rates of both fungal strains were found to increase in response to Cd stress,with the extent of this increase being influenced by the specific fungal species and the Cd level in the soil.Compared to the non-inoculation treatment,single inoculation with fungal strain resulted in enhanced biomass,root development,and nutrient contents in P.tabulaeformis seedlings under Cd stress.Furthermore,a synergistic effect was observed when these seedlings were co-inoculated with S.granulatus and Pseudopyrenochaeta sp.,as indicated by significantly greater measurements in various indicators compared to both the single and non-inoculation treatments.Fungal inoculation effectively regulated the antioxidant defense responses and photosynthesis of P.tabulaeformis seedlings subjected to Cd stress,particularly in the co-inoculation treatment.In addition,fungal inoculation facilitated the Cd accumulation in P.tabulaeformis,suggesting a promising potential for the implementation of bioremediation strategies in the areas contaminated with heavy metals.The findings from this study indicate that the utilization of root symbiotic fungi obtained from stress environments could potentially enhance the growth performance and tolerance of P.tabulaeformis towards heavy metals,and co-inoculation of both fungal groups may result in even more pronounced synergistic effects on the overall fitness of the plant.

Mycorrhizal fungi mitigate nitrogen losses of an experimental grassland by facilitating plant uptake and soil microbial immobilization

Nitrogen(N)is one of the most limited nutrients of terrestrial ecosystems,whose losses are prevented in tightly coupled cycles in finely tuned systems.Global change-induced N enrichment through atmospheric deposition and application of vast amounts of fertilizer are now challenging the terrestrial N cycle.Arbuscular mycorrhizal fungi(AMF)are known drivers of plant-soil nutrient fluxes,but a comprehensive assessment of AMF involvement in N cycling under global change is still lacking.Here,we simulated N enrichment by fertilization(low/high)in experimental grassland microcosms under greenhouse conditions in the presence or absence of AMF and continuously monitored different N pathways over nine months.We found that high N enrichment by fertilization decreased the relative abundance of legumes and the plant species dominating the plant community changed from grasses to forbs in the presence of AMF,based on aboveground biomass.The presence of AMF always maintained plant N:phosphorus(P)ratios between 14 and 16,no matter how the soil N availability changed.Shifts in plant N:P ratios due to the increased plant N and P uptake might thus be a primary pathway of AMF altering plant community composition.Furthermore,we constructed a comprehensive picture of AMF’s role in N cycling,highlighting that AMF reduced N losses primarily by mitigating N leaching,while N_(2)O emissions played a marginal role.Arbuscular mycorrhizal fungi reduced N_(2)O emissions directly through the promotion of N_(2)O-consuming denitrifiers.The underlying mechanism for reducing N leaching is mainly the AMF-mediated improved nutrient uptake and AMF-associated microbial immobilization.Our results indicate that synergies between AMF and other soil microorganisms cannot be ignored in N cycling and that the integral role of AMF in N cycling terrestrial ecosystems can buffer the upcoming global changes.

Isolation and identifi cation of antimicrobial metabolites from sea anemone-derived fungus Emericella sp.SMA01

Marine symbiotic fungi represent an intriguing source of discovery of novel secondary metabolites with various biological activities.Sea anemones are benthic marine invertebrates,however,the cultivable symbiotic fungi residing in the sea anemones are paid few attentions compared to those derived from their cnidarian counterparts.Here we show the identification of antimicrobial secondary metabolites from the sea anemone-derived symbiotic fungi.Out offive isolated fungal strains,only the strain SMA01 showed strong antimicrobial activities,which was assigned into the genus Emericella based on the morphological characteristics and the ITS sequencing.Media swift from liquid fermentation to solid rice medium presented little influence on its antibacterial activity.A chemical investigation of the ethyl acetate extract of the Emericella sp.SMA01 led to discovery of the primary antibiotic metabolite phenazine-1-carboxylic acid.The IC_(50) values of the phenazine-1-carboxylic acid against Phytophthora capsici,Gibberella zeae,and Verticillium dahliae were determined to be 23.26-53.89μg/mL.To the best of our knowledge,this was the first report of Emericella sp.in sea anemones.The current study may benefit understanding of the defensive chemical interactions between the symbiotic fungi and their host sea anemones.

Characterization of two chimeric sesterterpene synthases from a fungal symbiont isolated from a sesterterpenoid-producing Lamiaceae plant Leucosceptrum canum

Two chimeric sesterterpene synthases(Aa TPS1 and Aa TPS2)were functionally characterized from Alternaria alternata MB-30 isolated from the leaves of a sesterterpenoid-producing Lamiaceae plant Leucosceptrum canum.Aa TPS1 generated a 5/8/6/5 tetracyclic sesteraltererol(1)and its absolute stereochemistry was determined by X-ray crystallographic analysis of its derivative 10,11-epoxysesteraltererol(2),which enabled revision of the absolute configuration of C7 of sesterfisherol produced by Nf SS and PTTS014 characterized previously and its derivative 10,11-epoxysesterfisherol.Aa TPS2 produced a 5/15 bicyclic preterpestacin I(3).Site-directed mutagenesis suggested that F192 in Aa TPS1 was likely involved in controlling of the hydroxylation of C12,and eight amino acids were important for the enzyme activity of Aa TPS1 and Aa TPS2.The engineered Escherichia coli and Saccharomyces cerevisiae strains were constructed for the productions of compounds 1 and 3,and the highest titer of compound 1 reached 62.3 mg/L in shake-flask culture.Both compounds 1 and 2 showed anti-adipogenic activity.