Mycorrhizal Synthesis Involving Boletus edulis and Pinus massoniana

Mycorrhizae involving Boletus edulis and Pinus massoniana have been successfully established and their morphological characteristics described.Optimal conditions for mycorrhizal synthesis were determined using orthogonal testing of three parameters:substrate,age of host plant and inoculation procedure.Following integration analysis of plant survival rates,diameter of seedling rootstock,height of seedling shoot,number of mycorrhizae,ratio of shoot/root biomass,number of mycorrhizae,and the ratio of infected/non-infected root tips,we concluded that a double-layered growth substrate that simulated natural soil structure was superior for mycorrhizal synthesis compared with the more commonly used single-layered peat moss and vermiculite substrate,especially when sepiolite powder was also added.

Relationship Between Substrate Water Content and Weight Loss,Mycelium Culture Time and Hypsizygus marmoreus Growth Yields

Changes in the weight and water content of the growth substrate at different cultivation times significantly affected Hypsizygus marmoreus growth yields.Highest growth yields(132.6~146.3 g per culture) were achieved when the mycelium culture and maturation period was between 80 and 90 days,and the substrate water content was 73%~74%.Physiological maturation of H.marmoreus mycelium was correlated with the substrate water content.

Isolation,Purification and Immunomodifying Activity of Polysaccharides from Phellinus igniarius

Phellinus igniarius polysaccharides were obtained by hot water extraction of mushroom fruit bodies followed by ethanol precipitation,and further purified by anion exchange and gel filtration chromatography.Immuno-enhancing activity was evaluated by determining the effect of different polysaccharide fractions on mouse spleen lymphocyte proliferation in vitro.Three crude polysaccharide preparations,designated PIP30,PIP60 and PIP80 according to the percentage concentration of ethanol required for precipitation from hot aqueous extracts,promoted lymphocyte proliferation to varying degrees.Purification of polysaccharide present in PIP30 and PIP60 was carried out using anion exchange chromatography.Proliferation rates in samples treated with 200 μg/mL of fractions P30U,P30W and P30S1(obtained following anion exchange chromatography of PIP30) were approximately 5-,4-,and 3.5-fold higher,respectively compared with negative controls.Similarly,marked increases(3.4-and 2.8-fold) in cell proliferation rates compared with negative controls were observed with 200 μg/mL concentrations of P60W and P60S2(obtained following anion exchange chromatography of PIP60),respectively.Two purified polysaccharide preparations,P60W1-1 and P60S1-1,were obtained following gel filtation chromatography of fractions P60W1 and P0S1,respectively.Fraction P60W1-1(10-100 g/mL) enhanced splenocyte proliferation 0.5-1.4-fold compared with negative controls,whereas no effects were observed with P60S1-1.

Cloning of a Putative Pheromone Receptor Gene Fragment from Lentinula edodes

A 508 bp fragment of a putative pheromone receptor gene from Lentinula edodes was amplified by PCR using a pair of degenerate primers designed according to conserved sequences of pheromone receptor genes reported in other Basidiomycetes.The fragment encodes 117 amino acids and contains three introns,56 bp,51 bp and 50 bp in size.Alignment analysis revealed that the deduced amino acid sequence of the L.edodes fragment exhibited 65% identity and 82% similarity with the corresponding segments of pheromone receptors reported in Coprinus cinereus and Schizophyllum commune.

Analysis of Bacterial Communities Present in Agaricus bisporus Phase Ⅱ Compost Using Amplified Ribosomal DNA Restriction Analysis(ARDRA)(Ⅱ)——Analysis of Bacterial Communities

Changes in the bacterial communities inhabiting Agaricus bisporus compost during Phase Ⅱ treatment were monitored using amplified rDNA restriction analysis(ARDRA).Sequence analysis data indicated that the bacterial communities associated with the compost samples were far richer in composition when determined by ARDRA compared with traditional methods based on bacterial isolation.Furthermore,the composition of the communities,especially in terms of the dominant bacteria during each of the four treatment stages(beginning of Phase Ⅱ,after pasteurization,5 days aeration,end of Phase Ⅱ,respectively),were very different.Restriction endonuclease digestion of mainly bacterial clones from four 16S rDNA libraries,C1,C3,C6 and C7,previously constructed on the basis of 16S rDNA fragments amplified from the four different stages of the Phase Ⅱ composting process,revealed the presence of unique microbial types.Thermophilic bacteria belonging to the Bacilli,and previously unreported strains of Trichococcus,Planococcus,Caryophanon,and subclass γ-Proteobacteria,were identified among the clones from the compost sampled during the aeration period(C6).Bacteria belonging to Thermus thermophilus and subclass α-Proteobacteria were detected in C1 and C7 composts sampled at the beginning and the end of Phase Ⅱ,respectively.Clones of some uncultured bacteria were also scored.