Optimization of submerged culture conditions involving a developed fine powder solid seed for exopolysaccharide production by the medicinal mushroom Ganoderma lucidum.

To facilitate Ganoderma lucidum submerged cultivation and achieve high productivity, four fine powder solid substrates incorporated with different nitrogen-rich supplements were utilized to grow the fungus and as solid seed for its submerged culture. Of the four solid seeds, the soybean meal solid seed gave the highest biomass (10.73 g/L) and exopolysaccharide (EPS) (1.22 g/L), higher than those (8.36 g/L biomass and 0.44 g/L EPS) obtained with mycelial liquid seed. The optimal level of soybean meal supplementation was 20% (w/w) for production of the solid seed. Following single factor experiments, levels of three selected process variables were optimized as: the moisture content of solid seed, 70%; inoculum size, 0.8 g/flask; and rotary speed, 160 rpm. These conditions were validated experimentally with improved EPS yield of 1.33 g/L. The developed solid seed can be conveniently used for G. lucidum submerged culture with improved EPS productivity.

Development of a Novel Spawn (Block Spawn) of an Edible Mushroom, Pleurotus ostreatus, in Liquid Culture and its Cultivation Evaluation.

Mushroom cultivation has gained increased attention in recent years. Currently, only four types of spawn, including sawdust spawn, grain spawn, liquid spawn, and stick spawn, are commonly available for mushroom cultivation. This limited spawn diversity has led to difficulty in selecting suitable inoculum materials in some cultivation. In this study, three small blocks of lignocellulosic agro-wastes and one block of a synthetic matrix were prepared as support for growing Pleurotus ostreatus in liquid medium. Mycelium-adsorbed blocks were then evaluated for their potential as block spawn for fructification. Our results indicated that the edible fungus was adsorbed and abundantly grew internally and externally on loofah sponge and synthetic polyurethane foam (PUF) supports and also has the ability to attach and grow on the surface of sugarcane bagasse and corncob supports. The mycelia of P. ostreatus adhered on corncob exhibited the highest metabolic activity, while those on the PUF showed the least activity. Mycelial extension rates of block spawns made of agro-waste materials were comparable to that of sawdust spawn, but the block spawn of PUF showed a significantly lower rate. No significant differences in cropping time and yield were observed among cultivations between experimental block spawns and sawdust spawns. Moreover, the corncob block spawn maintained its fruiting potential during an examined period of 6-month storage. The developed block spawn could be practically applied in mushroom cultivation.

Hyperproduction of exopolysaccharides by submerged mycelial culture of Ganoderma lucidum using a solid seed grown in fine-powder of wheat bran and in vitro evaluation of the antioxidant activity of the exopolysaccharides produced.

To facilitate Ganoderma lucidum submerged culture and obtain high productivity, a fine powder of wheat bran was used to grow the fungus for solid-state fermentation and as solid seed for its submerged cultures. The results indicated that the optimal inoculum size was low, being 0.75 g in 250 mL-sized flasks containing 80 mL medium. The maximal exopolysaccharide concentration and biomass produced was 0.74 and 14.71 g/L, respectively, which is considerably higher than that obtained with the commonly used mycelial pellet liquid seed (0.47 and 8.56 g/L, respectively). The EPS and biomass productivity of the solid seed cultures decreased only slightly, even after a 6-month storage period. EPS produced showed higher antioxidant activity compared with that produced in the liquid seed cultures. The developed solid seed can serve as a ready-to-use inoculum for long-term use in G. lucidum submerged culture for the hyperproduction of highly bioactive EPS and biomass.

Investigating the Effect of Tissue Size on Mycelial Growth of Seven Mushroom Species by Using a Novel Device for Precise Tissue Isolation.

Tissue isolation from mushrooms is frequently practiced by both researchers and growers to isolate new and improved strains. In the present study, we designed a simple and convenient device for precise tissue isolation and therefore investigated the effect of tissue size on mycelial growth of seven mushroom species. The developed device consists of a cutting needle and a transfer needle. The cutting needle was used to obtain circular tissue plugs having a height up to 3 mm and variable diameters (2-5 mm) from mushroom fruit bodies. The transfer needle was a stainless steel round rod (1.5 mm in diameter) with a blade-like end. It can be used for collecting mushroom tissue when the cutting needle fails to extract it. With the aid of these devices, precise tissue isolation was achieved. Plate cultures demonstrated that tissue size had little effect on mycelium extension for Lentinula edodes (the winter shiitake), Hypsizygus marmoreus, and Agrocybe aegerita, but influenced the aerobic mycelium density. For Pleurotus ostreatus, Pleurotus eryngii, and Volvariella volvacea, large tissue plugs produced faster mycelial growth and higher aerobic mycelium density compared with small ones. On the contrary, small plugs from the tissue of the flower shiitake and Agaricus bisporus favored mycelial growth. The present study revealed that the preferable tissue size for mycelial growth varies among mushroom species, and the developed device is expected to greatly facilitate the isolation of new and improved mushroom strains.

[Identify the original plant of Forsythia suspensa by nucleotide sequence].

OBJECTIVE: To investigate the means of distinguishing the original plant of Forsythia suspensa from confusion. METHODS: To amplify the chloroplast matK gene by PCR using a consensus primer set and determine their nucleotide sequence by PCR direct sequencing. The ITS sequences were gained from NCBI. The characteristic analysis of matK and ITS sequences were generated using Clustal aligned. RESULTS: There were 30 bp and 8 bp unique nucleotide in ITS and matK sequence in Forsythia suspensa. The matK gene and ITS sequences might be good molecular marker to be used to identify the original plant of Forsythia suspensa. CONCLUSION: The sequence analysis of matK gene ITS sequences might become the mean to identify the original plant of Forsythia suspensa.