Noble strain of Sparassis latifolia produces high content of β-glucan

Objective: To classify four new Sparassis strains(CLM1, CKM1, CKM2, and KJM1) using the internal transcribed spacer sequence and to elucidate their β-glucan content and mycelial growth.Methods: Two different microbiological media were used to determine growth rate. The β-glucan contents were analyzed using the Megazyme Mushroom and Yeast Beta-Glucan kit. To determine the genetic relationships, phylogenetic trees were constructed using ClustalX. Multiple sequence alignments were printed and shaded with the BOXSHADE 3.21 program. Results: In this study, four new Sparassis strains were isolated from the southern region of the Korea Peninsula. They were all classified into the Sparassis latifolia clade as a monophyletic group based on the internal transcribed spacer sequence. Mycelial growth rate of the CLM1 strain was highest in potato dextrose agar and potato dextrose agar larch. The β-glucan content of the CLM1 strain was highest at 29.5%(w/w). A high degree of sequence divergence was detected in the RNA polymerase second largest subunit II gene(RPB2) within Sparassis spp. tested. The putative amino acid sequences of the RPB2 had a distinct sequence. The nucleotide sequences of the RPB2's intron were also divergent among Sparassis spp., even though their nucleotide length was well conserved within Sparassis latifolia. Conclusions: These results indicate that the nucleotide sequences and the amino acid sequences of RPB2 can be used to identify individual Sparassis sp. The Sparassis strain CLM1 may be best for developing a remedy to prevent or treat cancer and other chronic diseases.

Vegetative Development of <i>Sparassis crispa</i>in Various Growth Conditions and Effect of Electric Pulse Simulation on Its Fruit Body Production

This study was carried out to detect the favorable mycelial growth conditions and effect of electric pulse stimulation on fruit body production of SC-1 strain of Sparassis crispa. The optimum growth in PDA was found at 25°C (67 mm colony diameter in 28 days) followed by 20°C (63 mm colony diameter in 28 days). There was no mycelial growth at 35°C. The most favourable initial media pH range was found to be 5 - 7 and colony diameter measurements were not statistically different among these pH values (p > 0.05). However, the highest growth was obtained at pH 6 (57 mm colony diameter in 28 days). A basal medium composed of 0.05 g MgSO4, 0.46 g KH2PO4, 1.0 g K2HPO4, 120 μg thiamine-HCl, 20 g agar and 1000 ml of distilled water was used to investigate growth response of S. crispato different carbon and nitrogen sources. In 28 days, fructose and glucose exhibited best growth (49.4 mm and 31.6 mm colony diameters respectively) and there was no growth on the basal medium supplemented with galactose. Basal medium supplemented with glycine and alanine as nitrogen sources resulted in best growth, 54.4 mm and 50.5 mm colony diameters respectively. There was no mycelial growth in culture medium supplemented with ammonium acetate, ammonium phosphate, arginine and histidine. Electric pulse stimulation improved the fruiting body production. The yields obtained from all bottles in which electric pulse was applied were significantly higher than the yields from the bottles in which electric pulse was not applied. The percent increases of fresh weight yield from control on 100, 120, 130, and 170 kilovolts were 36%, 44%, 75% and 81% respectively. As regard to dry weight yield, the percent increases from control on 100, 120, 130, and 170 kilovolts were 27%, 54%, 63% and 67% respectively.