The present situation of studies on the fruit-body formation of the mycorrhizal edible mushroom fungi, Lyophyllum shimeji and Tricholoma matsutake

The ectomycorrhizal fungi Tricholoma matsutake and Lyophyllum shimeji are famous and delicious edible mushrooms from Japan. Since they form mycorrhiza on the fine roots of living plants, their growth depends facultatively on living plants. To cultivate these fungi for in a pure culture, glucose and a few other monosaccharides must be used as carbon sources. In 1994, it was reported that L. shimeji can form mature fruit-bodies on a medium that consists mainly of barley grains; in addition, they form without a host plant. These observations indicate that some mycorrhizal mushroom fungi have the ability to utilize barley starch. When fungi form fruit-bodies, large amounts of mycelia may be needed either to store nutrients or to transport nutrients to the fruit-bodies, or for both purposes. However, it is very difficult to cultivate large amounts of mycelia using monosaccharides in a pure culture. The addition of low molecular weight substances in high concentrations increases the osmotic pressures of a medium, which in this case would suppress the growth of the mycelium. Then, amylase productions during the vegetative mycelial growth of T. matsutake (No. 114 and Z-1 strain), which hydrolize starch as a substrate for the growth of the fungi were examined using a somewhat modified matsutake liquid medium. Amylase activity (a - and gluco-amylase) in the culture filtrates had relatively high values at 40 days after the inoculation. Maximum activity was attained 80 days after inoculation. a - Amylase showed more activity than glucoamylase in both strains. Amylase activity in the culture filtrate of T. matsutake was assayed by using several kinds of starch as the substrate; the starches were purified from barley (5 kinds), corn, sweet potato, and potato. The enzyme was most active in the presence of the starch prepared from the "Amagi" and "Ichibanboshi" strains in the "Nonwaxy" barley group among the starches. This activity was were observed at a circa 2.0 - fold higher value than that of the control (potato). The addition of potato and yam to the culture medium increased the dry weight of mycelia 4.8-5.6 times, as compared to the control (without addition); moreover, the value of amylase activity was 1.9 - 2.6 times that of the control. A positive correlation between vegetative mycelial growth and amylase activity was detected. To elucidate the properties of extracellular amylases in the T. matsutake Z-1 strain, the amylase fractions obtained from the DEAE - Toyopearl column chromatography were identified. One type of a-amylase, glucoamylases and a - glucosidases showed activity upon analysis by thin layer chromatography using hydrolyzates.