Handbook Part 2 - International Mycological Association

PS7-604-0337
Isolation of monokaryon from asexual spore of Taiwanofugus camphoratus and breeding of high
triterpenoid strains.
Fang-Mo Chang, Ching-Hua Su
Taipei medical university, Taipei, Taiwan
Taiwanofugus camphoratus Wu et al. contained high triterpenoids and other physiologically active compounds were
used as a precious herb in Taiwan. The culture of T. camphoratus was grown in PD agar plate, and the agar-slab
containing mycelium and asexual spore was spread over agar plates. After 5~8 days, the germinating asexual spore
was pick up by a aseptic syringe needle under dissect microscope. The germiling of the asexual spores was transferred
to PD agar plate for further growth. A small portion of the mycelium with small piece of agar-slab was transferred to a
slide and incubated for another 7-10 days under 25?.After the mycelium was spread over the slide, the agar-slab was
removed and the mycelium on the slide was stained by HCl-giemsa. It is obvious that the clamp-connection was
disappeared through the hyphae of the single-asexual-spore isolates and all the septate hypha containing only one
nucleus. It suggested that the isolates were monokaryons. The mating reaction of isolates are now under investigation.
To obtain high triterpenoid strains, cultures of dikaryon and monokaryons were grown on cellophane PD agar for 21
days at 25?. The mycelium part and medium were separately dried. Triterpenoid contain was analyzed by TLC and
HPLC. The study will continue to detect the triterpenoid contain in monokaryon and their mated dikaryons, thus high
production strain triterpenoid was able to be selected.
PS7-605-0358
Cultivation properties of termite mushroom
Y Tamai 1, J-Y Cha1, I.G.P Wirawan 2, M Terazawa 1
1 Hokkaido University, Sapporo, Hokkaido, Japan, 2 Udayana University, Denpasar, Bali, Indonesia
Termitomyces spp. are widely distributed in the tropics of Asia, Africa and South Pacific. They are well known as highly
valued edible mushrooms in the regions, but cultivation technology has not been developed yet. They have
symbiotic relationships with the fungus-growing termites, and mainly decompose the plant material of the nests. The
termites consume the decomposed materials and fungal biomass. Therefore, the fruit bodies arise only from nests
deep underground. In the present paper, we introduce a few preliminary cultivation properties of Termitomyces sp..
Fruit bodies were collected from Tenganan province in Bali Island, and tested strains were isolated from the tissue and
maintained as stock culture. Accoding to the morphologic characteristics, these specimens were identified as
Termitomyces eurrhizus (Berk.) Heim: Pileus 7-10cm wide, grayish brown, campanulate then almost to plane with
umbonate perforatorium, surface smooth and viscid when moist. Stipe 3-5.5 x 1.3-2.5cm, surface white, attenuated
towards the apex. Context fleshy, white. Gills white to pinkish gray. Spores 7-9 x 5-6 um, ellipsoid.
The pseudorrhiza was connected to living termite nest, and many workers, soldiers and larvae were observed inside.
The host termites were identified as Odontotermes sp.. The white tufts of Termitomyces and grayish green tufts of
Xylaria were observed on the surface of nests. The C/N ratio and pH of the fungus garden (termite nest) were 22.5-
33 and 3-5 respectively.
Mycelial growth speed was very low on the PDA medium. Moderately good growth occurred on Hamada medium,
and grayish-white colony with little aerial hyphae was presented. For optimum growth, initial pH of the medium was
adjusted to pH5.0, not changed during the incubation. The optimum temperature was ranged in 25-30C. Light
irradiation depressed the mycelial growth. Ammonium tartrate and urea were well utilized as nitrogen source in
synthetic medium. No conspicuous differece in the growth and density of mycelium could be observed between the
natural additives (wood or grass oriented).
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