Mixed-culture Cultivation of 6. Different growth parameters: T.

fuciformis growth parameters

are controlled to produce the desired phases (sexual vs asexual
Tremella fuciformis reproductive phases) during its life cycle .

on Synthetic Logs 7. Unstable mixed cultures or spawn: go back to mother culture

as necessary.
Alice W. Chen, Ph.D.
Specialty Mushrooms SUBSTRATE FORMULATION
1730 Penfield Rd., #41
Penfield, NY 14526 Supplemented sawdust-bran substrates are generally used.
Voice: 716-381-0804 Other agricultural wastes may also be used (Table 1). Cotton-
seed hull has been reported to give higher yields.
Nian Lai Huang
Director of Saming Mycological Institute PRODUCTION OF MIXED MOTHER CULTURE (OR
Saming, Fujian Province, China MOTHER CULTURE SPAWN):

INTRODUCTION Production of the mixed mother culture requires that T. fuciformis

be started first, then H. acheri is added into the same test tube.
The white jelly mushroom, Tremella fuciformis, was discovered in T. fuciformis is not fastidious and can be cultured on most labora-
China. It is an important and popular medicinal and gourmet tory media. First, subculture T. fuciformis into the desired number
mushroom in Southeast Asia where it is best known by its of agar slants. Incubate at 25oC. When the colony of T. fuciformis
Chinese name, (Yin Er) or silver ear. reaches 1 cm in diameter, inoculate a minute amount (a few
Cultivation on natural logs began in China in 1894 during the hypha) of H. acheri into each of the tubes to create a mixed cul-
Ching Dynasty. Successful cultiva- ture (T. fuciformis: H. acheri =
tion on synthetic logs was first 1000:1 approximately). When
achieved in Gutein County, Fujian Table 1. Substrate Formulation these two fungi have grown
Province, China, in 1978. together, the mixed cultures are
Cultivation in substrate bags has Supplemented Sawdust-bran Substrate A: ready to be sold as mother-cul-
been used almost exclusively for sawdust, hardwood 70 kg ture spawn or used to produce
the production of T. fuciformis in rice or wheat bran 19 kg the primary spawn.
China since the 1980s. However, gypsum or lime 1 kg
natural logs are still used in Taiwan. sucrose 1 kg Alternatively, H. acheri can be
The discussion below is primarily water, approximately 140 kg started in the test tube first. After
based upon Chinese methodology. the Hypoxylon mycelia has
Supplemented Sawdust-bran Substrate B: become established, yeast-like
Due to some unusual characteris- sawdust, hardwood 77 kg conidia of T. fuciformis, in mas-
tics of T. Fuciformis, some back- bran 18 kg sive amounts, can then be
ground information is essential. sucrose 1 kg smeared onto the substrate sur-
Prior mushroom cultivation experi- gypsum 1.5 kg face. A larger Tremella to
ence will also be helpful. calcium superphosphate 1 kg Hypoxylon ratio is required in this
soybean powder 1.5 kg approach. It can be tricky to use
BACKGROUND water 140 kg the yeast-like conidia of T. fuci-
INFORMATION formis for the production of mixed
Supplemented Cotton-seed Hull and Bran Substrate: culture, since not all the conidia
1. Carbohydrate metabolism : T. cotton-seed hull 100 kg germinate.
Fuciformis can not degrade and use bran 25 kg
either cellulose nor lignin efficiently, soybean powder 3 kg Recently Paul Stamets (2000a, b)
if at all. sucrose 1.5 kg of Fungi Perfecti produced the
calcium superphosphate 1.5 kg mixed mother culture spawn of
2. Mixed culture: for growth on com- water 120-140 kg Yin Er (T. fuciformis) by using liq-
plex substrate containing cellulose uid cultures. 24 drops of H. acheri
and lignin it is necessary to use a mixed culture of Tremella fuci- were added to 2 liters of T. fuciformis (liquid or submerged fer-
formis, a basidiomycete, and Hypoxylon acheri, an ascomycete, mentation). The ratio of mixing is larger than 1:1,000,000,000.
to insure that nutritional needs will be met.

3. Specificity of pairing T. fuciformis and H. acheri: The two fungi PRODUCTION OF MIXED PRIMARY SPAWN
must be isolated, in the wild, from the same log in the same eco-
logical niche. Strains of these two fungi of different origins should Use generous amount of mother-culture spawn to inoculate the
not be mixed. substrate for primary-spawn, usually supplemented sawdust-bran
substrate with 65% moisture content in bottles. Drier substrate
4. Reproduction: The life cycle of T. fuciformis involves both favors the growth of Tremella. Use the entire mixed culture from
basidia in sexual reproduction and yeast-like conidia in asexual a mother-culture agar slant in a tube for 1-4 bottles of primary
reproduction. spawn. Incubate at 25oC until the formation of primordia.
Primordia resemble 1/2 a grain of rice. Formation of primordia
5. Differential mycelial growth rate: mycelial growth of T. fuci- ensures that the primary spawn contains both T. fuciformis and H.
formis is much slower than the mycelial growth of H. acheri. acheri and is capable of producing Tremella fruiting bodies.
 tion to bags after 3 to 5 days to favor the growth of Tremella
PRODUCTION OF SPAWN mycelia. After 10-15 days of spawn run, the bags are moved to
the fruiting room. Refer to Table 2 for the control of growth param-
Screen primary spawn carefully for the production of spawn, on eters.
the basis of :
-formation of vigorous primordia MUSHROOM GROWTH (fruiting-body formation)
-deep mycelial penetration of T. fuciformis into the substrate
-presence of robust feather-like mycelia of H. acheri Formation of white mycelial globules and yellowish exudate at the
These are also the criteria to determine the number of subcul- inoculating sites signals the onset of primordia formation, in order
tures of spawn bottles
to be made from each
chosen primary spawn Table 2. Growth Parameters
bottle.
Spawn Run Primordia Fruiting-Body
Remove and discard Induction Development
the primordia from the
primary spawn, mix the Temperature 25-28oC, 3-5 days 18-23oC 23-24oC
mycelia thoroughly (to favor Hypoxylon)
under aseptic condi-
tions. Keep the mois- 23-25oC, 7-10 days 20-27oC
ture content of the (to favor Tremella) Oei, 1996 (woodlogs)
spawn substrate at 65-
70%. Wetter substrate Relative tolerance of drought 85-95% R. H. 85-95% R. H.
facilitates the formation Humidity 95-100% R. H.
of yeast-like conidia (for medium to large fruitings)
which can be easily dis-
tributed in liquid Light - - 50-600 lux
droplets into the deeper
layer of substrate. This No light required
offers better substrate according to Oei, 1996
utilization and can com-
pensate for lack of Ventilation - increase increase to
depth of mycelial pene- 3-5 times daily
tration by Tremella. (3-10 min each)

Incubate the spawn at Duration 10-15 days 15-20 days 35-40 days
26oC for 18-22 days. (16 days) after spawning
Readiness of the following the
spawn is indicated by appearance of white
the appearance of mycelial globules
white mycelial globules
(balls). Two to three
days after the formation of such globules, the spawn is ready for words, the readiness of synthetic logs to fruit. Drain if excessive
use. Use only the upper portion of the spawn since the lower part amount of exudate is produced. For the regulation of growth
contains less mycelia. Mix mycelia thoroughly before spawning. parameters following the formation of the white mycelial globules,
Use 1 bottle of spawn for approximately 20 synthetic logs in bags, see Table 2.
unless the spawn is of exceptional quality and well mixed. The
spawn may be further incubated for 3-5 days after mixing to INDUCTION OF PRIMORDIA
enhance success. (critical period during fruiting)

SPAWNING (INOCULATION) Mycelial metabolism reaches the peak during the period from the
formation of the whitish mycelial globules to the development of
Use only the top 3 cm of the mixed mycelial spawn. Before primordia. A tremendous amount of heat is generated.
spawning, crumble and mix the culture thoroughly to insure a Management of growth parameters should be closely monitored,
proper combination of Tremella and Hypoxylon. Narrow and long particularly to maintain the proper temperature and gradual and
synthetic bags, 12 cm x 60 cm, are used in China. Four inoculat- careful increase of ventilation to prevent abortion of primordia.
ing holes, 1.5 cm in diameter and 1.2-1.5 cm deep are made with
a mechanical tool. Commonly available wide plastic adhesive Primordia are developed approximately 16 days (15-20 days)
tapes are used to seal the inoculation holes. With the availability after the appearance of the white mycelial globules. Mature fruit-
of polypropylene and polyethylene bags with microfilter windows, ing bodies ready for harvest are produced in 35-40 days after
these can be adopted for the cultivation of silver ears. spawning. A second flush is possible.

SPAWN RUN CONCLUSION

Incubate at 25-28oC for 3 to 5 days to facilitate the growth of Tremella fuciformis is one of the major medicinal mushrooms and
Hypoxylon around inoculation sites and to prevent contamination. a treasured mushroom delicacy in China. Cultivation of this mush-
The Chinese continue incubation at 23-25oC and increase aera- room is a challenge for the most enthusiastic and the advanced
growers. Adoption of the basic methodology, as developed in
detail in China, should be a very worthy step forward. The practi-
cal knowledge of how to eat the mushroom will be a topic for
future discussion.

REFERENCES

Chang, S. T. and P. G. Miles, 1989. Edible mushrooms and their

cultivation. CRC Press, Boca, Raton, FL.

Chen, Alice W. 1998. Notes: studies on Tremella fuciformis in

China.

Chen, M. M., 1997 Fungi treasures. Booklet published by the

author, Berkeley, CA.

Cheung, PCK, 1996. The hypocholesterolemic effect of two edi-

ble mushrooms: Auricularia auricula (tree-ear) and Tremella
fuciformis (white jelly-ear) in hypercholesterolemic rats.
Nutr. Res. 16, 1721.

Hobb, C. 1995. Medicinal mushrooms, an exploration of tradition,

healing and culture. Botanica Press, Santa Cruz, CA.

Huang, Nian Lai (ed.), 1987. Self-study guide on edible fungi.

Nanjing University Press, Nanjing, China (in Chinese).

Huang, Nian Lai (ed.), 1993. Encyclopedia on edible fungi.

Agricultural Press, Beijing, China (in Chinese). Figure 2: Prolific production of whitish Tremella fuci-
formis fruiting bodies by mixed culture cultivation on
Huang, Nian Lai (ed.), 1998. Colored Illustrations of macrofungi synthetic logs (substrate blocks in bags)
(mushrooms) of China. China agricultural Press, Beijing,
China (in Chinese).

Oei, Peter, 1996. Mushroom cultivation (with special emphasis on

appropriate techniques for developing countries). Tool
Publications, Leiden, the Netherlands.

Stamets, Paul, 2000a. Personal communication.

Stamets, Paul, 2000b. New edition of growing gourmet and

medicinal mushrooms July,
2000.

Figure 1: Stock cultures on

agar slants. Tremella fuciformis
(right) is whitish and Hypoxylon
archeri (left) is greenish. Mixed
culture cultivation of the two
species is used for the produc-
tion of Tremella fruiting bodies.

© 2000 by the Authors and The Mushroom Growers’

Newsletter - Reprinted from the May 2000 issue.
The Mushroom Growers’ Newsletter
P.O. Box 5065 Figure 3: Delicate fruiting bodies of Tremella fuciformis,
Klamath Falls, OR 97601 USA whitish and gelatinous
On the world-wide-web at: www.mushroomcompany.com