In vitro boletes
rwinder at PFC.Forestry.CA
Wed May 24 12:26:49 EST 1995
A number of people have requested further information from me regarding
my in vitro culture of B. edulis primordia. So, I hope I am not flogging
this to death, but I decided that the best thing to do would be to post
a synopsis here. This is meant to be a fun, quick report and not a
comprehensive overview- there's probably lots of information out there that
I've missed. So, if you have anything to add, please be my guest!
In vitro culture of Boletus edulis primorida
Richard S. Winder
Pacific Forestry Centre
506 W. Burnside Rd.
Victoria, B.C. V8Z 1M5 CANADA
Mention of trade names does not imply endorsement of a particular
product, nor does it imply criticism of any similar products not
named. This posting may be distributed freely by anyone, as long as
this disclaimer remains in place. -RSW
Although they are ectomycorrhizal, it is possible to grow members of
the boletaceae in axenic culture and produce their associated
fruiting bodies or primordia (Table 1). The usual method involves
the use of Hagem-Modess medium (Modess, 1941) or some modification of
it (McLaughlin, 1970), in containers ranging in size from test tubes
to jars. This report outlines an in vitro method for generating
primodia of Boletus edulis using potato dextrose agar (PDA), a more
standard mycological medium.
Table 1. Some reports of in vitro production of fruiting bodies for
Fungi Citation Result
Boletus spp. Pantidou, 1961a Primordia
McLaughlin, 1970 Mushroom
Boletus edulis Karpi'nski, 1961 Mushroom
Boletus rubinellus McLaughlin, 1970 Mushroom
Yoon & McLaughlin, 1979 Mushroom
Leccinum spp. Pantidou, 1961a Primordia
McLaughlin, 1970 Mushroom
Phlebopus lignicola Pantidou, 1962 Mushroom
Phlebopus sulphureus Pantidou, 1961b Mushroom
Pulveroboletus spp. Pantidou, 1961a Primordia
Tylopilus spp. Pantidou, 1961a Primorida
McLaughlin, 1970 Mushroom
Xerocomus spp. Pantidou, 1961a Primordia
Xerocomus badius Pantidou, 1964 Mushroom
Xerocumus illudens Pantidou, 1964 Mushroom
-Materials and methods
Dehydrated potato dextrose agar (No. 6013-01-4 Bacto agar, Difco
Laboratories, Detroit, Michigan, U.S.A.) was combined (39 g/L) with
tap water (pH 6.9), autoclaved (15 p.s.i., 121 C) for 20 minutes, and
poured into 9 cm-diam. Petri plates to solidify. A sporophore of
Boletus edulis (white pore stage) growing in association with grand
fir (Abies grandis) was collected in Sooke, British Columba in
September, 1995. After peeling back the cuticle, tissue from the
sporophore was excised and aseptically transferred to 10 PDA plates.
The plates were sealed with wax film (Parafilm, American National
Can, Greenwich, CT, USA) and incubated cover-side-up in a controlled
environment chamber at 20 degrees C for eight months. The colony
diameters were recorded as well as the number and diameter of
primorida forming in each plate.
After eight months, the medium lost about 50% water content as
measured by the thickness of the agar. Five plates were discarded
due to contamination. The colonies did not colonize the entire
plate, colony diameter (largest dimension) ranged from 37-70 mm (mean
49 mm). Colony edges were highly irregular and diffuse, hyphae were
white. Sporophore primorida of various sizes formed in the remaining
plates. The primordia where chalky white in color, with a few drops
of brownish liquid present on some surfaces. Hyphae close to the
primordia tended to be rhizomorphic. The primordia were composed of
parenchyma-like cells which appeared to be somewhat organized into
layers. Most of the larger primordia developed abortive tubes on
their upper surfaces. In some cases, smaller globular primordia
appeared on top of ther larger primordia. Primordia were 7-32 (mean
19) mm in diameter, with an average of 7.1 primordia (all types) per
A variety of simple undefined media could probably be used to
produce primordia of the boletaceae in vitro. The method could
probably be improved by using a larger container for the agar. While
it is premature to speculate that this method could be used to
generate mature sporocarps in any quantity, it could provide an
easily accessbile starting point for testing various ammendments and
other other factors which might influence sporocarp production.
Karpi'nski, J. 1961. Investigation results (stage I) pertinent to the
cultivation of fruiting bodies of Boletus edulis Bull., in
artificial culture medium under laboratory conditions [In Polish,
English summary]; Sylwan 105:55-59.
McLaughlin, D. 1970. Environmental control of fruitbody developement
in Boletus rubninellus in axenic culture. Mycologia 62:1970.
Modess, O. 1941. Zur Kenntnis der Mykorrhizabildner von Kiefer und
Fichte. Symb. Bot. Ups. 5:1-147.
Pantidou, M. 1961(a). Cultural studies of boletaceae: Gyrodon
meruloides and four species of Boletinus. Can. J. B.
Pantidou, M. 1961(b). Carpophores of Phlebopus sulphureus in culture.
Can. J. Bot. 39:1163-1166.
Pantidou, M. 1962. Cultural studies of boletaceae: carpophores of
Phlebopus lignicola in culture. Can. J. Bot. 40:1313-1319.
Pantidou, M. 1964. Cultural studies of boletaceae: carpophores of
Xerocomus badius and Xerocomus illudens in culture. Can. J. Bot.
Yoon, K. and D. McLaughlin. 1979. Formation of the hilar appendix in
basidiospores of Boletus rubinellus. Am. J. Bot. 66:870-873.
RICHARD WINDER Title: Research Scientist
Canadian Forest Service Phone: (604) 363-0773
Victoria, B.C. Internet: RWINDER at A1.PFC.Forestry.CA
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