Note: Descriptions are shown in the official language in which they were submitted.
~047~
- 1 - 28113-3
BACKGROUND OF THE INVENTION
Sapstain of unseasoned lumber is a cosmetic defect that
is considered objectionable by many buyers. These discolorations,
caused by a variety of microfungi, are a serious problem on lumber
stored in lumber yards after sawing, but prior to planing and
chemical treatment, and also on untreated lumber that is exported
abroad. For the Canadian forest products industry, hem-fir
products, spruce-pine-fir or white pine products are particularly
prone to sapstain.
Several phenomena combine to create the discolorations.
Sapstaining fungi generally discolour the wood brown, grey or
black. The stain is caused by the pigmented fungal hyphae that
accumulate in the cells of the sapwood, particularly in the rays.
There is also evidence that dense accumulations of unpigmented
hyphae in the wood tissue can cause similar discolorations. Some
microfungi discolour wood by the production of coloured spores or
sporulating structures. In addition, some species discolour wood
red, purple, green or yellow by producing extracellular pigments
that diffuse into the wood tissues.
Sapstaining fungi are primary colonizers of wood that
20 subsist mainly on soluble nutrients. Although they cause little
structural damage, they are perceived as forerunners of decay
fungi by many consumers, and thus the objection to sapstain dis-
coloration may have a more practical basis than just aesthetics.
Many different chemicals have been used to control
sapstain. In Canada, the most widely used chemical formulations
incorporate the chemicals, 2( thiocyanomethyl)benzothiazole,
2 ~ ~ 7 ~ ~ ~
- 2 - 28113-3
copper-8-quinolinolate, borax or didecyldimethyl ammonium
chloride. The Canadian lumber industry has stated its intention
of eLiminating the use of toxic chemicals for sapstain control.
Biological control is a relatively new concept in forest
products. In biological control, a "harmless organism", in this
case one that does not decay or discolour wood, is deliberately
added to a product in order to prevent, retard or stop the growth
of undesirable organisms. The most widely used biological control
product is the bacterium Bacillus thuringiensis, better known as
BT, which is used to control spruce budworm in Ontario and parts
of Quebec. The bacterium produces a toxic crystal that is eaten
by the budworm as it eats the leaves, eventually killing the pest.
Approximately half a dozen biological control systems have been
marketed for agricultural use, for example DeVineTM, a fungal
control of parasitic vines in citrus orchards. Below, some exam-
ples of biological control related to wood products pathology are
reviewed.
The best known example of biological control in forest
products relates to the control of decay in wooden transmission
poles by the injection into the wood tissue of a dart containing
spores or mycelium of so-called "immunising commensals", as
described by J. Ricard in Canadian patents nos. 963387 and
1106201. Ricard claims a wide range of applications for his
invention, but all claims of said invention relate to the
possibility of inoculating biological control agents into wood
tissue, and none of the claims relate to the prevention of sap-
stain.
~'
20~7~
- 3 - 28113-3
Several research teams around the world have published
results of screening programs for biological control organisms for
the prevention of wood decay. The concept of Ricard, using mix-
tures of Trichoderma spp. and Scytalidium sp. to control decay in
transmission poles, has been investigated by other research teams
in the United Kingdom, the ~nited States and the Federal Republic
of Germany. Antifungal metabolites of Scytalidium sp. have been
isolated and chemically characterised, along with antifungal
metabolites from Hyalodendron sp. and Cryptosporiopsis sp., and
these metabolites have been applied to wood in an attempt to
prevent decay.l
Another application of biological control organisms is
to inhibit decay in round wood in storage. Shields2 reported that
decay by Bjerkandera adusta, Coriolus hirsutus and C. versicolor
was inhibited in wood blocks precolonized with Trichoderma
haræianum or an unidentified strain (now known to be Scytalidium
lignicola). The strain of T. harzianum was later used in a field
test on birch bolts3 where a conidial suspension was sprayed onto
freshly cut ends of birch bolts. After a two week precolonization
period, Blerkandera adusta was inoculated onto the bolts. After
six months, very little B. adusta was re-isolated from the bolts.
Stilwell4 isolated a strain of Cryptosporiopsis sp. from
yellow birch that inhibited the growth of 31 decay fungi in agar
interactions. Decay of blocks by Fomes fomentarius was inhibited
in precolonization experiments. In a field test, decay was
reduced in peeled birch logs inoculated with a water suspension of
Cryptosporiopsis sp., but no significant difference was noted in
20~74~5
- 4 - 28113-3
unpeeled logs. Culture filtrates of Cryptosporiopsis also inhibi-
ted growth of F. fomentarius. The antibiotic metabolite was puri-
fied, characterized and given the name cryptosporiopsin.5
Decay of wood chips during storage was also considered
as a possible target for antagonistic microorganisms. Bergman and
~ilsson6 tested several mould fungi isolated from wood chips for
their ability to inhibit chip decay in laboratory experiments, and
found that most decay fungi were inhibited. Gliocladium viride, a
mycoparasite frequently isolated from chips, was tested on spruce
chips in the field, and inhibited decay at temperatures less than
30C, but failed at higher temperatures.
Conifer chips inoculated with an antibiotic-producing
ryptosporiopsis sp. and stored outdoors for 12-15 months yielded
an improved quality of pulp although decay was not completely
inhibited.7 The results of trials using the antibiotic as a
chemical preservative, and of a proposed field test, have not been
published.
Bacteria were also tested as biological control agents
in chip piles. Some bacteria isolated from hardwood chips were
inhibitory to selected decay fungi in agar interactions, but the
antagonism was only effective on wood when the bacteria were in-
oculated onto the wood several weeks before the decay fungi.8
The results of a planned field trial were not published.
The possibility of controlling sapstain by using antago-
nistic organisms has also received some attention. The early work
of Stilwell and his colleagues9 demonstrated the antagonism of
some microorganisms towards some sapstain fungi. Stranks10 found
2~7~5
- 5 - 28113-3
that 0.25% and 0.50% solutions of the antibiotic hyalodendrin,
applied to white pine blocks by dipping, were effective at
preventing sapstain by Graphium sp., while cryptosporiopsin was
ineffective. Seifert et al.ll screened a variety of microfungi
for their abilities to prevent sapstain precolonization experi-
ments, and identified Nectria cinnabarina, Gliocladium roseum,
Trichoderma spp. and Tympanis sp. as promising candidates.
Russian workersl2 have demonstrated in vitro inhibition
of sapstain fungi by unidentified bacteria, but have not demons-
trated efficiency on wood. Bernier and colleaguesl3 showed that
an isolate of Bacillus subtilus prevented sapstain when wooden
-
blocks dipped into a cell suspension were placed on agar plates
inoculated with sapstaining fungi, but subsequent work at Forintek
Canada Corp. in Ottawa, Canada with the same culture showed that
it did not inhibit sapstain when the wood was not placed on agar.
The bacterium colonized wood very poorly and this prevented
effective biological control. Benkol4 has recently screened many
bacteria for antagonism towards sapstain fungi in agar inter-
actions, and has selected some strains of Pseudomonas for further
study.
Some innovative approaches towards biocontrol of saps-
tain have also been tried. Johnsonl5 studied polyoxin, an anti-
biotic that inhibits the synthesis of chitin, a major component of
fungal cell walls. The eight sapstain and mould species tested
were sensitive to this compound but at concentrations too high to
be economical on a commercial scale. Benkol6 demonstrated that
crude culture extracts of some antibiotic producing mycorrhizal
- 6 - 28113-3
fungi prevented growth of several sapstaining fungi on blocks of
pine.
FIELD OF THE INVENTION
The present invention consists of a method of protecting
unseasoned softwood lumber against unwanted sapstain by inocula-
tion of said lumber with the unique biological control microorga-
nisms from the genus Gliocladium, (fungi: Hyphomycetes), that
either prevents the growth of undesirable sapstaining organisms,
or prevents the formation of discolouration by these organisms.
The biological control organism is a fungus that does
not itself decay or discolour the wood to any objectionable
extent, and comprises one or more of the following strains:
Giocladium aureum (Forintek Culture Collection, FTK 784A),
Gliocladium roseum (FTK 321A, 321M), Gliocladium solani
(FTK 810A), Gliocladium viride (FTK 623E), Gliocladium virens
(FTK 258C, FTK 258D).
SUMMARY OF THE INVENTION
The present invention relates to a method for the
protection of wood or wood products against unwanted discoloration
caused by sapstain fungi. It is a feature of the present inven-
tion to provide a method of controlling sapstain in wood and wood
products comprising treating the wood or wood product with an
inoculum comprising one or more fungi of the genus Gliocladium.
20~7~
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The inoculum is of sufficient concentration and vigour to allow
rapid colonization of the wood tissue by the inoculated biological
control fungus. The actively growing and metabolizing biological
control fungus does not itself damage or discolour the wood, but,
likely through antibiotic facilities or mycoparasitism, protects
against discoloration of the wood by undesirable organisms already
present in the wood tissue, or that may be introduced to the wood
tissues during handling of the lumber. Because the biological
control fungus must subsist only on nonstructural wood
carbohydrates, the duration of control is of necessity finite, and
can be expected to become less effective as easily assimilable
nutrients are depleted, perhaps up to one year after inoculation
of the wood with the biological control agent. The wood or wood
product is preferably unseasoned softwood lumber such as conifer
wood.
The invention also relates to a wood or wood product
treated with a Gliocladium sp. Accordingly, it is another feature
of the present invention to provide a wood or wood product treated
with a fungus of the genus Gliocladium that is essentially free of
sapstain as the result of the activity of said fungus.
The invention is intended for use primarily in situa-
tions where sapstain is prevalent, but for which chemical protec-
tion is impractical or impossible. Suggested applications include
the protection of freshly sawn timber during seasoning, prior to
planing and subsequent chemical treatment, protection of export
lumber where chemical treatment, or specific chemical treatments,
are forbidden by the importing countries, or treatment of wood
~0~'74~
- 8 - 28113-3
chips during storage.
The genus Gliocladium is a biologically diverse group of
moulcls (Hyphomycetes) that includes species that are parasites of
other fungi (mycoparasites), parasites of slime moulds, parasites
of plant roots, and species that grow in soil and on wood.
Gliocladium roseum is commonly isolated from soil in many parts of
the world and is one of the most aggressive mycoparasites known.
All the tested isolates are effective biological control agents.
The isolates that we have employed are identified below. All of
these were collected independently from each other as follows:
0 FTK 784A: Gliocladium aureum Rader, isolated by W.E. Rader from
stored root of Daucus carota (carrot), received from H.
Bruckner, (Germany) (Centraalbureau voor schimmel-
cultures 226.48, American Type Cult~re
Collection 10406).
FTK 623E: Gliocladium viride Matr., isolated by M. Hawara from
hardwood chip, Thurso, Quebec, May 30, 1988, identified
by K.A. Seifert.
FTK 321A: Gliocladium roseum Bainier, isolated by C.K.J. Wang
from soil debris, Natural Bridge, N.Y. July 11, 1980.
0 FTK 321M: Gliocladium roseum Bainier, isolated from yellow poplar
stump, West Virginia, 1953/HL Barnett 914 (University
of Alberta Microfungus and Herbarium 419).
FTK 810A: Gliocladium solani (Harting) Petch, isolated by T.
Benedek. identified by W. Gams, (Centraalbureau voor
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- 9 - 28113-3
schimmel cult~res 187.29).
FTK 258C: Gliocladium virens. Miller et al., from W.H. Weston T-l
(American Type Culture Collection 9645).
FTK 258D: Gliocladium virens. Miller et al. from Dr.S. Gyorgy,
Budapest, Hungary, identified by J. Bisset (All-Union
Collection of Non-Pathogenic Organisms, Institute of
Microbiology, USSR Academy of Sciences - 1117).
Cultures of the isolates referred to above are available
upon request made to Forintek Culture Collection of Forintek
Canada Corp., 800 Montreal Road, Ottawa, Ontario, Canada, KlG
3Z5. Please note that any culture described in the specification
that is available from the Forintek Culture Collection possesses a
number that is preceded by the letters, FTK.
DETAILED DESCRIPTION OF THE INVENTION
The examples below describe the effectiveness of Glio-
cladium spp. in preventing or inhibiting sapstain or sapstain
fungi, and demonstrate that Gliocladium spp. does not itself
damage wood.
EXAMPLE 1
An inoculum of the biological control agent is prepared
by removing plugs of agar from stock cultures and growing them on
agar in petri dishes. The growth medium employed is typically
DifcoTM malt extract with 2~ agar (2% MA) for the Gliocladium
spp. and for the sapstaining fungi.
Stock cultures are maintained at 4C on agar media
20~7~
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containing 2% DifcoTM malt extract as a nutrient source.
All other incubations described in this example are at 27C, 75%
relative humidity, in the dark.
The fungi used as biological control agents in this
example are selected from the following strains maintained in the
Forintek culture collection of wood-inhabiting fungi: Gliocladium
roseum 321M, Gliocladium aureum 784A, Gliocladium solani 810A
Sapstaining fungi employed are: Ophiostoma piceae FTK 387I, O.
piliferum (Fr.) H. & P. Sydow FTK 55F, FTK 55H and Ophiostoma sp.
FTK C28.
After 1-3 weeks, a plug from the colony of the biologi-
cal control agent is placed on one side of 2% MA in a 9 cm petri
dish, and a plug from the colony of the sapstaining fungus is
placed on the opposite side of the petri dish, such that the two
fungi will grow together near the centre of the plate. Each
possible combination of biological control agent and sapstaining
fungus is set up. The plates are examined periodically and the
interactions between the organisms are observed.
In all cases, the biological control agents inhibit the
growth of the sapstaining fungus when the two colonies make
contact. None of the Gliocladium isolates inhibit the sapstaining
fungi before contact, suggesting that diffusible antifungal
metabolites are not produced in this experimental design.
EXAMPLE 2
Inocula of the biological control fungi are prepared by
transferring plugs of stock cultures onto DifcoTM potato
dextrose agar in 6 cm petri dishes as in Example 1. Stock culture
2 OD~ l~fl ~ ~a
- ll - 28113-3
maintenance and incubation conditions are as in Example 1.
The biological control agents are selected from the
following strains: Gliocladium roseum 321A, 321M, Gliocladium
viride 623E, Gliocladium aureum 784A, and Gliocladium solani 810A,
and Gliocladium virens 258C, 258D.
The wood blocks used in this example are Jack Pine sap-
wood 3 cm long and l cm x 0.5 cm in cross section. These are
sterilized by gamma irradiation and placed in glass petri dishes,
eight blocks per dish, upon w-shaped glass bars fashioned from
3 mm glass tubing, that rest upon 2 sheets of filter paper in
which 5 mL sterile distilled water has been absorbed.
A spore suspension from 1-3 week old agar cultures is
prepared. The colonies from G. roseum 321A and 321M, G aureum
784A, and G. virens 258D are transferred into a sterile
WaringTM blender and homogenized for 30 seconds in 75 mL
sterile distilled water. Spore suspensions of G. viride 623E, G.
solani 810A and G. virens 258C are prepared by flooding the agar
plates with 6 mL of sterile distilled water and liberating the
spores with an L shaped glass rod.
The spore suspension of each Gliocladium strain is
squirted onto the surface of 64 blocks, (8 per petri dish) using a
sterile syringe such that the entire length of the block, though
not necessarily the entire width, receives some liquid.
The blocks are then incubated for l week. Staining
fungus inocula are grown in 6 cm petri dishes on appropriate agar
media for 1-2 weeks. Spore suspensions are prepared in the same
way as described for the biological control strains above. The
20~7~
- 12 - 28113-3
sapstaining fungi employed are: Ophiostoma piceae FTK 387I,
O. piliferum FTK 55F, FTK 55H and Ophiostoma sp. FTK C28, plus a
"80Up" mixture of Cephaloascus fragrans FTK 307I, Ophiostoma
piliEerum FTK 55H, Black Yeast FTK 86-010-1-1-1, Aureobasidium
pullulans FTK 132Q, Leptodontidium elatius FTK 268A, Cladosporium
cladosporioides FTK 273D, Ophiostoma populinum FTK 671A,
Ophiostoma perfectum FTK 703A, Phialophora botulispora FTK 707A,
Leptographium sp. FTK 2A2, Phoma sp. FTK 86-8-3-2-1, Alternaria
alternata FTK 2G and FTK 2H.
The sapstaining fungi spore suspensions are then inocu-
lated onto the surface of the wood blocks in a manner identical to
that used for the biological control fungi. Each sapstaining
fungi spore suspension is inoculated onto 2 groups of 8 blocks for
each Gliocladium strain.
The wood blocks are incubated a further four weeks. The
surface and interior of the wood blocks thus treated are free from
discolorations caused by the sapstaining fungi, while control
blocks inoculated at the same time with only sapstaining fungi
become darkly discoloured after only 1-2 weeks. The results of
this experiment are illustrated in Table 1.
EXAMPLE 3
In this example, the ability of the isolate Gliocladium
roseum 321A to cause weight loss in Jack Pine blocks is tested.
The standard ASTM soil block test (D1413-76) is used. In this
method, 200 g of a 3:1 soil-sand mixture are placed in a 500 mL
glass jar with 60 mL distilled water. A feeder strip made of red
pine sapwood, 41 x 29 x 3.0 mm, is placed on the surface of the
2047~
- 13 - 28113-3
soil. The jars are sterilized for one hour, cooled, then re-
sterilized for a second hour. The lids are then replaced with
sterile culture lids with a microbiological filter with a 0.2 ~ m
pore size fitted over a 5 mm hole in each lid. The soil is inocu-
lated with an agar plug from a growing colony of Gliocladium
roseum 321A and incubated for 3 weeks. Then, two sterilized 19 mm
cubes of Jack Pine sapwood of known dry weight are added to each
jar. After 12 weeks incubation, the blocks are dried and re-
weighed.
The weight loss caused by Gliocladium roseum 321A was
2~, and was not significantly different than the weight loss in
the blank control. A typical decay fungus, Poria carbonica
FTK 120AM, incubated under the same conditions, caused a weight
loss of 33%.
EXAMPLE 4
In this example, the ability of Gliocladium roseum 321A
to cause strength loss in wood is determined. Jack Pine wood
beams are incubated in a modified soil block test and the impact
bending strength is measured using the Toughness Testing (ISOD)
machine, as detailed below.
The ISOD impact bending machine measures the force
required to break a span of wood. A weight attached to a pendulum
is released using a foot pedal, which pulls a chain attached to a
vertical metal bar. The bar then is pulled into the sample (=
impact), and the wood is broken. The force required to break the
sample is determined by converting the value recorded by the
pendulum of the machine (in degrees and minutes) to inch-pounds.
2047~
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The [SOD machine was modified for the smaller wood beams by
reducing the weight on the pendulum and modifying the sample
holder.
The soil block test was modified from ASTM stand and
D-1413 to allow for the different block sizes. Rather than using
glass jars, 1 L NalgeneTM polypropylene jars are used. Each
jar contains 400 g of a 3:1 soil:sand mixture and 120 mL of
distilled water. Three red pine feeder strips,
4.5 x 2.5 x 0.5 cm, are placed side by side on the surface of the
soil. The jars are autoclaved for one hour, cooled overnight,
then autoclaved again the next day for one hour. The jars are
cooled in a biological safety hood, and the lids replaced with
culture lids. The culture lids are modified lids with a central
hole, 5/16 of an inch in diameter, covered on the inner surface
with a GelmanTM filter to allow air exchange.
The wood beams, 9.0 x 0.75 x 0.75 cm, are prepared from
green Jack Pine (Pinus banksiana) sapwood. The beams are sorted
into sets with more or less the same number of growth rings. Only
beams with the grain more or less parallel to the long axis are
selected. The beams are sterilized by gamma radiation and frozen
until use.
The inoculum for Gliocladium roseum 321A is a spore
suspension in water made from a 1-2 week old 2% MA culture. The
spore suspension is inoculated onto the wooden beams, and the
beams are preincubated for 1 week in a 1 L Nalgene polypropylene
jar. At the bottom of the jar is filter paper moistened with
distilled water, then alternating layers of glass rods and wood
~47~
- 15 - 28113-3
beams. All beams used in the experiment are from a matched set.
After the preincubation period, five test beams are aseptically
added to each soil jar. For controls, uninoculated test beams are
aseptically added to jars. Five jars are set up for each treat-
ment, for a total of 25 beams per treatment. All incubations are
at 27C.
After four weeks incubation, the beams are removed from
the jars and placed on screen racks. The samples are then placed
at a constant temperature and humidity and allowed to equilibrate
for 7 days. The force required to break each beam is then measur-
ed with the ISOD toughness testing machine. The values areconverted using the equation:
Toughness (inch-pounds) = pendulum weight x (cosA2-
cosAl) where Al is the initial angle, and A2 is the angle of the
pendulum at failure.
The readings given by the machine when no specimen was
present are converted using the same equation, and this value is
subtracted from the converted test values to give a corrected
value.
Wood specimens colonized with G. roseum require 12.76 +
0.32 inch-pounds to break. The control blocks, with no added
fungus, require 11.93 + 0.44 inch-pounds to break while the decay
control blocks required 8.87 + 0.45 inch-pounds. Statistical
analysis revealed that test blocks incubated with G. roseum 321A
do not undergo significantly more strength loss than uninoculated
Jack Pine test blocks under the conditions employed.
20~7~a
- 16 - 28113-3
EXAMPLE 5
In this example, the ability of the candidate strain
Gliocladium viride 623E to prevent decay by known decay fungi
Gloeophyllum trabeum FTK 47D and Merulius tremellosus FTK 52H is
determined. Retained strength, a measure of a candidate's decay-
prevention ability, is the ratio of impact bending strength of
test beams incubated with both the candidate biocontrol fungus and
the decay fungi to the impact bending strength of test beams in-
cubated with the candidate biocontrol fungi alone. This figure is
expressed as a percentage. The experimental set-up is the same as
the set-up in Example 4 except that the blocks containing the
biological control fungi were added to a NalgeneTM
jar/incubator which had been inoculated one week earlier with a
decay inoculum.
The decay inoculum utilized was prepared by blending 150
mL of sterile water and growing agar cultures of two 9 cm petri
plates from each of the aforementioned decay organisms. Five
drops of this inoculum were placed on Jack Pine feeder strips in
each ~algene jar/incubator. After an incubation period of one
week, the beams colonized with the biological control fungus were
added and allowed to incubate a further four weeks. The impact
bending strength for each test beam was determined (as in Example
4) and the retained strengths calculated.
The test beams inoculated with Gliocladium viride 623E
had a retained strength of 94%, compared to 87% for the control
test beams which contained no biological control fungi. G. viride
623E, therefore, prevented a significant amount of decay from
occuring in the test blocks.
20~7~
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EXAMPLE 6
This example examines the ability of the Gliocladium
8pp. to protect larger pieces of lumber from sapstaining organisms
in a small scale trial under conditions closely related to those
in field trials. The Gliocladium strain was grown on two 14 cm
agar plates (2% MA see Example 1) and allowed to sporulate. A
spore suspension of each strain was prepared by flooding the sur-
face of the plates with 50 mL of sterile distilled water, pooling
the suspensions and diluting them to 4 liters. The final spore
concentration was measured using a haemocytometer. This
concentration ranged from 2 x 104 to 1 x 106 spores/mL. Freshly
sawn 1" x 6" x 15" white pine lumber, 28 pieces per strain, was
dipped in these spore suspensions and placed in plastic bins, 14
pieces per bin. Control bins containing lumber with no biocontrol
organisms were also set up. Humidity was maintained in these bins
by the addition of lL of sterile distilled water. The bins were
covered with tight fitting lids equipped with ventilation holes
covered with 0.22 ~L GelmanTM filters. The bins were incuba-
ted in a ventilated temperature monitored shed at ambient tempera-
ture. After 6 and 14 weeks, each piece of lumber was rated for
the evidence of surface mold, decay and sapstain fungi. A visual
rating system for the prsence of the specified fungi is used. A
piece of lumber is rated as acceptable if it contains less than
10% surface area covered with stain. The results of these trials
are summarized in Table 2. A statistical analysis of this data
(based on the analysis of the Chi-squared test of homogeneity of
proportions) reveals that G. roseum 321A and 321M, G. solani 810A,
~74~
- 18 - 28113-3
and G aureum 784A afford the lumber a significant protection from
sapstain, mould and decay while wood treated with G. viride 623E
was not significantly different than the control.
2 0 ~ S
- 19 - 28113-3
Table 1
Results summarizing the ability of the candidate biological
control fungi to prevent sapstain on Jack Pine.
____._____________________________________________________________
Number of
Biocontrol Fungus Sapstainer blocks
(FTK No.) (FTK No.) stained
Gliocladium roseum (321A) Ophiostoma picea (387I) 0/8 , 0/8
Ophiostoma sp. (C28) 0/8 , 0/8
Aureobasidium pullulans (132Q)0/8 , 0/8
Alternaria alternata (2G) 0/8 , 0/8
__________________________________________________________________
Gliocladium aureum (784A) soup (see Example 2) 0/8 , 0/8
Ophiostoma piliferum (55H) 0/8 , 0/8
Phialophora botulispora (707A)0/8 , 0/8
Black Yeast (86-010-1-1-1) 0/8 , 0/8
__________________________________________________________ _______
Gliocladium roseum (321M) soup (see Example 2) 0/8 , 1/8
Ophiostoma piliferum (55H) 0/8 , 0/8
Phialophora botulispora (707A)0/8 , 0/8
Black Yeast (86-010-1-1-1) 0/8 , 0/8
__________________________________________________________________
Gliocladium solani (810A) soup (see Example 2) 0/8 , 1/8
Ophiostoma piliferum (55H) 0/8 , 0/8
Alternaria alternata (2H) 0/8 , 0/8
Black Yeast (86-010-1-1-1) 0/8 , 0/8
___________________________ ______________________________________
Gliocladium viride (623)E soup (see Example 2) O/4*
__________________________________________________________________
Gliocladium virens (258C) soup (see Example 2) 0/8 , 0/8
(258D) soup 0/8 , 0/8
__________________________________________________________________
*readings taken from wood chips
2~7~
- 20 - 28113-3
able 2. Summary of data from small scale field trials
detailing the number of acceptable pieces of lumber
after 6 and 14 week incubation periods.
Treatment Acceptable Pieces (~)
6 weeks 14 weeks
Control (no fungi) 71 32
Gliocladium aureum 784A 100 82
Gliocladium roseum 321A 79 71
Gliocladium roseum 321M 100 82
Gliocladium solani 810A 96 86
Gliocladium viride 623E 54 32
-.
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REFERENCES
1. Unligil, H.H. 1978. Decay resistance of wood treated with
fungal antibiotics: cryptosporiopsin, hyalodendrin, and
scytalidin. Wood. Sci. 11: 30-32.
2. Shields, J.K. 1966. Wood Pathology. Rept. For. Prod. Res.
Branch, Dept. For. Can., April 1964-March 1965: 48-52.
3. Shields, J.K. 1968. Role of Trichoderma viride in reducing
storage decay of birch logs. Bimonthly Res. Notes Dept. For.
Can. 24: 9-10; Hulme, M.A. and J.K. Shields. 1972. Effect
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