BIBLIOGRAPHY ALFREDO, NOEL VELASCO. ...
BIBLIOGRAPHY
ALFREDO, NOEL VELASCO. APRIL 2011. Characterization of Two
Mycorrhizal Mushroom Associated with Benguet Pine (Pinus kesiya). Benguet State
University, La Trinidad, Benguet.
Adviser: Bernard S. Tad-awan, Ph.D.
ABSTRACT
Boletus leonis and Tuber sp. were collected within pine-based forest of Tublay. At
an altitude of 1,300 meters above sea level, both Tuber sp. and Boletus leonis were
abundantly present. Tuber sp. has the higher number of basidiocarp at the whole surveyed
altitude compared to Boletus leonis. Boletus leonis was not found at altitude 1,200 masl
and 1,100 masl. However, the Tuber sp. was found at the altitude of 1,200 masl and 1,100
masl which it suggests a trend that the higher altitude, the higher the number of basidiocarp
and the lower altitude, the lower the number of basidiocarp.
Soil sample of truffle Tuber sp. have pH 5.45, 41.56% moisture content, 4.85%
organic matter, 1.88% Nitrogen, 10.2 ppm Phosphorus and11.5 ppm Potassium. The edible
Tuber sp. occurs in cluster, scattered or gregarious on the ground, pathways or roads and it
is more abundant during rainy season.
Boletus leonis mushroom has been found to grow on soil with pH of 5.4, 41.53%
moisture content, 6.4% organic matter, 1.9% Nitrogen, 10.36 ppm Phosphorus and 12 ppm
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Potassium. The Boletus leonis grows on areas covered with pine needles, mostly in clusters
and is abundant during rainy season.
The cap of Boletus leonis is yellow-orange to orange-brown, 8.0 to 12.0 cm wide
and slightly convex when young and flatly expanded when matured. The hymenophore is
tubular with tubes that are yellowish-brown and attachment of tube in the stipe is adnexed.
The stipe is 4.0 to 8.o cm long and basal part is thickening and tapering to a beet-like
extension. Spore print is yellowish to orange.
The cystidium is cylindrical, basidium is unicellular holobasidium with two
sterigmata, basidiospore is subglobose and colorless. The spores measure 4-7mm and 8.5
x 4µm.
The truffle Tuber sp. is 1.2-12 cm in diameter but could reach up to 15 cm. The
wall is dirty yellow to brown. Tuber sp. is semicircular, solid, hard and has a rough texture.
The spore is circular with a dedicate ornamentation. The spore is darkyellow with
a diameter of 10 mm and 10 x 5µm.
Stained roots of pine seedlings taken from their natural habitat in surveyed area in
Tublay developed ectomycorrhizal relationship with the mushroom. Likewise, the
inoculated pine tree seedlings developed ectomycorrhizal relationship as evidenced by
ocular inspection through a microscope. On the other hand, pine seedlings without
inoculation of suspension did not show such relationship.Pine seedlings taken from their
natural habitat had significantly higher mean plant height than those inoculated pine
seedlings with spore concentration of 6.7 x 104/ml and pine seedlings without inoculation.
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
RESULTS AND DISCUSSIONS
Presence of Mycorhizal Mushroom
At an altitude of 1,300 meters above sea level, both “Bu-o” and “Dappanan”
were abundantly present (Table 1). “Bu-o” has the higher number of basidiocarp at the
whole surveyed altitude compared to “Dappanan”which was not found at altitude of
1,200 and 1,100 m asl. However, the “Bu-o” was found at the altitude of 1,100 m asl to
1,300 m asl. At a higher altitude, there was higher number of basidiocarps compared to
lower altitude.
Table1. Abundance of “Dappanan” and “Bu-o” in Tublay
ALTITUDE NUMBER OF BASIDIOCARP WITH IN A 100 m² AREA
(masl) (“Dappanan”) (“Bu-o”)
1, 300
7
139
1, 200
0
39
1, 100 0 36
TOTAL
7
214
Characterization of “Dappanan” and
“Bu-o” Mushroom
Habitat. Both “Dappanan” and ”Bu-o” were found under pine trees associated
with grasses (Figure 1).
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
The edible “Bu-o” occurs in cluster, scattered or gregarious on the ground,
pathways or roads and it is more abundantly during rainy season (Figure 2).
The “Dappanan” grows on soil covered with pine needles, mostly in
cluster and abundantly during rainy season (Figure 3).
Analysis of the soil samples of “Bu-o” revealed: pH 5.45, 41.56% moisture
content, 4.85% Organic matter, 1.88% nitrogen, 10.2 ppm phosphorus and11.5 ppm
potassium. Soil analysis of “Dappanan: pH 5.4, 41.53% moisture content, 6.4%
Organic matter, 1.9% nitrogen, 10.36 ppm phosphorus and 12 ppm potassium. This
mushroom grows from August to October.
Figure 1. Surveyed area for the presence of “Dappanan” and “Bu-o” mushroom in
Tublay
Figure 2. Truffle “Bu-o” was specifically collected at the pine forest
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Figure 3. “Dappanan” mushroom in its habitat
Physical Description of “Dappanan” mushroom. The cap of this mushroom is
yellow-orange to orange brown, 8.0 to 12.0 cm wide and slightly convex when young,
flatly expanded when matured. The hymenophore is tubular, tubes are yellowish-brown
and attachment of tube in the stipe is adnexed. The stipe is 4.0 to 8.o cm long and basal
part is thickening and tapering to a beet-like extension (Figure 4a). Spore print is yellowish
to orange.
Microscopic Description. The Cystidium is cylindrical (Figure 4b), basidium is
unicellular holobasidium with two sterigmata (Figure 4c), basidiospore is subglobose
and colorless (Figure 4d). The spores measure 4-7mm and 8.5 x 4 um (Figure 4-e).
Based on the above descriptions as well as its edibility as identified by local folks,
the mushroom is identified as Boletus leonis. Physical and microscopic descriptions agree
with the description of Surcek and Kubicka (1980) and Phillips (1994).
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Figure 4a. Fruiting body of Boletus leonis found at the surveyed area
Figure 4b. Cystidia of Boletus leonis at 40x
Figure 4c. Basidium of Boletus leonis at 40x
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Figure 4d. Spores of Boletus leonis at 40x
Figure 4e. Length of the spore of Boletus leonis with 7µm at 100x
Physical Description of “Bu-o” mushroom. Size of fruiting body is 1.2- 12 cm
in diameter but could reach up to 15 cm. The wall is dirty yellow to brown; it is
semicircular, solid, hard and ha a rough texture (Figure 5a).
Microscopic Description of “Bu-o” mushroom. The spore is circular with a
dedicate ornamentation (Figure 5b). The spore is dark yellow and with a length of 10
mm, 10 x 5 µm (Figure 5c). This mushroom is edible and it was identified as Tuber sp.
supported by information from Simpson (1989).
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Figure 5a. Truffle Tuber sp. found at the surveyed area
Figure 5b. Spores of Tuber sp. at 10x
Figure 5c. Diameter of the spore of Tuber sp. with 10 µm at 100x
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Test for Mycorrhizal Relationship
on Pine Tree Seedlings
Stained roots of pine seedlings at the surveyed area showed presence of mycelia
of mycorrhizal mushrooms (Figure 6). Conversely, pine seedling without inoculum did
not show any presence of mycelia (Table 2 and Figure 11). Likewise, inoculated pine
seedlings showed the presence of mycelia of mycorrhizal relationship (Figures 7-10).
There were significant differences between pine seedlings inoculated with 25
ml, 75 ml and 100 ml compared to the pine seedlings taken from their natural habitat.
Pine seedlings inoculated with 25 ml suspension with 167.5 x 104 ml spores showed the
presence of mycelia indicating mycorrhizal relationship. Increasing the amount of
inoculum resulted to a more indicative mycorrhizal relationship.
Table 2. Presence or Absence of Mycelia of Tuber sp.
TREATMENTS MEAN
Pine seedling taken from their natural habitat
1a
Pine seedling without inoculation of suspension 0b
Pine seedling with 25 ml inoculation suspension 0.6ab
pine seedling with 50 ml inoculation of suspension 0.1b
pine seedling with 75 ml inoculation of suspension 0.4ab
pine seedling with 100 ml inoculation of suspension 0.6ab
*Significant differences exists at 0.05 level using DMRT
CV = 23.85%
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
a
b
Figure 6. Stained roots of pine seedlings taken from their natural habitat at 40x(a) and
10x(b) magnification (arrow indicates the mycelia of Tuber sp.)
a
b
Figure 7. Stained roots of pine seedlings inoculated with (167.5 x 104) / ml spore
suspension after 136 days at 40x magnification. (arrow indicates the mycelia
of Tuber sp.)
a
b
Figure 8. Stained roots of pine seedlings inoculated with (335 x 104) / ml spore
suspension after 136 days at 10x and 40x magnification. (arrow indicates
the mycelia of Tuber sp.)
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
a
b
Figure 9. Stained roots of pine seedlings inoculated with (502.5 x 104) / ml spore
suspension after 136 days at 40x and 10x magnification. (arrow indecates
the mycelia of Tuber sp.)
a
b
Figure 10. Stained roots of pine seedlings inoculated with (67 x 106) /ml suspension
after 136 days at 10x magnification. (arrow indicates the mycelia of Tuber
sp.)
a
b
Figure 11. Stained roots of pine seedlings without inoculation
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Figure 12. Pine (Pinus kesiya) seedling replicates for each treatments
Pine seedlings taken from their natural habitat had significantly higher mean plant
height than the inoculated pine seedlings. Significantly taller pine seedlings were from
those taken from natural habitat. This is followed by pine seedlings inoculated with 25 ml
and 100 ml of suspension. The results suggest that pine seedlings taken from their natural
habitat had significantly higher mean plant height than those inoculated regardless of
amount of inoculum. These differences may be due to the maturity of the pine seedlings
(figure 13).
Table 3. Height of Pine Seedlings
TREATMENTS HEIGHT (cm)*ᵧ
pine seedling taken from their natural habitat 11.75a
pine seedling without inoculation of suspension 8.98b
pine seedling with 25 ml inoculation of suspension 9.77ab
pine seedling with 50 ml inoculation of suspension 8.84b
pine seedling with 75 ml inoculation of suspension 8.78b
pine seedling with 100 ml inoculation of suspension 9.52b
TOTAL 57.64
*No significant differences at 0.05 level, significant differences at 0.10 level,
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
P-value=0.085
ᵧMeans with different letters signify significant differences using DMRT
CV=25.63%
Plant Height at 7, 21, 35, 49, 63, 77, 91, 105 and 119 Days After
Inoculation
25
20
m)
T1
15
(c
T2
ht
ig
T3
T4
10
lant He
P
T5
T6
5
0
7
21
35
49
63
77
91
105
119
Days after Inoculation
Figure 13. Plant Height at 7, 21, 35, 49, 63, 77, 91, 105 and 119 Days after inoculation
(DAI)
Alexopoulus et al., (1996) reported that mycorhizal fungi benefit their host by
increasing the plants ability to capture water and nutrients such as nitrogen,
phosphorous, potassium and calcium from the soil, increasing tolerance to drought, high
temperature and extremes of soil acidity and providing protection from certain plant
pathogens.
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
SUMMARY, CONCLUSIONS AND RECOMMENDATIONS
Summary
The study was conducted to characterized “Bu-o” and “Dappanan” mushroom
based on: habitat (type of tree cover where found, altitude, temperature, moisture and soil
pH) type of mycorrhizal relationship, morphological and anatomical features, abundance
of each species with altitude, relative humidity and temperature and determined the
mycorrhizal relationship between each mushroom and pine seedlings.
Microscopic studies were done at the Department of Plant Pathology, Benguet State
University from October 2010 to October 2011. Soil analysis of mineral content was done
at Department Soil Science, Benguet State University and Regional Soils Laboratory, San
Fernando, La Union.
At an altitude of 1,300 meters above sea level, both “Bu-o” and “Dappanan” were
abundantly present. “Bu-o” has the higher number of basidiocarp at this altitude, compared
to “Dappanan”. However at altitudes 1,200 to 1,100 m asl, only the “Bu-o” was found but
at a lesser. This indicates that at higher altitudes, a higher number of basidiocarps were
observed which decreased as altitude decreases.
The edible “Bu-o” occurs in cluster, scattered or gregarious on the ground,
pathways or roads and it is more abundant during rainy season.
Soil samples of Truffle “Bu-o” revealed: pH 5.45, 41.56% moisture content, 4.85%
Organic matter, 1.88% nitrogen, 10.2 ppm phosphorus and11.5 ppm potassium.
The “Dappanan” grows on covered with pine needles, mostly in cluster and
abundantly during rainy season.
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Soil analysis of “Dappanan” mushroom: pH 5.4, 41.53% moisture content, 6.4%
Organic matter, 1.9% nitrogen, 10.36 ppm phosphorus and 12 ppm potassium.
The cap of “Dappanan” is yellow-orange to orange brown, 8.0 to 12.0 cm wide
and slightly convex when young, flatly expanded when matured. The hymenophore is
tubular with tubes that are yellowish-brown and attachment of tube in the stipe is adnexed.
The stipe is 4.0 to 8.o cm long and basal part is thickening and tapering to a beet-like
extension. Spore print is yellowish to orange.
The Cystidium is cylindrical, basidium is unicellular holobasidium with two
sterigmata, basidiospore is subglobose and colorless. The spores measure 4-7mm and 8.5
x 4 µm.
The truffle “Bu-o” is 1.2- 12 cm in diameter but could reach up to 15 cm. The wall
is dirty yellow to brown; it is semicircular, solid, hard and has a rough texture.
The spore is circular with a dedicate ornamentation. The spore is dark yellow with
a length of 10 mm and 10 x 5 µm.
Based on physical and microscopic features, these mushrooms are edible.
“Dappanan” is identified as Boletus leonis and truffle “Bu-o” is identified as Tuber sp.
Stained roots of pine seedlings at the surveyed area showed presence of mycelia of
mycorrhizal mushrooms. Conversely, pine seedling without inoculum did not show any
presence of mycelia. Likewise, inoculated pine seedlings showed the presence of mycelia
of mycorrhizal relationship. Results of the analysis of variance test showed significant
differences between pine seedlings inoculated with 25ml, 75 ml and 100 ml against the
pine seedling taken from their natural habitat at the 0.05 level using DMRT.
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Also, pine seedlings taken from their natural habitat had significantly higher mean
plant height than those inoculated pine seedlings and pine seedlings without inoculation of
suspension at the 0.05 level using DMRT. The results suggest that pine seedlings taken
from their natural habitat had significantly higher mean plant height than those inoculated
regardless of amount of inoculum.
Conclusion
“Dappanan” and “Bu-o” were found under pine-based areas associated with grasses
in Tublay. Altitude, relative humidity, temperature, moisture content and soil pH provides
a habitat which favors the growth and abundance of many truffles and mushrooms. Based
on physical and microscope features, “Dappanan” is identified as Boletus leonis and truffle
“Bu-o” is identified as Tuber sp. These mushrooms developed ectomychorrhizal
relationship with its habitat.
Recommendation
It is recommended that further studies be conducted to identify other areas where
truffle “Bu-o“ and “Dappanan” may be found. In addition, studies to further understand
the ectomycorrhizal relationship between the mushroom and pine tree roots should be done
so that production could be explored. There is also a need to further verify the effects of
inoculum on the growth and development of the pine tree seedlings.
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
LITERATURE CITED
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Mycology. 4th Ed. John Wiley and Sons, Inc. New York, USA. Pp 514,524.
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Trinidad Benguet. Pp 1-4.
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DAMES, J.F., G.L. HAWLEY and A.F.S. TAYLOR. 2008. Ectomycorrhizas in
association with Pinus patula in Sabie, South Africa. Retrieved August 2010 from
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Champaign Urbana, Illinois. Pp. 279.
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production of extramatrical hyphae by two types of willow ectomycorrhizas at
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8137.1990.tb00451.x/pdf
IOTTI, M., E. BARBIERI, V. STOCCHI and A. ZAMBONELLI. 2005. Morphological
and Molecular characterization of mycelia of ectomycorrhizal fungi in pure culture.
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kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Fungal
Diversity
19:
51-68.
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from
http://www.fungaldiversity.fdp/sfdp/19-3.pdf.
KAUL, T. N. 1997. Introduction to Mushroom Science (systematic). Science Publishers,
Inc. Enfield, New Hampshire 03748, USA. P. 49.
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scleroderma_michiganense.html.
LUMYONG, S., R. SANMEE, P. LUMYONG and B. DELL. 2010. In vitro cultivation
and fruit body formation of the black bolete, Phlebopus portentosus, a popular
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1995. Identification of Tuber spp. and corresponding ectomycorrhizae through
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PHILIPS, R.1994. Mushrooms and other Fungi of Great Britain and Europe.London.
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Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
ALFREDO, NOEL VELASCO. APRIL 2011. Characterization of Two
Mycorrhizal Mushroom Associated with Benguet Pine (Pinus kesiya). Benguet State
University, La Trinidad, Benguet.
Adviser: Bernard S. Tad-awan, Ph.D.
ABSTRACT
Boletus leonis and Tuber sp. were collected within pine-based forest of Tublay. At
an altitude of 1,300 meters above sea level, both Tuber sp. and Boletus leonis were
abundantly present. Tuber sp. has the higher number of basidiocarp at the whole surveyed
altitude compared to Boletus leonis. Boletus leonis was not found at altitude 1,200 masl
and 1,100 masl. However, the Tuber sp. was found at the altitude of 1,200 masl and 1,100
masl which it suggests a trend that the higher altitude, the higher the number of basidiocarp
and the lower altitude, the lower the number of basidiocarp.
Soil sample of truffle Tuber sp. have pH 5.45, 41.56% moisture content, 4.85%
organic matter, 1.88% Nitrogen, 10.2 ppm Phosphorus and11.5 ppm Potassium. The edible
Tuber sp. occurs in cluster, scattered or gregarious on the ground, pathways or roads and it
is more abundant during rainy season.
Boletus leonis mushroom has been found to grow on soil with pH of 5.4, 41.53%
moisture content, 6.4% organic matter, 1.9% Nitrogen, 10.36 ppm Phosphorus and 12 ppm
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Potassium. The Boletus leonis grows on areas covered with pine needles, mostly in clusters
and is abundant during rainy season.
The cap of Boletus leonis is yellow-orange to orange-brown, 8.0 to 12.0 cm wide
and slightly convex when young and flatly expanded when matured. The hymenophore is
tubular with tubes that are yellowish-brown and attachment of tube in the stipe is adnexed.
The stipe is 4.0 to 8.o cm long and basal part is thickening and tapering to a beet-like
extension. Spore print is yellowish to orange.
The cystidium is cylindrical, basidium is unicellular holobasidium with two
sterigmata, basidiospore is subglobose and colorless. The spores measure 4-7mm and 8.5
x 4µm.
The truffle Tuber sp. is 1.2-12 cm in diameter but could reach up to 15 cm. The
wall is dirty yellow to brown. Tuber sp. is semicircular, solid, hard and has a rough texture.
The spore is circular with a dedicate ornamentation. The spore is darkyellow with
a diameter of 10 mm and 10 x 5µm.
Stained roots of pine seedlings taken from their natural habitat in surveyed area in
Tublay developed ectomycorrhizal relationship with the mushroom. Likewise, the
inoculated pine tree seedlings developed ectomycorrhizal relationship as evidenced by
ocular inspection through a microscope. On the other hand, pine seedlings without
inoculation of suspension did not show such relationship.Pine seedlings taken from their
natural habitat had significantly higher mean plant height than those inoculated pine
seedlings with spore concentration of 6.7 x 104/ml and pine seedlings without inoculation.
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
RESULTS AND DISCUSSIONS
Presence of Mycorhizal Mushroom
At an altitude of 1,300 meters above sea level, both “Bu-o” and “Dappanan”
were abundantly present (Table 1). “Bu-o” has the higher number of basidiocarp at the
whole surveyed altitude compared to “Dappanan”which was not found at altitude of
1,200 and 1,100 m asl. However, the “Bu-o” was found at the altitude of 1,100 m asl to
1,300 m asl. At a higher altitude, there was higher number of basidiocarps compared to
lower altitude.
Table1. Abundance of “Dappanan” and “Bu-o” in Tublay
ALTITUDE NUMBER OF BASIDIOCARP WITH IN A 100 m² AREA
(masl) (“Dappanan”) (“Bu-o”)
1, 300
7
139
1, 200
0
39
1, 100 0 36
TOTAL
7
214
Characterization of “Dappanan” and
“Bu-o” Mushroom
Habitat. Both “Dappanan” and ”Bu-o” were found under pine trees associated
with grasses (Figure 1).
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
The edible “Bu-o” occurs in cluster, scattered or gregarious on the ground,
pathways or roads and it is more abundantly during rainy season (Figure 2).
The “Dappanan” grows on soil covered with pine needles, mostly in
cluster and abundantly during rainy season (Figure 3).
Analysis of the soil samples of “Bu-o” revealed: pH 5.45, 41.56% moisture
content, 4.85% Organic matter, 1.88% nitrogen, 10.2 ppm phosphorus and11.5 ppm
potassium. Soil analysis of “Dappanan: pH 5.4, 41.53% moisture content, 6.4%
Organic matter, 1.9% nitrogen, 10.36 ppm phosphorus and 12 ppm potassium. This
mushroom grows from August to October.
Figure 1. Surveyed area for the presence of “Dappanan” and “Bu-o” mushroom in
Tublay
Figure 2. Truffle “Bu-o” was specifically collected at the pine forest
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Figure 3. “Dappanan” mushroom in its habitat
Physical Description of “Dappanan” mushroom. The cap of this mushroom is
yellow-orange to orange brown, 8.0 to 12.0 cm wide and slightly convex when young,
flatly expanded when matured. The hymenophore is tubular, tubes are yellowish-brown
and attachment of tube in the stipe is adnexed. The stipe is 4.0 to 8.o cm long and basal
part is thickening and tapering to a beet-like extension (Figure 4a). Spore print is yellowish
to orange.
Microscopic Description. The Cystidium is cylindrical (Figure 4b), basidium is
unicellular holobasidium with two sterigmata (Figure 4c), basidiospore is subglobose
and colorless (Figure 4d). The spores measure 4-7mm and 8.5 x 4 um (Figure 4-e).
Based on the above descriptions as well as its edibility as identified by local folks,
the mushroom is identified as Boletus leonis. Physical and microscopic descriptions agree
with the description of Surcek and Kubicka (1980) and Phillips (1994).
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Figure 4a. Fruiting body of Boletus leonis found at the surveyed area
Figure 4b. Cystidia of Boletus leonis at 40x
Figure 4c. Basidium of Boletus leonis at 40x
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Figure 4d. Spores of Boletus leonis at 40x
Figure 4e. Length of the spore of Boletus leonis with 7µm at 100x
Physical Description of “Bu-o” mushroom. Size of fruiting body is 1.2- 12 cm
in diameter but could reach up to 15 cm. The wall is dirty yellow to brown; it is
semicircular, solid, hard and ha a rough texture (Figure 5a).
Microscopic Description of “Bu-o” mushroom. The spore is circular with a
dedicate ornamentation (Figure 5b). The spore is dark yellow and with a length of 10
mm, 10 x 5 µm (Figure 5c). This mushroom is edible and it was identified as Tuber sp.
supported by information from Simpson (1989).
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Figure 5a. Truffle Tuber sp. found at the surveyed area
Figure 5b. Spores of Tuber sp. at 10x
Figure 5c. Diameter of the spore of Tuber sp. with 10 µm at 100x
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Test for Mycorrhizal Relationship
on Pine Tree Seedlings
Stained roots of pine seedlings at the surveyed area showed presence of mycelia
of mycorrhizal mushrooms (Figure 6). Conversely, pine seedling without inoculum did
not show any presence of mycelia (Table 2 and Figure 11). Likewise, inoculated pine
seedlings showed the presence of mycelia of mycorrhizal relationship (Figures 7-10).
There were significant differences between pine seedlings inoculated with 25
ml, 75 ml and 100 ml compared to the pine seedlings taken from their natural habitat.
Pine seedlings inoculated with 25 ml suspension with 167.5 x 104 ml spores showed the
presence of mycelia indicating mycorrhizal relationship. Increasing the amount of
inoculum resulted to a more indicative mycorrhizal relationship.
Table 2. Presence or Absence of Mycelia of Tuber sp.
TREATMENTS MEAN
Pine seedling taken from their natural habitat
1a
Pine seedling without inoculation of suspension 0b
Pine seedling with 25 ml inoculation suspension 0.6ab
pine seedling with 50 ml inoculation of suspension 0.1b
pine seedling with 75 ml inoculation of suspension 0.4ab
pine seedling with 100 ml inoculation of suspension 0.6ab
*Significant differences exists at 0.05 level using DMRT
CV = 23.85%
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
a
b
Figure 6. Stained roots of pine seedlings taken from their natural habitat at 40x(a) and
10x(b) magnification (arrow indicates the mycelia of Tuber sp.)
a
b
Figure 7. Stained roots of pine seedlings inoculated with (167.5 x 104) / ml spore
suspension after 136 days at 40x magnification. (arrow indicates the mycelia
of Tuber sp.)
a
b
Figure 8. Stained roots of pine seedlings inoculated with (335 x 104) / ml spore
suspension after 136 days at 10x and 40x magnification. (arrow indicates
the mycelia of Tuber sp.)
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
a
b
Figure 9. Stained roots of pine seedlings inoculated with (502.5 x 104) / ml spore
suspension after 136 days at 40x and 10x magnification. (arrow indecates
the mycelia of Tuber sp.)
a
b
Figure 10. Stained roots of pine seedlings inoculated with (67 x 106) /ml suspension
after 136 days at 10x magnification. (arrow indicates the mycelia of Tuber
sp.)
a
b
Figure 11. Stained roots of pine seedlings without inoculation
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Figure 12. Pine (Pinus kesiya) seedling replicates for each treatments
Pine seedlings taken from their natural habitat had significantly higher mean plant
height than the inoculated pine seedlings. Significantly taller pine seedlings were from
those taken from natural habitat. This is followed by pine seedlings inoculated with 25 ml
and 100 ml of suspension. The results suggest that pine seedlings taken from their natural
habitat had significantly higher mean plant height than those inoculated regardless of
amount of inoculum. These differences may be due to the maturity of the pine seedlings
(figure 13).
Table 3. Height of Pine Seedlings
TREATMENTS HEIGHT (cm)*ᵧ
pine seedling taken from their natural habitat 11.75a
pine seedling without inoculation of suspension 8.98b
pine seedling with 25 ml inoculation of suspension 9.77ab
pine seedling with 50 ml inoculation of suspension 8.84b
pine seedling with 75 ml inoculation of suspension 8.78b
pine seedling with 100 ml inoculation of suspension 9.52b
TOTAL 57.64
*No significant differences at 0.05 level, significant differences at 0.10 level,
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
P-value=0.085
ᵧMeans with different letters signify significant differences using DMRT
CV=25.63%
Plant Height at 7, 21, 35, 49, 63, 77, 91, 105 and 119 Days After
Inoculation
25
20
m)
T1
15
(c
T2
ht
ig
T3
T4
10
lant He
P
T5
T6
5
0
7
21
35
49
63
77
91
105
119
Days after Inoculation
Figure 13. Plant Height at 7, 21, 35, 49, 63, 77, 91, 105 and 119 Days after inoculation
(DAI)
Alexopoulus et al., (1996) reported that mycorhizal fungi benefit their host by
increasing the plants ability to capture water and nutrients such as nitrogen,
phosphorous, potassium and calcium from the soil, increasing tolerance to drought, high
temperature and extremes of soil acidity and providing protection from certain plant
pathogens.
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
SUMMARY, CONCLUSIONS AND RECOMMENDATIONS
Summary
The study was conducted to characterized “Bu-o” and “Dappanan” mushroom
based on: habitat (type of tree cover where found, altitude, temperature, moisture and soil
pH) type of mycorrhizal relationship, morphological and anatomical features, abundance
of each species with altitude, relative humidity and temperature and determined the
mycorrhizal relationship between each mushroom and pine seedlings.
Microscopic studies were done at the Department of Plant Pathology, Benguet State
University from October 2010 to October 2011. Soil analysis of mineral content was done
at Department Soil Science, Benguet State University and Regional Soils Laboratory, San
Fernando, La Union.
At an altitude of 1,300 meters above sea level, both “Bu-o” and “Dappanan” were
abundantly present. “Bu-o” has the higher number of basidiocarp at this altitude, compared
to “Dappanan”. However at altitudes 1,200 to 1,100 m asl, only the “Bu-o” was found but
at a lesser. This indicates that at higher altitudes, a higher number of basidiocarps were
observed which decreased as altitude decreases.
The edible “Bu-o” occurs in cluster, scattered or gregarious on the ground,
pathways or roads and it is more abundant during rainy season.
Soil samples of Truffle “Bu-o” revealed: pH 5.45, 41.56% moisture content, 4.85%
Organic matter, 1.88% nitrogen, 10.2 ppm phosphorus and11.5 ppm potassium.
The “Dappanan” grows on covered with pine needles, mostly in cluster and
abundantly during rainy season.
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Soil analysis of “Dappanan” mushroom: pH 5.4, 41.53% moisture content, 6.4%
Organic matter, 1.9% nitrogen, 10.36 ppm phosphorus and 12 ppm potassium.
The cap of “Dappanan” is yellow-orange to orange brown, 8.0 to 12.0 cm wide
and slightly convex when young, flatly expanded when matured. The hymenophore is
tubular with tubes that are yellowish-brown and attachment of tube in the stipe is adnexed.
The stipe is 4.0 to 8.o cm long and basal part is thickening and tapering to a beet-like
extension. Spore print is yellowish to orange.
The Cystidium is cylindrical, basidium is unicellular holobasidium with two
sterigmata, basidiospore is subglobose and colorless. The spores measure 4-7mm and 8.5
x 4 µm.
The truffle “Bu-o” is 1.2- 12 cm in diameter but could reach up to 15 cm. The wall
is dirty yellow to brown; it is semicircular, solid, hard and has a rough texture.
The spore is circular with a dedicate ornamentation. The spore is dark yellow with
a length of 10 mm and 10 x 5 µm.
Based on physical and microscopic features, these mushrooms are edible.
“Dappanan” is identified as Boletus leonis and truffle “Bu-o” is identified as Tuber sp.
Stained roots of pine seedlings at the surveyed area showed presence of mycelia of
mycorrhizal mushrooms. Conversely, pine seedling without inoculum did not show any
presence of mycelia. Likewise, inoculated pine seedlings showed the presence of mycelia
of mycorrhizal relationship. Results of the analysis of variance test showed significant
differences between pine seedlings inoculated with 25ml, 75 ml and 100 ml against the
pine seedling taken from their natural habitat at the 0.05 level using DMRT.
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
Also, pine seedlings taken from their natural habitat had significantly higher mean
plant height than those inoculated pine seedlings and pine seedlings without inoculation of
suspension at the 0.05 level using DMRT. The results suggest that pine seedlings taken
from their natural habitat had significantly higher mean plant height than those inoculated
regardless of amount of inoculum.
Conclusion
“Dappanan” and “Bu-o” were found under pine-based areas associated with grasses
in Tublay. Altitude, relative humidity, temperature, moisture content and soil pH provides
a habitat which favors the growth and abundance of many truffles and mushrooms. Based
on physical and microscope features, “Dappanan” is identified as Boletus leonis and truffle
“Bu-o” is identified as Tuber sp. These mushrooms developed ectomychorrhizal
relationship with its habitat.
Recommendation
It is recommended that further studies be conducted to identify other areas where
truffle “Bu-o“ and “Dappanan” may be found. In addition, studies to further understand
the ectomycorrhizal relationship between the mushroom and pine tree roots should be done
so that production could be explored. There is also a need to further verify the effects of
inoculum on the growth and development of the pine tree seedlings.
Characterization of Two Mycorrhizal Mushroom Associated with Benguet Pine (Pinus
kesiya)| ALFREDO, NOEL VELASCO. APRIL 2011
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