Antifungal Activity of Lichen-forming Fungi against Colletotrichum acutatum on Hot Pepper

Abstract

Antifungal activity of Korean and Chinese lichen-forming fungi (LFF) was evaluated against plant pathogenic fungus of Colletotrichum acutatum, causal agent of anthracnose on hot pepper. This is the first attempt to evaluate antifungal activity of LFF, instead of lichen thalli, against C. acutatum. Total 100 LFF were isolated from the lichens with discharged spore method or tissue culture method. Among the 100 isolates, 8 LFF showed more than 50% of inhibition rates of mycelial growth of the target pathogen. Especially, Lecanora argentata was highly effective in inhibition of mycelial growth of C. accutatum at the rate of 68%. Antifungal activity of other LFF was in the order of Cetrelia japonica (61.4%), Ramalina conduplicans (59.5%), Umbilicaria esculenta (59.5%), Ramalina litoralis (56.7%), Cetrelia braunsiana (56.5%), Nephromopsis pallescensn (56.1%), and Parmelia simplicior (53.8%). Among the tested LFF, 61 isolates of LFF exhibited moderate antifungal activity against the target pathogen at the inhibition rates from 30 to 50%. Antifungal activity of the LFF against C. acutatum was variable at the species level rather than genus level of LFF. This study suggests that LFF can be served as a promising bioresource to develop novel biofungicides.

Full text

Plant Pathol. J. 24(2) : 202-206 (2008) The Plant Pathology Journal
© The Korean Society of Plant Pathology
Antifungal Activity of Lichen-forming Fungi against Colletotrichum acutatum on
Hot Pepper
Xinli Wei1,2,†, Hae-Sook Jeon1,†, Keon Seon Han1, Young Jin Koh1 and Jae-Seoun Hur1*
1Korean Lichen Research Institute, Sunchon National University, Sunchon 540-742, Korea
2Key Laboratory of Systematic Mycology & Lichenology, Institute of Microbiology, Chinese Academy of Sciences, Beijing
100080, China
(Received on December 1, 2007; Accepted on March 26, 2008)
Antifungal activity of Korean and Chinese lichen-form-
ing fungi (LFF) was evaluated against plant pathogenic
fungus of Colletotrichum acutatum, causal agent of
anthracnose on hot pepper. This is the first attempt to
evaluate antifungal activity of LFF, instead of lichen
thalli, against C. acutatum. Total 100 LFF were isolated
from the lichens with discharged spore method or tissue
culture method. Among the 100 isolates, 8 LFF showed
more than 50% of inhibition rates of mycelial growth of
the target pathogen. Especially, Lecanora argentata was
highly effective in inhibition of mycelial growth of C.
accutatum at the rate of 68%. Antifungal activity of
other LFF was in the order of Cetrelia japonica (61.4%),
Ramalina conduplicans (59.5%), Umbilicaria esculenta
(59.5%), Ramalina litoralis (56.7%), Cetrelia braunsiana
(56.5%), Nephromopsis pallescensn (56.1%), and Parmelia
simplicior (53.8%). Among the tested LFF, 61 isolates of
LFF exhibited moderate antifungal activity against the
target pathogen at the inhibition rates from 30 to 50%.
Antifungal activity of the LFF against C. acutatum was
variable at the species level rather than genus level of
LFF. This study suggests that LFF can be served as a
promising bioresource to develop novel biofungicides.
Keywords :biofungicide, fungal growth inhibition, lichen-
forming fungi, novel bioresource, plant pathogenic fungus
Pepper anthraconose caused by Collectotrichum acutatum,
is one of the most important diseases in pepper cultivation
in Korea (Choi et al., 2008; Kang et al., 2005). The disease
was reported to be responsible for 10% annual yield loss of
total pepper production in Korea, equivalent to approxi-
mately $0.1 billion per year (Shin et al., 1999). Current
practice for controlling pepper anthracnose is mainly based
on use of synthetic pesticides. However, many synthetic
pesticides may lose their usefulness due to negative
consequences for human health and the environment and
development of resistance in pathogen populations (Russell,
1995). Recently, biological control has been developed as
an alternative to synthetic pesticide treatment. A variety of
microbial antagonists and their metabolites have been
reported to control several different pathogens on various
fruit and vegetables (Fravel, 2005).
Lichens are symbiotic organisms composed of a fungus
(mycobiont) and an algae (photobiont). They produce
characteristic secondary metabolites, lichen substances,
which seldom occur in other organisms. Antifungal activity
of lichen extracts and lichen acids against plant pathogenic
fungi was reported (Gulluce et al., 2006; Halama and Van
Haluwin, 2004; Oh et al., 2006). Nevertheless, the potential
fungal obligate symbionts in lichen have long been
neglected by mycologists and overlooked by agrochemical
industry because of its slow growth in nature and diffi-
culties in their artificial cultivation. The large-scale
industrial production of the lichen metabolites has never
been accomplished. However, use of lichen-forming fungi
can overcome the disadvantage of natural lichen extracts
for industrialization of their metabolites because of their
much faster growth and larger production of the metabolites
in culture than the natural thalli. This is the first attempt to
screen antifungal activity against C. acutatum with use of
large number of lichen-forming fungi, instead of natural
lichen thallus, for further development of industrial produc-
tion of natural fungicides.
The natural thalli of Korean lichens were collected from
20 mountain areas in South Korea during 2002 to 2004
(Hur et al., 2004). Chinese lichens were also collected from
highland areas of Yunnan Province, China (Hur et al.,
2005). Lichen specimens were air-dried for 1 week at room
temperature and stored −20oC until isolation of lichen-
forming fungi (LFF: mycobints). All the lichen materials
were deposited at the herbarium of the Korean Lichen
Research Institute, Sunchon National Univeristy, Korea.
Collectotrichum acutatum KACC40042 was obtained from
Korean Agricultural Culture Collection (KACC), Rural
Development Administration (RDA), Suwon, Korea.
The colonies of LFF were obtained using discharged
*Corresponding author.
Phone) +82-61-750-3383, FAX) +82-61-750-3308
E-mail) jshur1@sunchon.ac.kr
†These authors contributed equally to this work.
Note

Antifungal Activity of Lichen-forming Fungi Against Colletorichum acutatum 203
spore method (S in Table 1) and/or tissue culture method (T
in Table 1) (Yoshimura et al., 2002). Details of LFF iso-
lation using discharged spore method were previously
described (Oh et al., 2006).
The fungal components of sterile lichens, or those in
which isolation from discharged spore had not proved
successful, were isolated from thallus fragments by the
method of Yamamoto (Yoshimura et al., 2002). Small
pieces of thallus such as 1 cm in length of fruticose lichen
or 1 cm2 of foliose and crustose lichen were cut from apical
regions, washed in a turbulent flow of tap water for 1 h,
then in excess sterile water, and finally macerated in a
further 3-5 ml sterile water using a mortar and pestle. The
resultant suspension was sieved through 500 μm and 150
μm nylon mesh filters in sequence with sterile water.
Macerate retained on the 150 μm sieve was examined
under a dissecting microscope and the fragments free from
algae were picked out and transferred to agar media using
sterilized stainless steel syringe-needle and inoculated onto
the medium of 24 multi well (d=2 cm) plates. Total 94
inoculums from each lichen material were prepared.
Cultures were incubated at 18oC in the dark and examined
periodically during a week period. Thallus fragments that
remained free of contamination were transferred to fresh
medium. After 2-3 month’s growth, mycobionts produced a
compact mycelium 2-5 mm in diameter. These were
subcultured onto fresh medium for fungal mass production.
Species that continued to grow in subculture were recorded
as successfully isolated. The culture medium of mal-extract
agar was routinely used for isolation and growth of LFF
(Yoshimura et al., 2002).
Analysis of the ribosomal DNA sequence of ITS region
was attempted for molecular confirmation of the isolates
obtained by tissue culture method. The sequences of
resultant fungal mass of isolates and the original lichen
thallus used for isolation were analyzed and compared.
Fresh lichen thalli and fungal mass were fractioned with
cryo-tissue-crasher (SK200, Tokken, Japan). Total DNA
was extracted directly from whole thalli according to Ekman
(1999) with DNeasy Plant Mini Kit (QIAGEN, Germany).
10−1 dilution of the total DNA was used for PCR
amplification of the nuclear rDNA ITS and 5.8S genes.
Primers for amplification were: ITS4 (5'-TCCTCCGCTT-
ATTGATATGC-3'; White et al., 1990) and ITS5 (5'-GG-
AAGTAAAAGTCGRAACAAGG-3'; White et al., 1990).
Conditions for PCR amplification and cycle sequencing
have been described previously (Arup, 2002). PCR prod-
ucts were purified by PCRquick-spinTM PCR Product
Purification Kit (iNtRON Biotechnology, INC.) and then
sequenced using ABI 3700 automated DNA Sequencer in
NICEM at Seoul National University.
Freshly grown two mycelial masses (3 mm diam.) of the
isolated LFF were placed at the edge of malt extract agar
plate (6 cm diam.) at same distance from the plate center.
Due to slow growth of LFF, the isolates were incubated on
the agar medium at 18oC in dark condition 60 days before
inoculation of C. acutatum. Freshly grown mycelium agar
block (3 mm diam.) of C. acutatum was placed on the
center of the pre-incubated agar plate. The inhibition zone
of mycelial growth of the pathogenic fungus was rated 3 to
5 days after incubation at 18oC and compared with the
control plate. Five replicate plates were used for the
bioassay. Antifungal activity of the lichen-forming fungi
was compared using one-way analysis of variance (ANOVA).
Total 100 lichen-forming fungal isolates were obtained
from Korean and Chinese lichens using discharged spore
method and tissue culture method (Table 1). Many ascos-
pores were successfully discharged from a single apotheci-
um and generally germinated within 1 week. Several fungal
isolates was grown to visible size within 2 months. Lichen-
forming fungi were also induced from lichen thalli by tissue
culture method. Confirmation of lichen-forming fungi was
carried out by ITS sequences analysis. Isolation rates of
lichen-forming fungi from ascospore or lichen thalli were
approximately 30% in this study (data not shown).
Some isolates leached large amount of pigments into the
agar medium during the incubation. This suggested that
secondary metabolites of LFF were produced and diffused
into the medium. Compared with normal fungi, LFF grew
slowly and developed less than 1 cm diameter of mycelium
mass within 5 months. However, the growth rate can be
considered to be much faster than that of the natural lichen
thalli (Yamamoto et al., 1993).
Several LFF showed very strong antifungal activity against
the pepper anthracnose pathogenic fungus (Table 1). Among
the 100 isolates, 8 LFF showed more than 50% of inhibi-
tion rates of mycelial growth of the target pathogen. Especi-
ally, LFF of Lecanora argentata was mostly effective in
inhibition of mycelial growth of C. accutatum at the rate of
68%. Antifungal activity of other LFF was in the order of
Cetrelia japonica (61.4%) > Ramalina conduplicans (59.5
%) = Umbilicaria esculenta (59.5%) > Ramalina litoralis
(56.7%) ≥ Cetrelia braunsiana (56.5%) ≥ Nephromopsis
pallescensn (56.1%) > Parmelia simplicior (53.8%). Their
antifungal activity was significantly higher than other lichen-
forming fungi tested in this study (P < 0.05). Among the
tested LFF, 61 isolates of LFF exhibited moderate anti-
fungal activity against the target pathogen at the inhibition
rates from 30 to 50%. Among the rest of 31 LFF isolates,
only one LFF of Cladonia cervicornis showed less than
20% inhibition rate (17.8%) and others demonstrated
recognizable antifungal activity from 20 to 30%. Anti-
fungal activity of the LFF against C. acutatum was variable
at the species level rather than genus level of LFF. How-

204 Xinli Wei et al.
Table 1. Antifungal activity of various lichen-forming fungi isolated from Korean and Chinese lichens against hot pepper anthracnose
pathogen, Colletortichum acutatum in vitro
Lichen species Collection
number Locality Isolation
methoda
Mycelium growth inhibition
Diam. (mm)b% of control
Amandinea punctata (Hoffm.) Coppins & Scheid. 50623 Korea T 36.54 ± 1.58 60.9
Anaptychia palmatula (Michx.) Vain. 41078 Korea S 34.37 ± 0.11 57.3
Anaptychia palmatula (Michx.) Vain. 40180 Korea T 34.36 ± 0.86 57.3
Anzia opuntiella Müll. Arg. 40280 Korea S 43.97 ± 1.63 73.3
Bacidia schweinitzii (Tuck.) A. Schneid. 40162 Korea S 38.94 ± 1.75 64.9
Bryoria confusa (D.D. Awasthi) Brodo & D. Hawksw. CH050187 China S 44.18 ± 0.76 73.6
Bryoria himalayensis (Motyka) Brodo & D. Hawksw. CH050365 China S 45.43 ± 3.54 57.7
Caloplaca flavorubescens (Huds.) J.R. Laundon 50696 Korea T 40.76 ± 1.58 67.9
Cetrelia braunsiana (Müll. Arg.) W.L. Culb. & C.F. Culb. 40188 Korea T 26.67 ± 0.58 44.5
Cetrelia japonica (Zahlbr.) W.L. Culb. & C.F. Culb. 30397 Korea T 23.18 ± 1.61 38.6
Cetrelia braunsiana (Müll. Arg.) W.L. Culb. & C.F. Culb. 40425 Korea T 40.48 ± 1.05 67.5
Cladonia furcata (Huds.) Schrad. 40034 Korea T 45.96 ± 0.90 76.6
Cladonia gracilis subsp. turbinata (Ach.) Ahti 41582 Korea S 42.43 ± 3.50 70.7
Cladonia gracilis (L.) Willd. 41474 Korea S 35.46 ± 0.59 59.1
Cladonia metacorallifera Asahina. 41466 Korea S 40.79 ± 2.45 68
Cladonia coniocraea (Flörke) Spreng. 040634-1 Korea T 42.88 ± 1.37 71.5
Cladonia yunnana (Vain.) Abbayes. CH050136 China S 36.48 ± 1.10 60.8
Cladonia macilenta Hoffm. CH050214 China S 45.35 ± 2.47 75.6
Cladonia coccifera (L.) Willd. CH050255 China S 43.20 ± 1.71 72
Cladonia cervicornis (Ach.) Flot. CH050299 China T 49.31 ± 1.14 82.2
Cladonia pleurota (Flörke) Schaer. CH050056 China T 47.49 ± 3.83 79.2
Cladonia squamosissima (Müll. Arg.) Ahti CH050180 China T 43.84 ± 1.02 73.1
Cladonia rangiferina (L.) Weber ex F.H. Wigg. CH050184 China T 40.26 ± 1.00 67.1
Cladonia scabriuscula (Delise) Leight. 40481 Korea S 46.41 ± 1.68 77.4
Everniastrum cirrhatum (Fr.) Hale ex Sipman. CH050065 China S 45.96 ± 1.44 76.6
Gymnoderma coccocarpum Nyl. CH050374 China T 39.98 ± 1.66 66.6
Heterodermia japonica (M. Satô) Swinscow & Krog. CH050263 China T 43.89 ±2.93 73.2
Heterodermia hypoleuca (Mühl.) Trevis. 40067 Korea T 43.07 ± 3.55 71.8
Heterodermia hypoleuca (Mühl.) Trevis. 40598 Korea T 40.99 ± 0.81 68.3
Heterodermia microphylla (Kurok.) Skorepa 40196 Korea T 41.46 ± 1.87 69.1
Hypogymnia delavayi (Hue) Rass. 41338 Korea S 37.7 ± 0.36 62.8
Hypogymnia pseudoenteromorpha M.J. Lai. CH050143 China S 41.65 ± 1.51 69.4
Hypogymnia pruinosa J.C. Wei & Y.M. Jiang. CH050101 China S 38.24 ± 1.22 63.7
Hypogymnia hengduanensis J.C. Wei. CH050100 China T 44.35 ± 1.06 73.9
Hypotrachyna osseoalba (Vain.) Y.S. Park & Hale CH050261 China T 44.76 ± 1.90 74.6
Icmadophila ericetorum (L.) Zahlbr. CH050403 China S 37.26 ± 3.31 62.1
Lecanora argentata (Ach.) Malme 50657 Korea S 19.02 ± 0.12 31.7
Megalospora tuberculosa (Fée) Sipman. 50724 Korea S 41.03 ± 3.00 68.4
Melanelia olivacea (L.) Essl. 40371 Korea S 38.59 ± 1.11 64.3
Menegazzia terebrata (Hoffm.) A. Massal. 41300 Korea T 44.24 ± 0.80 73.7
Menegazzia pseudocyphellata Aptroot, M.J. Lai & Sparrius. CH050262 China S 43.99 ± 0.17 73.3
Myelochroa aurulenta (Tuck.) Elix & Hale. 40664 Korea T 39.13 ± 0.59 65.2
Myelochroa irrugans (Nyl.) Elix & Hale. 40974 Korea T 32.85 ± 2.05 54.8
Myelochroa irrugans (Nyl.) Elix & Hale. 40303 Korea T 33.82 ± 0.79 56.4
Myelochroa irrugans (Nyl.) Elix & Hale. 30730 Korea T 33.53 ± 1.84 55.9
Myelochroa galbina (Ach.) Elix & Hale. 40954 Korea S 41.93 ± 0.29 69.9
Myelochroa aurulenta (Tuck.) Elix & Hale. 40678 Korea S 40.03 ± 2.15 66.7
Myelochroa indica (Hale) Elix & Hale. 40002 Korea T 39.74 ± 1.97 66.2
Myelochroa entotheiochroa (Hue) Elix & Hale. 41602 Korea S 33.27 ± 2.03 55.5
Nephromopsis yunnanensis (Nyl.) Randlane & Saag. CH050123 China S 35.16 ± 1.61 58.6
Nephromopsis pallescens (Schaer.) Y.S. Park. CH050089 China S 26.96 ± 1.93 44.9
Nephromopsis ornate (Müll. Arg.) Hue. 41359 Korea T 38.21 ± 2.20 63.7

Antifungal Activity of Lichen-forming Fungi Against Colletorichum acutatum 205
Table 1. Continued
Lichen species Collection
number Locality Isolation
methoda
Mycelium growth inhibition
Diam. (mm)b% of control
Nephromopsis asahinae (M. Satô) Räsänen. 40500 Korea S 40.64 ± 2.15 67.7
Nephromopsis pseudocomplicata (Asahina) M.J. Lai CH040025 China S 37.34 ± 1.15 62.2
Parmelia simplicior Hale CH050260 China S 27.69 ± 4.02 46.2
Parmelia laevior Nyl. 40257 Korea S 41.46 ± 0.29 69.1
Parmelia adaugescens Nyl. 40810 Korea S 32.65 ± 2.38 54.4
Parmelia pseudolaevior Asahina. 50069 Korea S 34.71 ± 2.64 57.9
Parmelia omphalodes (L.) Ach. 40356 Korea T 38.11 ± 2.37 63.5
Parmotrema austrosinense (Zahlbr.) Hale. CH050252 China T 40.83 ± 2.79 68.1
Parmotrema ultralucens (Krog) Hale. CH050249 China T 40.83 ± 2.45 68.1
Pannaria leucosticta Tuck. 41227 Korea T 32.41 ± 0.54 54
Phaeophyscia limbata (Poelt) Kashiw. 40014 Korea S 43.20 ± 2.06 72
Phaeophyscia exornatula (Zahlbr.) Kashiw. 40202 Korea T 43.77 ± 1.35 73
Phaeophyscia melanchra (Hue) Hale. 40625 Korea S 41.87 ± 2.06 69.8
Physcia caesia (Hoffm.) Fürnr. 40912 Korea S 39.41 ± 1.05 65.7
Phaeophyscia exornatula (Zahlbr.) Kashiw. 40923 Korea T 43.73 ± 4.76 72.9
Phaeophyscia hirtella Essl. 41166 Korea S 47.47 ± 4.07 79.1
Physcia stellaris (L.) Nyl. 41642 Korea S 42.88 ± 2.06 71.5
Physcia stellaris (L.) Nyl. 50213 Korea S 44.69 ± 0.92 74.5
Phaeophyscia melanchra (Hue) Hale. 40089 Korea T 40.74 ± 0.16 67.9
Punctelia borreri (Sm.) Krog 40051 Korea T 39.00 ± 1.78 65
Pyxine endochrysina Nyl. 41658 Korea T 34.21 ± 0.40 57
Pyxine consocians Vain. 40935 Korea S 30.77 ± 1.07 51.3
Ramalina exilis Asahina. 30474 Korea T 45.67 ± 0.81 76.1
Ramalina intermedia Delise ex Nyl. CH050064 China S 37.45 ± 0.16 62.4
Ramalina pertusa Kashiw. 40628 Korea T 38.90 ± 1.50 64.8
Ramalina litoralis Zahlbr. 50319 Korea S 26.58 ± 1.31 44.3
Ramalina conduplicans Vain. 40402 Korea S 24.28 ± 2.41 40.5
Ramalina sp. CH040302 China S 39.15 ± 2.49 65.3
Ramalina complanata Ach. CH050099 China S 29.89 ± 0.92 49.8
Ramalina yasudae Räsänen. 30330 Korea T 35.69 ± 1.41 59.5
Ramalina almquistii Vain. 30237 Korea S 40.92 ± 0.62 68.2
Ramalina sinensis Jatta. CH040020 China S 45.94 ± 0.26 76.6
Ramalina roesleri (Hochst. ex Schaer.) Hue. CH050380 China T 42.62 ± 1.33 71
Rimelia clavulifera (Räsänen) Kurok. 50038 Korea S 36.80 ± 2.38 61.3
Rimelia clavulifera (Räsänen) Kurok. 50150 Korea S 41.95 ± 1.78 69.9
Rimelia reticulata (Taylor) Hale & A. Fletcher. CH050393 China T 42.61 ± 1.87 71
Rimelia reticulata (Taylor) Hale & A. Fletcher. 40068 Korea T 39.68 ± 3.04 66.1
Stereocaulon commixtum (Asahina) Asahina. 40659 Korea S 44.63 ± 1.92 74.4
Tephromela atra (Huds.) Hafellner. 40191 Korea S 42.84 ± 3.05 71.4
Tuckneraria pseudocomplicata (Asahina) Randlane & Saag 40516 Korea S 41.73 ± 3.15 69.6
Umbilicaria proboscidea (L.) Schrad. CH040077 China S 39.65 ± 0.72 66.1
Umbilicaria kisovana (Zahlbr. ex M. Satô) Kurok. 30221 Korea T 37.52 ± 0.28 62.5
Umbilicaria esculenta (Miyoshi) Minks. 40040 Korea T 24.31 ± 1.12 40.5
Umbilicaria yunnana (Nyl.) Hue. CH050097 China T 40.36 ± 2.10 67.3
Usnea longissima Ach. CH050148 China S 42.25 ± 0.65 70.4
Usnea orientalis Motyka. CH050316 China S 29.94 ± 1.68 49.9
Xanthoparmelia hirosakiensis (Gyeln.) Kurok. 50141 Korea S 38.42 ± 1.27 64
Xanthoria elegans (Link) Th. Fr. 41247 Korea S 34.31 ± 0.58 57.2
Control 60.00 −
aS; Discharged spore method, T; Tissue culture method.
bThe inhibition zone of mycelial growth of the pathogenic fungus was rated 3 to 5 days after incubation at 18ºC and compared with the control
plate. Data represent the mean and standard deviation of five replications. Bold letters indicate the lichen-forming fungi which showed signifi-
cantly higher antifungal activity against C. acutatum than other lichen-forming fungi tested in this study (P< 0.05, LSD) .

206 Xinli Wei et al.
ever, LFF isolated from lichen species belong to Cetrariod
genera (Cetrelia and Nephromposis) and Ramalina genus
possessed stronger antifungal activity against the target
pathogenic fungus than those isolated from other lichen
genera tested in this study. Mycelial growth of Botryo-
sphaeria dothidea, Botrytis cinerea, Pythium sp., Rhizo-
ctonia solani and Sclerotium cepivorum were completely
inhibited by LFF of Parmelia laevior in our previous study
(Oh et al., 2006). However, it was not the case for C.
acutatum in this study. This suggests that secondary
metabolites produced during LFF culture have differential
sensitivity to various plant pathogenic fungi and the
compounds acted in a species-specific manner.
Methanol or acetone extracts of several lichen thalli were
already proved to have strong antifungal activity against
various plant pathogenic fungi (Gulluce et al., 2006; Halama
and Van Haluwin, 2004). Unlike the previous results using
intact lichen thalli extracts, LFF exhibited different anti-
fungal activity against Colletotrichum fungus in this study.
It is well known that LFF in axenic cultures retain the
capacity to biosynthesize secondary products found in the
lichenized state (Culberson et al., 1992), but the metabolites
produced in the greatest abundance might differ from those
found in the lichen (Miyagawa et al., 1993). Therefore, it
will be very interesting to investigate the compounds
responsible for strong antifungal activity of the LFF in
cultures with comparison of natural lichen substances.
Mass cultivation of the LFF is now under progress in
laboratory conditions for chemical identification of anti-
fungal substances. In conclusion, the secondary metabolites
of LFF in cultures might be of potential use as antifungal
agents and LFF can serve as a novel bioresources to
develop new biofungicides alternative to current fungicides
to control C. acutatum, hot pepper anthracnose pathogenic
fungus.
Acknowledgements
This work was supported in part by Rural Development
Administration program (Grant 20070301-033-016-001-
02-00), Republic of Korea and by NON DIRECTED
RESEARCH FUND (2004), Sunchon National University,
Korea.
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