LICHENS POTENTIAL

SUBSTANCES

Presented to: Dr. Abdul Nasir khalid

Presentedby:
Presented to:Munazza
Dr. AbdulGhazanfar
Nasir khalid
Presented
Roll. by: Munazza Ghazanfar
No. BOT-03-F19
Roll.
M.S No. BOT-03-F19
Botany (Regular)
1
M.S Botany
University (Regular)
of Punjab, Lahore.
University of Punjab, Lahore.
 2

Contents:
 Introduction
 History of lichen substances
 Primary and Secondary metabolites
 Biosynthetic pathways
 Methods of determination of lichen substances
 Activities of lichen substances
 Conclusion
 References
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Lichens are symbiotic organisms

composed of a fungal partner
(mycobiont) in association with
one or more photosynthetic
partners (photobiont)

What are Lichens?

• About 18500 species

• (98%) are Ascomycota
• 40 genera as photosynthetic
partners (25 of algae and 15 of
cyanobacteria)
• Common photobionts are the
genera Trebouxia, Trentepohlia
and Nostoc
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Major differences in algae versus
cyanobacteria

Algae Cyanobacteria
Nature: Eukaryotic algae Nature: Oxygenic photosynthetic
bacteria

Genera: Trebouxia and Genera: Nostoc

Trentepohlia

Organelles: Chloroplasts, Organelles: Lack all those found

mitochondria, nucleus in algae

Metabolite transfer: Metabolite transfer: Glucose

Carbohydrates are sugar alcohols

Eukaryotic phycobionts makes Cynobionts makes 10% of lichens

90% of lichens
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Lichen thallus types:

Crustose
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Contin..
Foliose
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Contin..
Fruticose
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Contin..
Squamulose
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Contin..
Filamentous
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Contin..
Leprose
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Lichen substances

Fungus absorb
moisture and
nutrients from
Fungus utilizing these
atmosphere
carbohydrates
produces secondary
metabolites

Supplied to
algae

Supplied to
fungus
Algae due to
photosynthetic
apparatus produces
carbohydrates
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History of lichen substances:

 Bebert (1831) isolated vulpinic acid in 1831 from Citraria vulpina
 Alms (1832) isolated picrolichenic acid in 1832 from Pertusaria amara
 Knop (1844) isolated usnic acid in 1844
 W. Zopf (1846-1909) and O. Hesse (1835-. 1917) are known as the pioneers of
research of lichen substances in the early 20th century. Both published a series of
papers on lichen substances.
 Asahina and Shibata published another classic ‘Chemistry of Lichen
Substances’ in 1954. They not only described the structure of numerous
compounds but also described their synthesis and introduced the determination of
lichen substances by micro crystallization.
 A milestone in the field of lichen substances was the publication of Chicita F.
Culberson’s ‘Chemical and Botanical Guide to Lichen Products’, which
appeared in 1969. It summarizes the data of about 430 lichen substances.
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Types of metabolites:
Include proteins, amino
acids, polyols, carotenoids,
polysaccharides, and
Primary metabolites vitamins

sugar alcohols, aliphatic acids,

macrolytic lactones, monocyclic
aromatic compounds, quinines,
 Secondary chromones, xanthones,
dibenzofurans, depsides,
metabolites depsidones, depsones,
terpenoids, steroids, carotenoids,
and diphenyl ethers.
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Primary metabolites:
 Common intracellular products occurring in lichens
include proteins, amino acids, polyols, carotenoids,
polysaccharides, and vitamins, which are bound in the
cell walls and the protoplasts, are often water-soluble,
and can be extracted with boiling water .
 Some of these products are synthesized by the fungus
and some by the alga.
 Most of the intracellular products isolated from lichens
are nonspecific, and also occur in free-living fungi,
algae and in higher green plants
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Secondary Metabolites:
 The majority of organic compounds found in lichens are secondary
metabolites of the fungal component, which are deposited on the
surface of the hyphae rather than within the cells and are also called
as lichen acids.
 These products are usually insoluble in water and can only be
extracted with organic solvents.
 More than 800 secondary metabolites are known from lichens,
most are unique to these organisms
 These metabolites are not absolutely essential for the survival and
growth of lichens . However, it is important that they may help to
protect the thalli against herbivores, pathogens, competitors and
external abiotic factors, such as UV irradiation ( e.g. lichen
derieved phenolic compounds including depsides , depsidones,
usnic acid derivates are effective UV- absorbing compounds)
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Contin..
 In many cases of axenic cultures, mycobionts can
produce secondary substances that are different from the
secondary metabolites found in symbiosis (Brunauer et al
2007)
 For example natural lichen

Lecanora dispersa contains 2, 7-dichloroliche-xanthone as

the major secondary
compound but cultured spore
isolates growing without algae
produce pannarin instead.
(Leuckert et al. 1990)
Lecanora dispersa
lichen
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Biosynthetic pathways of Lichen

Secondary Metabolites:
Three major pathways :
1. Acetyle poly-malonyl Pathway:
 Includes most common lichen metabolites (80%)
 These metabolites are aromatic compounds formed by
bonding of 2 or 3 orcinol or β-orcinol type phenolic
units (e.g. depsides, depsidones, usnic acids, depsones)
 Other aromatic compounds produced by internal
cyclization of single or more polyketide chain
(chromomes, xanthones, anthraquinones)
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Cont…
2. Mevalonic acid Pathway:
 17% of total compounds are produced by this
pathway
 include carotenes (β-carotene, γ-carotene,
xanthophylls, violoxanthin), sterols (ergosterol,
fungisterol, β-sitosterol) and triterpenes.
3. Shikimic acid Pathway:
 Small portion of secondary metabolites by this.
 Includes terphenylquinones and pulvinic acid
derivatives
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Cont…
 The genera of lichens best known for their
production of pulvinic acid pigments  Stictaceae
family.
 Algal symbionts are frequently blue green rather
than green. Lichens with green algae generally
show very low nitrogen content unless they grow
on high nitrogen substrates. Although nitrogen
metabolism in lichens is poorly understood. In
contrast to green algae, many blue green algae
show much enhanced nitrogen content.
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Lichen secondary metabolites that are obtain from

different lichens:

Source: Identification of
lichen substances (A
book)
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Cont..
 The present study investigates nineteen lichen species
belonging to eleven genera (Flavopunctelia, Flavoparmelia,
Cladonia, Parmelia, Umbilicaria, Xanthoria, Ochrolechia,
Hyperphyscia, Hypogymnia, Dermatocarpon and
Parmotrema) of Himalayan region (Abbottabad) Pakistan. 
 Spot test results showed the presence of different lichen
substances (gyrophoric, lecanoric acid, umbilicaric acids,
usnic acid, atranorin, chloroatranorin, salazinic acid and
parietin)
(Hussain et al 2019)
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Methods for determining Lichen

Secondary Metabolites:
Different methods are used for identification
of lichen compounds:
 Paper chromatography:

One dimensional
the development of Two dimensional
chromatogram in only one development of
direction is quiet chromatogram using
satisfactory for the another solvent at right
separation of many angle to the first occurs
mixtures.
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Classic spot tests Micro crystallography

(10% aqueous KOH solution, aqueous Glycerol, ethanol and glacial acetic acids are
solution of NaOCl2 or and 0.5% alcoholic some of the chemicals that are used in
phenylene-diamine solution) combination to make the reagent.

GC-MS
Xanthones, anthraquinones, dibenzofurans,
terpenes and pulvinic acid derivatives which
lack thermolabile ester groups can be
studied by GCMS
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Cont...
 Thin layer Chromatography (TLC):
The best micro-chemical methods for the systematic
botanist due to its sensitivity, rapidity, general applicability
and simplicity. Lichen compounds identified using TLC
visualization and crystallography (Parizadeh et al 2017)
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Cont..
 High performance liquid chromatography (HPLC):
 used to identify and quantify characteristic substances in
oakmoss(a type of light green to green-black fungal
lichen) products.
 bonded reverse phase columns are used
 Samples are dissolved in methanol and injected into the
appropriate portion column through which an appropriate
solvent is passed under high pressure.
 UV detector
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Activities of Lichens Secondary

compounds

Antiviral,
Antimicrobial /
cytotoxic and Antioxidant
Antifungal
anti-tumour activity
activity
activity

Anti-herbivore
Antipyretic and
and insecticidal Allelopathy analgesic drugs
activity

Photoprotection Allergenic activity

 27

Antibacterial and Antifungal activity:

 Lichen metabolites such as altranorin, fumar-proto-
cetraric acid, gyrophoric acid, lecanoric acid, physodic
acid, static acid and usnic acid showed relatively
strong antimicrobial effects against bacteria and fungi,
among which were human, animal and plant
pathogens.

Parmelia sulcata Usnea hirta

(salzinic acid (Hirtusneanoside
secondary secondary
metabolite) metabolite)
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Antiviral activity:
 Anethyl acetate-soluble fraction of the crude
methanolic extract of Ramalina farinacea can be
used as a broad-spectrum antiviral agent against
RNA and DNA viruses.

Ramalina
farinacea
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 Usnic acid extracted from Ramalina celastri

showed specific antiviral activity against the
Junin virus (Arenaviridae), which cause
Argentine hemorrhagic fever in humans.
 Usnic acid:
 decreases proliferation of human breast cancer
cells
 decreases human lung cancer cells without any
DNA damage.
 used as strong hepatotoxic agent against
monogastric murine hepatocytes.

Ramalina
celastri
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Conti..
 Epanorin (EP) is a secondary metabolite of
the Acarospora lichenic species.
 Epanorin has:
 antimicrobial activity
 UV-absorption properties
 inhibits cancer cell growth

(Moreno et al 2019)

Acarospora lichenic specie
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Antioxidant activity:
 Antioxidants(Antioxidants are the substances which
interfere with the oxidation process by reacting with
free radicals, chelating free catalytic metals and also by
acting as oxygen scavengers.)
 Lichens synthesize secondary lichen metabolites
which show strong antioxidant activity.
 Some depsides, such as atranorin (extracted from
Placopsis sp.), divaricatic acid (extracted from
Protousnea malacea), depsidones, pannarin (extracted
from Psoroma pallidum) showed antioxidant activity.
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Conti..
 Methanol extracts of
Xanthoparmelia
camtschadalis and its
extracted lichen
compounds such as
salazinic acid, stictic
acid, and usnic acid
protected human
astrocytes from hydrogen
peroxide-induced damage
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Antiherbivour and insecticidal

activity:
 According to research, lichen metabolites are poisonous to
insects, snails, and nematodes. Lichen substances also exhibit
harmful effect on vertebrate herbivores. It was also reported
that caperatic acid and extracts of the lichens Flavoparmelia
baltimorensis and Xanthoparmelia cumberlandia exhibit
antiherbivore activities against the snail Pallifera varia.
Researchers studied the effect of some lichen substances
i.e. atranorin, pulvinic acid dilactone, calycin, parietin,
fumarprotocetraric, stictic, norstictic, salazinic, vulpinic,
rhizocarpic, and usnic acids affect on the growth and
development of the polyphageous insect Spodoptera littoralis
 34

Allelopathy:
 Lichen secondary metabolites affect the
growth and development of neighboring
lichens, mosses, algae and vascular plants, as
well as microorganisms and can function as
allelopathic agents called allelochemicals.
 It was reported that allelopathic compounds
are released into the environment and might
Growth
Growth of of mossess
mossess
influence photosynthesis, respiration, Hedwigia
Hedwigia ciliata
ciliata and
and
transpiration, protein and nucleic acid Anomodon
Anomodon attenu-atum
attenu-atum
synthesis, membrane ion transport, and and
and the
the liverwort
liverwort
permeability of other organisms. Porella
Porella platyphylla
platyphylla
found
found toto be
be inhibited
inhibited
by
by lichen
lichen Porpidia
Porpidia
albocaerulescens.
albocaerulescens.
 35

Antipyretic and Analgesic drugs:

 Relievepain effectively or reduce fever and
inflammation in various mammals.

Usnic acid, isolated from

Roccella montagnei,
showed significant anti-
inflammatory activity
and also acts as an
antipyretic agent against
lipopolysaccharide-
induced fever.
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Conti..
 Physciosporin, isolated from Pseudo-cyphellaria
coriacea ((Yang et al 2015), atranorin isolated from
Everniastrum vexans (Zhou et al 2017) identified as an
effective compound and showed significant inhibitory
activity in migration and invasion assays against human
lung cancer cells.

Pseudocyphellaria Everniastrum vexans

coriacea
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Cont..
 Sixteen compounds were isolated from lichen  Usnea
longissima (useanol, lecanorin, 3'-methylevernic acid,
evernic acid, barbatinic acid, dimethyldibenzofuran,
orcinol,, methyl orsellinate, methyl everninate,  2-
hydroxy-4-methoxy-3,6-dimethyl benzoic acid, ethyl
everninate,ethyl 2,4-dihydroxy
methylbenzoate.. etc)
(Paul et al 2019)

Usnea longissima
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Conclusion:
 Lichens usually are very slow growing organisms and
must be protected against lower and higher plants, the
lichen substances are active, protective and antibiotic
substances.
 Many lichens grow under extreme conditions
concerning change of temperature, humidity and
intensity of light. Recent studies has shown that plants
under stress conditions are able to synthesize so called
stress-metabolites and it is likely that lichens respond
to such extreme changes in their environment by the
synthesis of large amount of stress metabolites.
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 Aromatic lichen substances absorb UV light and

protect the algae against too intensive irradiation.
 Lichen substances affect the permeability of the
cell walls of phycobionts, and could play an
important role in symbiotic equilibrium.
 Certain aliphatic and aromatic lichen acids (e.g.
norstictic, iso-usnic and usnic acids) are strong
chelating agents and are essential for supplying the
lichen with minerals from the substrate
 Lichen substances act as antifeedents for insects
and other animals.
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References:
 Boustie, J., Grube, M., 2005. Lichens-a promising source of bioactive
secondary metabolites. Plant Genet Resour. 3 (2): 273-287.
 Dayan, F.E., Romagni, J.G., 2001. Lichens as a potential source of
pesticides. Outlook Pestic. 12: 229 – 232.
 Feuerer, T.,Hawksworth, D.L., 2007. Biodiversity of lichens, including a
world-wide Analysis of checklist data based on Takhtajan’s floristic regions.
Biodivers Conserv. 16: 85
 Lawrey JD (1986). Biological role of lichen- Bacillus subtilis, Proteus
vulgaris, Aeromonas substances. J Bryol. 89(2):111-122
 Muller K (2002). Pharmaceutically relevant metabolites from lichens.
Applied Microbiology and Biotechnology. 56: 9-16
 Ranković, B. and Kosanić, M., 2019. Lichens as a potential source of
bioactive secondary metabolites. In Lichen secondary metabolites (pp. 1-29)
 Rowe, J.G., Saenz, M.T., Garcia M.D., (1999). Some lichen products have
antimicrobial activity. Zeitschrift für Naturforschung. 54(7-8): 605-609.