Full text pdf - Tropical Bryology

Tropical Bryology 31: 33-42, 2010
Chemosystematics of selected liverworts collected in
Borneo
Agnieszka Ludwiczuk 1,2 & Yoshinori Asakawa 1
1
Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Yamashiro-cho; Tokushima 770-
8514, Japan (asakawa@ph.bunri-u.ac.jp)
2
Chair and Department of Pharmacognosy with Medicinal Plant Unit, Medical University, 1 Chodzki
Str., 20-093 Lublin, Poland (aludwiczuk@pharmacognosy.org)
Abstract: The GC/MS analysis of the volatile components present in diethyl ether extracts of 15
Marchantiophyta species collected in Borneo island indicated that each liverwort species, produce own
characteristic compounds. Most of the studied species elaborate a large quantity of sesquiterpenoids whereas
only a few synthesize monoterpenoids, diterpenoids and aromatic compounds. Sesquiterpenoids, such as
herbertanes, gymnomitranes, chiloscyphanes as well as eudesmane, germacrane and guaiane sesquiterpene
lactones, can be used as chemosystematics markers. Aromatic compounds like methyl benzoates with prenyl
ether group are characteristic of Trichocolea pluma. Diterpenoids belonging to labdane-, clerodane- and
fusicoccane-types are chemical markers of Pleurozia gigantea, while cembranes are characteristic for
Chandonanthus hirtellus and rearranged 7,8-secolabdane-type diterpenoids for Pallavicinia species.
Monoterpenoids, responsible for characteristic fragrance, occur mainly in thalloid liverworts belonging to the
order Marchantiales, here represented by Wiesnerella denudata and Dumortiera hirsuta.
Keywords: Borneo liverworts, chemosystematics, chemotypes, sesquiterpenoids, diterpenoids, aromatic
compounds
Introduction
Liverworts are diverse phylum of small, herbaceous,
terrestrial plants, estimated to comprise about 5,000
species (Crandall-Stotler et al. 2008). These spore
forming plants can grow in almost every available
habitat, most often in humid locations, although there
are desert and arctic species as well. Liverworts are
characterized by the presence of oil bodies, unique
organelles in which terpenoids and aromatic
compounds are accumulated (Flegel & Becker 2000;
Suire et al. 2000). Although the function of the oil
bodies is still controversial, these single-membranebound
organelles are restricted to liverworts and occur
in approximately 90% of taxa (Shaw & Renzaglia
2004). Many different oil bodies types have been
described and/or illustrated, and sometimes used to
organize taxonomic units. Variations that occur in oil
body size, shape, color, number and distribution are
taxonomically informative characters of liverworts
(Schuster 1992a; 1992b; Crandall-Stotler & Stotler
2000). Terpenoids and aromatic metabolites, which
constitute the oil bodies of the Marchantiophyta, are
also of value for taxonomic investigations. A survey
of the scientific literature allows us to find papers
concerning chemosystematics of liverworts in general
(Asakawa 1982; 1995; 2004) as well as
TROPICAL BRYOLOGY 31 (2010)
chemosystematics of liverworts families or genera
(Gradstein et al. 1985; Asakawa et al. 2000; Rycroft
et al. 2001; Rycroft 2003). Recently we reported that
the Greek Fossombronia angulosa (Dicks.) Raddi.
and Tahitian Chandonanthus hirtellus (Web.) Mitt.
produce exactly the same acetogenins as those found
in the brown algae, Dictyopteris species (Ludwiczuk
et al. 2008; 2009). The chemical identity of certain
liverworts with brown algae supports the position of
liverworts as the basal most group of extant land
plants, as has been shown by molecular analyses, e.g.
Qiu et al. (2006).
The southern hemisphere together with tropic region
is characterized by extraordinarily high liverwort
diversity. This area has more endemic liverwort
species than the northern hemisphere (Inoue 1988).
Recent studies concerning biodiversity hotspots have
shown that Madagascar and Sundaland (which
include Borneo island) together with Philippines have
been identified as the leaders among the eight hottest
hotspots in the world, while the Tropical Andes and
mentioned Sundaland are the top hotspots in terms of
endemic plants (Myers et al. 2000). In case of the
liverworts, New Zealand has a greater density of those
spore forming plants than any other country. There
are 606 species known in New Zealand and about half

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LUDWICZUK & ASAKAWA: CHEMOSYSTEMATICS OF SELECTED LIVERWORTS
of them are endemic (Engel & Gleny, 2008). In
tropical America there are an estimated 1350
liverworts species in 188 genera and 41 families
(Gradstein et al., 2001). On the other hand in Europe
there are 490 liverworts species and only 4 endemic
(Söderström & Séneca 2008). These differences could
be explained by different histories of both Earth
hemispheres. In the northern hemisphere the supercontinent
of Laurasia split into North America and
Eurasia which are relatively close today. East-west
winds could carry propagules and maintain at least
sporadic gene flow between distant populations. By
contrast the southern hemisphere history is dominated
by the break-up of Gondwana. This continent split
into Antarctica, Africa, India, Australasia and South
America, which move far apart from each other. A
gene flow between the different continents became
very difficult. From that stage the bryophytes in the
different Gondwanan continents evolved largely
independently of each other (Frahm 2008; Kürschner
2008).
Borneo is the third largest island in the world. Most of
the island is covered with dense tropical rainforest.
The Borneo rainforest is one of the most biologically
diverse habitats on Earth possessing extremely high
numbers of endemic plant species, including
liverworts. From the viewpoint of the differentiation
of the bryophytes species chemical analysis of the
liverworts growing in this region is very interesting.
In this paper, we report the volatile components of
selected liverworts collected in Borneo, with the focus
of their chemosystematics.
Materials and Methods
Plant material. 15 liverworts species belonging to the
Jungermanniales and the Marchantiales were
collected in Sabah, Borneo (East Malaysia) on May
1988. There are: Trichocolea pluma (Reinw., Blume
& Nees) Mont., Frullania serrata Gottsche, Lindb. &
Nees, Lepidozia borneensis Steph. and L. fauriana
Steph., Chandonanthus hirtellus (Web.) Mitt.,
Mastigophora diclados (Brid. ex F. Weber) Nees,
Pleurozia gigantea (F. Weber) Lindb., Scapania
javanica Gottsche, Heteroscyphus aselliformis
(Reinw., Blume & Nees) Schiffn., Dumortiera hirsuta
(Sw.) Nees, Wiesnerella denudata (Mitt.) Steph.,
Bazzania harpago (De Not.) Schiffn., B. spiralis
(Reinw., Blume & Nees) Meijer, B. praerupta
(Reinw., Blume & Nees) Trevis and an unidentified
Pallavicinia Gray species. All liverworts have been
identified by Dr. Sinske Hattori (Hattori Botanical
Laboratory, Miyazaki, Japan) and deposited in the
Faculty of Pharmaceutical Sciences, Tokushima Bunri
University, Japan.
Extraction and analysis. Each liverwort, after being
air-dried, was extracted with diethyl ether (Et2O) for
two weeks. Each extract was filtered through a short
glass column packed with silica gel (240-400 mesh)
and the solvent evaporated to give crude extract which
was analyzed by TLC and GC/MS. For thin-layer
chromatography (TLC) a precoated silica gel glass
plates (0.25 mm) F254 and n-hexane–EtOAc (1:1 and
1:4) as mobile phase have been used. Spots were
visualized by spraying with Godin reagent (Godin
1954) and heated at 100-110 o C. Gas chromatographymass
spectrometry (GC/MS) was performed on 1%
SE-30 glass column (2m x 2mm). Oven temperature:
50 o C with 3 min initial hold, and then to 250 o C,
temperature programmed at 5 o /min. and 10 min. at
250 o C. Injection temperature was 260 o C and helium
(30ml/min) was used as a carrier gas. The detector
was operated in electron impact mode (70eV with 3
scans/s and mass range m/z 40-500) at 270 o C.
Identification of components. Identification of each
peak appeared on gas chromatogram was carried out
by comparison of retention times and mass spectra of
those of authentic samples, reported mass spectra
(Joulain & König 1998; König et al. 2008 -
MassFinder 4.0) and our library databases.
Results & Discussion
In continuing our chemosystematics studies of the
Marchantiophyta we investigated 15 samples of
liverwort species collected in Malaysian part of
Borneo. The classification of investigated liverworts
according to Crandall-Stotler et al. (2008) is presented
in Table 1. Table 2 shows the distribution of detected
terpenoids and aromatic compounds in examined
Borneo liverworts.
The genus Trichocolea, which belongs to the order
Jungermanniales, is of nearly worldwide distribution.
It is best represented in the tropics while only a single
species is known from the North Temperate Zone
(Hatcher 1958). Some of the Trichocolea species
collected in Japan, Taiwan, New Zealand and Europe
have been already investigated chemically. Methyl
benzoates with prenyl ether group, diterpenoids and
flavonoids were detected in these liverworts species
(Asakawa et al. 1981; Perry et al. 1996; Lorimer et
al., 1997; Baek et al. 1998; Nagashima et al. 2003).
Methyl benzoates with prenyl ether group were
detected in all studied specimens and these
compounds are significant chemical markers of
Trichocolea genus. Trichocolea pluma (Reinw.,
Blume & Nees) Mont. investigated here also produces
these compounds. Trichocolein (1), tomentellin (2)
and isotomentellin (3) together with vanillic acid
methyl ester (4) have been detected in this liverwort.
Recently, compound 4 and prenylated mehyl
benzoates have been isolated from Tahitian specimen
(Ludwiczuk et al. 2009). Beside trichocolein (1), in
the Taiwanese T. pluma labdane-type diterpenene
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LUDWICZUK & ASAKAWA: CHEMOSYSTEMATICS OF SELECTED LIVERWORTS 35
Table 1. Classification of the analyzed Borneo liverworts according to Crandall-Stotler et al. (2008)
Code
Class: Marchantiopsida Gonquist, Takht & W. Zimm. 1
Subclass: Marchantiidae Engl.
Order: Marchantiales Limpr.
Family: Wiesnerellaceae Inoue; Wiesnerella denudata (Mitt.) Steph.
Dumortieraceae D.G. Long; Dumortiera hirsuta (Sw.) Nees
Class: Jungermanniopsida Stotler & Crand.-Stotl. 2
Subclass: Pelliidae He-Nygrén, Juslén, Ahonen, Glenny & Piippo
Order: Pallaviciniales W. Frey & M. Stech
Suborder: Pallaviciniineae R.M. Schust.
Family: Pallaviciniaceae Mig.; Pallavicinia Gray a
Subclass: Metzgeriidae Barthol.-Began
Order: Pleuroziales Schljakov
Family: Pleuroziaceae Müll. Frib.; Pleurozia gigantea (F. Weber) Lindb. b
Subclass: Jungermanniidae Engl.
Order: Porellales Schljakov
Suborder: Jubulineae Müll. Frib.
Family: Frullaniaceae Lorch; Frullania serrata Gottsche, Lindb. & Nees c
Order: Jungermanniales H. Klinggr.
Suborder: Lophocoleineae Schljakov
Family: Trichocoleaceae Nakai; Trichocolea pluma (Reinw., Blume & Nees) Mont.
Mastigophoraceae R.M. Schust; Mastigophora diclados (Brid. ex F. Weber) Nees
Lepidoziaceae Limpr.; Bazzania harpago (De Not.) Schiffn.
Bazzania spiralis (Reinw., Blume & Nees) Meijer
Bazzania praerupta (Reinw., Blume & Nees) Trevis
Lepidozia borneensis Steph.
Lepidozia fauriana Steph.
Lophocoleaceae Vanden Berghen; Heteroscyphus aselliformis (Reinw., Blume &
Nees) Schiffn.
Suborder: Cephaloziineae Schljakov
Family: Scapaniaceae Mig.; Chandonanthus hirtellus (Web.) Mitt.
Scapania javanica Gottsche
TROPICAL BRYOLOGY 31 (2010)
a
b
d
e
f 1
f 2
f 3
f 4
f 5
g
h 1
h 2

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LUDWICZUK & ASAKAWA: CHEMOSYSTEMATICS OF SELECTED LIVERWORTS
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LUDWICZUK & ASAKAWA: CHEMOSYSTEMATICS OF SELECTED LIVERWORTS
Table 2. Distribution of terpenoids and aromatic compounds in the studied Borneo liverworts. Wiesnerella
denudata (Wd), Dumortiera hirsuta (Dh), an unidentified Pallavicinia species (Psp), Pleurozia gigantea (Pg),
Frullania serrata (Fs), Trichocolea pluma (Tp), Mastigophora diclados (Md), Bazzania harpago (Bh); Bazania
spiralis (Bs); Bazzania praerupta (Bp), Lepidozia borneensis (Lb), Lepidozia fauriana (Lf), Heterescyphus
aselliformis (Ha), Chandonanthus hirtellus (Ch) and Scapania javanica (Sj). * code in Table 1
Liverwort Class & Family* 1a 1b 2a 2b 2c 2d 2e 2f1 2f2 2f3 2f4 2f5 2g 2h1 2h2
Compounds
Species
TROPICAL BRYOLOGY 31 (2010)
Wd Dh Psp Pg Fs Tp Md Bh Bs Bp Lb Lf Ha Ch Sj
Trichocolein (1) +
Tomentellin (2) +
Isotomentellin (3) +
Vanillic acid methyl ester (4) +
Frullanolide (5) +
Dihydrofrullanolide (6) +
Tulipinolide (7) + +
Dihydrotulipinolide (8) + +
-Elemene (9) + + +
Eremophilene (10) +
Spathulenol (11) + + + + + + +
-Barbatene (12) + + +
Gymnomitr-8(12)-en-9-one (13) +
Gymnomitr-8-en-12-ol (14) +
Gymnomitr-8(12),9-diene (15) +
Gymnomitr-8(12)-en-9-ol (16) +
Drimenol (17) +
Albicanol (18) +
ent--Selinene (19) + +
ent-8-Hydroxyeudesm-3-one (20) +
Anastreptene (21) + + +
Trinoranastreptene (22) +
-Bazzanene (23) +
Isobazzanene (24) +
Cuparene (25) + +
11,12-Dihydrochiloscyphone (26) +
Dihydrochiloscypholone (27) +
Eudesm-3-en-7-ol (28) +
Kolavelool (29) +
Fusicogigantone A (30) + +
2,5-Fusicoccadiene (31) +
Pleuroziol (32) +
8-epi-Sclareol (33) +
Herbertene (34) +
-Herbertenol (35) +
-Herbertenol (36) +
Herbertene-2,3-diol (37) +
Herbertene-1,2-diol (38) +
Isochandonanthone (39) +
Pallavicinin (40) +
Cubebol (41) +
-Maaliene (42) +
Cadina-3,5-diene (43) +
Bicyclosesquiphellandrene (44) +
Bornyl acetate (45) +
Borneol (46) +
Limonene (47) + + + +
Costunolide (48) +
8-Acetoxyzaluzanin D (49) +
Dumhirone A (50) +
37

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LUDWICZUK & ASAKAWA: CHEMOSYSTEMATICS OF SELECTED LIVERWORTS
alcohol, (12E)-3-hydroxylabda-8(17),12,14-triene
has also been detected by Chang & Wu (1987). We
did not confirm the presence of diterpenoids in the
specimen collected in Borneo island. Among other
Trichocolea species so far investigated diterpenoids
(isopimarane-type) have been detected only in the
New Zealand T. mollissima (Hook f & Taylor)
Gottsche (Lorimer et al. 1997; Nagashima et al.
2003).
The Borneo Frullania serrata Gottsche, Lindb. &
Nees like many other Frullania species belonging to
chemotype I (sesquiterpene lactone-type) produces
eudesmane-type sesquiterpene lactones. All of these
compounds, which possess an -methylene-butyrolactone
group, are allergy-inducing substances,
while -methyl--butyrolactones do not cause allergy
(Asakawa 1995; 2008; Asakawa et al. 2009).
Frullanolide (5) and dihydrofrullanolide (6) have been
detected in Frullania serrata along with germacranetype
lactones, tulipinolide (7) and dihydrotulipinolide
(8). The presence of germacrane-type sesquiterpene
lactones in this leafy liverwort is quite interesting,
since these compounds are chemical markers of
Wiesnerella denudata (Mitt.) Steph. placed in the
order Marchantiales. However germacranolide,
costunolide (48) have been previously detected in F.
monocera (Hook f & Taylor) Gottsche, Lindb. &
Nees, F. tamarisci (L.) Dum. ssp. tamarisci and F.
tamarisci ssp. nisquallensis (Sull.) Hatt. (Asakawa
1982; 1995). It is worth to mention that the presence
of compounds 7, 8 together with 48 have been also
confirmed by Asakawa et al. (1991) in different
Malaysian collection of Frullania serrata. Other
sesquiterpenoids, such as pentalene, silphin-1-ene, elemene
(9), eremophilene (10) and spathulenol (11)
have been also detected as minor components.
Bazzania species are known as rich sources of many
kinds of sesquiterpenoids (Asakawa 1982; 1995;
2004). The volatile components of three Bazzania
species from Borneo have been investigated.
Received results indicate that these species are not
related chemically to one another as each of studied
liverworts produce different compounds.
Gymnomitranes (=barbatanes) are characteristic of
Bazzania harpago (De Not.) Schiffn., eudesmanes of
B. spiralis (Reinw., Blume & Nees) Meijer and
drimanes of B. praerupta (Reinw., Blume & Nees)
Trevis.
B. harpago here investigated produces -barbatene
(12), gymnomitr-8(12)-en-9-one (13), gymnomitr-8en-12-ol
(14), gymnomitr-8(12),9-diene (15) and
gymnomitr-8(12)-en-9-ol (16). Gymnomitrane-type
sesquiterpenoids, among others components, have
been also found in the Japanese liverworts, B.
pompeana (Sande Lac.) Mitt. (Matsuo 1982) and B.
fauriana (Toyota & Asakawa 1988). Drimenol (17)
and albicanol (18) are characteristic compounds for
Borneo B. praerupta. These two compounds were
detected in this liverwort among other like limonene
(47), anastreptene (21), trinoranastreptene (22), ent-selinene
(19) and spahulenol (11). Drimanes,
especially drimane-type sesquiterpene esters are
characteristic components of Japanese B. fauriana
(Toyota & Asakawa 1988) and B. japonica (Sande
Lac.) Lindb. (Asakawa 1995) and also of Malagasy B.
decrescens (Lehm. & Lindenb.) Trevis.
(Harinantenaina & Asakawa 2007). Drimenol (17)
along with gymnomitranes, bazzananes and
calamenanes have been also detected in Japanese B.
trilobata (L.) Gray (Huneck et al. 1984). B. spiralis
collected in Borneo produces ent-eudesmane-type
sesquiterpenoids, ent--selinene (19) and ent-8hydroxyeudesm-3,11-diene
(20). Interestingly, the
Japanese B. fauriana elaborates 6-hydroxyeudesm-
3-one, whose configuration is opposite to that of
eudesmanes found in B. spiralis, but identical to that
of many eudesmanes found in higher plants (Toyota
& Asakawa 1988; Kondo et al. 1990). Besides 19 and
20, B. spiralis also produces -elemene (9), cubebene
and spathulenol (11), but neither
gymnomitranes nor drimanes.
In the ether extract of Lepidozia borneensis Steph. the
following sesquiterpenoids were identified: bazzanene
(23), isobazzanene (24), cuparene (25), chamigrene,
-bourbonene and guaia-6,9-dien-4-ol
among which -bazzanene (23) is the major
component. It is noteworthy that bazzananes and
cuparanes are widespread in Bazzania species
(Asakawa 1995), which belong to the same family as
Lepidozia, the Lepidoziaceae. In comparison to L.
borneensis Steph., the Borneo Lepidozia fauriana
Steph. biosynthesizes chiloscyphane-type
sesquiterpenoids; 11,12-dihydrochiloscyphone (26)
and dihydrochiloscypholone (27), which were
previously detected in the Taiwanese specimen (Paul
et al. 2001). Apart from Lepidozia species,
chiloscyphanes have been also found in the Japanese
liverworts, Jungermannia vulcanicola (Schiffn.)
Steph. and Chiloscyphus polyanthos (L.) Corda
(Asakawa 2004). The Borneo Lepidozia fauriana also
produces eudesm-3-en-7-ol (28). The eudesmanetype
sesquiterpenoids, including the mentioned
compound 28, have previously been isolated from L.
vitrea Steph. of Japanese origin (Toyota et al. 1996a).
Interestingly, compound 28 is also biosynthesized by
the Japanese Chiloscyphus polyanthus (Toyota et al.
1999). The results indicated that some Lepidozia
species: L. fauriana and L. vitrea (Lepidoziaceae) are
chemically very closely related to the Chiloscyphus
species (Lophocoleaceae).
Both Heteroscyphus and Chiloscyphus, belong to the
family Lophocoleaceae (Crandall-Stotler et al. 2008),
but there is no chemical affinity between
Heteroscypus species and the other genus of this
family, which were investigated chemically (Asakawa
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LUDWICZUK & ASAKAWA: CHEMOSYSTEMATICS OF SELECTED LIVERWORTS 39
2004). Genus Heteroscyphus is known as a rich
source of diterpenoids. Previously investigated H.
planus (Mitt.) Schiffn. of Japanese origin
biosynthesized ent-clerodane- and cyathane-type
diterpenoids together with ent-2,3secoaromadendrene-
and calamenene-type
sesquiterpenoids (Hashimoto et al., 1995). ent-
Clerodanes have also been isolated from the other
Japanese species, H. coalitus (Hook. f.) Schiffn.
together with halimanes and aromadendrane-type
sesquiterpenoids (Toyota et al. 1996b). H. aselliformis
(Reinw. Blume & Nees) Schiffn. originated from
Borneo produces mainly sesquiterpenoids.
Anastreptene (21), cuparene (25) and-elemene (9)
have been detected.
The genus Pleurozia (Pleuroziaceae), formerly
classified in the leafy liverworts (order
Jungermanniales), in the new classification of the
Marchantiophyta is placed into the order Pleuroziales
within subclass Metzgeriidae (simple thalloid
liverworts) (see Table 1) (Crandall-Stotler et al.
2008). The East Malaysian Pleurozia gigantea (F.
Weber) Lindb. produces mainly diterpenoids. In the
diethyl ether extract only one sesquiterpenoid,
spathulenol (11) has been identified. The main
compound detected in this liverwort is kolavelool
(29), which is a member of clerodane-type
diterpenoids, and also occurs in Jungermannia species
(Asakawa 2004). Beside clerodanes P. gigantea also
produces fusicoccane- and labdane-type diterpenoids.
Fusicogigantone A (30), 2,5-fusicoccadiene (31),
pleuroziol (32) and 8-epi-sclareol (33) have been
detected in ether extract of this liverwort. The present
data are in accordance with those, reported previously
by Asakawa et al. (1990), except for the absence of
dolabellane-type diterpenoids in the present
collection.
The members of the genus Mastigophora
(Mastigophoraceae) are chemically very closely
related to the Herbertus (Herbertaceae) species, since
both produce herbertane-type sesquiterpenoids as
major constituents (Asakawa 1995; 2004;
Harinantenaina & Asakawa 2007). GC/MS
examination of the ether extract of the Borneo M.
diclados (Brid. Ex F. Weber) Nees showed the
presence of herbertene (34), -herbertenol (35), herbertenol
(36), herbertene-2,3-diol (37) and
herbertene-1,2-diol (38). In a previous collection of
the East Malaysian M. diclados, apart from
herbertanes 34-38, herbertane dimers,
mastigiphorenes A-D have been found (Asakawa et
al., 1991). However, the West Malaysian species does
not produce any herbertanes; instead trachylobanetype
diterpenoids have been isolated (Leong &
Harrison, 1997). A Japanese collection elaborates
herbertene (34) and -herbertenol (35) together with
cyclic chlorinated bis-bibenzyls, whereas no
diterpenoids and herbertane dimers have been
TROPICAL BRYOLOGY 31 (2010)
detected (Hashimoto et al., 2000). These data point
out that there are at least three geographical races of
M. diclados in Asia; bis-bibenzyl-type in Japan,
mastigophorene-type in Borneo (East Malaysia), and
pimarane and pimarane-derived trachylobane
diterpenoid type in Taiwan and West Malaysia.
Interestingly, all three types have been found in
Malagasy M. diclados (Harinantenaina & Asakawa,
2004).
Many of the previously investigated Scapania species
produce various kinds of sesquiterpenoids which are
ubiquitous in other liverworts (Asakawa 1982; 1995;
2004). The most common sesquiterpene in all
investigated species is anastreptene (21). Among
volatile components occurring in ether extract of
Scapania javanica Gottsche collected in Borneo, barbatene
(12) and spathulenol (11) have been
identified. However, the presence of anastreptene (12)
has not been confirmed. This compound has been
detected in Chandonathus hirtellus (Web.) Mitt.
among other constituents like limonene (47),
sesquisabinene, spathulenol (11), fusicogigantone A
(30) and isochandonanthone (39). The genus
Chandonanthus is known to produce cembrane-type
diterpenoids, which are significant chemical markers
of this liverwort (Asakawa 1995, 2004; Shy et al.
2002). Chandonanthus hirtellus originated from
Borneo elaborates isochandonanthone (39), which has
previously been detected in a Chinese collection (Shy
et al. 2002). Interestingly, the Borneo C. hirtellus also
produces a fusicoccane-type diterpenoid,
fusicogigantone A (30). Apart from this collection,
fusicoccanes have been detected only in Tahitian C.
hirtellus (Ludwiczuk et al. 2009). This is the second
report of such compounds in Chandonanthus
hirtellus, however, dolabellanes, which are
biogenetically related to fusicoccanes have been
isolated from West Malaysian species (Wang et al.
2009).
Pallavicinia species are simple thalloid liverworts.
The chemical analysis of several Pallavicinia species
afforded a number of common sesquiterpenoids,
sterols and carbohydrates (Asakawa 1982; 1995). The
unidentified Pallavicinia species collected in Borneo
elaborates sesquiterpenoids. Cubebol (41), -maaliene
(42), cadina-3,5-diene (43) and bicyclosesquiphellandrene
(44) have been detected. Among volatile
components pallavicinin (40), the rearranged 7,8secolabdanoid
has also been identified. This
compound has been previously isolated from Japanese
and Taiwanese P. subciliata (Aust.) Steph. and
Chinese P. ambigua (Mitt.) Steph. (Wu et al., 1994;
Toyota et al., 1998; Liu & Wu, 1999; Li et al., 2005).
The investigated Borneo Pallavicinia species is
closely related chemically to P. subciliata and P.
ambigua.

40
LUDWICZUK & ASAKAWA: CHEMOSYSTEMATICS OF SELECTED LIVERWORTS
The main compound of Wiesnerella denudata (Mitt.)
Steph. collected in Borneo is bornyl acetate (45) in
contradiction to the same specimen collected in Japan,
which produce a large amount of neryl acetate
(Ludwiczuk et al. 2008). The Borneo W. denudata
also produces borneol (46) and limonene (47). All of
the compounds mentioned above are responsible for
unique fragrant odor of W. denudata, whose chemical
profile is almost identical to that of Conocephalum
conicum (L.) Dum. belonging to bornyl acetate
chemotype (Toyota et al. 1997). Beside
monoterpenoids the Borneo W. denudata elaborates
germacrane- and guaiane-type sesquiterpenoids.
Costunolide (48), tulipinolide (7), dihydrotulipinolide
(8) and 8-acetoxyzaluzanin D (49) have been
detected in ether extract. There are three different
chemical races of W. denudata: I – costunolideguaianolide
type, II – costunolide type and III –
guaianolide type (Asakawa 2004). The present
specimen belongs to type I in comparison to the
Japanese specimen, which is placed in type III
because of lacking of costunolide (Ludwiczuk et al.
2008).
In addition to limonene (47), -curcumene, barbatene
(12) and spathulenol (11), the Borneo
Dumortiera hirsuta (Sw.) Nees produces dumhirone
A (50) (phenylethyl cyclohexadienone) previously
reported by Xie et al (2007) in a Chinese specimen. It
is known that some of the Dumortiera species
biosynthesized lunularin (Gorham 1977; Asakawa
1995), which could be precursor of dumhirone A. In
contrast, D. hirsuta from Argentina and Japan
produce dumortane- and germacrane-type
sesquiterpenoids as the main components,
respectively (Bardon et al. 1999; Ludwiczuk et al.
2008). These data indicated that there are several
chemotypes of D. hirsuta.
Since the classification of the Matchantiophyta
especially that of leafy liverworts is extremely
difficult, studies of their chemical constituents are
necessary from a chemosystematics point of view.
Many liverworts produce their own peculiar
constituents and some of them such as the
sacculatane- and pinguisane-type terpenoids have not
been found in higher plants, fungi or marine
organisms (Asakawa 1982; 1995). The pattern of
terpenoids and aromatic compounds depends on the
type of environment in which each liverwort plant
normally lives. Sometimes liverwort species or genus
collected from different locations produces
compounds characteristic only for one environment
e.g. the Taiwanese Trichocolea pluma, which
produces labdane diterpene alcohol (Chand & Wu
1987), and this compound does not occur in Tahitian
and Borneo specimens. Despite these differences all
mentioned specimens produce characteristic
prenylated methyl benzoates, which are significant
chemical markers of Trichocoleaceae family. There
are many more such examples among liverworts
(Asakawa 2004). It is also worth to mention about
cases when the analysis showed the significant
differences in composition within one liverwort
species. The good example is Conocephalum
conicum. The analysis of the flavonoids composition
of the European and American collections showed the
occurrence of the geographical races of this liverwort
(Markham et al. 1976). The differences in the
composition of the volatile components of the
Japanese C. conicum from different collections sites
provided to description of three chemically different
types of C. conicum in Japan (Toyota et al. 1997).
However, these differences are in agreement with
molecular analysis, which have shown that C.
conicum is a complex of a few cryptic species
(Odrzykoski & Szweykowski 1991).
GC/MS analysis of the ether extract of several
Marchantiophyta species collected in Borneo
indicated that each liverwort species, genus or family
produce own characteristic compounds which could
be used as taxonomic indicators. The present
liverworts are rich sources of sesquiterpenoids with a
variety of carbon skeletons, whereas only a few
monoterpenoids, diterpenoids and aromatic
compounds have been found.
Among sesquiterpenoids, the Borneo liverworts
produce mainly eudesmanes, drimanes, herbertanes,
gymnomitranes, aromadendranes, cuparanes,
chiloscyphanes, germacranes, guaianes, bazzananes
and elemanes. Some of them can be used as chemical
markers, like herbertanes for Mastigophora diclados,
gymnomitranes for Bazania harpago, eudesmanolides
for Frullania serrata, and germacrane- and guaianetype
sesquiterpene lactones for Wiesnerella denudata.
Among other terpenoids identified in the Borneo
liverworts, the monoterpenoids occurred in thalloid
liverworts belonging to order Marchantiales are
responsible for characteristic fragrance. Diterpenoids
and aromatic compounds are also important from a
chemosystematics point of view. Diterpenoids
belonging to ladane-, clerodane- and fusicoccanetypes
are chemical markers of Pleurozia gigantea
while cembranoids are characteristic for
Chandonanthus hirtellus and rearranged
secolabdanoids for some Pallavicinia species. Methyl
benzoates with prenyl ether group are significant
chemical markers of the Trichocoleaceae
(Trichocolea pluma).
Acknowledgements: The authors thank Dr. S. Hattori, The
Hattori Botanical Laboratory, Miyazaki, Japan for his
identification of the liverworts. This work was supported in
part by Grant-in-Aid for Open Research and by Grant-in-
Aid for Senryaku Research (to A.L.) from the Ministry of
Education, Culture, Sports, Science and Technology, Japan.
TROPICAL BRYOLOGY 31 (2010)

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