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ISSN: 1412-033X
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Journal of Biological Diversity

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B I O D I V E R S IT A S
Volume 13, Number 2, April 2012
Pages: 53-58
ISSN: 1412-033X
E-ISSN: 2085-4722
The conservation of mitochondrial genome sequence in Leucadendron

(Proteaceae)
1
MADE PHARMAWATI1, GUIJUN YAN2, PATRICK M. FINNEGAN2
Department of Biology, Faculty of Mathematic and Natural Sciences, Udayana University, Kampus Bukit Jimbaran, Bali, Indonesia.
Tel.: (0361) 703137, Fax.: (0361) 703137, email: pharmawati@hotmail.com
2
School of Plant Biology, Faculty of Natural and Agricultural Sciences, The University of Western Australia, 35 Stirling Highway, Crawley WA,
6009, Australia
Manuscript received: 26 February 2012. Revision accepted: 27 March 2012.
ABSTRACT
Pharmawati M, Yan G, Finnegan PM. 2012. The conservation of mitochondrial genome sequence in Leucadendron (Proteaceae).
Biodiversitas 13: 00-00. Mitochondrial DNA (mtDNA) is useful for developing molecular markers and for studying plant phylogeny.
However, its usefulness depends on the degree of detectable sequence variation. In seven species of the genus Leucadendron, PCRRFLP failed to reveal any polymorphisms in seven separate regions of the mtDNA. Sixty-two primer pair - enzyme combinations were
used to assay at least 248 restriction sites, resulting in the direct sampling of a minimum of 992 bp across 17,500 bp of mt DNA. The
highly conserved nature of the mtDNA sequence in the genus Leucadendron was confirmed by the absence of sequence variation in the
1434 bp mtDNA nad1/B-C intron across these species. Mitochondrial DNA sequences are more highly conserved than the chloroplast
DNA sequences in Leucadendron and the mtDNA sequences in many other plant genera. Phylogenetic analysis using this intron
sequence was consistent with other phylogenetic analyses in regard to the position of Proteaceae.
Key words: Leucadendron, mtDNA, nad1/B-C, PCR-RFLP
INTRODUCTION
Leucadendron is a genus of South African Proteaceae
that comprises 85 species and 11 subspecies (Williams
1972). Members of the genus have been classified into two
sections (Alatosperma and Leucadendron) by Williams
(1972) based on fruit characteristics. Leucadendron are
successfully cultivated in a number of countries including
the United States of America, Australia and New Zealand.
In Australia, these plants are popular garden plants and are
grown commercially for both the domestic and export cutflower industries. The availability of molecular markers for
assessing variation and relatedness of Leucadendron
species would greatly aid the development of new hybrids.
A thorough understanding on the phylogenetic
relationships within Leucadendron requires information
from all three genomes. Leucadendron phylogeny has been
inferred from sequence analysis of the internal transcribed
spacer (ITS) regions of the nuclear rRNA genes (Barker et
al. 2004). Chloroplast DNA (cpDNA) variation has also
been used to evaluate interspecific relationships in
Leucadendron (Pharmawati et al. 2004). Both ITS- and
cpDNA- derived phylogenies disagree substantially with
the previous morphological classification of Leucadendron
species (Williams 1972). The phylogenies based on
molecular evidence also differ from one another. To further
define the phylogeny of Leucadendron, information from
mtDNA would be beneficial.
The use of cytoplasmic DNA sequences as breeding,

phylogenetic or phylogeographic markers requires
knowledge of the mode of inheritance and the level of
sequence variation. In the genus Leucadendron, the
chloroplast genome is maternally inherited and contains
useful amounts of sequence variation (Pharmawati et al.
2004). In contrast, the mode of inheritance of mtDNA in
Leucadendron, as well as the phylogenetic information
contained within it, has not yet been examined.
In this study, universal primers specific to land plant
mitochondrial genomes (Demesure 1995; Dumolin 1997)
were tested for their ability to amplify specific mtDNA
fragments from seven Leucadendron species. The resulting
fragments were subjected to RFLP analysis to evaluate the
applicability of this method to the detection of mtDNA
sequence variation in Leucadendron.
MATERIALS AND METHODS
Plant material
Leaf tissue from L. discolor Buek ex Meisn, L.
eucalyptifolium Buex ex Meisn, L. gandogeri Schinz ex
Gandoger, L. laureolum (Lam.) Fourcade, L. procerum
(Salisb. ex Knight) Williams, L. salignum Berg and L.
uliginosum R.Br were collected from the living collection
of the Leucadendron Breeding Program, University of
Western Australia (Perth, Australia). Voucher specimens
were lodged to Australian National Herbarium, Centre for
54
B I O D I V E R S IT A S 13 (2): 53-58, April 2012
Australian National Biodiversity Research with voucher

number CANB668317, CANB668319, CANB668324,
CANB668332, CANB668337, CANB668342 respectively
to samples stated above.
DNA extraction and electrophoresis
DNA was extracted from 0.1 g of leaf tissue using a
commercial kit (DNeasy plant mini kit, Qiagen, Clifton
Hill, VIC, Australia) according to manufacturers
instructions. Genomic DNA and DNA fragments were size
fractionated by electrophoresis on agarose gels and
visualized by ethidium bromide staining (Sambrook et al.
1989). To determine DNA concentration, the ethidium
bromide staining intensity of specific bands was compared
to that of a range of co-fractionated lambda DNA mass
standards (MBI, Fermentas, Hanover, MD, USA).
PCR-RFLP analysis
Each 25 L polymerase chain reaction (PCR) assay
contained 20 ng DNA template, 16 mM (NH4)2SO4, 67 mM
Tris-HCl, pH 8.8, 0.01% (v/v) Tween-20, 1.5 mM MgCl2,
0.3M each primer (Table 1, synthesized by Invitrogen
Life Technologies, Mount Waverley, VIC, Australia), 200
M each standard dNTP and 1 unit of Taq DNA
polymerase (Bioline, Alexandria, NSW, Australia). The
program for the thermal cycler (iCycler, Bio-Rad, Regents
Park, NSW, Australia) had an initial denaturation step of 4
min at 94oC, followed by 30 cycles of 45 sec at 94oC, 45
sec at 47oC to 59oC (depending on the primers used, Table
1) and 2 to 4 min (depending on the product length, Table
1) at 72oC, with a final extension step of 10 min at 72oC.
Aliquots of each amplification product were digested
singly with AluI, CfoI, HaeIII, MaeI, NdeII, RsaI, TaqI,
ThaI (Promega, Annandale, NSW, Australia), HindfI, MspI
or MvaI (Roche Diagnostic, Castle Hill, NSW, Australia),
but not every fragment was digested with every enzyme.
Digestion was for 3 h at 37oC in 10 l 1 x buffer (supplied
by enzyme manufacturer) containing 2 units of enzyme.
DNA sequencing and analysis

The PCR-amplified nad1/B-C intron was sequenced
directly with dideoxynucleotide chain termination
chemistry (BigDye v3.1, ABI, Western Australia) using
nad1 exon B-C primer pair (Table 1). The products were
separated and analysed by The West Australian Genome
Resource Centre. The sequences were deposited in
Genebank (Accession numbers L. discolor, DQ250042;
L. eucalyptifolium, DQ250043; L. gandogeri, DQ250044;
L. laureolum, DQ250045; L. procerum, DQ250046; L.
salignum, DE250047; L. uliginosum, DQ250048). The
sequencing data was assembled using AssemblyLIGN
(Accelrys, Sydney, Australia) and aligned using ClustalW
(MacVector 8.0, Accelrys) software. Multiple sequence
alignments generated with ClustalW were used to generate
phylogenetic trees using DNApars-PHYLIP (Felsenstein
1989).
Sequence diversity was calculated using MEGA4
(Tamura et al. 2007). For comparison, diversity of nad1/BC sequences from Actinidia species was calculated by
extracting sequences from GeneBank (AJ536471.1;
AJ536472.1; AJ536470.1; AJ536469.1; AJ536467.1;
AJ536468.1).
RESULTS AND DISCUSSION
Seven universal PCR primer pairs designed to amplify

mtDNA fragments from land plants (Demesure et al. 1995;
Domulin-Lapegue et al. 1997) were used in this study. The
mtDNA regions evaluated were six introns from the genes
encoding subunits 1, 4, 5 and 7 of NADH dehydrogenase
and the intergenic region between the rps14 and cob genes
(Table 1). After optimizing the reaction conditions, each
primer pair robustly amplified a single fragment (Table 1)
of 1,500 to 4,500 bp, depending on the primer pair, from
Leucadendron genomic DNA. Amplification products from
seven Leucadendron species using the nad7/2-3r mtDNA
primer pair are shown in Figure 1.
The
fragments
amplified
from
Leucadendron were similar in size to those
Table 1. Primers and primer pairs used in this study
amplified from other plants using these
primer pairs (Demesure et al. 1995;
Approximate
Annealing Extension
Dumolin-Lapegue et al. 1997; Vaillancourt
product size
Primer pair temperature Time
References
et al. 2004). The seven amplified fragments
in
this
study
(oC)a
(min)a
totaled about 17,500 bp of Leucadendron
(bp)
nad1 exon B
57.5
2
1700
Demesure et al. 1995
mtDNA, but no length polymorphisms were
nad1 exon C
detected among the seven species.
nad4 exon 1
59
2
2000
Intergenic and intronic regions were
nad4 exon 2
chosen for analysis because these regions of
rps14
59
2
1500
mtDNA are less constrained than their
cob
adjacent exons for both overall number of
nad5/1
57.5
3
2600
Dumolin-Lapegue et al.
substitutions per site and indels (Laroche et
nad5/2r
1997
al. 1997), and therefore demonstrate higher
nad1/4
47
3
3500
rates of polymorphism than the very well
nad1/5r
1997
nad7/2
57
2
1700
conserved exonic regions (Duminil et al.
nad7/3r
1997
2002). It was therefore surprising that
nad4/2c
53.5
4
4500
polymorphisms were not detected.
nad4/3r
Note: aDetermined empirically
1997
1 kb ladder
L. uliginosum
L. discolor
L. procerum
L. gandogeri
L. eucalyptifolium
L. salignum
L. laureolum
55
2000b
1p500b
p
Figure 1. Amplification of the PCR products from seven
Leucadendron species using the nad7/2-3r mtDNA primer pair.
The PCR products were separated on a 1.2% agarose gel
electrophoresis and stained with ethidium bromide. The lane
containing a 1 kb ladder (Promega) is indicated, as are the sizes of
selected marker fragments.
100 bp ladder
L. laureolum
L. salignum
L. eucalyptifolium
L. gandogeri
L. procerum
L. discolor
L. uliginosum
100 bp ladder
L. procerum
L. discolor
L. uliginosum
L. eucalyptifolium
L. gandogeri
L. salignum
L. laureolum
In an attempt to reveal mtDNA sequence polymerphisms across species, each PCR product was digested with
a battery of restriction endonucleases having four-base
recognition sequences. Sixty-two primer pair - enzyme
combinations were tested, revealing a total of 248
discernable restriction sites for each species, directly
assaying variation at 992 positions within the mtDNA
sequence of each species. Despite this level of sampling, no
sequence variations or length polymorphisms were
detected. The monomorphic profiles of the mtDNA
fragments obtained with the nad1 exon B-C - MvaI and
nad7/2-3r - HaeIII primer pair - enzyme combinations are
representative (Figure 2).
The extreme conservation of Leucadendron mtDNA
sequence is in sharp contrast to the cpDNA in this genus. In
the same species examined here, polymorphisms were
500bp
300bp
200bp
detected in cpDNA with 14 of 33 primer pair - enzyme

combinations when a sequence space of 16,800 bp was
sampled (Pharmawati et al., 2004). At least 120 restriction
sites representing 480 bp of cpDNA sequence were
evaluated across the 16,800 bp region, resulting in the
identification of seven restriction site polymorphisms and
33 indels. Across the seven species, this is a site
polymorphism rate of 1 per 480 bp assayed.
In contrast, at least 248 restriction sites, representing
992 bp of sequence across a sequence space of
approximately 17,500 bp, were examined in Leucadendron
mtDNA by PCR-RFLP. The lack of polymorphism
suggests that mtDNA sequences are highly conserved
among Leucadendron species, a conclusion that is
supported by no polymorphisms within the 1,434 bp
nad1/B-C sequence that was determined directly for all
seven species of this study. This contrasts strongly to the
situation found in other plants. For example, PCR-RFLP
analysis of mtDNA intronic regions has been used
successfully to detect interspecific polymorphisms in
Actinidia (Testolin and Cipriani, 1997), Elymus (Sun,
2002), Musa (Nwakanma et al. 2003), Vasconcellea (Van
Droogenbroeck et al. 2004) and Houttuynia (Wei et al.
2005), and even intraspecific polymorphisms in Quercus
robur (Dumolin-Lapegue et al. 1995), Actinidia deliciosa
(Testolin and Cipriani, 1997), Picea abies (Grivet et al.,
1999), Solanum tuberosum (Bastia et al., 2001), Eucalyptus
globulus (Vaillancourt et al. 2004) and Buchlo dactyloides
(Gulsen et al. 2005). In a study of eight genotypes of
Eucalyptus globulus, a mtDNA polymorphism was
detected within 7960 bp of sequence space after screening
only 36 primer pair-enzyme combinations (Vaillancourt et
al. 2004). The number of polymorphisms is expected to be
higher when mtDNA evaluation is done across species. For
500bp
300bp
200bp
Figure 2. PCR-RFLP patterns of mtDNA from seven Leucadendron species. Patterns were generated using the primer pair - enzyme
combinations of nad1/B-C and MvaI (A), and nad7/2-3r and HaeIII (B). The digestion products were separated on a 3% agarose gel
electrophoresis and stained with ethidium bromide. Sizes of selected fragments of the 100 bp ladder (Promega) are indicated.
Pages: 53-58
ISSN: 1412-033X
E-ISSN: 2085-4722
56
Pogonophora
Macaranga
Lasiocroton
Havea
Pimelodendron
Crinodendron
Sloanea
Brunellia
Citrullus latanus
Eucryphia
Balanops
Heewoodia
Coriaria
Laurus
Platanus
Liriodendron
Sabia
Meliosma
1000
979
Clusia
983
994
843
Acridocarpus
Thryallis
664
Centroplacus
Leucadendron Lamprocapnos
Daphniphyllum
Liquidambar
Dissiliaria
Podocalyx
Corylopsis
Vitis
Dillenia
Actinidia
Ixonanthes
Endospermum
Omphalea
Irvingia
Klainedoxa
Galearia
Croton
Neoscortechin
Durandea
Hugonia
Humiria
Vantanea
Citrullus
Dapania
Averrhoa
Suregada
Panda
Tetrorchidium
Figure 3. Phylogenetic relationships among plants derived from mtDNA NADH dehydrogenase subunit 1 intron 2 sequences. The most parsimonious unrooted tree for 50 plant genera, including
Leucadendron was constructed using DNApars-PHYLIP. Bootstrap values are indicated above the relevant branches based on 1000 reiterations
PHARMAWATI et al. Mitochondrial genome of Leucadendron
example, among 13 Elymus species analysed,

polymorphisms were detected within 1600 bp of mtDNA
sequence space with two of seven primer-enzyme
combinations tested (Sun, 2002). The complete inability to
detect site variation or indels among seven Leucadendron
species with 62 primer pair-enzyme combinations indicates
the extreme conservation of mtDNA sequence in this
genus. Since mtDNA polymorphisms were not found, the
inheritance of mtDNA could not be determined by
examining the progeny produced by interspecific crosses.
To further investigate the level of sequence
conservation in Leucadendron mtDNA, the DNA sequence
of the 1.5 kb fragment amplified using the nad1 exon B-C
primer pair was determined for each of the seven species.
All seven sequences were identical, demonstrating that the
mtDNA sequence is very highly conserved across
Leucadendron species. The Leucadendron sequence was
95% identical to that of the intron located between exons 2
and 3 of the mitochondrial NADH dehydrogenase subunit 1
gene of Lamprocapnos spectabilis (AY674682),
confirming the sequence identity of nad1/B-C. Comparison
of the Leucadendron and Lamprocapnos spectabilis
sequences revealed that the length of the intron in the seven
Leucadendron species tested was 1,434 bp. Each
Leucadendron nad1/B-C intron had characteristics typical
of plant group II introns. There was a GCGCG motif near
the 5 splicing site and a domain V sequence
(GAGCCACATGCAGGGAAACTTGCACGTGTGGTT)
near the 3 splicing site (Chapdelaine and Bonen, 1991).
Direct sequencing of the nad1/B-C intron showed
complete conservation among the seven Leucadendron
species examined. The nucleotide diversity () among
samples was 0 and the over all main distance between
samples was also 0. In contrast, the mtDNA nad1B/C
sequences from Actinidia species extracted from GeneBank
showed nucleotide diversity of 0.312 and the over all main
distance between samples was 0.313. The mtDNA nad1/BC intron showed sufficient sequence variation to
distinguish among the Cucurbita species and allow the
construction of a phylogenetic tree (Sanjur et al. 2002).
Moreover, intraspecific sequence variation was detected
between C. pepo subspecies, as well as within C. moschata
and C. sororia (Sanjur et al. 2002).
Leucadendron is a genus which is much younger than
other genera in Proteacee. Using molecular dating, it was
reported that Leucadendron evolved 40 millions year ago.
(Barker et al. 2007). Actinidia is dated even younger,
approximately 20-26 million year ago (Qian and Yu 1991).
However, polymorphism in mtDNA of Actinidia was
higher. Several repeated motifs within nad1B/C were
detected (Chat et al. 2004). This was predicted due to
hibridisation or introgression event as well as high
poliploidisation in Actinidia (Chat et al. 2004). Extensive
studies on mtDNA of Leucadendron have not been
available. This report is the first report on exploration of
mtDNA in Leucadendron which shows extreme
mitochondrial DNA sequence conservation.
The Leucadendron nad1/B-C sequence was compared
to that of other plants (Figure 5.3). The most parsimonious
phylogenetic tree shows that Leucadendron (Proteaceae) is
57
most closely related to Lamprocapnos spectabilis

(Fumariaceae) and Liriodendron tulipifera (Magnoliaceae).
The most parsimonious phylogenetic tree obtained
using the nad1/B-C intron sequence showed that
Leucadendron is closely allied to Lamprocapnos
spectabilis (Fumariaceae) and Liriodendron tulipifera
(Magnoliaceae) and is part of a clade that contains the
Platanaceae (Platanus occidentalis). Phylogenetic analyses
based on rbcL cpDNA sequence data (Chase et al. 1993)
and combined rbcL and atpB cpDNA sequences (Drinnan
et al. 1994) revealed that Proteaceae was sister to
Platanaceae. The result from this study demostrated that
Leucadendron and Platanus are not sister to one another.
However, Leucadendron and Platanus are part of the same
clade and consensus tree of bootstrapped trees showed that
the grouping of Leucadendron to a clade consisting of
Laurus, Platanus and Liriodendron was 979.4 times out of
999.99 tress. Thus, this preliminary analysis of phylogeny
based on mtDNA sequence data supports the suggestion
that the Proteaceae is placed in a clade with Platanaceae as
suggested by Chase et al. (1993) and Drinnan et al. (1994).
This placement of the Proteaceae differs from the
classification by Johnson and Briggs (1975) based on
morphological, anatomical and developmental studies
which suggested that the Proteaceae were not closely
related to any of the major dicotyledonous orders. Further
studies using other markers are required to resolve the
position of Proteaceae within the angiosperm lineage.
CONCLUSION
Evaluation of organellar DNA variation showed that no
variation could be detected in mtDNA of seven
Leucadendron species tested using PCR-RFLP and
sequence analysis. This indicates that mtDNA is extremely
well conserved. Comparison with nad1/B-C equences from
other plants with high homology to that of Leucadendron
resulted in a phylogenetic tree which shows that
Leucadendron is clade to Platanus.
ACKNOWLEDGEMENTS
The first author thanks AUSAID for providing a Ph.D.
scholarship. We thank Dr. Ralph Sedgley and Ben
Croxford (The University of Western Australia) for
assistance in plant material collection.
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Pages: 59-64
ISSN: 1412-033X
E-ISSN: 2085-4722
Isolation and phylogenetic relationship of orchid-mycorrhiza from

Spathoglottis plicata of Papua using mitochondrial ribosomal large
subunit (mt-Ls) DNA
SUPENI SUFAATI, VERENA AGUSTINI, SUHARNO
Department of Biology, Fakulty of Mathematics and Natural Sciences, University of Cenderawasih (UNCEN), Jayapura 99358, Papua. Tel. +62-967572116. email: penisufaati@yahoo.com; verena_agustini@yahoo.com, harn774@yahoo.com
Manuscript received: 26 June 2011. Revision accepted: 16 April 2012.
ABSTRACT
Sufaati S, Agustini V, Suharno. 2012. Isolation and phylogenetic relationship of orchid-mycorrhiza from Spathoglottis plicata of Papua
using mitochondrial ribosomal large subunit (mt-Ls) DNA. Biodiversitas 13: 59-64. All terrestrial mycorrhiza have mutual symbiotic
with mycorrhizal fungi in order to gain nutrient from surrounding environment. This study was done to isolate and to identify
mycorrhiza orchid that associates with Spathoglottis plicata and were collected from Cagar Alam Pegunungan Cycloops (CAPC),
Jayapura. Isolation of mycorrhizal orchid came after the modified method of Manoch and Lohsomboon (1991). The result showed that
based on the morphological characteristic, there was presumably 14 isolations. However, only 2 isolations have been known, namely
Rhizoctonia sp. and Tulasnella sp., while the rest were not identified yet. Among them, the DNA of the 11 isolations were able to be
extracted for further analysis. The constructed phylogenetic tree performed that those species could be grouped into 4 major clusters.
Two species, Rhizoctonia sp. and Tulasnella sp. were in different clusters.
Key words: Spathoglottis plicata, orchid-mycorrhiza, Rhizoctonia sp., Tulasnella sp., Papua
INTRODUCTION
Spathoglottis plicata Blume (ground orchid) is one of
the orchid species that is widely distributed in the world,
including in the area of New Guinea (Papua New Guinea,
PNG and Papua, Indonesia). According to Agustini et. al.
(2008, 2009) Cycloop Mountains Nature Reserve (CAPC)
is one of the conservation area in Papua. This species of
orchid is also found here. One of buffer zone areas of this
nature reserve is a University of Cenderawasih (Uncen)
campus forest in Waena, Jayapura. It has plenty of S.
plicata orchids. This orchid is easy to grow and has purplewhite flowers and bloom throughout the year.
In nature, for the germination and early growth of
orchid seed and its development, it needs an association of
orchid roots and mycorrhizal fungi (orchid-mycorrhiza)
(Warcup 1981; Cameron et al. 2006; Suarez et al. 2006;
Agustini et al. 2009). This is because orchid seeds are very
small, smooth and soft and have no cotyledon which is a
food reserve in the early growth of seeds (Sarwono 2002;
Agustini and Kirenius 2002). The association to
mycorrhizal fungi helps the orchid in providing the
nutrients needed for the process of germination of its seeds
(Agustini and Kirenius 2002; Agustini 2003). The hyphae
of mycorrhizal fungi penetrates to the cell walls of roots
and then will be developed in the root cortex as dense
coiled called peloton. Mycorrhizal hyphae are very delicate
therefore they will absorb water and certain minerals

(Suarez et al. 2006; Agustini et al. 2009).
The role of mycorrhizal fungi is very interesting in the
life cycle of orchids. In the early growth, all orchid species
are heterotrophic, so it requires association with
mycorrhizal fungi to obtain the needed nutrients (Brundrett
et al. 2003; Taylor et al. 2004; Wu et al. 2010). Orchids
that have low levels of heterotrophic are dependants on the
presence of mycorrhizae. Based on morphological
characteristics, most of the fungi associated with orchids
are members of the form of anamorphic (asexual)-genus
Rhizoctonia (Athipunyakom et al. 2004) and generally
come from families of Basidiomycetes. Several studies of
terrestrial orchid-mycorrhiza in temperate-climate areas
have been done a lot, but the studies on the tropics are very
few (Otero et al. 2002). In Papua, researches and
publications on ground orchid-mycorrhiza, especially S.
plicata are very small in number. Therefore, there is a need
for the study on the diversity of orchid-mycorrhiza
associated with ground orchids S. plicata. Furthermore,
according to Kristiansen et al. (2001) to find out the
phylogenetic of the obtained isolates, the DNA sequencing
analysis is required. Therefore, the purpose of this study
was to isolate and to find out the phylogenetic relationship
of the orchid-mycorrhiza of S. plicata species using
mitochondrial sequences of ribosomal large subunit (mtLs) DNA.
60

Time and area study
The research was carried out for 10 months in 2008.
The isolation of orchid-mycorrhiza was done in the laboratory of Plant Tissue Culture and Laboratory of Microbiology, Department of Biology, Faculty of Mathematics
and Natural Sciences, University of Cenderawasih (Uncen),
Jayapura, Papua, Indonesia. The maintenance and regeneration of the isolates were carried out at the same place.
Isolation of orchid-mycorrhiza
According to Manoch and Lohsomboon (1991),
isolation of orchid-mycorrhiza was done by the modified
method of Masuhara and Katsuya (1989). The roots of S.
plicata was cut for about 1 cm, and sterilized with 10% of
Clorox for 30 seconds, and then washed with aquadest and
rinsed with distilled water, then sterilized again with 70%
alcohol. The sterilized pieces of roots were sliced
transversely using a sterile knife (razor blade) in the LAF
(Laminar Air Flow). The thickness of the slice was 200300 m or 3-4 pieces in 1 mm slice. The slices then were
planted in the media of potato dextrosa agar (PDA) in petri
dish. Each dishes were filled with 1-3 pieces of roots and
then incubated at 28C. Mycorrhizal mycelia start to appear
within 2-3 days.
After mycorrhizae grew, isolation was conducted if,
through a morphology observation, there was a difference
in shapes and growth patterns, to separate each of these
groups, because there possibly was more than one species
of media. In the same medium, the isolation was carried out
again if in the growing up process, the different shape and
growth patterns can still be found, until finally a really pure
and free from contamination-free isolate was obtained
(each mycorrhizal was cultured separately). In 2-3 times of
isolation, the isolates obtained were really pure.
Identification of orchid-mycorrhiza
The identification of the species of orchid-mycorrhiza
was done by looking at the morphological and microscopic
characteristics. Morphological features, including the
diameter of colony growth, mycelia morphology and other
characteristics. Microscopic characteristics, including
presence or absence of septum and spores shape or conidia.
Some of the literatures used in the identification of orchidmycorrhiza were Athipunyakom et al. (2004) and KarlFranzens (2006).
Isolates of orchid-mycorrhiza and phylogenetic
relationship
The isolates culture and DNA extraction
The mycelia of each isolate was dissolved in 2 L of
aquadest and transferred into 0.5 mL Eppendorf tubes
containing 6 L of dd H2O and 2 L of 10X PCR
(polymerase chain reaction) buffer (0.35 M Sorbitol, 0.1 M
Tris, 5 mM EDTA pH 7.5). The solution was then heated at
95C for 10 minutes in a water bath. The DNA of these
isolates was extracted using the method of hexadecyltrimethylammonium bromide (CTAB) (Gardes and Bruns
1993). The DNA quality was checked by electrophoresis.
DNA amplification by PCR

Of the extracted Isolates, 8 L was used as a template
for PCR amplification. ML5/ML6 regions of the genes of
mitochondrial large submit (mt-Lr) RNA was amplified with
a PCT100 thermocycler (MJ Research Inc.., MA). PCR
solution which was used for each isolates contains 2 L of
10X PCR Buffer, 2 L (mM) for each primer, 6 L ddH2O,
8 L dNTPs (2 mM) and 0.1 L Taq polymerase. PCR was
conditioned according to the method of Gardes and Bruns
(1993): initial denaturation at temperature of 95C for 1
minute 35 second, followed by 13 cycles of denaturation
for 35 seconds, temperature 94C, primer annealing for 55
seconds at a temperature of 55C, and polymerization for
45 seconds at 72C. The addition of nine cycles of the
polymerization was done by extending to 2 minutes. the 9
cycles consisting of 3 min of extension, and ending with 10
min of polymerization at a temperature of 72C. Primer used
was ML5 (CTCGGCAAATTATCCTCATAAG), MLIN3
(CGACACAGGTTCGTAGGTAG) and MI6
(CAGTAGAAGCTGCATAGGGTC).
DNA sequencing
The results of PCR amplification was purified with QIA
Quick PCR extraction kit (Qiagen GmbH GE). DNA was
dissolved in 30 L of sterile H2O and was being sequenced
with a primer ML5/ML6 using ABI 377 automatic
sequencer (Perkin Elmer) (Kristiansen et al. 2001). The
sequencing uses the dideoxy chain termination procedure
(Sanger et al. 1977).

Isolation of orchid-mycorrhiza
The results of orchid-mycorrhiza isolation associated
with S. plicata on eight sites are 13 isolates. Of these 13, 2
of them morphologically are identified as Rhizoctonia sp.
and Tulasnella sp., while 11 other species of orchidmycorrhiza, namely sp.1, sp.2, sp.3, sp.6, sp.7, sp.8, sp.9,
sp.10, sp.11, sp.12, and sp.13 isolates are associated with S.
plicata which is only from 1 different location (Table 1)
not all of them can be identified based on differences in
morphological features (Figure 1). However, the results of
phylogenetic analysis (Table 2, Figure 2) shows that there
are 4 clusters that are likely encountered two other species
of orchid-mycorrhiza.
The results of this isolation adds the data of isolation
result of the initial survey conducted by Agustini et al.
(2009) in the campus of Faculty of Mathematics and
Natural Sciences, Cendrawasih University, located in the
buffer zone of CAPC. In her research, she discovered two
species of orchid-mycorrhiza, namely Rhizoctonia sp. and
Tulasnella sp which are isolated from orchids S. plicata
and several other species of orchids. In this research, at
least there are two unknown species of orchid-mycorrhiza.
Both species are different from Rhizoctonia sp. and
Tulasnella sp.
SUFAATI et al. Orchid-mycorrhiza from Spathoglottis plicata
Figure 1. Morphology of some isolates of orchid mycorrhiza isolated from S. plicata.

A-B: Isolates sp7., C-D: Isolate sp.2., E-F: Tulasnella sp., and G-H: Rhizoctonia sp.
Table 1. Species of orchid-mycorrhiza from S. plicata in the Campus area
Cendrawasih University, Waena, Jayapura.
Code of sample location
Isolates of
orchid-mikoriza L1
L2
L3
L4
L5
L6
L7
L8
Rhizoctonia sp
Tulasnella sp
sp.1
sp.2
sp.3
sp.6
sp.7
sp.8
sp.9
sp.10
sp.11
sp.12
sp.13
Total
5
2
1
1
3
4
2
1
Note: = found; - = not found; L1 to L8 = sites in Waena, Jayapura
61
In a study, Athipunyakom et al.

(2004) isolated orchid-mycorrhiza of S.
plicata obtained from Chiang Mai,
Chanthaburi, Nakhon Ratchasima and
Bangkok and found three genera and
four species of orchid-mycorrhiza which
has successfully been identified based on
morphological characteristics, namely
Epulorhiza
repens,
Rhizoctonia
globularis,
Rhizoctonia
sp.,
and
Sebacina sp.
Table 1 show that S. plicata can be
associated with more than one species of
mycorrhiza-orchid. Kristiansen et al.
(2001) showed that the orchid can be
associated with more than one species of
mycorrhizal
fungi.
In
addition,
Handayanto and Hairiah (2007) also said
that some mycorrhizal fungi can
colonized one root of the plant. This is
because identification of orchidmycorrhiza fungi in conventional way is
still difficult to conduct. According to
Rasmussen (2004), Hollick et al. (2004)
and Kristiansen et al. (2001) the
characteristic of the fungus which never
reach sexual phase or sterile in a culture
condition made it difficult for
identification process so that the
identification with molecular techniques
needs to be done.
Description of orchid-mycorrhiza
species
Orchid-mycorrhiza orchids isolated
from
S.
plicata,
showing
the
development of a variety. The most
rapid growth of the colony diameter
reached by isolates sp.2 with a diameter
of 9 cm in 3 days, followed by
Rhizoctonia sp., Tulasnella sp., sp.12
and sp.1. While other species of orchidmycorrhiza with colony growth of more
than 15 days showed a very slow
growth, whereas the optimum diameter
of colony growth for orchid-mycorrhiza
is 15 days. The growth rapidity of this
colony affects the speed of nutrients
absorption in nature. According to Smith
and Read (2008), the growth rapidity of
mycelia/hyphae potentially increased the
extent of nutrients absorption surface in
the soil during symbiosis process with
the plant root system. Thus, the plants
quickly obtained nutrients needed
through hyphae.
62
Table 2. Description of the morphology of orchid-mycorrhiza species associated with S. plicata.

Mycorrhiza
Growth on
(isolates)
Morphology
PDA
species
Rhizoctonia The growth of the colony very fast, within 4 days. Branched hyphae, less dense
sp.
Mycelia are translucent white and thin. There is a At the end of hyphae branch, it is formed small sphere-shaped
set of dark green granules forming a round shape
granules, which in turn form a collection of granules. The
in the middle.
presence of globulus oil is found around it.
Tulasnella
sp.
Growth of colonies is rather slow, 8 days.

Mycelia are white and thicker.
Branched hyphae with dense septate.

The presence of conidia, in a group or scattered, and with a
magnification of 1000x, the conidia seems like bananas, in
sectional.
sp.1
Colony growth is slow.

Achieving a diameter of 9 cm in 12 days.
Mycelia are whitish-green brown.
Mycelia are thick and round.
Branched hyphae, more than 1core, dense septate.

Spores are brown and on the tip of hyphae.
The hyphae are long and round and in the shape of chain, and
the conidia are round, semi round, ramiform (like kidney
shapes), allantoidal (like cashew nut shape with no-sharp
edge), and crystal look-like.
sp.2
The colony growth is very fast, within 3 days

Blurred white mycelia
Mycelia are rather thick and the granules are yellow to green
and are scattered in a circle in the middle and edge.
Microscopically, hyphae are branched with long septet. There
are dispersed small oval-shaped spores.
sp.3
The colony growth is very fast, within 5 days

The hyphae are septate and dense. At the tip of the hyphae,
Mycelia are white to yellow and have tree-like
there are round and brown spores, and also in V like shape
branches when mature.
in brown color with parallel lines in it.
sp.6
Colony growth is very slow, reaching only 9 cm Branched hyphae with dense septate.
in 22 days
Spores are clustered and dispersed small round shape. Some
Mycelia are yellowish green, thick and form concentric
tips of hyphae form many branches covered by the collection
zones. On the edge side, mycelia are thin and white.
of spores.
sp.7
Slow colony growth, 15 days

The hyphae are branched with non-close septate.
Mycelia are white with dark brown sides that form Conidias shape are like crystals and resembles a figure of number
a circle in the middle and thick.
eight are scattered and there is a chain composed ellipse.
sp.8
Colony growth is slow, 17 days

Hyphae are long with septate.
Mycelia are white with brown color in the middle, Conidia are round and crystals-shaped with elongated dots
and appear thicker
which are arranged like a chain.
The presence of large round-shaped spores is found on the tip
of hyphae.
sp.9
Colony growth is very slow 20 days

branched hyphae with non-close septate. With 400x
At the beginning, Mycelia grow clearly white and
magnification, the hyphae look like spiral.
thin. As the growth continues, white mycelia Some hyphae appear thickened and the presence of spiny
appear thicker in the middle.
conidia is visible and scattered round.
sp.10
Growth of colony is very slow, 24 days.

There are branched hyphae and close septate.
Mycelia are white with blackish brown in the The shapes of conidia are like crystals, and there is much
middle and it looks thicker.
globular oil which is attached to the hyphae.
sp.11
Slow growth colony, 16 days.

Hyphae are septate and branched.
Mycelia are thick and white forming concentric Some hyphae appear thickened and there are many globular
zone.
attached to the hyphae. Among hyphae, there are conidia in
crystal shape.
sp.12
Colony growth is rather slow, 8 days

Mycelia are blackish green and thick.
sp.13
Growth of colonies is very slow; it needs more Hyphae have long septate and a lot of branches.
than a month to reach a diameter of 9 cm.
There are many scattered elliptical conidia and are attached to
Mycelia are white and thick.
the hyphae.
In addition, there is presence of globular attached to the hyphae.
Hyphae are branched with non-close septate.

Many spores are black round-shaped and are found on the tip
of hyphae. Some hyphae are coil-shaped.
SUFAATI et al. Orchid-mycorrhiza from Spathoglottis plicata
63
Table 3. Phylogenetic relationship of 11 isolates of orchid orchid-mycorrhiza based on molecular approach (DNA) isolated from orchid
S. plicata.
*
Isolate
1
2
3
4
5
6
7
8
9
10
11
1
****
0.4055
1.0986
1.0986
1.0986
0.4055
0.0000
0.4044
0.4055
1.0986
1.0986
2
0.6667
****
0.4055
0.4055
0.4055
0.0000
0.4055
0.0000
0.0000
0.4055
0.4055
3
0.3333
0.6667
****
1.0986
0.0000
0.4055
1.0986
0.4055
0.4055
1.0986
0.0000
4
0.3333
0.6667
0.3333
****
1.0986
0.4055
1.0986
0.4055
0.4055
0.0000
1.0986
5
0.3333
0.6667
1.0000
0.3333
****
0.4055
1.0986
0.4055
0.4055
1.0986
0.0000
Phylogenetic relationship of orchid-mycorrhiza of

Spathoglottis plicata
In Table 2, the isolates number of mycorrhizal orchid
which were isolated from Spathoglottis plants are 13
species, but only 11 species that can be isolated their DNA.
Therefore, phylogenetic analysis (Table 3) would only be
done on these 11 species that have been successfully
analyzed. The results of DNA analysis shows that there are
four major groups (clusters) of 11 species of orchidmycorrhiza isolated from S. plicata (Figure 2). With the
method of UPGMA (unweighted pair group method with
arithmetic mean), cluster 1 shows the phylogenetic ties
among sp.1 and sp.7 isolates. Cluster 2 shows the
relationship among the four species, namely, sp.2, sp.6,
sp.8 and sp.9 isolates. Cluster 3 shows that there are 3
isolates having a very close relationship, namely sp.3
isolates, Tulasnella (Tul) and sp.11 isolates. Whereas
cluster 4 shows that there are two species of isolates having
a close relationship ties i.e. the species of Rhizoctonia (Rhi)
and sp.10 isolates. And also, the results show that each
isolates of those four clusters have the same index value of
relationship. Rhizoctonia has a relationship index equal to
100% to isolates sp.10, Tulasnella to isolates sp.3 and
sp.11, so it can be said that these isolates have the same
species.
The growth rate of the colonies did not affect the
nearby distant phylogenetic relationship in one cluster. It
can be seen from the Tulasnella, which grows slowly (8
days/9 cm), is different from sp.11, which has a diameter of
colony growth on PDA medium for up to 16 days/9 cm,
although morphologically the relationship among species
and isolates within a cluster is very close. It is similar with
Rhizoctonia case, morphologically, it grows very fast (4
days) on PDA medium, whereas sp.10 grows very slow and
it takes about 24 days to reach a diameter of 9 cm.
However, microscopically both are similar in its
morphology (Table 1). The same thing occurs in clusters 1
and 2. For the species group of the two clusters, the DNA
relationship is closer to cluster 3 of Tulasnella sp. and is
futher to Rhizoctonia sp. group.
6
0.6667
1.0000
0.6667
0.6667
0.6667
****
0.4055
0.0000
0.0000
0.4055
0.4055
7
1.0000
0.6667
0.3333
0.3333
0.3333
0.6667
****
0.4055
0.4055
1.0986
1.0986
8
0.6667
1.0000
0.6667
0.6667
0.6667
1.0000
0.6667
****
0.0000
0.4055
0.4055
9
0.6667
1.0000
0.6667
0.6667
0.6667
1.0000
0.6667
1.0000
****
0.4055
0.4055
10
11
0.3333
0.6667
0.3333
1.0000
0.3333
0.6667
0.3333
0.6667
0.6667
****
1.0986
0.3333
0.6667
1.0000
0.3333
1.0000
0.6667
0.3333
0.6667
0.6667
0.3333
****
Figure 2. Dendrogram showing the phylogenetic relationship of

several species and isolates of orchid-mycorrhiza isolated from S.
plicata with UPGMA method.
CONCLUSION
Based on morphological and microscopic character,
there were 13 isolates of orchid mycorrhizal fungi has been
isolated from S. plicatas root. Among them, isolates has
been known as Rhizoctonia sp and sp Tulasnella. The
constructed phylogenetic tree of 11 isolates performed that
those species could be grouped into 4 major clusters. Two
species, Rhizoctonia sp. and Tulasnella sp. were in
different clusters.
ACKNOWLEDGEMENTS
The authors gratefully thank Maria Imco, Ira Ardila
Putri, and Dewi Katemba, which has helped the research at
the Laboratory of Biological Science University of
Cenderawasih, Papua; to the General Director of Higher
Education that has funded this research through
64
Fundamentals Grant of 2008. To Prof. Zamir K. Punja

(Simon Fraser University, Burnaby, Canada), thanks for the
help during the discussion of this matter.
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New Phytol 87 (2): 371-381.
BIODIVERSITAS
Pages: 65-71
ISSN: 1412-033X
E-ISSN: 2085-4722
Diversity of macrofungal genus Russula and Amanita in Hirpora

Wildlife Sanctuary, Southern Kashmir Himalayas
SHAUKET AHMED PALA, ABDUL HAMID WANI, RIYAZ AHMAD MIR
Section of Mycology and Plant Pathology, Department of Botany, Faculty of Biological Sciences, University of Kashmir, Hazratbal, Srinagar 190006,
Jammu and Kashmir , India. Tel.: +99- 9419010336; Fax.: +99-942421357; email: sapala29@gmail.com; ahamidwani@yahoo.com
Manuscript received: 11 January 2012. Revision accepted: 30 March 2012.
ABSTRACT
Pala SA, Wani AH, Mir RA. 2012. Diversity of macrofungal genus Russula and Amanita in Hirpora Wildlife Sanctuary, Southern
Kashmir Himalayas. Biodiversitas 13: 65-71. The Hirpora Wildlife Sanctuary that extends over an area of 114 km2 lies in the Pir Panjal
range at a distance of 70 km in south-west of summer capital Srinagar. It is rich in biodiversity including macrofungal diversity. The
Sanctuary has been subjected to high ecological and anthropogenic disturbance due to the construction of Mughal road which is major
threat for its biodiversity. Since there is hardly any report of documentation of macrofungi from this sanctuary. In this back drop a
survey was carried out during the year 2010 and 2011 to explore and invetorise macrofungal diversity of the sanctuary. During the
survey a no of macrofungi were documented, among which Amanita and Russula were dominant genus represented by 7 species each.
All the 14 species viz. Amanita ceciliae (Berk. & Broome) Bas. Amanita flavoconia G.F. Atk., Amanita muscaria var. formosa Pers.,
Amanita pantherina (Fr.) Krombh., Amanita phalloides (Fr.) Link., Amanita vaginata (Bull. ex Fr.) Vitt., Amanita virosa (Fr.) Bertillon,
Russula aeruginea Fr., Russula atropurpurea (Krombh.) Britz., Russula aurea Pers., Russula cyanoxantha (Schaeff.) Fr., Russula
delica Fr. Russula emetica (Schaeff. ex Fr.) Gray. and Russula nobilis Velen. are ectomycorrhizal in nature and among them Russula
aeruginea Fr. is reported first time from the Kashmir.
Key words: Russula, Amanita, Hirpora, macrofungi, Kashmir
INTRODUCTION
Mushroom is regarded as a macrofungus with a
distinctive fruiting body that can be either epigeous or
hypogeous and large enough to be seen with the naked eye
and to be picked by hand (Chang and Miles 1992).
Mushrooms belong to the kingdom fungi, which constitutes
the most diverse group of organisms after insects on this
biosphere. Defining the exact number of fungi on the earth
has always been a point of discussion and several studies
have been focussed on enumerating the worlds fungal
diversity (Crous 2006). Current studies estimate that out of
1.5 million species of fungi existing on this biosphere,
140,000 species may be considered as mushrooms, but only
14,000 species are known to man, which accounts for 10%
of the estimated mushroom species (Chang and Miles
2004). Only a fraction of total fungal biodiversity has been
subjected to scientific scrutiny and mycologists continue to
unravel the unexplored, hidden and fascinating fungal
biodiversity as many macro-fungi are becoming extinct or
facing threat of extinction because of habitat destruction
and global climate change (Swapana et al. 2008). There are
about 7750 macrofungal species known to have
ectomycorrhizal nature (Rinaldi et al. 2008). The genus
Amanita contains about 500 species, including some of the
most toxic known mushrooms found worldwide (Zhang et
al. 2004). This genus is responsible for approximately 95%
of the fatalities resulting from mushroom poisoning, with
death cap accounting for about 50% on its own. There are
around 750 worldwide species of mycorrhizal mushrooms

which compose the genus Russula (Miller et al. 2006). The
state Jammu and Kashmir is rich in macrofungal diversity
due to wide agro-climatic variations diverse physiography
and undulating topography, but understanding of the
macro-fungal flora of the Kashmir is still in an exploratory
stage and undoubtfully there are many more species to be
recorded (Watling and Abrahim 1992).
The present communication describes the brief
morphological description, macro and microscopic details,
seasonal occurrence and edibility of the 14 species of
macrofungi belonging to genus Russula and Amanita
collected from Hirpora wild life sanctuary.

Regular field trips were carried to document the
macrofungal diversity of Hirpora wildlife sanctuary (Figure
1) dominated by conifer forests. Usually 4-5 trips were
carried out per month to cover as much species as possible.
These field trips were organised according to the method
given by RE Hailling (1996). Standard method of
collection, preservation, macro and microscopic studies
were followed (Kumar et al. 1990; Atri et al. 2003) and the
shape, size, colour of fresh specimen, time of collection
and their edibility were recorded on the field notebook
before brought to the laboratory for further observations
B I O D I V E R S I T A S 13 (2): 65
5-71, April 2012
6
66
Kashmir
S
Srinagar
Figure 1. Map showing the collection

F
c
site, Hirpora Wildliife Sanctuary, Southern
S
Kashm
mir Himalayas of Jammu and
d Kashmir Statte,
I
India.
aand preservatiion for herbarrium purposess. The spore prints

p
w
were
taken acccording to thhe guideliness given by Michel
M
K (2001), thhen their morrphology, suchh as shape, siize of
Kuo
s
spores,
surface features of spores, preseence or absennce of
o drops andd starch grannules were reecorded undeer the
oil
t
trinocular
miicroscope at USIC (Uniiversity Scieentific
I
Instrumentatio
on centre) in Kashmir Unniversity. Reaagents
u
used
for prepaaration of spoore slides weree 3% KOH, cotton
c
b
blue,
lactophenol and Melzer,s
M
reageent. To elicitt the
n
necessary
infoormation regarrding their edibility local people
w
were
consulteed. Photograpphs were takken in field using
C
Cyber
shot Sony
S
10.1 megapixel
m
Cam
mera. The fungal
fu
s
specimens
weere also preseerved in form
maline solutioon for
herbarium purpooses, in funngal collectio

on of KASH
H
Herb
barium of Plant
P
Taxonoomy, Divisio
on of Botanny
Kash
hmir Universitty, Kashmir.
RES
SULTS AND DISCUSSION
During
D
the survey
s
to diifferent placees of Hirporra
Wild
dlife Sanctuaary, 14 ecttomycorrhizall macrofungaal
species belongingg to genus A
Amanita and Russula werre
colleected and identified
i
(F
Figure 2). Their
T
detaileed
description along with photograaphs is as:
PALA et al. Russula and Amanita of Hirpora, Kashmir
Amanita ceciliae (Berk. & Br.) Bas

Synonym (s). Amanita inaurata Secr., Amanita
stragulata (Fr.) Quel., Agaricus ceciliae Berk & Br.
Common name (s). Snakeskin grisette
Description
Cap. 5-10 cm in diameter, initially convex, expanding
to planoconvex or flat with a shallow umbo, brown in color
with slightly darker centre, usually covered with
grayish patches, margins deeply lined at maturity.
Gills. Free, crowded, entire, white in color.
Stipe. 6-15 cm long, 0.7 1.5 cm thick, tapers towards
the apex, whitish, finely hairy, brownish colored volva or
patches of volval remnants dotted around stipe base.
Annulus (ring) is absent.
Flesh. Thick, soft, white, not changing when sliced.
Spores. Globose, smooth, non-amyloid, 9-12 microns;
Spore print white.
Habit and habitat. Mycorrhizal, growing alone or
scattered under conifer trees.
Season. Summer
Edibility: Inedible, considered to be poisonous.
Amanita flavoconia G.F. Atk.
Synonym (s). Amplariella flavoconia (Atk.) Gilbert,
Venenarius flavoconius (Atk.) Murill
Common name (s). Orange Amanita or Yellow-dust
Amanita
Description
Cap. 3 to 8 cm in diameter, initially ovoid, but with
maturity becomes convex and finally flattened, orange to
bright yellow-orange in color. The young specimens are
covered with chrome yellow warts that may be easily
rubbed off or washed away with rain,
Gills. Free, crowded, white to cream colored and
initially covered with a yellowish partial veil.
Stipe. 4-12 cm long, 0.5 to 1.3 cm thick, equal or
slightly tapered upward (bulbous), white to yellowish
orange in color, mostly smooth sometimes covered with
small scales. The persistent skirt-like ring (annulus) is
present on the upper portion of stipe 1-2 cm below the cap
and partially underground powdery yellow volva is present
at the base.
Flesh. Thick, brittle,white in color.
Spores. Elliptical to globose, smooth, amyloid, 7-9 x 58 microns; Spore print white.
Habit and habitat. Mycorrhizal, growing either solitary
or in groups under conifer trees.
Season. Summer and early autumn
Edibility. Inedible (poisonous)
Amanita muscaria var. formosa Pers.
Synonym (s). Amanta muscaria var. guessowii Veesely
Common name (s). Fly agaric
Description
Cap. 5-11 cm in diameter, initially oval to convex or
campunulate, but fattens or curve upward with maturity,
yellowish to tannish with concentrically arranged white
scales on entire cap surface
Gills. Free, broad, crowded, white in color.
67
Stipe. 4-12cm long, 1-2cm thick, white, bulbous with a

ring on the upper half. Volva consisting of 2-3 concentric
rings is present at the base of stipe.
Flesh. Thick, white in color.
Spores. Oval, smooth, non-amyloid, 9-13 x 6.5-9
microns; Spore print white.
Habit and habitat. Mycorrhizal, growing solitary or
Season. Late summer
Edibility. Inedible
Amanita pantherina (Fr.) Krombh.
Synonym (s). Amanitaria pantherina (DC.) Krombh,
Agaricus pantherinus DC.
Common name (s). Panther cap
Description
Cap. 4-12 cm in diameter, initially hemispherical but
turns convex to plano-convex at maturity, color dark brown
to yellowish brown, veil remnants forming pointed white
warts on upper surface.
Gills. Free, crowded, white in color.
Stipe. 6-10 cm long, 0.8-2 cm thick, unequal, tapers
towards the tip, white in color. The partial membranous
veil is leaving a white ring at the upper portion of stipe and
universal veil forms a single roll or collar on the basal bulb.
Flesh. Thick, white not discoloring on exposure or
bruising.
Spores. Globose, smooth, non-amyloid, 8-12 x 6-8
microns; Spore print white.
Habit and habitat. Mycorrhizal, growing singly,
scattered, or gregariously under pine trees.
Season. Summer and early autumn especially in rainy
season.
Edibility. Inedible, poisonous but not deadly poisonous.
Amanita phalloides (Fr.) Link.
Synonym (s). Agaricus phalloides Fr., Amanita viridis
Pers., Amanitina phalloides (Fr.) Gilbert
Common name (s). Death cap
Description
Cap. 4-14 cm broad, initially oval, becoming convex,
then broadly convex to flat with age, smooth, sticky when
wet, shiny when dry; color ranging from olive-brown to
yellowish-brown
Gills. Free, crowded, moderately broad, white to cream
colored.
Stipe. 4-15 cm in length, 1-2 cm thick, more or less
equal or tapering towards apex, smooth, white or with tints
of the cap color; ring present in the upper part but is often
lost; white volva encases the base.
Flesh. Thick near the disc, soft, white in color.
Spores. Globose, amyloid, 7-12 x 6-9 microns; Spore
print white.
Habit and habitat. Mycorrhizal, growing singly or in
small groups on the ground under coniferous trees.
Season. Summer and early autumn
Edibility. Deadly poisonous. It has been estimated that
30 gms or half a cap of this mushroom is enough to kill a
human (Gopinath et al. 2011).
68
Amanita vaginata (Bull. ex Fr.) Vitt.

Synonym (s). Amanitopsis vaginata (Bull. Ex Fr.) Roze,
Amanita vaginata var. plumbea (Bull.) Quel. & Bataille,
Agaricus plumbeus Schaeff.
Common name (s). Grisette
Description
Cap. 4-10 cm broad, initially oval then convex and
eventually flattens as it matures, sticky when wet, gray to
grayish brown in color, mostly with a few scattered white
to grayish patches on the surface. The margin is
prominently lined or grooved that duplicate the gill pattern
underneath.
Gills. Free or slightly attached to stipe, crowded, edges
are minutely fringed, white in color.
Stipe. 7-14 cm long, 1-2. cm thick, more or less equal or
bulbous, smooth or with a few greyish scales, ring absent.
White sack-like volva encloses the base of the stem.
Flesh. Thin, white, and does not change color upon
bruising or injury.
Spores. Globose, smooth, non-amyloid, 8-12 microns;
Spore print white.
Habit and habitat. Mycorrhizal, growing singly or
scattered in coniferous trees as well as on road side of
Mughal road under broad leaved trees like Populous, Salix
etc.
Season. Late spring and summer.
Edibility. Edible
Amanita virosa (Fr.) Bertillon
Synonym (s). Agaricus virosus Fr.
Common name (s). Destroying angel
Description
Cap. 5-10 cm across, initially oval, becoming convex
then expanded flat with age, white, smooth, margins not
lined, viscid when moist.
Gills. Free, crowded, white in color
Stipe. 6-18 cm long, 0.6-2 cm thick, tapering towards
the tip, white with surface often disrupted into shaggy
fibrils, white sac like volva encases the base or collapses at
early stage, ring collapses very early.
Flesh. Thick, soft, white in color
Spores. Globose, smooth, amyloid, 8-10 microns; Spore
print white.
Habit and habitat. Mycorrhizal, growing, singly or
scattered under broad leaved trees near road sides of
Mughal road.
Season. Late spring and summer.
Edibility. Deadly poisonous, its poisonings are common
in the area because of its resemblance with Agaricus sps
Russula aerugenia Fr.
Synonym (s). Russula graminicolor (Gillet) Quel.
Common name (s). Grass-green Russula
Description
Cap. 4-8 cm in diameter, initially convex, becoming
broadly convex to flat with a shallow depression, smooth,
grayish green to yellowish green to pale green, margin
often lined at maturity.
Gills. Almost free, crowded, forked, creamy to light
yellow.
Stem: 4-6 cm long, 0.8-1.8 cm thick, cylindrical,
smooth, white
Flesh. Thick, brittle, white in color, color does not
change on bruising.
Spores. Globose, warty, 6-8 microns; Spore print
creamy to light yellow.
Habit and habitat. Mycorrhizal, growing scattered, or
gregariously under conifer trees or under Salix near
roadsides of Mughal road.
Season. Summer to early autumn
Edibillity. Edible
Russula atropurpurea (Krombh.) Britz.
Synonym (s). Russula krombholzii Krombh., Russula
undulate Velen.
Common name (s). Blackish purple Russula or Purple
brittlegill
Description
Cap. 4-9 in diameter, convex to flat with a shallow
depression in centre or turns upward, smooth, purple violet
in color with dark centre.
Gills. Adnexed, crowded, cream or pale straw in color.
Stipe. 2-7 cm long, 0.8-2 cm thick, cylindrical, brittle,
smooth, white often becoming gray with age.
Flesh. Thick, brittle, white, with an odour of apple fruit.
Spores. Globose, warty, 6-8 microns; Spore print
creamy to light yellow.
Season. Summer
Edibility. Inedible
Russula aurea Pers.
Synonym (s). Agaricus auratus With., Russula aurata
(With.) Fr.
Common name (s). Golden Russula or Gilded brittlegill
Description
Cap. 5-9 cm in diameter, initially convex but flattens
with age with a slight depression in the centre, reddish
orange with yellow tinge near the margin, smooth, margin
sulcate when mature, cuticle peeling halfway to center.
Gills. Free to adnexed, broad, fairly distant, yellow in
color.
Stipe. 3-7 cm tall, 1-2 cm thick, cylindrical, smooth,
light yellow in color.
Flesh. Thick, brittle, yellow in color.
Spores. Echinate, 7-9 microns; Spore print ochraceous.
Habit and habitat. Mycorrhizal, growing solitary or
scattered under pine trees.
Season. Summer to autumn.
Edibility. Edible.
Russula cyanoxantha (Schaeff.) Fr.
Synonym (s). Agaricus cyanoxanthus Schaeff., Russula
cutefracta Cook, Russula furcata Sensu Auct.
Common name (s). Charcoal burner
Description
Cap. 5-12 cm in diameter, initially globose, becomes
convex to flat at maturity with a central depression in the
centre, greyish purple to dark violet, slimy when moist.

Cuticle can be separated upto half to the centre.
Gills. Subdecurrent to adnexed, intermediately spaced,
flexible, greasy to touch, white.
Stipe. 4-7 cm long, 0.7-1.5 cm thick, cylindrical or
slightly bulbous, smooth, white.
Flesh. Thick, brittle, white.
Spores. Ellisoid, warty, 7-8 x 6-7 microns; spore print
white.
Habit and habitat. Mycorrrhizal, growing scattered to
gregariously under conifer trees.
Season. Summer
Edibility. Edible
Russula delica Fr.
Synonym (s). Agaricus exsuccus (Pers.) Anon.,
Lactarius exsuccus (Pers.) Sm.
Local name (s): Milk-white brittlegill
Description
Cap. 5-13 cm in diameter, initially convex with a
depression in the centre or infundibuliform at maturity,
margins inrolled, whitish often developing brownish
discolorations, mostly contains leaf debris on the upper
surfaces. Cuticle cannot be peeled off.
Gills. Decurrent, crowded, often forked, white.
Stipe. 2-4 cm long, 1.5-3 cm thick, cylindrical, smooth,
white.
Flesh. Thick, brittle, white, not changing color on bruising.
Spores. Ovoid, warty, 8-11 x 7-9 microns; Spore print
white.
Habit and habitat. Mycorrhizal, growing scattered on
ground under Pine trees.
Season. Early autumn.
Edibility. Edible
Russula emetica (Schaeff. ex Fr.) Gray.
Synonym. Agaricus emeticus Schaeff.: Fr., Agaricus
linnaei var. emeticus (Schaeff.) Fr., Russula clusii Fr.
Common name (s). Sickener
Description
Cap. 3-8 cm in diameter, convex when young latter on
flattened, sticky, bright scarlet to cherry red or blood red in
color with finely ridged margins. The cuticle is readily
peeled from the cap.
Gills. Free, crowded, cream or pale straw in color.
Stipe. 2-9 cm in length, cylindrical, brittle, smooth, white.
Flesh. Thick, brittle, white in color.
Spores. Roughly spherical, covered with small spines,
6-8 microns; Spore print white
Season. Summer
Edibility. Inedible
Russula nobilis Velen.
Synonym (s). Russula mairei Singer, Russula emiticus
Schaeff., Russula fagetorum Bon
Common name (s). Beechwood sickener
Description
69
Cap. 4-9 cm in diameter, initially globular, becoming

flattened and finally slightly depressed or sometimes turns
upward, red to pink colored, sticky, very often damaged by
slugs at places.
Gills. Adnexed, intermediately spaced, broad, creamy
white
Stipe. 3-6 cm long, 1-1.5 cm thick, cylindrical, smooth,
white
Flesh. Thick, brittle, white but pink beneath the cuticle.
Spores. Ovoid with warts, 7-8 x 6-7 microns; Spore
print white
Habit and habitat. Mycorrhizal, scattered near the
roadsides of Mughal road under broad leaved trees.
Season. Summer and autumn
Edibility. Poisonous, but some people use after
cooking.
Discussion
A wide variety of ectomycorrhizal fungi are symbionts
of many tree species in temperate climatic zones. In the
recent years, however, anthropogenic activity has made
different countries all over the world to show serious
concern
about
the
dwindling
biodiversity
of
ectomycorrhizal macrofungal species being last at the rate
never known before. The present study confirms the
remarkable species richness of the macrofungal genus
Amanita and Russula in Hirpora Wild Life
sanctuary. Fourteen species, seven of each Russula and
Amanita were recorded from the selected area. Watling and
Gregony (1980) recorded 119 taxa of macrofungi from
Kashmir including many species Russula and Amanita.
Watling and Abrahim (1992) reported about 77 species of
ectomycorrhizal macrofungi from coniferous forests of
Kashmir with Amanita and Russula as dominant genus.
Dar et al. (2009, 2010) pointed out that coniferous
forest of jammu and Kashmir harbours some 260 species of
macrofungi. They reported Russula delica, Russula aurea,
Russula atropurpurea and Russula paludosa first time from
coniferous forests of Kashmir. Lakhanpal (1996, 1997);
Atri et al. (2000); Beigh (2008) have also documented
many species of Russula and Amanita from conifer forests
of Kashmir. Gardezil and Ayoub (2003a) described eight
species of Russula first time from Azad Kashmir. Gardezil
and Ayoub (2003b) reported Amanita muscaria var. alba,
A. cecillae, A. pantherina and Amanita virosa from Azad
Kashmir. Boda (2009) reported 44 different species of
mushrooms from southern part of the Kashmir including
Russula emetica, and Amanita mascaria.
Russula and Amanita represent two major genera
having multifarious medicinal properties besides their
mycorrhizal role. They have been found to live in
symbiotic association with a wide variety of coniferous and
deciduous trees. The compounds derived from these
mushrooms avert diseases and boost up immune system
thereby improving human health (Wasser 2002). Different
species of Russula are known to possess antiviral,
antibacterial, antiparasitic, anti-inflammatory, antioxidant,
hepatoprotective, antidiabetic and anticancer activities
(Turkoglu et al. 2007; Wasser 2010).
70
Figure 2. Diversity of Russula and Amanita in Hipora, Kashmir. A. Amanita ceciliae (Berk. & Br.) Bas, B. Amanita flavoconia G.F.
Atk., C. Amanita muscaria var. formosa Pers, D. Amanita pantherina (Fr.) Krombh., E. Amanita phalloides (Fr.) Link., F. Amanita
vaginata (Bull. ex Fr.) Vitt., G. Amanita virosa (Fr.) Bertillon, H. Russula aerugenia Fr., I. Russula atropurpurea (Krombh.) Britz., J.
Russula aurea Pers., K. Russula cyanoxantha (Schaeff.) Fr., L. Russula delica Fr. M. Russula emetica (Schaeff. ex Fr.) Gray., N.
Russula nobilis Velen.
CONCLUSION
Since the wild macrofungi play an important ecological
role for the healthy maintenance of the ecosystem
particularly that of forest ecosystems, besides their
tremendous medicinal value, therefore it becomes quite
necessary to explore, document and conserve this natural
wealth. Also the area is ecologically fragile, but has been
subjected to high magnitude of disturbance due to
construction of National highway through the wild life
sanctuary which is a major threat for its biodiversity;
therefore it becomes quite imperative to document its
biodiversity. The present communication reports the
fourteen species of ectomycorrhizal macrofungus from the
area among which Russula aeruginea Fr. is reported first
time from the Jammu and Kashmir.
ACKNOWLEDGEMENTS
The authors are highly thankful to Head Department of
Botany, University of Kashmir, Prof. Zaffar Ahmed Reshi
for providing necessary support to carry out the work and
Prof. N.S. Atri from Punjabi University, Patiala, India for
their help in identification of specimens.
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Pages: 72-78
ISSN: 1412-033X
E-ISSN: 2085-4722
Effect of plantations on plant species diversity in the Darabkola,

Mazandaran Province, North of Iran
1
HASSAN POURBABAEI1,, FATEMEH ASGARI1, ALBERT REIF2, ROYA ABEDI1
Department of Forestry, Faculty of Natural Resources, Somehsara, University of Guilan, Islamic Republic of Iran. P.O. Box 1144, Tel.: +98-1823220895, Fax.: +98-182-3223600, email: H_pourbabaei@guilan.ac.ir
Department of Vegetation Classification, Waldbau Institute, Faculty of Forest and Environment, Albert-Ludwigs University, Freiburg, Germany
Hassan Pourbabaei, Associate Professor at Department of Forestry, Faculty of Natural Resources, Somehsara, P.O.Box 1144
Manuscript received: 18 December 2011. Revision accepted: 2 April 2012.
ABSTRACT
Pourbabaei H, Asgari F, Reif A, Abedi R. 2012. Effect of plantations on plant species diversity in the Darabkola, Mazandaran Province, North
of Iran. Biodiversitas 13: 72-78. In this study, the effect of plantations on plant species diversity was investigated in Darabkola,
Mazandaran province, north of Iran. To conduct the study, a natural mixed forest, a broadleaved plantation (Alnus subcordata-Acer
velutinum) and a coniferous plantation (Cupressus sempervirens var. horizontalis-Pinus brutia) were selected. 35 sampling plots were
taken in systematic random method in each area. Data analysis was carried out using Simpson, Hill's N 2, Shannon-Wiener and Mc
Arthur's N1 diversity indices, Smith and Wilson evenness index and species richness. Results revealed that there were 32 plant species in
natural forest and 30 plant species were found in each plantation. Rosaceae and Lamiaceae were the main families in the studied areas.
Diversity and evenness indices of all vegetation layers had the most values in the natural forest. Richness of woody plants had the
highest value in the natural forest, while herbaceous richness was the highest in coniferous plantation. Mc Arthur's N1 had the highest
value among diversity indices and followed by Hill's N2, Shannon-Wiener and Simpson indices, respectively. In addition, results showed that
there were significant differences among diversity, evenness and richness indices in all vegetation layers in the three studied areas.
Key words: plantation, plant species diversity, Darabkola, Mazandaran.
INTRODUCTION
Afforestation and replanting programs have helped
reverse the decline of forest cover. Currently 3% of the
worlds forests are plantations, comprised of 60 million
hectares in developed nations and 55 million hectares in
developing nations (Hartley 2002). Forest plantations that
achieve yields corresponding to site potential are part of the
economic growth of forest resources; such economic growth
should not be hampered by a lack of ecological information.
Conservation and enhancement of biological diversity is a
key component of sustainable forest management in
vegetation communities (Jobidon et al. 2004). Conservation
of ecological services, prevent a lack of special species and
aesthetics values, attention to principles of forest
management, promote social and commercial of medical
and industrial plants are the most important components of
biodiversity conservation in forest plans (Pilehvar 2000).
Biodiversity has been an important objective of forest
management, because it provides a broader array of
ecosystem services (Wang and Chen 2010).
There is a common belief that forest management
negatively influences biodiversity (Wagner et al. 1998).
Plantations are often viewed unfavorably compared to
natural forests by the public and conservation biologists,
because of the lack of biodiversity (Perley 1994; Potton
1994; Freedman et al. 1996). Plantations usually include
exotic and non-native species, or native species in pure
stands. Plantations contribute to biodiversity conservation

variously. Most directly, plantations can contain substantial
components of biotic diversity across many taxa (Ferns et
al. 1992; Allen et al. 1995; Michelsen et al. 1996; Chey et al.
1997; Estades and Temple 1999), including rare species in
some cases (Norton 1998; Tucker et al. 1998; Wilson and
Watts 1999). Even exotic plantations can help to restore
native biota to degraded sites by stabilizing soil and
creating site conditions suitable for native animals and
plants to recolonize (Lugo 1997). Plantations are most
likely to contribute positively to biodiversity conservation
when used to reforest degraded or deforested areas (Moss
et al. 1979; Evans 1982; Moore and Allen 1999). In
addition, plantations can benefit landscape composition
(Estades and Temple 1999). It may seem that the best use
of all plantations would be to maximize fiber production
while minimizing costs (Moore and Allen 1999), but this
assumes that plantations will increase the amount of natural
forests that are taken out of production or harvested
minimally. Replacement of native forest with exotic tree
plantation could cause important changes in diversity and
composition of community in local and regional scale
(Brockerhoff et al. 2001).
The most disputed of plantation management is
extensive use of exotic species in plantations (Potton 1994;
Tucker et al. 1998). Most studies suggested that polyculture plantations have abundant and diverse flora and
fauna more than monocultures (Baguette et al. 1994;
POURBABAEI et al. Plant species diversity in plantation forest of North Iran
Donald et al. 1997; Khanna 1997; Twedt et al. 1999;

Humphrey et al. 2002; Carnus et al. 2006), especially
where native species are planted. Poly-culture plantations
are generally host of many animal species because of the
strong relationship between native plant diversity and
animal diversity within a divers forest stand (Braganc et al.
1998; Donald et al. 1998). Using native fast growing
species such as Alnus subcordata and Acer velutinum
increase the yield potential in short rotations caused by
decreasing timber harvesting in natural forest in the north
of Iran; on the other hand, these forests could play their
environmental roles. Also, using native species in plantations could decrease the concern of adaptation and being
infected the pests and diseases. The study on plantation in
Iran and the other part of the world was extended in recent
decades (Abdy and Mayhead 1992; Allen et al. 1995;
Menalled et al. 1998; Ferris et al. 2000; Coroi et al. 2004;
Lindenmayer and Hobbs 2004; Yamashita et al. 2004; Lee
et al. 2005; Lemenih and Teketay 2005; Pourbabaei et al.
2005; Corney et al. 2006; Ginsberg 2006; Mosayeb Neghad
et al. 2007; Pourbabaei and Roostami Shahraji 2007;
Poorbabaei and Poorrahmati 2009).
The study on herbaceous species diversity in Lajim,
Mazandaran province in Iran showed that herbaceous
species diversity in natural broadleaved forest was
significantly more than coniferous plantation (Ghelichnia
2003). In addition, comparison of plant biodiversity in
Alnus subcordata and Acer velutinum-Fraxinus excelsior
plantations in Guilan province of Iran indicated that
diversity indices (Shannon-Wiener and Mc Arthur's N1),
evenness index and species richness in Acer velutinumFraxinus excelsior was more than Alnus subcordata
plantation and there was no significant difference between
plantations in diversity and evenness indices, but there was
a significant difference in richness (Pourbabaei et al. 2005).
The investigation of biodiversity indices (Simpson,
73
Menhinick richness and Peets evenness) of woody species

in mixed coniferous stand of Pinus nigra-Picea abies and
natural broad-leaved coppice stand revealed that the most
number of native species was recorded in natural broadleaved coppice stand, but richness and evenness indices
had lower value in natural forest (Memarian et al. 2007).
Comparison of vegetation diversity in forest floor and
fauna diversity in coniferous and broadleaved plantations
showed that flora and fauna diversity was lower in
coniferous plantation. Therefore, the forest floor diversity
could be a criterion to realize the effect of plantation on
wildlife diversity (Magurran 1996).
Considering the necessity of plantation and importance
of biodiversity conservation in all life forms (tree, shrub,
herb and regeneration) in the north forests of Iran, the objective of this study was the investigation and comparison of
the effects of plantation on plant species diversity in
Darabkolas region, Mazandaran province, north of Iran.
Study area
Three areas were selected including broadleaved
plantation consist of Acer velutinum and Alnus subcordata
(parcel No. 30), coniferous plantation of Cupressus
sempervirens var. horizontalis and Pinus brutia (parcel No.
40) and natural broad-leaved mixed forest (parcel No. 29),
which located in district No. 1 of Drarabkola region in
Mazandaran province, north of Iran and had 11, 14 and 15
ha area, respectively. These areas located in 36 28 00 N
latitude and 52 31 00 E longitude in watershed No.74.
Average altitude is 300 m a.s.l. in broad-leaved plantation,
270 m a.s.l. in coniferous plantation and 450 m a.s.l. in
natural forest. Average slope in the most parts of all regions
was 30 and general aspect was northern ( Figure 1).
Figure 1. Location of the study area, Darabkola, Sari district, Mazandaran Province in northern Iran. Parcel no. 29. Natural stand, parcel
no. 30. Broad leaved plantation, parcel no. 40. Coniferous plantation.
74
Procedures
The sampling method was systematic random. 35
sampling plots were surveyed in each area. All tree species
(diameter at breast height, DBH 10 cm) were identified
and measured, and shrub species and seedlings (DBH < 10
cm) were identified and counted in 400 m2 (20 m 20 m)
sampling plots. Cover percentage of herbaceous species
were estimated according to Braun-Blanquet criterion in 64
m2 (8 m 8 m) circular plots obtained minimal area
method (Poorbabaei and Poorrahmati 2009; Eshaghi Rad et
al. 2009).
Simpson (1-D) and Hill's N2 diversity indices were used
due to more sensitivity to the most frequent plant species.
Shannon-Wiener (H) and Mc Arthur's N1 diversity indices
were used due to more sensitivity to frequency of rare
species. Smith and Wilson evenness index (Evar) was used
to study the distribution of individual among species.
Diversity and evenness indices of each plot were calculated
using Ecological Methodology software. Species richness
(S) was number of species per plot (Krebs 1999). Finally,
Jacard's similarity index was used to find similarity among
regions (Pourbabaei 2004):
JI
a
abc
JI: Jacard's index, a: number of common species in

samples or communities, b: number of species that exist
just in first sample or community, c: number of species that
exist just in second sample or community.
Three studied areas were compared using one-way
ANOVA and Tukeys test.
Results
Results showed that natural forest has 32 plant species,
which consist of 10 trees, 4 shrubs and 18 herbaceous
species. 30 plant species were recorded in Alnus
subcordata-Acer velutinum broadleaved plantation
including 7 trees, 5 shrubs and 18 herbaceous species, and
30 plant species including 8 trees, 4 shrubs and 18
herbaceous species were presented in Cupressus
sempervirens var. horizontalis-Pinus brutia coniferous

plantation (Table 1). Values of biodiversity indices in tree,
shrub, herbaceous and regeneration layers had higher
values in the natural forest (Table 2).
Natural forest had the highest value of evenness in all
vegetation layers and coniferous plantation had the lowest
value (Table 3). Natural forest had the highest value of
species richness in all vegetation layers other than
herbaceous layer (this layer had the most value in broad
leaved plantation) (Table 4). ANOVA test indicated that
there were significant differences among diversity,
evenness indices and richness in all vegetation layers in the
three studied areas (P < 0.05) (Table 5).
In tree layer, the results of Tukey test (P < 0.05)
revealed that there was no significant difference between
natural forest and broadleaved plantation in Simpson,
ShannonWiener and evenness indices. While, natural
forest and coniferous plantation had significant difference
in all diversity indices. Also, broadleaved and coniferous
plantations had significant differences in all diversity
indices in this layer.
In shrub layer, Tukey test revealed that there was no
significant difference between natural forest and broad
leaved plantation in all diversity indices. In addition,
broadleaved and coniferous plantations had significant
difference in species richness. There was significant
difference between natural forest and coniferous plantation
in all indices in this layer.
In herbaceous layer, Tukey test indicated that there was
no significant difference between natural forest and broad
leaved plantation, but there was significant difference
between natural forest and coniferous plantation in all
indices. In addition, broadleaved and coniferous
plantations had significant differences in all indices except
evenness index.
In regeneration layer, Tukey test showed that there was
no significant difference between natural forest and broad
leaved plantation, but natural forest and coniferous
plantation and also two plantations had significant
differences in all indices.
Jacard's similarity index revealed that natural forest and
broadleaved plantation had the most similarity in woody
and herbaceous layers, and the lowest value was between
natural forest and coniferous plantation (Table 6).
Table 2. Mean and standard errors values of diversity indices in different vegetation layers in the studied areas
Vegetation layers
1-D
N2
H'
N1
Tree
0.32 0.03
1.47 0.10
0.76 0.07
1.68 00.11
Shrub
0.21 0.03
0.91 0.13
0.46 0.07
1.03 0.15
Herb
0.56 0.04
2.94 0.25
1.96 0.12
3.53 0.27
Regeneration
0.40 0.03
1.75 0.08
0.92 0.06
1.94 0.08
Tree
0.24 0.04
1.01 0.16
0.52 0.08
1.07 0.17
Broadleaved plantation
Shrub
0.15 0.03
0.66 0.13
0.33 0.07
0.73 0.15
Herb
0.55 0.03
2.73 0.19
1.61 0.09
3.34 0.22
Regeneration
0.34 0.03
1.50 0.13
0.81 0.08
1.68 0.14
Tree
0.05 0.01
0.31 0.08
0.12 0.03
0.34 0.09
Coniferous plantation
Shrub
0.09 0.02
0.35 0.09
0.17 0.04
0.37 0.10
Herb
0.18 0.04
0.81 0.18
0.47 0.10
0.94 0.21
Regeneration
0.19 0.04
0.80 0.17
0.41 0.09
0.87 0.18
Note: 1-D = Simpson diversity index, N2 = Hill's diversity index, H = Shannon-Wiener diversity index, N1 = Mc Arthur's diversity index
Natural forest

Table 3. Mean values of evenness in different vegetation layers in
the studied areas
Vegetation
layers
Tree
Shrub
Herbaceous
Regeneration
Natural forest
0.56 0.05
0.35 0.05
0.55 0.04
0.62 0.03
Broadleaved
plantation
0.43 0.07
0.28 0.06
0.42 0.04
0.51 0.05
Coniferous
plantation
0.11 0.03
0.15 0.04
0.25 0.05
0.28 0.06
Table 4. Mean values of species richness in different vegetation

layers in the studied areas
Vegetation
layers
Tree
Shrub
Regeneration
Herbaceous
Natural
forest
2.26 0.13
1.83 0.16
2.51 0.12
5.43 0.44
Broadleaved
plantation
1.71 0.13
1.37 0.14
2.43 0.21
6.17 0.49
Coniferous
plantation
1.71 0.07
0.88 0.14
1.46 0.23
1.57 0.34
Table 5. Results of ANOVA analysis of diversity, evenness

indices and richness in different vegetation layers
Vegetation Biodiversity
Mean
df
F
P-Value
layers
indices
square
Tree
1-D
0.665
2
17.021
0.000
N2
12.061
2
18.419
0.000
H
3.609
2
17.918
0.000
N1
15.890
2
21.286
0.000
Evar
1.906
2
15.474
0.000
S
10.314
2
23.483
0.000
Shrub
1-D
0.130
2
2.914
0.059
N2
2.814
2
4.179
0.018
H
0.783
2
4.146
0.019
N1
3.782
2
4.739
0.011
Evar
0.381
2
3.164
0.046
S
8.267
2
10.890
0.000
Herb
1-D
1.666
2
26.453
0.000
N2
48.317
2
19.534
0.000
H
16.312
2
27.282
0.000
N1
73.398
2
23.602
0.000
Evar
0.823
2
8.307
0.000
S
213.438 2
33.905
0.000
Regeneration 1-D
0.426
2
7.432
0.000
N2
8.453
2
8.663
0.000
H
2.486
2
7.957
0.000
N1
10.946
2
9.751
0.000
Evar
1.072
2
9.391
0.000
S
12.067
2
9.185
0.000
Note: 1-D = Simpson diversity index, N2 = Hill's diversity index,
H = Shannon-Wiener diversity index, N1 = Mc Arthur's diversity
index, Evar = Smith and Wilson evenness index, S = species richness.
Table 6. Percentage of Jacard's similarity index of woody and
herbaceous species in the studied areas
HerbaWoody
Study area
ceous
species
species
Natural forest-Broadleaved plantation
0.63
0.50
0.37
0.40
Natural forest-Coniferous plantation
Broadleaved plantation-Coniferous
0.50
0.38
plantation
75
Discussion
There is no single or simple answer to the question of
whether planted forests are good or bad for biodiversity.
Plantations can have either positive or negative impacts on
biodiversity of the tree, stand or landscape level (Hartley
2002; Zerbe 2002; Lindenmayer and Hobbs 2004;
Ginsberg 2006; Paritsis and Aizen 2007). It has been
argued that plantation may protect natural biodiversity
indirectly by enabling greater wood production from
smaller, intensively managed areas, thus sparing remaining
natural forests harvesting pressure (Carnus et al. 2006).
In our study, the number of tree species recorded in the
natural forest, broadleaved and coniferous plantations
were 10, 8 and 7, respectively. Grazing, collection of litter
and dry branches may be most likely reduced in woody
species number in plantations (Yirdaw and luukkanen
2003), and forest managers should pay attention to the
natural composition of forest communities (Eshaghi Rad et
al. 2009). In this study, species of shrubs was higher in
broadleaved plantation. Plantation management studies
have shown that shrub species are more resistant and
recover more easily than tall tree species (Nagaike 2002)
We found that number of plant species in natural forest
were significantly higher than plantations. Single species
plantations have often been criticized for being associated
with a low level of biodiversity in the ecosystems
(Montagnini et al. 1995; Lindenmayer and Hobbs 2004).
High biological diversity at the landscape level could bring
about many benefits from forests including wood
production, water and environmental conservation, carbon
stocking, education and science recreation. To achieve this
objective, it is essential to retain some natural forests in the
reforestation area, avoiding large scale clear-cutting (Kamo
et al. 2002).
Tree species richness and evenness were the lowest
values in coniferous plantation. It seems that due to lack of
attention to the mix structure and presence of many
individuals of two species include Cupressus sempervirens
var. horizontalis and Pinus brutia reduced evenness in this
site. It was stated that species richness and evenness were
the most in natural forest and the least in Acer velutinum
and Pinus taeda plantations in the north of Iran (Baktash
2003).
Also, we found that diversity of herbaceous species in
natural forest is more than plantations. Ghelichnia (2003)
showed the same results in the comparison of diversity in
natural hardwood stand and softwood plantation in
Mazandaran province, north of Iran.
Investigation on the relation of plant diversity, planting
distance and soil types in native and exotic pine plantations
showed that more planting distance caused more richness,
less woody species abundant and more coverage of
herbaceous species. Also, light absorption of pine's needles
and their fallings had negative impact on plant diversity
(Newmaster et al. 2006). Broadleaved deciduous species
increase the organic matter of soil and caused soil
productivity (Jalali et al. 2007).
Results of this study revealed that seedling had been
established in broadleaved plantation. The growth of
under storey is influenced by various factors, including
76
competition for light and nutrients,

Table 1. List of plant species in the studied areas
pattern tree regeneration, soil and
Broad
microclimatic effects and past stand
Natural Coniferous
Scientific name
Family name
leaved
conditions (disturbance) (Kamo et al.
forest plantation
plantation
2002). It remains to be determined
Acer velutinum Boiss.
Aceraceae
+
+
which factors were most important in
Alnus subcordata C. A. Mey
Betulaceae
+
+
our study stands. Many studies
Artemisia annua L.
Asteraceae
+
considered that plantations should be
Asplenium trichomanes L.
Aspleniaceae
+
+
+
managed
to
produce
natural
Atropa belladonna L.
Solanaceae
+
+
regeneration instead of clear cut
Bromus benekenii (L.)Triman
Poaceae
+
(Kamo et al. 2002; Nagaike 2002;
Carex stenophylla Wahlenb.
Cyperaceae
+
+
+
Carpinus betulus L.
Betulaceae
+
+
+
Nagaike et al. 2006; Koonkhunthod
Chenopodium album L.
Chenopodiaceae
+
et al. 2007). Increasing structural
Cornus australis C.A. Mey.
Cornaceae
+
complexity could attract seed
Crataegus microphylla C.Koch
Rosaceae
+
+
+
dispersing wildlife and thus increase
Cupressus sempervirens var.
Cupressaceae
+
seed inputs from neighboring native
horizontalis G.
forest (Koonkhunthod et al. 2007).
Diospyros lotus L.
Ebenaceae
+
Natural forests as a seed source, the
Equisetum sp.
Equisetaceae
+
forest
plantations
should
be
Fagus orientalis Lipsky.
Fagaceae
+
established contiguous to natural
+
Geum urbanum L.
Rosaceae
+
Gleditschia caspica Desf.
Caesalpinaceae
+
+
forest stands or, if that is not
Hedera pastuchovii Woren.ex. Grossh. Araliaceae
+
possible, plantation corridors may be
Hypericum perforatum L.
Hypericaceae
+
+
established (Yirdaw and luukkanen
Lamium album L.
Lamiaceae
+
+
+
2003). Different plant and animal
Lamium amplexicaule L.
Lamiaceae
+
species
that
guaranty
the
Mentha pulegium L.
Lamiaceae
+
+
+
environmental health, adapted to
Mespilus germanica L.
Rosaceae
+
+
+
native trees thus native tree species
Morus nigra L.
Moraceae
+
should accompany exotic tree species
Nepeta micrantha Bge.
Lamiaceae
+
in plantations (Hartley 2002).
Oplismenus undulatifolius (Ard) P.Beauv. Gramineae
+
+
Oxalis corniculata L.
Oxalidaceae
+
+
+
Investigation of stand structure and
Parrotia persica (DC.) C.A. Mey.
Hammamelidaceae
+
+
+
species diversity of Notofagus
Phylitis scolopendrium(L.) Newm.
Aspleniaceae
+
+
+
dombeyi and pine plantations showed
Picris
pauciflora
Willd.
Asteraceae
+
that plantation of exotic pine species
Pinus brutia Ten.*
Pinaceae
+
had significant effect on reduction of
Plagemnium cuspidatum L.
Mniaceae
+
+
biodiversity and species richness and
Potentilla reptans L.
Rosaceae
+
changed vegetation structure (Paritsis
Poterium sanguisorba M.
Rosaceae
+
and Aizen 2007).
Prunus divaricata Ledeb.
Rosaceae
+
+
+
+
Natural forest had the most value
Pteris cretica L.
Pteridaceae
+
+
Punica granatum L.
Punicaceae
+
of richness in regeneration layer in
Quercus castaneaefolia C. A. Mey
Fagaceae
+
+
+
our
study,
and
minimum
Robinia pseudoacacia L.*
Papilionaceae
+
regeneration was considered in
Rubus
hyrcanus
Juz.
Rosaceae
+
+
+
coniferous plantation because of
Rumex acetosa L.
Polygonaceae
+
closed canopy cover and litter in
Ruscus hyrcanus Woron.
Liliaceaea
+
+
forest floor. Also, silvicultural
Salvia nemorosa L.
Lamiaceae
+
treatments (especially releasing in
Scutellaria nepetifolia Benth.
Lamiaceae
+
broadleaved plantation) caused the
Smilax excelsa L.
Liliaceaea
+
+
growth of invasive species such as
Sorghum sp.
Poaceae
+
Ulmus glabra Hudson.
Ulmaceae
+
Rubus hyrcanus that prevented
Veronica persica Poir.
Scrophulariaceae
+
regeneration growth in broadleaved
Viola alba L.
Violaceae
+
+
+
plantation, but this problem was less
Zelkova carpinifolia (Pall.) Dipp.
Ulmaceae
+
+
+
in natural forest due to the canopy
Note:
(+:
presence,-:
absence,
*:
Exotic
species)
coverage of seed trees that are the
best shelter of lighting conditions to
survival of natural regeneration.
Nevertheless, total number of regeneration was low in plant diversity (Jobidon et al. 2004). Plantation
natural forest because of the forest degradation and the less management practices, such as weeding, salvage logging
abundance of regeneration of Fagus orientalis and Quercus and thinning effectively set the plant community back to a
castaniefolia, in contrast regeneration of Parrotia persica previous stage of succession (Nagaike et al. 2003)
There were higher similarity between natural forest and
was high. It seems that thinning could help increase
broadleaved
plantation in woody and herbaceous species
structural diversity. Then, structural diversity maintains
layers (63 and 50% respectively), while coniferous
plantation and natural forest have the lowest similarity in

woody species layer (37%). Poorbabaei and Poorrahmati
(2009) considered high similarity in species composition
between plantation and adjacent natural forest due to the
natural forest was the main source of seed in plantation.
Neighboring of plantation and natural forest has been
resulted in dispersion of hardwood trees seeds within the
plantation.
CONCLUSION
In conclusion, we found that the natural mixed forest
had the most plant diversity and plantations reduced
species diversity in this area. Also, we found that
coniferous species could diminish the biodiversity
especially in herbaceous layer more than broadleaved
species in plantations.
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Pages: 79-85
ISSN: 1412-033X
E-ISSN: 2085-4722
Determination of long-tailed macaques (Macaca fascicularis)

harvesting quotas based on demographic parameters
1
YANTO SANTOSA1, KUSMARDIASTUTI2, AGUS PRIYONO KARTONO1, DEDE AULIA RAHMAN1,
Wildlife Ecology Laboratory, Department of Forest Resource Conservation and Ecotourisme, Faculty of Forestry, Bogor Agricultural University, IPB
Campus at Darmaga, Bogor 16680, P.O. BOX 168, West Java, Indonesia, Tel. +62-251-8629150, Fax. +62-251-8629150, e-mail : dede_fahutanipb@mail.com
2
Department of Forestry, Natural Resources Conservation Center of Central Java, Semarang, Central Java, Indonesia.
Manuscript received: 15 November 2011. Revision accepted: 28 Maret 2012.
ABSTRACT
Santosa Y, Kusmardiastuti, Kartono AP, Rahman DA. 2012. Determination of long-tailed macaques (Macaca fascicularis) harvesting
quotas based on demographic parameters. Biodiversitas 13: 79-85. Harvesting quota of long-tailed macaques for breeding purpose
should be set up based on demographic parameters. The objectives of this research were to determine demographic parameters affecting
the set up of harvesting quota and the sustainable harvesting quotas of long-tailed macaca in Indonesia. This study was expected to
provide useful information for consideration of setting up harvesting quotas for long-tailed macaque in Indonesia. This study was
conducted in November 2009-Januari 2010 using the equation of Q = Nt-MVP. The results showed varied harvesting quotas for
different age classes of long-tailed macaque with an average number of 5 for infant males, 3 for infant females, 5 for juvenile males, 4
for juvenile females, 6 for sub-adult males, 8 for sub-adult females and 2 for adult males. The dominant variable determining quota was
survival rate.
Key words: fecundity, harvesting quota, long-tailed macaque, survival.
INTRODUCTION
Indonesia is one of the largest exporters of longtailed macaques in the world, besides Malaysia and the
Philippines (MacKinnon 1983). The demands of the
species are growing due to the development of technology
and products used by humans, where long-tailed macaque
has significant role as animal model for biomedical
research (Eudey 2008). While most wildlife is traded
locally and the majority nationally (that is within the
political borders of a country or state), long tailed-macaque
is also traded internationally (Stoett 2002; Blundell and
Mascia 2005; Schlaepfer et al. 2005; Nijman and Shepherd
2007). Long-tailed macaques may be taken from the wild
to meet the needs of domestic market, while export
demands should come from captive breeding. This is in line
with the Decree No.26/Kpts-II/94 concerning the
utilization of long-tailed macaque (Macaca fascicularis)
and southern pig-tailed monkey (Macaca nemestrina) for
export, that exported animals must come from captive
breeding. To ensure the sustainable population of longtailed macaque, harvesting quota was used. The number of
long-tailed macaques captured from the wild should follow
the catch quotas issued by the Director General of Forest
Protection and Nature Conservation (PHKA), Ministry of
Forestry as the Management Authority in accordance to the
recommendation from the Indonesian Institute of Sciences
(LIPI) as the Scientific Authority. For example the number
of harvesting quota for long-tailed monkeys from the wild
in 2006 was 2.000, which increases to 4.100 in 2007, 5.100
in 2008, 15.100 in 2009 but dropped to 5,000 in 2010

(Ditjen PHKA 2011).
Special Province of Yogyakarta (DIY) is one of the
locations where long-tailed macaques were harvested. Until
2009, long-tailed macaques were captured from Paliyan
Wildlife Sanctuary and Kaliurang Recreation Forest. The
harvesting quotas for long-tailed macaque in these areas
were 200 animals in 2006 and 2007, zero in 2008 and 1200
animals in 2009 (BKSDA DIY 2007-2010). Previously,
based on the export statistics compiled by the Directorate
of Protection and Nature Conservation (BKSDA DIY
2002), a total of more than 86,332 long-tailed macaques
were exported between the periods of 1992-1997.
Currently, the harvest quotas issued by PHKA contain only
the figures for allowable catch, and have not considered
detailed classification of the speciess sex and age classes.
This has raised a concern that such quotas will threaten the
sustainability of the macaques population (Santosa and
Desi 2010).
Numerous studies have concluded that regulation of
wildlife trade laws within Asia, be it in relation to
international or domestic trade, are insufficient (Davies
2005; Lee et al. 2005; Giles et al. 2006; Nijman 2006;
Nekaris and Nijman 2007; Shepherd and Nijman 2007a, b;
Eudey 2008; Zhang et al. 2008; Chen et al. 2009),
furthermore, wildlife in Southeast Asia such as a long
tailed-macaque is under attack due to habitat loss and
degradation, global climate change, commercial hunting
and competition with introduced species (McNeely et al.
2009; Sodhi et al. 2004). Therefore, there is an urgent need
80
for a more effective wildlife trading regulatory mechanisms

f, particularly in Indonesia.
Santosa (1996) says that harvesting quota is determined
by demographic parameters including birth rates, mortality,
sex ratio and size of population. These parameters are
important components for the development of wildlife
population. Thus, the planning and determination of
harvesting quotas requires information on demographic
parameters of the population in question (Santosa 1993).
Based on the above, research on the determination of longtailed macaque harvesting quotas based on demographic
parameters was conducted in order to determine the harvest
quota of long-tailed macaques based on number and sex
ratio, in Paliyan Wildlife Sanctuary and Kaliurang
Recreation Forest, both in Yogyakarta, Indonesia. It is
expected that such results would become a model for the
determination of long-tailed macaques harvesting quotas in
other places and identification of variables that affect the
demographic parameters of quotas.
f =
f
Xi
Bi
Px
Lx
Lx +1
Lx
Xi
Bi
= crude birth rate/fecundity
= number of infants in ith group
= number of reproductive adult females in i-group
= Lx+1
= 1 -Mortality
= number of individuals in X +1age class
= number of individuals in age class
Population growth
To determine the population growth (Nt +1) in each
group was used a modified of matrix Leslie
not adrift density (Priyono 1998) where only the female
part of the populations would be considered, while the male
populations were determined using sex ratio. The
calculation was made using Powersim 2.0. The matrix
formula for the calculation was:
M x Nt = Nt+1

Time and location
The study was conducted from November 2009 to
January 2010 in Yogyakarta Special Province, precisely in
Paliyan Wildlife Sanctuary of Gunungkidul District which
is a karts region with an area of 434.60 hectares and in
Kaliurang Recreation Forest of Sleman District which is a
montane forest.
Data collection
Data on demographic parameters of long-tailed
macaque (Macaca fascicularis) were collected using
concentration count method based on preliminary
information. Observations were carried out twice daily, in
the morning (6-9 a.m) and afternoon (3-6 p.m). Data
recorded include number of individuals in each group,
number of individuals based on sex, age and class. The
numbers of individuals were recorded based on the
encountered/directly observed individuals within the
observation trail. Individuals were classified based on age
as infants, juvenile, sub-adult and adults after Napier and
Napier (1967), i.e. infant (0-18 months/ still breastfeeding),
juvenile (18 months-4 years old), sub-adult (4-9 years old),
and adult (9-21 years). Classification of age classes based
on Napier and Napier (1967) was more likely to be
applied in
a
study
in nature,
related
to the
difficulty to distinguish individual age classes if age is
classified into infant, weaning, juvenile, sub-adult and
sexual maturity (male and female) by Rowe (1996) which
further allows for a controlled study such as in captivity.
Data analysis
Demographic parameters
Numbers of individuals in each group, as well as
numbers of individuals based on sex and age classes were
used to calculate the crude birth rate / fecundity, and
survival rate for each age class. The formula used to
calculate fecundity is as follows:
M=
Fxm
Fxd
Xd
0
P1
1
P2
2
P2
3
Fm
Fd
N0,t
P1
P2
N2,t
P2
N3,t
Nt =
N1,t
= sub-adult fecundity
= adult fecundity
= adult age class
= proportion of infant age class
= probability of infant survival
= proportion of juvenile age class
= probability of juvenile survival
= proportion of sub-adult age class
= probability of sub-adult survival
= proportion of adult age class
Minimum Viable Population (MVP)

Minimum viable population (MVP) is the smallest
population size that will ensure a species survival in the
long term (Shaffer 1981). MVP is important when
decisions are made which can alter the available habitat,
and consequently the species population and risk of
catastrophic extinction (Harcourt 2002). Similar to the
population growth, MVP was also calculated for each age
class and sex based on females population only, whereas
males populations were obtained based on sex ratio. The
analysis used two algebraic equations, i.e. B = D and Nt,
where the intersection value between the two equations
determined the MVP value. The formula used was:
B=D
B
D
= birth (birth)
= death (mortality)
Further described by the following equation:
SANTOSA et al. Harvesting quotas of Macaca fascicularis
Fxm. Xm + Fxd. Xd = mb. Xb + ma. Xa + mm. Xm +

md.Xd. ..... ( i)
Fxm
Xm
Fxd
Xd
mb
Xb
ma
Xa
mm
Xm
md
Xd
= number of individuals of sub-adult age class
= adult fecundity
= number of individuals of adults age class with
maximum breeding age of 12 years
= infant mortality
= number of individuals of infant age class
= juvenile mortality
= number of individuals of juvenile age class
= sub-adult mortality
= proportion of adult mortality
= number of individuals of adult age class
Nt value used N1 value as constant for Powersim, thus

if translated in algebraic form became:
Nt = (Xm + Fxd Fxm.. Xd + b. Xb) + (XB + a Pxb.. Xa)
+ (Xa + m Pxa.. Xm) + (Xm + d Pxm.. Xd) ..... .. (ii)
Nt
Fxm
Xm
Fxd
Xd
b
Xb
Pxb
a
Xa
PXA
m
Xm
Pxm
d
Xd
= population size in year-t

= adult fecundity
= number of individuals adult age class with maximum
breeding age of 12 years
= proportion of infants
= number of individuals of infant age class
= infant survival rate
= proportion of juvenile
= number of individuals of juvenile age class
= juvenile survival rate
= proportion of sub-adult
= number of individual of sub-adult age class
= sub-adult survival rate
= proportion of adult
= number of individual of adult
The two equations above were combined to produce the

following intersection:
Fxm. Xm + Fxd. Xd-mb. Xb + ma. Xa + mm. Xm +
md.Xd = Nt-Fxm. Xm + Fxd. Xd + b. Xb) + (XB + a
Pxb.. Xa) + (Xa + m Pxa.. Xm) + (Xm + d Pxm..
Xd)......... ( iii)
Nt
Fxm
Xm
Fxd
Xd
mb
Xb
ma
Xa
mm
Xm
md
Xd
b
Pxb
a
= population size in year-t

= adult fecundity
= number of individuals of adult age class with maximum
breeding age of 12 years
= infant mortality
= number of individual of infant age class
= juvenile mortality
= the number of individuals of juvenile age class
= sub-adult mortality
= proportion of adult mortality
= number of individuals of adult age class
= proportion of infants
= infant survival rate
= proportion of juvenile
PXA
m
Pxm
d
81
= juvenile survival rate

= proportion of sub-adult
= sub-adult survival rate
= proportion of adult
Value of harvesting quota

Value of harvesting quota represented the difference
between the numbers of existing individuals with the
minimum viable population (MVP). The harvesting quota
was calculated for each age class and sex, using the
following formula:
Qij
= Ntij-MVPi
Qi
= harvesting quota of i-age class to j-sex
Ntij
= number of individuals of i-age class with j-sex on year-t
MVPij = minimum viable population of i-age class with j-sex
Analysis of demographic variables that determine quota

Based on the matrix equation of population growth and
MVP, the variables affecting demographic parameters were
survival quota (Px) and fecundity. To determine the
dominant variables affecting the quotas, a regression test
was done using the following equation:
Y
= b1X1 + b2X2 +
b1
1
X2
= regression coefficient
= average fecundity
= average survival rate
Sensitivity analysis of dominant quota determinant

variables
Sensitivity test was conducted for the dominant
variables determining the quota, which was done by
addition and subtraction of 10% to 30% of the dominant
variables to observe the extent of the impacts on the value
of the quota.

Long-tailed macaque group characteristics
During the observation periods, three groups of longtailed macaques were encountered in Paliyan Wildlife
Sanctuary and four groups in Kaliurang forest. Although
Paliyan Wildlife Sanctuary has been disturbed by human
activities, nevertheless the number of individuals within
each group was higher than those in Kaliurang forest.
Group size referred to the number of individuals contained
in a group (Priyono 1998). In Kaliurang forest, the group
size ranged from 20-45 animals, whereas in Paliyan
Wildlife Sanctuary it ranged between 48-68 animals (Table
1). This was in line with Bismarks (1986) finding where
formation and size of groups varies according to habitat
types. In primary forest, one group of long-tailed macaque
comprised of 10 individuals, in mangrove forest about 15
individuals, and more than 40 individuals in disturbed
forest.
Adult
M
F
8
8
6
16
17
9
3
3
4
11
5
7
68
57
48
5
5
4
2
9
9
7
3
3
4
3
2
5
7
4
3
36
45
30
20
There were four groups of long-tailed macaque groups

observed in Kaliurang forest with a total of 131 individuals,
which indicated an increased from previous research
results. Ningrum finds as many as 63 individuals in 2002,
which later developed into 2 sub-groups with 80
individuals (in Fallah 2005) and again into 3 sub-groups in
2009 (Raharjo 2009).
Demographic parameters
Age structure and sex ratio
Age structure is the ratio of individuals of a population
in each age class. Table 2 showed the age structure of longtailed macaques in Paliyan Wildlife Sanctuary and
Kaliurang forest based on age classification by Napier and
Napier (1967).
Table 2. Age structure of long tailed macaque group
Group size
Number of individuals
(individuals) Infant: juvenile : sub-adult : adult
Paliyan Wildlife Sanctuary
1
68
12 : 18 : 24 : 14
2
57
10 : 14 : 25 : 8
3
48
9 : 13 : 15 : 11
Kaliurang Forest
1
36
6 : 8 : 14 : 8
2
45
9 : 13 : 14 : 11
3
30
5 : 7 : 11 : 7
4
20
4:6:5:5
Group
7
6
5
4
3
2
1
0
Group1
Group2
Group3
Agegroup
Adult
Subadult
Juvenil
Group4
Infant
Numberofindividu
In almost all groups, the highest number of individuals

was found within the sub-adult age class and the lowest
within the infant age class. These results were similar to
those found in Musi Hutan Persada Forest Concession
(Priyono 1998). The weakness of a qualitative grouping is

that time interval between age classes was not the same and
that there would be accumulation of individuals in
particular age class. Priyono (1998) states that this
condition would result in the emergence of a declining
population structure (regressive population). The
development of population in such age structure would
continue to decline if the environmental conditions stayed
the same, which after some time, would result in the
extinction of the population (Peres 2001).
To overcome the inequality of age interval of age class,
a population structure was developed within the same age
class by dividing the population into age-class intervals.
This was done to obtain an increased age structure where
the infant age class would be higher than other age classes
(progressive populations). Figure 1 showed the graph for
group size of each age class with its range of class.
Age structure can be used to measure the success of
wildlife development and hence to assess its sustainability
prospects. Figure 1 indicated sustainable populations of
each group of long-tailed macaque in both Paliyan Wildlife
Sanctuary and Kaliurang forest shown by the greater
number of infants as compared to adults, thus possible
future regeneration could occur properly.
Sex ratio is the ratio of males to females. Sex ratio of
each group of long-tailed macaques in Paliyan Wildlife
Sanctuary and Kaliurang forest were given in Table 3. In
general, the long-tailed macaque groups in both Paliyan
Wildlife Sanctuary and Kaliurang forest had a sex ratio of
1: 2 (Table 3), which was similar to research results by
Sularso (2004) in Alas Purwo National Park and by
Tangguh (2007) in Pangandaran Forest Recreation Park.
Tabel 3. Sex ratio of each group of long-tailed macaque
Group
Number of males

1
18
2
17
3
14
Kaliurang Forest
1
11
2
13
3
9
4
7
Number of
females
Sex ratio
38
30
25
1 : 2.11
1 : 1.76
1 : 1.79
19
23
16
9
1 : 1.73
1 : 1.77
1 : 1.78
1 : 1.29
9
8
7
6
5
4
3
2
1
0
Group1
Group2
Group3
Adult
Total
Sub-adult
M
F
Subadult
Juvenile
Group
M
F
1.
12
7
11
2.
10
6
8
3.
9
4
9
Kaliurang Forest
1.
6
3
5
2.
9
4
7
3.
5
2
5
4.
4
3
3
Juvenil
Infant
Infant
Table 1. Long-tailed macaques groups characteristics
Numberofindividu
82
Agegroup
Figure 1. Age structure of long-tailed macaque group in (A) Paliyan Wildlife Sanctuary and (B) Kaliurang recreation forest
Natality and mortality

Natality is the ratio between the number of infants with
the number of productive females in sub-adult and adult
age classes. The natality of all groups in both study sites
showed similar values, i.e., between 0.44-0.67. These
suggested that on the average, half of the productive
females were able to produce offspring under the
assumption that the number of birth is 1 individual
(Lavieren 1982).
Mortality or death rate is an important determinant of
wildlife preservation. High mortality would threaten the
wildlife sustainability. Estimation of population mortality
rate of long-tailed macaques in the wild was based on the
survival rate of each age class. The survival rate of each
age class and birth rate of each long-tailed macaque groups
in the study sites were tabulated in Table 4.
Table 4. Natality and survival rates of each long-tailed macaques
group
Group
Natality
Pxb_a
Pxa_m
Pxm_d

1.
0.44
0.90
0.67
0.24
2.
0.45
0.84
0.89
0.13
3.
0.56
0.87
0.58
0.31
Kaliurang Forest
1.
0.43
0.80
0.87
0.24
2.
0.56
0.73
0.64
0.33
3.
0.45
0.84
0.79
0.27
4.
0.67
0.90
0.42
0.32
Note: Pxb_a : survival rate of infant to juvenile age class; Pxa_m :
survival rate of juvenile to sub-adult age class; Pxm_d : survival
rate of sub-adult to adult age class
The lowest survival rate was found in sub-adult and

adult age classes. The assumption made was based on the
presence of competitions for social status that caused the
release of individual adult male resulting in fewer numbers
of adult males. Meanwhile, infant deaths were usually
caused by accidents or eaten by predators (Priyono 1998).
However, in general, natural death of long-tailed macaque
was very rare. Death was often caused by hunting,
capturing to meet quotas and killing by local people as
long-tailed macaques are considered as pests in some areas
(Thoisy et al. 2000).
Population growth
The results of analysis using Powersim 2.0 on
population growth showed that each long-tailed macaque
group experienced an average increase of 20-30% annually.
Group 1 in Paliyan Wildlife Sanctuary had an initial
number of 68 individuals, and during year 1 increased to 89
and in year 2 to 108 individuals; group 2 initially had 57
individuals, then increased to 71 in year 1 and to 89 in year
2; group 3 with initial total individuals of 48 and increased
to 72 in year 1 and to 69 in year 2. Analysis on population
growth in Kaliurang forest resulted the following figures:
group 1 with an initial total number of 36 individuals,
increased to 44 in year 1 and to 53 in year 2; group 2 with
an initial number of 45 individuals, increased to 57 in year
83
1 and 69 in years 2; group 3 started with 30 individuals,

became 36 in year 1 and 44 in year 2; group 4 with starting
number of 20, increased to 23 in year 1 and 29 in year 2.
Minimum Viable Population (MVP)
The results showed that the MVP values varied between
each long-tailed macaque groups. The highest MVP was
found in group 1 of Paliyan Wildlife Sanctuary with 86
individuals and the lowest was found in group 3 of
Kaliurang forest with 37 individuals (Table 5).
Table 5. The MVP values of each group of long-tailed macaque.
Infant
Juvenile
Sub-adult
Adult
Total
Male Female Male Female Male Female Male Female
1.
10
19
7
14
5
10
7
14
86
2.
7
14
9
17
3
5
6
12
73
3.
8
3
3
6
16
6
6
11
59
Kaliurang Forest
1.
4
8
5
9
2
4
4
7
43
2.
5
10
4
8
4
8
5
10
54
3.
3
6
2
4
3
5
5
9
37
4.
6
6
4
4
5
5
9
9
48
Group
Harvest quota
Analysis of both, population growth and MVP values
suggested that MVP for each group should be obtained in
year 1 so that harvesting could be done in year 2. The
harvest quota itself was calculated for each age class and
sex to avoid over-exploitation of certain sex and age class
that could threaten groups sustainability. The value of the
harvest quota was determined based on consideration of
sex ratio, where a male can mate with more than 2 females,
even up to 1: 7 for the purpose of reproduction (Rowe
1996).
Results of this research showed that harvests for subadult age class were found greater in group 2 of Paliyan
Wildlife Sanctuary, which comprised of 11 sub-adult males
and 18 sub-adult females, while the lowest was found in
group 4 of Kaliurang forest that comprised of only 2 subadult males and 1 sub-adult female. For infant age class,
the largest harvesting quota was found in group 2 of
Paliyan Wildlife Sanctuary and Kaliurang forest, while the
largest harvesting quota for juvenile age class was found in
group 3 of Paliyan Wildlife Sanctuary. On the other hand,
the largest harvesting quota for adult age class was found in
group 1 of Paliyan Wildlife Sanctuary. The average harvest
quotas for the seven groups in year-2 comprised of 5 infant
males, 3 infant females, 5 juvenile males, 4 juvenile
females, 6 sub-adult males, 8 sub-adult females and 2 for
adult males. The harvesting quotas for each age class and
sex in each group studied were tabulated in Table 6.
Based on the calculation, for the next 11 year,
harvesting quotas for each age class and sex would
experienced annual increase with an average of 20% per
year, as shown in Figure 2. This figure showed the increase
in harvest quotas for female infant age class. The
calculation was made based on population growth data
without harvesting quota. With the increasing number of
population, the harvest quota would also increase.
B I O D I V E R S I T A S 13 (2): 79
9-85, April 2012
8
84
T
Table
6. Harvessting quotas forr each long-taileed macaque grooup
nfant
Juvvenile
In
Male Female Malle Female
P
Paliyan
Wildlifee Sanctuary
1
1.
18
S
Size
111 22
9
14
19
2
M
MVP
3
4
3
7
Q
Quota
8
2
2.
12
S
Size
100 19
6
17
M
MVP
2
14
3
0
5
3
Q
Quota
7
3
3.
15
13
8
S
Size
7
6
16
1
M
MVP
3
9
0
7
Q
Quota
4
K
Kaliurang
Foresst
1
1.
7
11
4
S
Size
6
9
8
2
M
MVP
2
0
3
2
Q
Quota
4
2
2.
12
16
6
S
Size
8
8
10
2
M
MVP
2
4
6
4
Q
Quota
6
3
3.
8
7
4
S
Size
4
4
6
1
M
MVP
1
4
1
3
Q
Quota
3
4
4.
11
8
6
S
Size
4
4
6
1
M
MVP
1
7
2
5
Q
Quota
3
4
3
5
A
Average
5
G
Group
Sub-aadult Adult
Male Female Male Female
F
11
2
9
21
10
11
6
2
4
11
14
0
12
1
11
23
5
18
3
2
1
5
12
0
6
1
5
12
6
6
4
2
2
7
11
0
6
1
5
12
4
8
3
1
2
5
7
0
6
2
4
12
8
4
4
2
2
7
10
0
5
1
4
9
5
7
2
2
0
4
9
0
3
1
2
6
6
5
1
8
2
2
0
2
3
9
0
0
Analysis of demoographic variables determ

mining quota
The
T survival rate and fecuundity were analyzed
a
usinng
lineaar regression. The averagee value of surrvival rate waas
foun
nd to be 0.60, while the aaverage fecun
ndity was 0.166.
Regrression resultts showed thhat the dom
minant survivaal
variaable determiniing the harvesst quotas was the equation Y
= 0.0
0885 X + 0.77873, meaningg that indepen
ndent variablees
(X) significantly affected deppendent variab
bles (Y). This
was indicated byy the analyssis of varian
nce regressioon
equaation, which had a valuee of R2 = 0.8174, whicch
signiified that oveerall, all indeppendent variaables (X) werre
signiificantly affeccting dependeent variables (Y) and morre
than 81% of the quota frequeency was desscribed by thhe
existting variabless. Harvesting quota would
d increase witth
the in
ncreased valuue of survival rrate.
Senssitivity analysiis of dominan
nt variables aff
ffecting quotass
Sensitivity
S
annalysis (Tablee 7) of surviv
val probabilitty
show
wed that an increase
i
and a decrease by
b 10% woulld
affecct the size of quotas, by ann increase and
d decrease of
15%
%.
Tablle 7. Sensitivityy analysis survivval chance on quota.
q
No.
1
2
3
4
5
6
7
S
Scenarios
Early survival rate
Survival rate decreased
d
by 100 %
d
by 200 %
d
by 300 %
Survival rate increased by 100 %
Av
verage quota
(iindividuals)
26
22
19
16
30
35
39
Table
T
7 signified that incrreasing surviv
val rate woulld
increease the harveest quotas. Onne effort to in
ncrease survivaal
rate was througgh populatioon managem
ment such as
a
harvesting (Novaaro et al. 22000) apart from habitaat
manaagement throuugh planting of dietary vegetation, coveer
and providing sufficient sppace. This will facilitatte
proteection of prrimates and provide opp
portunities foor
imprroving the suustainable usse of other forest speciees
(Tho
oisy et al. 20055).
CONCLUS
SION AND R
RECOMENDA
ATIONS
Figure 2. The size

F
s of harvest quota
q
for infannt females up too year1
11
Harvestingg is one of sttrategy in maiintaining a naatural

bbalance. Accoording to Bailley (1984), poopulations thaat are
n harvested rarely showeed increased in
not
i population,, thus
h
harvest
increassed the populaation growth.
The
T
size of harvesting qquotas for eaach long-taileed
macaaque group varied dependiing on the su
urvival rate annd
fecun
ndity. The average harvest quotas for th
he seven groupps
studiied in year 2 composed oof 2 infant males,
m
3 infannt
femaales, 2 juvenille males, 4 juuvenile females, 4 sub-aduult
malees and 8 sub-aadult females. The determiinant dominannt
variaable of the harrvesting quotaa was survivall rate.
As
A a scientific study, resultss of this researrch can be useed
for considerationns in settingg up quota in Indonesia,
specifically withinn the Speciall Region of Yogyakarta.
Y
T
To
balan
nce the growtth in demandss for long-taileed macaques, it
is necessary
n
to establish iintensive ex--situ breedinng
prog
gramme to redduce the pressuure of wild-caaught animals.
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Pages: 86-91
ISSN: 1412-033X
E-ISSN: 2085-4722
The population condition and availability of feed of cuscus in the Arfak

Mountain Nature Reserve, West Papua
1
ANTON SILAS SINERY1,, CHANDRADEWANA BOER2, WARTIKA ROSA FARIDA3,
Faculty of Forestry, State University of Papua, Jl. Gunung Salju, Amban, Manokwari 98314, West Papua, Indonesia. Tel. +62-986-211065, Fax. +62986-211065, email: anton_sineri@yahoo.com
2
Faculty of Forestry, Mulawarman University, Samarinda 75123, East Kalimantan, Indonesia.
3
Zoology Division, Research Center for Biology, Indonesian Institute of Sciences (LIPI), Cibinong-Bogor 16911, West Java, Indonesia. email:
wrfarida@indo.net.id
Manuscript received: 21 February 2011. Revision accepted: 28 Maret 2012.
ABSTRACT
Sinery AS, Boer C, Farida WR. 2012. The population condition and availability of feed of cuscus in the Arfak Mountain Nature Reserve,
West Papua. Biodiversitas 13: 86-91. The cuscus is a pouched marsupial grouped in the Phalangeridae family, which is nocturnal,
arboreal, herbivore, and in most cases the tail is prehensile. The animals are legally protected due to low reproduction, limited
distribution area, and high rate of illegal hunting. The illegal hunting happened not only in the production forest areas but also in the
reserve areas such as Nature Reserve of Arfak Mountain, directly or indirectly, affects the life quality of the ecosystem, mainly cuscuses
population. Therefore, it is necessary to do efforts to have a better management of the region to ensure the sustenance of many
components in it. This research is aimed to know the population density of cuscus in Arfak Mountain Nature Reserve and carried out for
two months. The method used was descriptive by using direct and indirect observation. The result shows that cuscuses existing in the
Arfak Mountain conservation area were northern common cuscus (Phalanger orientalis), ground cuscus (Phalanger gymnotis) and
common spotted cuscus (Spilocuscus maculatus). The biggest individual number is of P. orientalis with 39 individuals consisting of 18
males and 21 females, the second is of P. gymnotis with 10 individuals consisting of 4 males and 6 females, and the smallest is of S.
maculatus with 9 individuals consisting of 4 males and 5 females. From the total of 58 cuscuses, there are 38 adult and 20 young
cuscuses. There are 20 forest plant species identified as feed resources of cuscus in Arfak Mountain Nature Reserve. The parts of forest
plant consumed by cuscus are fruits and young leaves. P. gymnotis also consumes small insects such as grasshopper. The cuscuses
spread from lowland forest to highland forest (2,900 m asl)
Key words: cuscus, population, forest plant species, Arfak Mountain
INTRODUCTION
Papua and West Papua as an integral part of Indonesia
has the region's natural wealth and tremendous biodiversity
in South East Asia. The diversity includes the richness of
fauna such as mammals and the terrestrial ecosystem
diversity from the coastal to the high mountain ecosystems.
Approximately 200 species of land mammals have been
found in this region, and 154 species of which form a large
population including the endemic species and the
introduction one (Petocz 1987).
This abundance is a human life support but its existence
cannot be confirmed for sure, because of the lack of
information, the uneven distribution and excessive
exploitation. According to the International Union for the
Conservation of Nature and Natural Resources (IUCN),
Indonesian wildlife threatened with extinction are 128
species of mammals, 104 species of birds, 19 species of
reptiles, 60 species of fishes, and 29 species of
invertebrates (Anon 2004). When linked to deforestation
rate, which according to Greenpeace was 2.8 million
hectares per year, it is expected to lead to more extinction
in the future.
Cuscus is a marsupial mammal (marsupials) which is

arboreal, nocturnal, and herbivore. Menzies (1991), Petocz
(1994) and Flannery (1994), each states that the
deployment of cuscus includes the islands of Indonesia
(Papua, Sulawesi, Maluku, and Timor Island), Papua New
Guinea, New Britain, Solomon Islands, Cape York
Australia, and Queensland. In New Guinea (PNG and
Papua) there are 11 species of cuscus from the genus of
Spilocuscus (spotted cuscus) to the genus of Phalanger. In
Papua, there are seven species of cuscus, namely common
spotted cuscus (Spilocuscus maculatus), black spotted
cuscus (S. rufoniger), waigeo cuscus (S. papuensis),
northern common cuscus (Phalanger orientalis), ground
cuscus (P. gymnotis), silk hair cuscus (P. vestitus), and
hilly forest cuscus (P. permixtio).
Cuscus is protected by the Decree of the Minister of
Agriculture No. 247/KPTS/UM/4/1979 and Government
Regulation No. 7 of 1999 on the preservation of plants and
animals. Globally this species is listed in Appendix II of
Convention on International Trade in Endangered Species
of Wild Fauna and Flora (CITES).
The Arfak Mountain Nature Reserve is one of the
conservation area in Papua with an area of 68.325 ha
SINERY et al. Cuscus of the Arfak Mountain Nature Reserve, West Papua
according to the Minister of Forestry Decree No. 783/KptsII/1992 with a high biodiversity (Anonymous, 2003). The
explorations done by Lesson (1824-1827) and d'Albertis
and Beccari (1872-1873) found at least 320 species of
birds, 350 species of butterflies, and 110 species of
mammals. Aside from being a center of diversity of
butterfly wings (Ornithoptera spp), it also serves as a
habitat for many plants such as matoa, nyatoh, gaharu,
rattan, bamboo, various species of orchids and a number of
endemic animals like cendrawasih, tree kangaroos,
porcupines, skunks and various other species of animals
(Laksono et al. 2001).
According to Menzies (1991) and Flanery (1994),
Arfak Mountains is one of distribution areas of spotted
cuscus. According to Sinery (2010), in the southern part of
Arfak Mountains exist some populations of cuscus. Petocz
(1994) mentioned that unspotted cuscus have a large
population allegedly spreads in almost all mainland of New
Guinea, including Arfak Mountains. The results of
interviews with one of the staff of Natural Resources
Conservation Center of West Papua and several
communities around the nature reserve of Arfak Mountains
show that there are at least three species of cuscus in the
region. It is also known that cuscus is often hunted in the
surrounding region as a source of food, but scientific things
about the population and other aspects of the animals have
not been widely known.
This study aims to determine the condition of cuscus
population and the carrying capacity of habitat based on the
availability of food in Arfak Mountains Nature Reserve.
The results are expected as one of the information and
consideration for all parties in wildlife management efforts
both in-situ and ex-situ and the development of the Arfak
Mountains Nature Reserve area in the future.
The research was conducted in the area of Arfak
Mountains Nature Reserve, Manokwari, West Papua
Province. The method used in this research is descriptive
with the technique of direct and indirect observations and
interviews. The duration of the study was 2 months from
July to August 2007.
Equipments used in the study were the Global
Positioning System (GPS), binoculars, dissecting set,
thermo-hygrometer, clinometers, thirst meter, compass,
scales, tape measure, camera trapping, phi band, scissor
cuttings, gloves, flashlights, timer, stopwatch, machetes,
cage/plastic bags, plastic bracelets, tally sheet, a key of
Identification of the type of cuscus, camping equipment,
location map (scale 1: 350,000) and 2004 Landsat imagery.
The materials used consisted of chloroform/diethyl ether,
cotton, alcohol 70% and plastic rope.
The plot of the study was determined in accordance
with the presence of cuscus from the previous survey
results, namely the information from the regular cuscus
hunters. The area of the study site was 420 ha with a
baseline parallel to the shoreline and transect direction
parallel to the contours which is perpendicular to baseline.
87
The study began by creating a baseline that was

pioneering parallel of 1,200 m along the shoreline. The
baseline was divided into six point transects perpendicular
to the baseline and proportionally placed at the point of 01,200 m with the distance transects of 200 m. Each transect
length is 3,500 m so that the length of transect was 21,000
m and the width corresponding to the minimum sight
distance (50 m).
The data collected consists of primary data that is the
data of observations and field interviews and the secondary
data that is data obtained from the relevant authorities.
Primary data collected consists of: cuscus population (the
description of the population and the type of cuscus),
species of feed and the general condition of cuscus habitat.
Secondary data collected includes data on climate and the
general state of the study site.
The morphological data of the cuscus were tabulated
from the measurement results, direct identification, and
documentation via camera trapping. Cuscus population
density analysis was made using the equation according to
Sugianto (1994) which was as follows:
n(2n 1)A
2L r
N = density of population,
n = number of individuals encountered, A = Area of
the region (plot observations),
L = length of line / transect,
r = Distance of the points where the cuscus were
found with a track point / transect
Determination of the diversity of feed types as
indicators of habitat carrying capacity to the existence of
cuscus is done by using species diversity indices (H) with
the formula from Shannon and Wiener, (1949) in Odum,
(1993):
ni ni
H log
N N
H = diversity index (Shannon Index),

ni = number of individuals of each species
N=Number of individuals of all species
Determination of the individuals which are more
concentrated in one or several species in the study site was
conducted by using the dominance index (D) according to
the formula (Simpson 1949 in Odum 1993) as follows:
ni
C
N
C = dominance index, ni = number of individuals of

each species, N = Number of individuals of all species
Determination of individual distribution of each species
in the region carried out by using the evenness index (e) of
Pielou (1966) in Odum (1993) with the following formula:
88
H
logS
E = the evenness index,

H = the diversity index
S = number of species that was present
Data related to the climatic condition of the study site
consisting of rainfall, temperature, and humidity will be
analyzed in the tabulation.
RESULTS AND DISCUSSIONS
The results show that there are 3 species of cuscus in
the Nature Reserve of Arfak Mountains which were
captured, namely northern common cuscus (P. orientalis),
ground cuscus (P. gymnotis) and the common spotted
cuscus (S. maculatus) (Figure 1).
Communities surrounding the Arfak Mountains Nature
Reserve group cuscus into several distinct species based on
morphological characters such as coat color, body size, and
habitat. The Hatam people who were the majority in the
Arfak Mountains Nature Reserve recognizes three species
of cuscus in the region, namely mengrep (plain brown
cuscus, that is P. orientalis), minyam (ground cuscus, that
is P. gymnotis) and mbrat and mifan (common spotted
cuscus, males and females, that is S. maculatus).
Description of species and number of cuscus
The identification of six samples of P. orientalis from
Arfak Mountains Nature Reserve shows that this type of
male has a body length and weight that range from 400 mm
to 445 mm and 2,200 grams to 2,800 grams. Females body
size is smaller than the males, namely body length ranges
from 385 mm to 425 mm and body weight ranges from
2,000 grams to 2,500 grams.
The males and females of P. orientalis have similar hair
dominated by brown, along the middle dorsal stripe that
extends from the base of the nose (anterior) through the
inter-parietal bone, dorsal and to the rear (posterior) to the
base of the tail. The females fur is dark (dark brown) when
compared with males who have lighter fur (grayish brown).
The head of both is more elongated with a protruding ear
condition.
The identification of six samples of the captured P.
gymnotis of both male and female adults from the area of
Arfak Mountains Nature Reserve showed that males of this
species has body length and body weight ranged from 405
mm to 446 mm and 2,200 grams to 2,800 grams. The size
of the female bodys length and weight ranges from 385
mm to 425 mm and 2,000 grams to 2,500 grams. There are
white patches behind the ear which is clearly visible
because it is not covered with feathers in it. The color of
the dorsal of the males is like the head which is brown with
silvery effect on the tip of the feather. Finer hairs similar to
wools spread from the dorsal to ventral and ends on the
outer side of the wrist and leg.
Based on the observation on S. maculatus in the region
of Arfak Mountains Nature Reserve, there are two
variations of cuscus, namely the plain white and spotty

brown. The identification of six examples of this type of
the captured cuscus showed that males from this region
have body length and body weight ranged from 515 mm to
555 mm and 4,000 grams to 4,800 grams. The female
bodys length and weight range from 485 mm to 525 mm
and 3,000 grams to 4,100 grams. The head of the male is
yellowish brown and spread out from the base of the nose
through the inter-parietal bone toward the back (posterior).
The dorsal feathers with yellowish brown spots are spread
from the base toward the back of the head spotted with the
darker color (dark brown). This color is spread toward the
side of the body until the outer part of arms and legs and
ventral borders.
Estimated population of cuscus
Based on the results of monitoring, it is known that in
the area of 14,780 m2 or 1.4 ha there are as many as 58
individuals. From that number, 39 individuals or 67.2% is
the type of P. orientalis with the highest individual spread
compared with the other two cuscus species found at the
sites. P. gymnotis consist of 10 individuals or 17.2% and S.
maculatus consist of 9 individuals, or 15.5%. P. orientalis
has a larger population so its existence is predicted to be
able to continue even improved in the future when
compared to P. gymnotis and S. maculatus. This is because
P. orientalis has a higher reproductive capacity than the
two other species (Farida et al. 2001; Sinery 2006, 2010).
But this is not true in general, because to this date on their
population has not been known.
The results of observation indicate that two of the three
samples found each having 2 and 1 young aged about 1 to 3
months in the pouch. Body length of the young ranges 5085 mm. Based on these results it can be concluded that the
litter size of P. orientalis is one or more infants. This is
consistent with the statement of Menzies (1991) stating that
the genus of Phalanger usually gives more than one infant
in one birth so this type of cuscus has a large population
compared to the genus of Spilocuscus. But the statement
did not say how many individuals of cuscus were in a
specific scale area. According to Sinery (2010), in their
natural habitat and in the captivity cuscus deliver 1 or 2
infants in each birth with the reproductive phase of 1-2
times per year. The life span of cuscus in the wild ranges
from 10 years to13 years. The young cuscus aged less than
a week is not covered with fur. The body length of young
cuscus just weaned ranges from 100 mm to125 mm. Its
body is covered with fine hairs evenly on the dorsal,
ventral, head, hands and feet.
Based on the number of individuals in the population
documented, there are 44.83% male and 55.17% female.
These conditions indicate the existence of equilibrium in
reproduction pair because cuscus is not monogamous. This
fact indicates that the regeneration cuscus population in the
region is expected to continue in the future.
Table 1 shows that total population of cuscus in an area
of 105 ha is 95 individuals. Population distribution of
cuscus is as follows: P. orientalis 67 individuals, P.
gymnotis 17 individuals, and S. maculatus 11 individuals.
The estimation of cuscus population at the study site is 317
individuals in the area of 420 ha or approximately one

individual per ha. Therefore, the cuscus population is
potential to be pursued as one of the objects in the
development of the area of Arfak Mountains Nature
Reserve in the future.
Tabulated results show effective population size of
cuscus can be managed, because population density
reached 317 individuals, or approximately 105 cuscuses for
each species. The number of 317 is considered to comply
the effective population size of the threat of local extinction
in the northern region of Arfak Mountains Nature Reserve.
According to Franklin (1980) in Maturbongs (1999) it is
required at least 50 individuals to maintain genetic
diversity in captivity. The number is determined
accordance to the experience that the stock of animals in
89
captivity can be maintained if the loss of biodiversity as

much as 2-3% per year, while for the 50 individuals will
only lose 1% of genetic diversity.
Table 1. Population density of cuscus in the Arfak Mountains
Nature Reserve
The species of Number of The distance between the Population
cuscus
individuals
cuscus and the transect
density
(ni)
(r)
(N)
P. orientalis
39
1131
67
P. gymnotis
10
276
17
S. maculatus
9
341
11
Total
58
1748
95
Note: r = meter2; transects length (L) = 21 000 m transects area
(A) = 105 ha
Figure 1. A. Phalanger orientalis, B. Phalanger gymnotis, C. The male (left) and D. female (right) of Spilocuscus maculatus found in
the Arfak Mountains Nature Reserve, Manokwari.
90
Based on these assumptions, the size of the open

population such as cuscus population in Arfak Mountains
Nature Reserve will be able to survive in the future because
of the flexibility activities (feeding, mating, and other
activities) and the abundant of feed resources. The
estimation results if connected with home range per
individual cuscus which is between 1,225 m2 and 2,400 m2,
then the population of 317 cuscuses will require a
minimum areal of 388,325 m2 (38 ha) and a maximum of
760,800 m2 (76 ha). The analyzes indicate that the carrying
capacity of cuscus habitat at the site of this study is
sufficient because the cuscus in the region has a larger
effective area compared to its population.
The food of the cuscus
The species of forest plants consumed by cuscus as a
feed resource in Arfak Mountains Nature Reserve is quite
varied, consisting of fruit and leaves. There are 17 types of
forest plants consumed by cuscus in the region. The
analysis showed that the pole stage vegetation has the
largest number of 87 individuals consisting of 16 species.
Ten species of them are the main feed of cuscus which are
dominant and spread evenly over the study site. They are
Sterculia parkinsonii, Dracontomelon edule, Pometia sp.,
Myristica sp., Syzygium sp., Ficus spp., Lansium domesticcum, Hernandia peltata, Cerbera floribunda and Intsia
bijuga, respectively. On the other side, sawtimber stage
vegetation has 81 individuals consisting of 15 species. Ten
species of them are the main feed of cuscus which are
dominant and spread evenly on the observed location. They
are Cerbera floribunda, Intsia bijuga, Gnetum gnemon,
Hernandia peltata, Ficus spp., Syzygium sp., Lansium
domesticum, Myristica sp., Pometia sp., and Sterculia
parkinsonii, respectively. Sinery (2002, 2006) states that
any species of cuscus is fruit eater (frugivores). However
Phalanger gymnotis is also carnivores, especially against
insects and other small animals.
Analysis of the level of diversity, dominance, and
evenness showed that species diversity index was 0.301.
Regarding the criteria of diversity index or the degree of
species diversity, Shannon in Sugianto (1994) considered
that species diversity is high when H > 3, moderate when 1
< H < 3, and low when H < 1. Therefore, Arfak Mountains
Nature Reserve has a low diversity of vegetation species
for the cuscus feed (H < 1). The high diversity of species
within a vegetation community is characterized by an
abundance of many species with the same or nearly the
same.
The analysis of the dominance level shows that the
dominance index of the vegetation which is cuscus feed at
the study site was 0.501. This condition illustrates that the
distribution of cuscus feed according to each type is low.
Distribution of individuals of each species is not balanced,
and only a few species have the number of individuals that
is dominant. The analysis of the level of evenness shows
that evenness index of vegetation which are cuscus feed in
the study site was 0.253. This figure illustrates that the
distribution of cuscus feed according to the species is not
well spread. According to Santosa (1995), evenness index
indicates the proportion size of individuals number in each
species found in a particular community. When each type

has the same number of individuals then the community is
said to have a maximum value of evenness index. The
value of species evenness according to Krebs (1989) in
Sugianto (1994) ranges from 0 to 1.
Estimated availability of cuscus feed as an illustration
of habitat carrying capacity is as follows: the basic
assumption that the average feed requirement is 0.5 to 1 kg
per head per day. Individual density is 1 head per ha. When
a tree (e.g. Ficus septica or Syzygium sp.) produces 30-50
kg of fruit per season with a frequency of at least 2 times a
year then the estimated availability of food is abundant at
the sites. According to Sinery (2010), of all species of
cuscus feed in the forest, Ficus septica or Syzygium sp. is a
key species for wildlife frugivore like cuscus because it can
bear fruit throughout the year.
CONCLUSION
Of the seven species of cuscus found in Papua and West
Papua, three species are found in the area of Arfak
Mountains Nature Reserve, especially in the southern
region. The three species of cuscus are northern common
cuscus (Phalanger orientalis), ground cuscus (P.
gymnotis), and the common spotted cuscus (Spilocuscus
maculatus). The number of cuscus found during the study
period was 58 with an estimated density of 95 cuscus per
105 ha, then it is predicted only one cuscus is found in 1
ha. P. orientalis has the biggest number of individuals with
the highest level of dominance compared to others species
because this species has higher reproductive capacity.
There are 17 species of forest plants consumed by cuscus
as feed resources consisting of 16 species of pole stage
vegetation and 15 species of sawtimber stage vegetation. In
term of quantity, the amount of feed available is abundance
for the needs of cuscus in the region.
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Pages: 92-97
ISSN: 1412-033X
E-ISSN: 2085-4722
Vegetation stands structure and aboveground biomass after the shifting

cultivation practices of Karo People in Leuser Ecosystem, North Sumatra
1
T. ALIEF ATHTHORICK1,2,, DEDE SETIADI3, YOHANES PURWANTO4, EDI GUHARDJA3
Department of Biology, Faculty of Mathematics and Natural Sciences, North Sumatra University (USU), Medan 20155, North Sumatra, Indonesia.
Jl. Bioteknologi No.1 Kampus USU Medan, Tel.: 061-8223564, Fax.: 061-8214290, email: talief@lycos.com
2
Post Graduate School, Bogor Agricultural University (IPB), Bogor 16680, West Java, Indonesia
3
Department of Biology, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University (IPB), Bogor 16680, West Java, Indonesia
4
Laboratory of Ethnobotany, Research Center for Biology, Indonesian Institute of Sciences (LIPI), Cibinong-Bogor 16911, West Java, Indonesia
Manuscript received: 8 February 2012. Revision accepted: 29 March 2012.
ABSTRACT
Aththorick TA, Setiadi D, Purwanto Y, Guhardja E. 2012. Vegetation stands structure and aboveground biomass after the shifting
cultivation practices of Karo People in Leuser Ecosystem, North Sumatra. Biodiversitas 13: 92-97. Vegetation stands structure and
aboveground biomass after the shifting cultivation practices of Karo People in Leuser Ecosystem, North Sumatra. Shifting cultivation
has been practiced by Karo People in Leuser Ecosystem for a very long time and caused a mosaic of patches that shift over time
between traditional agriculture and secondary forest. The objectives of this study were to investigated the recovery of vegetation stands
structure and aboveground biomass in four age classes of secondary forest, i.e. 5-years old, 10-years old, 20-years old, 30-years old and
primary forest as a control. In total, 496 subplots were surveyed. Saplings contributed 62.82% of basal area in 5-years forest and still
important in 10 and 20-years forest, but density decreased in 30-years and primer forest whereas tree stands dominated in 30-years and
primary forest and shared basal area of 96.36% and 97.03%, respectively. Aboveground biomass of trees achieved its highest values in
primary forest, i.e. 659.22 t/ha and contributed to total aboveground biomass of 99.38%.
Key words: leuser ecosystem, traditional agriculture, secondary forest
INTRODUCTION
Agricultural encroachment by shifting cultivation
occupies a central position in the debate on tropical
deforestation. Shifting cultivators are often seen as the
primary agents of deforestation in developing countries;
estimates of their share range as high as 45% (UNEP 1992)
to 60% (Myers 1992). Shifting cultivation could be
considered as an early stage in the evolution of agricultural
systems. The system is based on cutting and burning the
vegetation in the dry season, and planting crops in the wet
season. The field eventually grows into secondary forest,
before the cycle is repeated. The length of this fallow
period varies considerably 5-20 years is common (FAO
1974). Fallow duration and cultivation periodicity may be
influenced by multiple factors, including ecological factors
such as precipitation, soil conditions and topography, as
well as socio-economic factors (Mertz, 2002). Abandoned
fields are distributed worldwide and therefore allow
comparison of secondary succession from various
geographical regions (Osboronova et al. 1990). A number
of studies on these old-field successions have been
conducted in many countries (Osboronova et al. 1990;
Wilson and Tilman 1991; Lee, 2002).
Secondary forests, which comprise a large area of
tropical forests (ITTO, 2002), are forests in the process of
recovery following natural or anthropogenic disturbance,
such as agriculture, logging, or ranching (Brown and Lugo
1990; Chazdon, 2003). Secondary forests can serve as

carbon sinks (Fearnside and Guimaraes 1996), as well as
enhance regional biodiversity, environmental services, and
forest-based economies (Brown and Lugo 1990; FAO,
2005; Finegan 1996). Forests at different stages of
succession differ in total biomass, net primary production,
and species composition, which affects their relative
contribution to regional and global carbon cycles
(Fearnside and Guimaraes 1996). As the population
depending on shifting cultivation increases, the system
increasingly fails to satisfy the requirements for higher
production per unit area. This may result in shorter fallow
and longer cropping periods, initiating an accelerating and
self-reinforcing process of land degradation (FAO 1974).
Studies of succession after cessation of shifting cultivation
in tropical region have also indicated that the diversity of
woody species gradually increases with time since
abandonment of fallow (Lawrence, 2004; Lebrija-Trejos et
al. 2008).
Tropical secondary forests play an essential role in the
global carbon cycle and in determining a countrys carbon
storage for the REDD (reducing emissions from
deforestation and degradation in developing countries)
scheme (Gibbs et al. 2007) due to the degradation of large
areas of tropical rain forest (Brown and Lugo 1990; Wright
2005). Accurate estimation of biomass changes in
secondary forests after degradation contributes to
calculating forest carbon storage in the region, because
ATHTHORICK et al. Vegetation stand of Karo people homeland
uncertainties in the rate of biomass accumulation in

secondary forests create critical data gaps limiting our
understanding of the role of tropical forests as sources and
sinks of atmospheric carbon (Kauffman et al. 2009). In
Southeast Asia, tropical secondary forests constituted 63%
of the total forest cover in 2005 (Kettle 2010). However,
knowledge of biomass changes after forest degradation in
Southeast Asia is still limited compared with that of the
neotropical region, particularly for belowground components (Brown and Lugo 1990). Quantifying the initial few
decades of biomass changes in secondary forests after
degradation will decrease these uncertainties since biomass
accumulation during the initial stage is usually very large
and shows complex changes (Brown and Lugo 1990). For
example, many tropical secondary forests show rapid rates
of aboveground production during the initial stage of
succession (Ewel 1971; Ewel et al. 1983; Uhl and Jordan
1984; Lugo 1992; Jepsen 2006; Kendawang et al. 2007).
Shifting cultivation has been practiced by Karo People
in Leuser Ecosystem for a very long time and caused a
mosaic of patches that shift over time between traditional
agriculture and secondary forest. As such, many forests in
the Leuser Ecosystem are secondary forests at various
stages of succession following crop cultivation. However,
little attention has been given to long term forest recovery
after abandonment of shifting cultivation so little is known
93
regarding how abandoned fallow fields have changed over

time. The objectives of this study were to investigated the
recovery of vegetation stands structure and aboveground
biomass in four age classes of secondary forest, i.e. 5-years
old, 10-years old, 20-years old, 30-years old and primary
forest as a control.
Study area
Our study was carried out in secondary forests
abandoned after traditional shifting cultivation by the Karo
People at Telaga village in Leuser Ecosystem of North
Sumatra (Figure 1). Vegetation in the study area is
generally of the submontane forest type (800-1400 m asl)
as proposed by Laumonier (1997). The area has a moist
tropical climate. The average annual rainfall is 2777 mm,
ranging from 2044 to 4022 mm over a 10-year period
(measured from 2000 to 2009). The last cultivated crop and
stand age of each site were recorded based on information
from elderly villagers who were born in the village. Information on stand age, however, was credible until for 20years old, whereas 30-years old villagers did not remember
the exact year when they had opened up the field.
Figure 1. Study area at Telaga village in Leuser Ecosystem of North Sumatra. Dotted circle = study area
94
Data collection and analysis

In this study, we analyzed forest stands and aboveground biomass for sapling with diameters at breast height
(DBH) 2-9.99 cm and tree with DBH 10 cm. Formulas
and definitions were compiled from Mueller-Dombois and
Ellenberg (1974), Greig-Smith (1983) and Schreuder et al.
(1993). The number of subplot for each stage of secondary
forest (5, 10, 20 and 30 years old) was 32 subplots for tree
(10x10 m) and sapling (5x5 m) whereas primer forest was
comprised 120 subplots for both stages. In total, 496
subplots were surveyed. Height, stem height, density, DBH,
basal area and biomass for each stand age were measured
with the purpose of characterizing forest stands in a
quantitative basis. The analysis of aboveground biomass
used two allometric equations. For trees, the equation given
by Brown (1997) was used: Y = exp{-2.134+2.530*ln
(D)}. For saplings, the equation given by Honzak et al.
(1996) was used: Y = exp[-3.068 + 0.957 ln (D2 * H)];
where Y = biomass (t/ha), D = diameter and H = height.
Finally, analysis of variance (ANOVA) was used as a
statistical technique designed to determine whether or not a
particular classification of the data is meaningful.
Stands structure
Saplings
In 5 years secondary forest, species was dominated by
pioneers such as light-demanding herbaceous plants,
grasses, vines, seedlings, and saplings. These species have
a short life cycle, high growth rate and high reproductive
resource allocation (Gomez-Pompa and Vasquez-Yanes
1981). An important characteristic of this stage is the much
higher density (1,425.00 individuals/ha) compared to the
density of trees (64.06 individuals/ha). High sapling
competition was expressed by its highest basal area
compared to tree stands. Saplings contributed 62.82% of
basal area in this stage indicated that saplings were very
important at this stage of re-growth. In 10 and 20 years
forest, saplings are still important for the stand as a whole
(density of 2,175 and 2,725 individuals/ha, respectively)

but density decreased in 30 years and primer forest (Table 1).
DBH of saplings in all classes age are constant
relatively causing the classification of age stages is not
meaningful (p<0.41). These indicated the closer values of
DBH from 5 years forest to primer forest and mainly at the
succession stages. For saplings, although total height did
not increase distinctly, the difference between all variables
for all classes is statistically significant (p<0.00).
Figure 2 illustrate the distribution of stand structure
variables in density, DBH, basal area and total height of
saplings in all classes. DBH in 5 and 10 years forest have
many outliers indicating the variance of DBH in many
individuals. This phenomenon was found too in total height
especially in 10 years forest. These indicated the high
competition in vertical and horizontal growth in saplings
phase. Density and basal area have the similar trend of
distribution, the values increase from 5 years up to 20 years
forest and then decrease to primer forest. Ecologically,
these trends are explained by the competition for light
within the vegetation community. The growth of saplings
was limited in 30 years and primer forest caused by
covering canopy layers.
Trees
Density of trees in 5 years forest is lower (64.06
individuals/ha) indicating the early vegetation has soon
recovered. In general, the density of trees increased from 5
years up to primary forest, except in 20 years forest where
its density (218.75 individuals/ha) is lower than in 10 years
forest (279.69 individuals/ha). This phenomenon also was
applied to basal area, where 10 years forest has 10.49
m2/ha, whereas 20 years forest has 9.34m2/ha. This result
indicated the recovery process in 10 years forest more
intensive compared to 20 years forest. Besides that, 20
years forest is close to village and forest was often
disturbed by the people for harvesting the construction
materials and fire woods. DBH increased significantly from
5 years up to primary forest indicating the success
competition of horizontal growth.
Table 1. Stand structure variables for each age classes of forest in study area
Stand structure variables
Density of saplings (individuals/ha)
Density of trees (individuals/ha)
DBH of saplings (cm)
DBH of trees (cm)
Basal area of saplings (m2/ha)
Basal area of trees (m2/ha)
Total basal area (m2/ha)
Saplings contribution to basal area (%)
Trees contribution to basal area (%)
Total height of saplings (m)
Total height of trees (m)
Biomass of sapling (t/ha)
Biomass of trees (t/ha)
Total biomass
Sapling contribution to biomass (%)
Tree contribution to biomass (%)
5
years
1,425
64.06
4.28
15.80
2.45
1.45
3.9
62.82
37.18
4.25
7.57
5.79
10.73
16.52
35.05
64.95
10
years
2,175
279.69
4.15
20.79
3.57
10.49
14.06
25.39
74.61
4.03
14.66
8.24
113.27
121.51
6.78
93.22
Forest type
20
30
years
years
2,725
687.50
218.75
329.69
4.00
3.97
21.17
26.78
4.18
.96
9.34
25.43
13.52
26.39
30.92
3.64
69.08
96.36
4.65
4.58
14.19
18.52
10.88
2.43
86.62
289.33
97.50
291.76
11.16
0.0083
88.84
99.17
Primary
forest
850
544.17
4.29
25.40
1.45
47.42
48.87
2.97
97.03
5.10
22.30
4.10
659.22
663.32
0.0062
99.38
F
19.37
68.94
0.99
6.61
14.32
35.91
13.22
65.70
10.69
26.09
-
Sig
0.00
0.00
0.41
0.00
0.00
0.00
0.00
0.00
0.00
0.00
-
10
9
4000
DBH of Saplings (cm)
Density of Saplings (individual/ha)
5000
95
3000
2000
7
6
5
4
3
1000
2
0
1
5 Years
10 Years
20 Years
30 Years
Primer Forest
5 Years
10 Years
20 Years
30 Years
Primer Forest
5 Years
10 Years
20 Years
30 Years
Primer Forest
12
10
6
Height of Saplings (m)
Basal Area of Saplings (m2/ha)
5
4
3
2
0
5 Years
10 Years
20 Years
30 Years
Primer Forest
900
160
800
140
700
120
600
DBH of Trees (cm)
Density of Trees (individuals/ha)
Figure 2. Distribution of density, DBH, basal area and total height of saplings in all classes.
500
400
300
100
80
60
200
40
100
20
0
10 Years
20 Years
30 Years
Primer Forest
5 Years
120
60
100
50
80
Height of Trees (m)
Basal Area of Trees (m2/ha)
5 Years
60
40
20
10 Years
20 Years
30 Years
Primer Forest
30 Years
Primer Forest
40
30
20
10
0
5 Years
10 Years
20 Years
30 Years
Primer Forest
5 Years
Figure 3. Distribution of density, DBH, basal area and total height of trees in all classes
10 Years
20 Years
96
Tree stands dominated in 30 years and primary forest

and shared basal area of 96.36% and 97.03%, respectively
but saplings still have a higher density (687.50 and 850
individuals/ha). A closer canopy alters the microclimate,
improving conditions for shade-tolerant tree species and
creating an unsuitable environment for pioneer species.
This reality sets the path to a more advanced stage of
vegetation re-growth.
Figure 3 illustrates the distribution of stand structure
variables in density, DBH, basal area and total height of
trees in all classes. The trend increased in all variables from
5 years forest up to primary forest, except 20 years forest.
Same with saplings, DBH and total height of trees stage
have many outliers indicating the big variance in many
individuals, especially in primary forest. Total height of
individuals is an important parameter indicating the stage
of recovery. Height described the competition for light
within the vegetation community.
Primary forest indicates the greatest range for all
variables distinctly and has long whisker compared to the
other classes age. This explains the recovery process of
trees from 5 years forest to primary forest was a long way
off. Saldarriaga et al. (1988) estimated that 190 years
would be taken by a previously cultivated site to reach
mature forest basal area and biomass values. Also, the
number of tree species present after 40 years of succession
is less than half the number in mature forests. In general
terms, soil fertility and land-use history emerge as the
critical factors influencing the rate of forest re-growth
(Tucker et al. 1998). Uhl et al. (1982) found that the time
of recovery depends on land use following removal. Slashburn-agriculture-abandon cycles have increasing secondary
succession duration. Large cleared patches, where seed
sources are far away, may take hundreds of years to return
to primary forest.
Aboveground biomass
Aboveground biomass of saplings increased from 5
years to 20 years forest, but decreased for 30 years and
primary forest, due to the lower importance of saplings in
closed tropical forest environments. As a function of DBH
and height, the trends of aboveground biomass from 5
years to primary forest are affected by those variables.
Aboveground biomass of trees achieved its highest values
in primary forest, i.e. 659.22 t/ha and contributed to total
aboveground biomass of 99.38% (Table 1). However, this
result indicated that aboveground biomass of primary forest
in this study was higher than in primary rain forests in
Southeast Asia, which ranged from approximately 300 t/ha
to 500 t/ha (Yamakura et al. 1986; Laumonier et al. 2010;
Niiyama et al. 2010).
CONCLUSION
In the early stage of vegetation recovery, saplings have
the important role expressed by the higher density and
basal area compared to trees. In the next stage (in 30 years
time), the growth of saplings was limited caused by
covering canopy layers of trees. Trees taken over the role
and forest developed to reach mature forest basal area and

biomass values. Although vegetation recovery process was
taking place in the study area, secondary forest need long
time to develop to primary forest. The time of recovery
depends on land use type, duration of cycles and large
cleared patches.
ACKNOWLEDGEMENTS
We would like to express gratitude to Irwansyah
Sembiring, the Head of Perteguhan Subvillage, Telaga
Village, Sei Bingei Subdistrict, Langkat District, North
Sumatra for fully assistance along the fieldwork.
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Pages: 98-106
ISSN: 1412-033X
E-ISSN: 2085-4722
The wild plants used as traditional medicines by indigenous people of

Manokwari, West Papua
OBED LENSE
Faculty of Forestry, State University of Papua, Jl. Gunung Salju, Amban, Manokwari 98314, West Papua, Indonesia. Tel. +62-986-211065, Fax. +62986-211065, email: obedlense@yahoo.com
Manuscript received: 12 December 2010. Revision accepted: 24 April 2012.
ABSTRACT
Lense O. 2012. The wild plants used as traditional medicines by indigenous people of Manokwari, West Papua. Biodiversitas 13: 98-106.
The aims of the research were to identify the main plant species which are used as traditional medicines by native people in Manokwari
District, West Papua Province and also to describe the method of preparation and uses of some of the medicinal plants. This research
was conducted in seven sub-regencies, ie. Manokwari, Ransiki, Kebar, Wasior, Mimyambouw, Merdey and Anggi-Sururey sub-District.
Information recorded including methods of diagnosis and treatment of diseases, tribal name of a plant they used for treating disease (s),
part of the plant used, preparation and mode of application, and whether the plant is used alone or in combination with other plants.
Results indicate that the indigenous people in Manokwari District have been using at least 99 plant species (93 genera and 59 families)
as sources of medicines. Most of these traditional medicinal plants are commonly gathered from the local tropical rainforest
communities. At least 40 kind of sickness and injuries such as malaria, fever, and wounds can be treated by using traditional medicinal
plants from Manokwari District. Reserach also found that all parts of plants used, but leaf extracts are the most common part of the plant
used for treating medical condition.
Key words: wild plants, traditional medicines, indegineous people, Manokwari.
INTRODUCTION
As modern worldviews and lifestyles reach rural
indigenous communities through technology and personal
contact, centuries-old traditional cultures are changing.
Change takes place daily, nothing remains the same. Every
day the world is becoming smaller due to the development
of travel and communication technologies, and hardly a
group of peoples on the planet remain untouched by forces
of progress. However, through this process a great store
of knowledge held by native peoples is threatened with
extinction. Historically, modern societies have regarded
indigenous people and traditions as less progressive and, as
a result, many groups of indigenous peoples, especially
their younger generations, are encouraged to devalue their
native culture and to adopt new lifestyles and technologies.
The knowledge of traditional medicinal plants,
accumulated over centuries, may disappear in only a couple
of generations if the current pace of cultural change
continues to occur amongst the tribes in Manokwari
District. Preliminary field visits (interviews) have indicated
that transferring the traditional knowledge of the use of
plant-based preparations in the primary health care of these
people is under threat. There were a small number of young
people (3 younger than 45) who have inherited a traditional
knowledge of medicinal plants from their old generation in
each village visited. The process of transferring traditional
knowledge appears to be the main factor leading to the
decline of knowledge of traditional medicine. There is no
formal school or traditional institution involved in passing

on this knowledge. Transferring knowledge only happens
amongst family groups when they are engaged in other
activities. At that time, many young people are not
interested in following their parents, and the number of
people who have a good knowledge of traditional
medicinal plants is declining. It is possible that in next
couple of decades, the knowledge of medicinal plant within
these ethnic groups may disappear completely.
The aims of this research were to identify the plant
species that are used as traditional medicines by the native
people of Manokwari District, West Papua, and to describe
their methods of preparation and use of the medicinal
plants. The study represents the first step to documenting
significant aspects of the local medicinal plant knowledge
before it disappears.
The location of this project was in Manokwari District
in the province of Papua, Indonesia. This research was held
in seven sub-regencies, ie. Manokwari, Ransiki, Kebar,
Wasior, Mimyambouw, Merdey and Anggi-Sururey subDistrict.
The plants were collected for botanical identification
from several location/villages (Mandopy, Merdey, Sururey,
Jandurau, Dembek, Siwi, Wasior, Tandia, Minyambouw,
Indabri, and Inambuari) and each plant allotted a TMHM
LENSE The wild medicinal plants of Manokwari, West Papua
(Traditional Medicine Herbarium Manokwariense). Plant

specimens were labelled based on the date, locality,
altitude, latitude, tribal name, collector, and collection
number. In addition, as plants are located and identified
their use and method of preparation were documented and
photographs were taken. The herbarium specimens were
identified with the assistance of Marthen Jitmau and lodged
in the Herbarium Manokwariense (MAN), Manokwari,
West Papua, Indonesia, and were preserved for reference
voucher at the Traditional Medicine Research Unit.
Several aspects of the medicinal plants were recorded
including methods of diagnosis and treatment of medical
conditions; tribal name of a plant used for treating the
conditions; part of the plant used; preparation and mode of
application; whether the plant is used alone or in
combination with other plants.
Interviews were conducted in order to record relevant
ethnobotanical data. These interviews were conducted, as
recommended by Chhabra and Mahunnah (1994). In each
village two older persons whose empirical knowledge was
respected by everyone in the area, and two traditional
healers who prescribed local herbs were interviewed.
Interview data were recorded in an ethnobotanical
notebook (Martin 1995).
Plants used for traditional medicine
In the Manokwari District, 99 plant species were
documented as traditional medicinal plants. They are
widely distributed among 94 genera and 59 families, ten of
which are members of the Araceae family, widespread in
the rainforests of the District. This indicates the diversity of
traditional medicinal plants in Manokwari District. Except
for cultivated species such as Cocos nucifera L., Carica
papaya L. and Musa x paradisiaca L., traditional medicinal
plants were most commonly gathered from the local
vegetation communities.
Figure 1. Plant families most commonly used for traditional

medicine by Indigenous people from Manokwari District.
99
Plants used to treat more than 40 different medical

conditions were grouped into several categories according
to use by indigenous people: gastrointestinal disorders,
dermatological conditions, illnesses associated with pain
and/or fever, respiratory illnesses, womens medicines,
plants used to counteract bites by venomous animals, eye
remedies, wounds and burns, and other uses (Ankli et al.
1999).
The category with the largest number of species was
that used to treat illnesses associated with pain and/or
fever, and the next largest group consisted of plants used to
treat gastrointestinal disorders, whereas only one species
was documented as being used to counteract venomous
animal bites. The cause of some of the sickness and injuries
were attributed to supernatural powers. Even minor
accidents to such as cuts and body pains were sometimes
attributed by tribal people such outside influences.
The number of medicinal plants found in the present
study was higher than that found in previous studies in
West Papua, and other regions of eastern Indonesia: the
Dani people in the Baliem Valley in Jayawijaya District
used 30 plant species to treat a number of local diseases
(Purwanto and Waluyo 1992), and the Tangma people in
Kurima Sub-District used 26 medicinal plants in their daily
health care (Alhamid and Sumarliani 1996). Roemantyo
and Wiriadinata (1991) reported that indigenous people in
Kupang, West Timor, recognize and use 37 species as
medicinal plants. However, the number of species found in
the present study is less than the 164 traditional medicinal
plants used by indigenous people in Tanimbar-Kai Island,
south-east Maluku, Indonesia (Purwanto and Waluyo
1992).
Unpublished data from local health authorities suggest
that illnesses associated with pain and/or fever (frequently
malaria, ear pain, and headaches) and gastrointestinal
disorders (diarrhoea, dysentery, and stomach-aches) are the
major health problems in Manokwari District. Infected
wounds, inflammatory skin diseases, and chronic and
infectious eye diseases are also common. Although bites
from poisonous snakes are feared, only a few cases have
been recorded.
The numbers of traditional medicinal plant species,
which are used simultaneously to treat a medical condition
appears to depend on the nature of the condition. Tribal
people will use several medicinal plant species to treat a
particular illness if they consider the illness to be
dangerous. The people of the present study used at least ten
plant species to treat malaria, since malaria is one of the
primary medical conditions that often result in death.
However, to cure influenza the people often use one only
medicinal plant species.
The people at all the study sites believed that there are
several reasons why someone suffers from certain illnesses.
These medical conditions include diarrhoea, dysentery,
influenza, and malaria, all known to be caused by
microorganisms. For these conditions, a combination of
modern medicines and traditional medicinal plants are very
popular choices. The local people also believe that certain
ailments may be caused by a person or a group entering an
area which does not belong to the group. The symptoms of
100
this illness are stomach-pains, swollen navel, diarrhoea,

pale appearance, and a feeling of general debility. They
believe that these conditions are related to the
supernatural and can be healed using traditional
medicinal plants only. Native people in Manokwari
District, especially people from the Big Arfak tribe (Sough,
Hatam, Meyah, and Moile sub-tribes), also believe that
their members may suffer from certain medical conditions
due to a curse from an ancestor. The main symptoms of this
illness are swollen parts of the body such as the eyebrows,
eyelids, and stomach, and they believe that both hands and
legs shrink and may not be able to move easily. Medical
conditions, caused by suanggi (a member of the group
who is able to kill another member using magic; tribal
communities consider this magical) are though to be
incapable being healed by either modern or traditional
medicines. The major symptoms of this sickness are
darkening of the entire body of the affected person and
immobility of hands, legs, and fingers. Based on the field
interviews, the only way to treat conditions caused by both
an ancestor curse and suanggi is by applying traditional
medicinal plants. However, this medicine cannot cure the
illness completely, as the person who suffers from the
illness will die at the later stage. During the present study,
no information has been recorded regarding the species of
medicinal plants which may be used to treat medical
conditions caused by both curses from ancestors or
suanggi.
The study also indicated that there were some
similarities and differences amongst the tribes in using
medicinal plants in their daily health care. Some species
used as traditional medicines were used to treat similar
medical conditions throughout the region. Alstonia
scholaris, widely distributed throughout Manokwari
District, has been used by the native people in Siwi and
Dembek village (Ransiki), Jandurau village (Kebar),
Tandia village (Wasior), and Mandopi village in
Manokwari sub-District to treat malaria and fever. Field
observations indicated that Laportea interrupta, Alstonia
scholaris, Pipturus repandus, Costus speciosus, and
Cordyline fructicosa occurred at all of the study sites.
Particular species of traditional medicinal plants were
found in two or more study sites to treat different medical
conditions. For example Cordyline fructicosa was used by
the community in Ransiki sub-District to treat dysentery,
whereas in Minyambouw sub-District the species was used
to treat menstruation problems. There were also some
similarities in medicine preparation and in the ways to
apply the potions. It may be that frequent visits of the
members of particular tribes, including the older person or
traditional healers, to meet their extended families or to
attend the traditional ceremonies may be one possible
factor that produced these similarities.
Plants used for pain and/ or fever
Fourty species were documented for illnesses
associated with pain and/or fever in which fever and
malaria were the frequent conditions (Table 1). In general,
traditional medicines were prepared as decoctions or
infusions, or sometimes applied or rubbed on to the part of
the body affected. For example, in treating fever, a potion

may be prepared as an infusion to be drunk and the solid
parts applied to the forehead.
Malaria and fever are the most frequently treated
medical conditions. This group includes diseases associated
with chest pain, headaches, muscular pain, and influenza.
Of the 40 species recorded to treat pain and/or fever, 21
species were used by the people in the District specifically
to treat malaria (Table 1). Alstonia scholaris and Pipturus
repandus were the most common plants used in four
different sub-regencies to treat fever. Cerbera manghas,
Casuarina equisetifolia, Flagellaria indica, Freycinetia
sp., Lansium domesticum, Loranthus sp., Senna alata, and
Solanum sp. were used in only one sub-District. However,
these species are widely distributed and seen in all four
areas. Nevertheless, each tribe in the region has their own
traditional plants to treat local medical conditions. Nettles,
Laportea interrupta (L.) Chew. are widely recognised by
almost all communities in the region as a medicinal plant to
combat muscular pains and fatigue. The method of use was
to rub the fresh, hairy leaves on the skin to produce a very
hot and itchy feeling.
Several species found in the present study to treat pain
and fever are also used by traditional communities around
the world to treat similar medical conditions. People in
Aceh (Erdelen et al.1999) and East Lombok, Indonesia
(Hadi and Bremner 2001), Malaysia (Salleh 1997), and
Karnataka Province, India (Shankar et al. 1999) use the
species Alstonia scholaris to combat malaria and fever.
Carica papaya L. (leaves, stem, roots, and flowers) is used
to treat malaria in West Lombok, Indonesia (Hadi and
Bremer 2001). The leaves of Bidens pilosa L. are made into
a decoction and then used as a gargle in Dominican
Republic and Papua New Guinea (Morobe Province) for
treating toothaches (Taylor 1998; Woodley 1991). The
Fijians use this plant as a traditional medicine but for
different diseases: the young shoots are used as an internal
remedy for influenza, and the leaves are used to treat
infective hepatitis (Cambie and Ash 1994).
Some species recorded to treat pain and fever are also
known to contain phytochemical compounds. Some of
these compounds have been tested in order to establish
their efficacy in treating particular medical conditions.
Bidens pilosa has been the subject of recent clinical studies
which have supported many of its uses in herbal medicine
(Taylor 1998). As early as 1979, scientists demonstrated
that specific chemicals found in this species were
phototoxic to bacteria and fungi (Wat et al. 1979; Arnason
et al. 1980). Subsequently, Swiss scientists isolated several
known phytochemical compounds with anti-microbial and
anti-inflammatory properties which led them to believe that
the presence of these compounds "may rationalise the use
of this plant in traditional medicine in the treatment of
wounds, against inflammation and against bacterial
infection of the gastrointestinal tracts (Geissberger and
Sequin 1991). In the same year, scientists in Egypt
documented the antimicrobial activity of Bidens pilosa L.
(Sarg et al. 1991), and another research group reported that
the species has anti-inflammatory properties (Chih et al.
1995). New bioactive phytochemicals were also discovered
101
Table 1. Manokwari traditional medicinal plants used for illnesses associated with pain and/or fever.
Plant name
Alpinia purpurata (Vieill.) K.Chum.
Alstonia scholaris R.Br.
Bidens pilosa L
Blumea saxatilis Zoll. & Mor.
Carica papaya L.
Casuarina equisetifolia L.
Cerbera manghas L.
Cinnamomum culilawan Blume
Coelogyne asperata Lindl.
Costus speciosus Sm.
Cyathea contaminans L.
Cyrtosperma sp.
Diplazium esculentum (Retz.) Sw.
Dysoxylum arborescens Miq.
Dryopteris filix-max (L.) Scott.
Ficus sp1.
Ficus sp2.
Flagellaria indica L.
Freycinetia sp.
Homalomena aromaticum (Roxb.) Schott.
Kalanchoe pinnata Pers.
Lansium domesticum Jack.
Laportea interrupta (L.) Chew.
Loranthus sp.
Macaranga mappa Muell. Arg.
Macaranga tanariius Muell. Arg.
Mucuna novo-guineensis Scheff.
Octomeles sumatrana Miq.
Palmeria sp.
Pentapalanqium pachycarpum A.C. Smith
Philodendron sp.
Pimelodendron amboinicum Hassk.
Pipturus repandus (Bl). Wedd.
Pisonia sp.
Platycerium sp.
Ricinus communis L.
Scindapsus hederaceus Schott.
Senna alata L.
Smilax sp.
Solanum sp.
Family
Zingiberaceae
Apocynaceae
Asteraceae
Asteraceae
Caricaceae
Casuarinaceae
Apocynaceae
Lauraceae
Orchidaceae
Zingiberaceae
Cyatheaceae
Araceae
Athyriaceae
Meliaceae
Dryopteridaceae
Moraceae
Moraceae
Flagellariaceae
Pandanaceae
Araceae
Crassulaceae
Meliaceae
Urticaceae
Loranthaceae
Euphorbiaceae
Euphorbiaceae
Fabaceae
Dasticaceae
Monimiaceae
Clusiaceae
Araceae
Euphorbiaceae
Urticaceae
Nyctaginaceae
Polypodiaceae
Euphorbiaceae
Araceae
Caesalpiniaceae
Smilacaceae
Solanaceae
Medical conditions
Earaches
Fever, malaria
Toothaches
Cold, influenza
Malaria
Malaria
Fever
Toothaches and muscular pains
Chest pain
Ear pain
Fever
Chest pain
Headaches
Fever and Malaria
Fever
Fever
Toothaches
Fever
Fever
Muscular pain
Fever
Malaria
Muscular pain
Fever in babies
Chest pain
Chest pain, malaria
Malaria, fever
Fever
Back pains
Joint pain
Rheumatic and joint pains
Headaches
Fever
Headaches
Malaria, fever
Malaria
Colds of babies
Fever
Headaches
Malaria
Plant parts used

Stem
Bark
Leaves
Leaves
Leaves
Bark
Latex
Bark
Bulbs
Stem
Stem
Tubers
Leaves
Bark
Leaves
Bark, shoot
Leaves, roots
Stem juices
Stem juices
Bulbs
Leaves
Bark
Leaves
Leaves
Leaves
Leaves
Stem
Bark
Stem
Bark
Stem
Leaves, Latex
Bark
Roots
Leaves
Leaves
Leaves
Leaves
Stem
Leaves
Table 2. Manokwari traditional medicinal plants used for gastrointestinal disorders.

Plant name
Acorus calamus L.
Adenanthera microsperma Teisjsm.&Binn.
Aquilaria malacensis Lam.
Artocarpus altilis (Park.) Fosb.
Canarium sp.
Canna indica L.
Cinnamomum culilawan Blume
Cocculus carolinus (L.) DC.
Commelina diffusa Burm. F.
Cordyline fructicosa A. Cheval.
Costus speciosus Sm.
Crinum asiaticum L.
Homalanthus nutans Guill.
Homalonema aromaticum (Roxb.) Schott.
Horsfieldia sp.
Intsia palembanica L.
Morinda citrifolia L.
Mucuna nova-guineensis Scheff.
Piper sp.
Planchonella sp.
Polygonum sp3.
Pothos scandens L.
Pterocarpus indicus Willd.
Senna alata L.
Wollastonia biflora DC.
Family
Araceae
Mimosaceae
Thymelaeaceae
Moraceae
Burseraceae
Cannaceae
Lauraceae
Menispermaceae
Commelinaceae
Dracaenaceae
Zingiberaceae
Amaryllidaceae
Euphorbiaceae
Araceae
Myristicaceae
Caesalpiniaceae
Rubiaceae
Fabaceae
Piperaceae
Urticaceae
Sapotaceae
Polygonaceae
Araceae
Fabaceae
Caesalpiniaceae
Asteraceae
Medical conditions
Dysentery
Diarrhoea
Dysentery
Diarrhoea/dysentery
Liver problems
Dysentery
Stomach-aches
Stomach-aches
Dysentery
Dysentery
Stomach-aches, food poisoning
Stomach-aches
Stomach-aches
Stomach-aches
Stomach complaint
Dysentery
Stomach-aches
Diarrhoea
Stomach-aches
Diarrhoea
Dysentery
Dysentery
Diarrhoea
Dysentery
Stomach-aches
Diarrhoea
Plant parts used

Rhizomes
Leaves
Leaves
Sap, bark
Bark
Stem
Bark
Water from stem
Leaves
Leaves
Stem
Tubers
Leaves
Bulbs
Bark
Bark
Leaves
Stem
Leaves
Leaves
Bark
Leaves
Leaves
Bark
Leaves
Leaves and flowers
102
in 1996 which indicated that B. pilosa was effective against

normal and transformed human cell lines (Alvarez et al.
1996). The plant extract was shown to possess
prostaglandin-synthesis inhibitory activity, a process linked
to headaches and inflammatory diseases (Jager et al. 1996).
Subsequently, a research group in Taiwan documented its
hepatoprotective (liver protecting) activity, and showed
that the species can protect liver injuries from various
hepatotoxins, and suggested that it has the potential as a
broad-spectrum anti-hepatic agent (Chin et al. 1996). In
addition, Rabe and Staden (1997) reported that the species
showed antibacterial activity against gram-positive
bacteria.
Plants used for gastrointestinal disorders
Twenty-six species from the Manokwari District were
documented for treating gastrointestinal disorders (Table
2). Majority of the species of traditional medicinal plants
recorded in this group were used to treat stomach-aches
and dysentery; other illnesses treated included diarrhoea,
liver diseases and poisoning. Unpublished data from the
Manokwari District community health centre show that
stomach-aches and dysentery are important medical
conditions throughout the region and they have caused
many deaths amongst these communities. The way of
life, the officer said, was the primary reason. Treatments
consist mostly of circular massages of the medicinal plants
around the navel as well as drinking a decoction or infusion
made from various plant parts.
Some species used to cure these medical conditions
were used by more than one tribe. Homalanthus nutans was
widely used by the sub-tribes in four different subregencies (Ransiki, Anggi-Sururey, Wasior, and Kebar) to
treat stomach-aches. The shrubs are easy to access,
growing mostly in secondary forest and previously
cultivated area surroundings the villages. The tribes also
used similar methods of preparation as a decoction or cold
infusion, followed by rubbing the prepared plants on the
affected areas.
Some of the species found under this section have also
been reported as treatments for similar medical conditions
in different parts of the world. Artocarpus altilis is
recognized in Java and other Indonesian areas, (root bark,
sap, and sometimes stem-bark), Samoa and Tonga (roots),
and in Papua New Guinea (latex) to treat diarrhoea and
dysentery (Perry 1980; Dittmar 1998). Another study also
reported that a decoction of leaves and rhizomes of

Cordyline fructicosa is used to cure diarrhoea and
dysentery in Central Lombok (Puyung), Indonesia (Hadi
and Bremner 2001), and Samoa (Dittmar 1998). In
Malaysia, Acorus calamus (Jerangau) is used to cure
fevers, dysentery, and to improve the appetite (Salleh
1997). A decoction of bark of Artocarpus altilis and the
leaves and bark of Morinda citrifolia were also used in the
Philippines and Tonga to treat stomach-aches (Perry 1980;
Singh 1984). A phytochemical study of the latex of
Artocarpus altilis has shown that it contains cardenolides
(Qujano and Arango 1979; Wong 1976) and cerotic acid
(Perry 1980), but no pharmacological information relating
to indigenous uses is available.
Plants used for dermatological conditions
In this group, eight plant species from the Manokwari
District have been reported to be used to treat a variety of
dermatological conditions such as scabies and abscesses
(Table 3). Treatments mostly consist of preparing a
decoction or cold diffusion and applying it to the affected
skin. Bark and roots were the most common parts of the
plant used. To treat measles, the roots of Imperata
cylindrica and Metroxylon rumphii were boiled, filtered,
cooled, and the solution is swalloed twice per day until the
patient is healed.
The native people in Wasior, Kebar, and Merdey have
used the bark of the stem of Ficus sp. and Leea aculeata as
well as the leaves of Polygonum sp. to treat abscesses.
Treatment is mainly by drinking a decoction or cold
inffusion of the potion followed by application of prepared
plants whereas to cure ringworm, these communities
applied the crushed bark and leaves directly to affected
skin.
Some of the species found under this category have
been using as medicinal plants worldwide. In West
Lombok, Indonesia, Cocos nucifera (leaves, stem, and
roots) is used for fever and dysentery (Hadi and Bremner
2001). People in the Marshall Islands used the leaf sheath
of this species to support broken limbs (Spennemann
2000). Elsewhere in Indonesia, the roots of Imperata
cylindrica are used to treat blood pressure, fever, coughs,
and hepatitis (Erdelen et al. 1999). In Sri Lanka, a
decoction of rhizomes is used to relieve the retention of
urine and passing of blood in the urine (Jayaweera 1999).
Table 3. Manokwari traditional medicinal plants used to treat dermatological conditions.

Plant name
Cocos nucifera L.
Ficus sp1.
Imperata cylindrica L.
Leea aculeata Blume
Lithocarpus brassii Soepadmo
Metroxylon rumphii Mart.
Polygonum sp1.
Polygonum sp2.
Family
Arecaceae
Moraceae
Poaceae
Leeaceae
Fagaceae
Arecaceae
Polygonaceae
Polygonaceae
Medical conditions
Measles
Abscesses
Measles
Abscess
Ringworm
Measles
Scabies
Abscesses
Plant parts used

Milk from young coconut
Bark, shoot
Roots
Bark
Bark
Roots
Roots
Leaves
103
Table 4. Manokwari traditional medicinal plants used for respiratory illnesses.

Plant name
Alstonia scholaris R.Br.
Endospermum mollucanum Becc.
Euodia sp.
Horsfieldia sp.
Family
Apocynaceae
Euphorbiaceae
Rutaceae
Myristicaceae
Medical conditions
Coughs, asthma
Bronchitis
Asthma
Asthma
Plant parts used

Bark, roots
Bark
Bark
Bark
Table 5. Manokwari traditional medicinal plants used as womens medicines.

Plant name
Ageratum conyzoides L.
Biophytum petersianum Klotzsch
Centella asiatica L.
Colocasia sp.
Musa x paradisiaca L.
Nauclea orientalis L.
Physalis angulata L.
Ricinus communis L.
Family
Asteraceae
Oxalidaceae
Umbelliferae
Araceae
Dracaenaceae
Cyatheaceae
Musaceae
Rubiaceae
Solanaceae
Euphorbiaceae
Medical conditions
Eases birth, decoction after delivering a baby
Fertility
Problems of menstruation
Childbirth
Easy birth
Easy birth
Prevent pain during menstruation
Decoction before delivering a baby
Plants used for respiratory illnesses

Four species of Manokwari medicinal plants were
locally used for coughs, bronchitis, and asthma (Table 4).
Bark from the stem was usually prepared as a decoction
and infusion to treat these medical conditions in Merdey,
Ransiki, Kebar, and Manokwari sub-regencies. Results
indicated that species Alstonia scholaris was used widely in
different tribes in Ransiki, Kebar, Wasior, and Manokwari
to combat coughs and asthma.
The number of species recorded to treat these medical
conditions was lower than the number of species found to
treat diseases in any other category. This may be because
respiratory illnesses are not considered as primary medical
conditions in the Manokwari District. Some of the species
have been reported to be used for similar medical
conditions in different countries. For example, the bark of
Alstonia scholaris was used to treat diseases from malaria
and epilepsy to skin conditions and asthma (Shankar et al
1999); also, in Malaysia, the species is used for cases of
fevers and coughs (Salleh 1997).
Plants used as womens medicines
Plants used during delivery and menstruation problem
are the most prominent group in this category (Table 5).
Treatments for these medical conditions were mostly
prepared as decoctions and were sometimes followed by
applying or rubbing the prepared plants on to the stomach,
so the mother would not feel pain during the delivery of a
baby.
The indigenous people in Minyambouw sub-District
used at least six species of traditional medicinal plants to
treat childbirth and menstruation problems. The bark and
leaves were the most preferred plant parts. However, for
species Biophytum petersianum, the whole plant was used
as fertility medicine. Based on the personal interviews of
the present study, the indigenous people of Kebar subDistrict believed that a decoction of this species can
increase the fertility of a couple, but clinical investigations
Plant parts used

Leaves
Whole plant
Leaves
Bulbs
Leaves
Stem
Stem
Bark
Leaves
Leaves
are needed in order to support such a view. The species

also has been traded locally. Sometimes people from
outside the tribe buy a couple of kilograms of the whole
plant of Biophytum petersianum as a fertility-enhancing drug.
There have been no previous reports of the species
listed in Table 6 being used for medical conditions
associated with women elsewhere in Indonesia, but some
of them are used in other countries. The species Ageratum
conyzoides L. is used in most African countries as a
contraceptive, whereas in Trinidad, the species was used as
an abortifacient (Durodola 1977). A similar use was
reported for the species Physalis angulata, in Papua New
Guinea (Kurtachi, Northern Bougainville) where the seeds
of this species are used as a contraceptive in women
(Cambie and Brewis 1997). People in Central America and
Jamaica have used a tea prepared from the whole plant of
Physalis angulata to prevent an abortion after a fall during
pregnancy (Cambie and Brewis 1997).
Plants used for eye conditions
In this group, six species have been documented as
being used to treat eye complaints including inflammation,
irritation, and infection of the surface of the eye (Table 6).
Treatments generally consist of dropping the potions into
the eye. Table 7 shows that native people in Wasior subDistrict used several plant species to treat these medical
conditions, whereas people in Kebar used one species only
to treat similar complaints.
Often drops are prepared by extracting liquids from the
squashed leaves and/or stems of the plants and they are
applied topically. When people in the Wasior sub-District
use Calophyllum inophyllum to treat eye problems, the
leaves are first soaked in a bucket of water for about 30-45
minutes prior to washing the eyes with the extracted water
for approximately 1-2 minutes; the eyes are opened and
closed several times during this process. Leaves were the
preferred plant part used to treat eye ailments, possibly
because leaves are easier to prepare as drops.
104
Table 6. Manokwari traditional medicinal plants used for eye complaints

Plant name
Calamus spp.
Calophyllum inophyllum L.
Dischidia sp.
Ficus sp2.
Family
Asteraceae
Arecaceae
Clusiaceae
Dracaenaceae
Asclepidaceae
Moraceae
Medical conditions
Irritated eyes
Inflammed eyes
Inflammed eyes
Irritated eyes
Irritated eyes
Eye infection
Plant parts used

Leaves
Stem liquid
Leaves
Leaves
Leaves
Leaves
Table 7. Manokwari traditional medicinal plants species used to treat wounds and burns.
Plant name
Cyperus rotundus L.
Diplazium esculentum (Retz.) Sw.
Bambusa vulgaris Schrad.
Melastoma malabathricum L.
Merremia peltata Merr.
Mucuna novo-guineensis Scheff.
Paspalum conjugatum L.
Pometia pinnata Forst.
Axonopus compressus (SW.) P.
Spathoglottis papuana F.M. & Bailey
Family
Asteraceae
Moraceae
Cyatheaceae
Cyperaceae
Athyaceae
Poaceae
Melastomataceae
Convolvulaceae
Fabaceae
Poaceae
Sapindaceae
Araceae
Orchidaceae
Medical conditions
Wounds
Wounds
Burns
Wounds
Wounds
Wounds
Wounds
Wounds
Old wounds
Wounds
Burns and wounds
Wounds
Wounds
Plant parts used

Leaves
Sap, bark
Juvenile stem
Leaves
Leaves
Outer bark
Leaves
Leaves, twigs
Stem
Leaves
Juvenile bark
Leaves
Bulbs
Table 8. Manokwari traditional medicinal plants used for other medical conditions
Plant name
Amomum sp.
Canarium sp.
Drimys beccariana L.S. Gibbs.
Ficus benjamina L.
Gnetum gnemon L.
Litsea sp.
Palaquium sp.
Pimelodendron amboinicum Hassk.
Platycerium sp.
Pometia pinnata Forst.
Epipremnum pinnatum (L.) Engl.
Schismatoglottis calyptrata Zoll. & Mor.
Selaginella Palisot de Beauvois
Spathodea campanulata Beauv.
Family
Zingiberaceae
Moraceae
Burseraceae
Winteraceae
Moraceae
Gnetaceae
Lauraceae
Sapotaceae
Euphorbiaceae
Urticaceae
Polypodiaceae
Sapindaceae
Araceae
Araceae
Selaginellaceae
Bignoniaceae
Medical conditions
Sexually transmitted diseases in men
Gonorrhoea
Hepatitis
Lethargy
Bone fracture
Hepatitis
Gonorrhoea
Epilepsy
Hepatitis
Lethargy
Hepatitis, sexually transmitted diseases in men
Bone fracture
Broken legs
Tonic
Similar traditional uses of some species recorded in this

group have also been reported in different countries. In
Samoa, the leaves of Cordyline fructicosa was used to cure
eye inflammation (Dittmar 1998), and in Tonga, leaves
were used for eye complaints, eye infections, as well as
toothache, gum infections, and gum abscesses (Weiner
1971). The leaves of this species contain thymidine and the
flowers contain chelidonic acid (Wong 1976).
Plants used for wounds and burns
Extracts from 13 species of Manokwari medicinal
plants (Table 7) were used to treat wounds and burns. Cuts
and wounds may be bathed with potions of certain
medicinal plants made simply by crushing the leaves, stem,
or bulbs of specific plants, or by heating those parts before
crushing and applying the juices. Sap of the stem of
Artocarpus altilis may also be applied. Excessive bleeding
may be stopped by applying the outer bark of Bambusa
vulgaris or the juices of Ageratum conyzoides.
Plant parts used

Leaves
Sap
Bark
Bark
Bark
Bark
Bark
Latex
Leaves, bark
Leaves
Leaves
Bark
Leaves
Leaves
Stem juices
Bark
Burns may be treated by applying crushed stems of

Cyathea contaminans and masticated juvenile bark of
Pometia pinnata. The potion of the Cyathea contaminans
was prepared by crushing the juvenile part of the stem to a
gel and applying directly on burns. P. pinnata is commonly
used throughout the Manokwari District.
Furthermore, the medicinal plants recorded in this
group also have been reported to be used to treat similar
medical conditions worldwide. Ageratum conyzoides is
used in Java, Indonesia, to cure similar ailments. In
Malaysia and the Philippines it is used to treat cuts, boils,
and wounds, and is thought to have anti-tetanus properties
(Salleh 1997). In Central Africa, Cameroon and Congo,
and elsewhere in Africa (Durodola 1977), A. conyzoides is
used to cure pneumonia, but the most common use is to
heal wounds and burns. The species is reputed to be a quick
and effective cure for burns and is recommended by the
Brazilian Drugs Central as an antirheumatic (Ming 1999).
Pharmacological investigations by Ming (1999) have
confirmed that Ageratum conyzoides has an effective

analgesic action in rats when an aqueous extract of leaves
(100 to 400 mg/kg) is used. Trials in Kenya, using aqueous
extracts of the whole plant, have demonstrated musclerelaxing activities, confirming its popular use as an
antipasmotic (Achola et al. 1994). Similar results were
obtained in experiments conducted by State University of
Campinas and Paraiba Federal University, Brazil. In
clinical trials on patients with arthritis who were given an
aqueous extract of the whole A.. conyzoides plant, 66% of
patients reported a decrease in pain and inflammation and
24% reported increased mobility; no side effects were
apparent after a week of treatment (Ming 1999). In Samoa
and Hawaii, the twigs and fruits of Artocarpus altilis are
used as a cure for wounds (Dittmar 1998), whereas the
people in Langkawi, Malaysia used the juvenile stem of
Cyathea contaminans L. (Salleh 1997;Woodley 1991).
Other uses (OTH)
The 16 species in this group have diverse applications.
Medical treatment ranges from treating sexual diseases of
men, lethargy, hepatitis, and bone fractures (Table 8). Of
these, only a few species stand out as being of some
importance.
Medical conditions related to sexually transmitted
diseases in men are the most common conditions treated.
Preparations of medicinal plants are applied to affected
areas. Table 8 indicates that many of the species listed can
produce exudates (latex or sap) that are applied directly,
whereas the bark of Artocarpus altilis, Litsea sp.,
Palaquium sp., and Pimelodendron amboinicum was
crushed and then applied. There is no supporting
information regarding the efficacy of these species in
treating similar conditions in different regions. Hence,
further phytochemical studies, as well as clinical
investigations, are needed in order to prove their efficacy.
The species Drimys beccariana (akuai) has become a
very popular medicinal plant in Minyambouw area and
surroundings. The stem-bark of this species is normally
used by the indigenous people, especially the Hatam tribe,
as a tonic. In general, the bark is chewed or some it can be
prepared as a decoction to drink to strengthen people and to
provide energy for long distance travel, such as visiting
family in different villages or visiting the capital city of
Manokwari District. Crushed bark of Ficus benjamina is
used for broken bones as a hard gypsum-like plaster
bandage.
Parts of plants used and preparation
The indigenous people in Manokwari District have been
shown to use almost all the parts of medicinal plants for
treating a wide range of sickness, accidents, and injuries.
Leaves and leaf extracts are the most common parts of the
plant used by traditional healers and all the materials are
collected from the rainforest (Figure 2). In most cases plant
preparation is minimal: crushing fresh material and
applying directly to the affected part of the body; or
brewing a tea or infusion for drinking. For example, for
treating coughs and malaria, the bark of Alstonia scholaris
is scraped, boiled, cooled, filtered, and then drunk. A small
105
amount of cold or hot water is usually added to the

concoction, especially to make liquid medicine more
palatable. Some traditional medicines are applied directly
to the patient without initial preparation. For example, to
cure wounds and sexually transmitted diseases in men, the
latex of Artocarpus altilis is applied to the wounds or
genital organs.
Figure 2. Plant parts are commonly used for traditional medicine

by indigenous people from Manokwari District.
Although each traditional plant has its own dosage and

frequency of use, most indigenous people in Manokwari
District will continue to use the traditional medicines twice
a day until the patient is totally healed. They do not
recognise contra-indications during the healing process,
except where the plants are used to heal particular diseases,
which are related to the supernatural.
Local preservation of plant knowledge
Recently, with greater movement of young people away
from the villages, a significant decrease in the plant
knowledge of the younger generations has been noticed in
many parts of the region. While one of the original
intentions of this study was to determine the extent to
which a similar trend is occurring in the Manokwari
District area, this was difficult to assess because of the
relatively small number of permanent residents less than 60
years of age. However, it was found that when members of
the younger generation returned to visit their villages,
many spent time with their parents and older relatives
asking about plants, animals, and cultural traditions.
Knowledge of the medicinal uses of plants is an
individually developed skill that is regarded as a person's
particular interests or gift. Certain individuals are
recognised as special authorities on healing, and are
consulted on a regular basis for remedies for serious and/or
persistent illnesses. It appears that most sick people first
attempt to cure themselves, and if unsuccessful, they will
then visit a traditional healer. If the healer's remedies also
fail, only then will the person seek the help of a distant
medical doctor. In most cases, when the indigenous people
106
of the Manokwari District felt malaise or have aches, they

called upon the services of a traditional healer.
Quite often returning villagers who have established
permanent residences elsewhere also turned to the
traditional healers for natural herbal remedies. Many
villagers expressed strong distrust and/or disbelief in
[western] medical knowledge, and only when desperate
would they visit the area doctor in another village or travel
to a city hospital. Many people simply refused to consult
doctors at all. The main factors contributing to such
attitudes seem to be the equating of pills of undetermined
origin and content with potent poisons, and to an
abhorrence for invasive surgical procedures. Such beliefs
have arisen from failed medical treatment of close relatives
who have suffered a chronic or serious disease or have
died. Undoubtedly, the frequency of such failed treatments
may be due to the fact that most diseases were in advanced
stages before patients subjected themselves to treatment.
Cost is another factor, and traditional healers were readily
accessible, whereas medical treatment often involved great
financial expense and arduous travel.
CONCLUSIONS
The indegenous people in Manokwari have been using
at least 99 plant species as sources of medicines; the plats
are widely dustributed among 93 genera and 59 families.
Most of these traditional plants are commonly gathered
from the local rainforest communities. The plants have
been used by the native people to treat medical conditions
grouped into several categories namely gastrointestinal
disorders, dermatological conditions, illnesses associated
with pain and/or fever, respiratory ilnesses, womens
medicines, plants used to counteract bites by venomous
animal, wounds, burn, and eye remedies, and other uses.
The indegenous people in Manokwari have used almost
part of the plants for treating several medical conditions,
but leaf extract are the most common part used by
traditional healers. Plant parts are prepared either by
crushing down or apply fresh material and by direct
application to the affected part of the body or brewing a tea
of infusion for drinking.
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GUIDANCE FOR AUTHORS

Aims and Scope Biodiversitas , Journal of Biological Diversity or
Biodiversitas encourages submission of manuscripts dealing with all
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(Boonkerd 2003a, b, c; Sugiyarto 2004; El-Bana and Nijs 2005; Balagadde et
al. 2008; Webb et al. 2008). Extent citation as shown with word cit should
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should not appear in the list but should be cited in the text only (e.g., Rifai
MA 2007, personal communication; Setyawan AD 2007, unpublished data).
In the reference list, the references should be listed in an alphabetical order.
Names of journals should be abbreviated according to the ISSN List of Title
Word Abbreviations (www.issn.org/2-22661-LTWA-online.php).
APA style in double space is used in the journal reference as follow:
Journal:
Saharjo BH, Nurhayati AD (2006) Domination and composition structure
change at hemic peat natural regeneration following burning; a case study
in Pelalawan, Riau Province. Biodiversitas 7: 154-158.
Book:
Rai MK, Carpinella C (2006) Naturally occurring bioactive compounds.
Elsevier, Amsterdam.
Chapter in book:
Webb CO, Cannon CH, Davies SJ (2008) Ecological organization, biogeography and
the phylogenetic structure of rainforest tree communities. In: Carson W,
Schnitzer S (eds) Tropical forest community ecology. Wiley-Blackwell,
New York.
Abstract:
Assaeed AM (2007) Seed production and dispersal of Rhazya stricta. 50th
annual symposium of the International Association for Vegetation
Science, Swansea, UK, 23-27 July 2007.
Proceeding:
Alikodra HS (2000) Biodiversity for development of local autonomous
government. In: Setyawan AD, Sutarno (eds) Toward mount Lawu
national park; proceeding of national seminary and workshop on
biodiversity conservation to protect and save germplasm in Java island.
Sebelas Maret University, Surakarta, 17-20 July 2000. [Indonesian]
Thesis, Dissertation:
Sugiyarto (2004) Soil macro-invertebrates diversity and inter-cropping plants
productivity in agroforestry system based on sengon. [Dissertation].
Brawijaya University, Malang. [Indonesian]
Information from internet:
Balagadde FK, Song H, Ozaki J, Collins CH, Barnet M, Arnold FH, Quake
SR, You L (2008) A synthetic Escherichia coli predator-prey ecosystem.
Mol Syst Biol 4: 187. www.molecularsystemsbiology.com
ISSN: 1412-033X
E-ISSN: 2085-4722
GENETIC DIVERSTY
The conservation of mitochondrial genome sequence in Leucadendron (Proteaceae)
MADE PHARMAWATI, GUIJUN YAN, PATRICK M. FINNEGAN
Isolation and phylogenetic relationship of orchid-mycorrhiza from Spathoglottis plicata
of Papua using mitochondrial ribosomal large subunit (mt-Ls) DNA
SUPENI SUFAATI, VERENA AGUSTINI, SUHARNO
53-58
59-64
ECOSYSTEM DIVERSTY
Diversity of macrofungal genus Russula and Amanita in Hirpora Wildlife Sanctuary,
Southern Kashmir Himalayas
SHAUKET AHMED PALA, ABDUL HAMID WANI, RIYAZ AHMAD MIR
Effect of plantations on plant species diversity in the Darabkola, Mazandaran Province,
North of Iran
HASSAN POURBABAEI, FATEMEH ASGARI, ALBERT REIF, ROYA ABEDI
Determination of long-tailed macaques (Macaca fascicularis) harvesting quotas based
on demographic parameters
YANTO SANTOSA, KUSMARDIASTUTI, AGUS PRIYONO KARTONO, DEDE AULIA RAHMAN
The population condition and the food availability of cuscus in the Arfak Mountains
Nature Reserve, West Papua
ANTON SILAS SINERY, CHANDRADEWANA BOER, WARTIKA ROSA FARIDA
ETHNOBIOLOGY
Vegetation stands structure and aboveground biomass after the shifting cultivation
practices of Karo People in Leuser Ecosystem, North Sumatra
T. ALIEF ATHTHORICK, DEDE SETIADI, YOHANES PURWANTO, EDI GUHARDJA
The wild plants used as traditional medicines by indigenous people of Manokwari,
West Papua
OBED LENSE
65-71
72-78
79-85
86-91
92-97
98-106
Front cover:
Leucadendron salignum
(PHOTO: SEAN PRIVETT)
Published four times in one year
PRINTED IN INDONESIA
ISSN: 1412-033X
E-ISSN: 2085-4722

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ISSN: 1412-033X

Journal of Biological Diversity

13 Number 2 April 2012

1412-033X (printed edition)

EDITORIAL BOARD (COMMUNICATING EDITORS):

English Editors: Suranto; Technical Editor & Banking: Solichatun (solichatun_s@yahoo.com)

Ahmad Dwi Setyawan (unsjournals@gmail.com)

The Society for Indonesian Biodiversity

Solichatun, BNI KC Sebelas Maret, Acc. No. 0033691646

EXPERTISE AND CORRESPONDING EMAIL OF THE COMMUNICATING EDITORS:

Society for Indonesia

Sebelas Maret University

The conservation of mitochondrial genome sequence in Leucadendron

MADE PHARMAWATI1, GUIJUN YAN2, PATRICK M. FINNEGAN2

The use of cytoplasmic DNA sequences as breeding,

B I O D I V E R S IT A S 13 (2): 53-58, April 2012

Australian National Biodiversity Research with voucher

DNA sequencing and analysis

Seven universal PCR primer pairs designed to amplify

detected in cpDNA with 14 of 33 primer pair - enzyme

B I O D I V E R S IT A S 13 (2): 53-58, April 2012

PHARMAWATI et al. Mitochondrial genome of Leucadendron

example, among 13 Elymus species analysed,

most closely related to Lamprocapnos spectabilis

B I O D I V E R S I T A S 13 (2): 53-58, April 2012

Chapdelaine Y, Bonen L. 1991. The wheat mitochondrial gene for subunit

Johnson LAS, Briggs BG. 1975. On the Proteaceae-the evolution and

Isolation and phylogenetic relationship of orchid-mycorrhiza from

therefore they will absorb water and certain minerals

B I O D I V E R S IT A S 13 (2): 59-64, April 2012

MATERIALS AND METHODS

DNA amplification by PCR

RESULTS AND DISCUSSION

SUFAATI et al. Orchid-mycorrhiza from Spathoglottis plicata

Figure 1. Morphology of some isolates of orchid mycorrhiza isolated from S. plicata.

In a study, Athipunyakom et al.

B I O D I V E R S IT A S 13 (2): 59-64, April 2012

Table 2. Description of the morphology of orchid-mycorrhiza species associated with S. plicata.

Growth of colonies is rather slow, 8 days.

Branched hyphae with dense septate.

Colony growth is slow.

Branched hyphae, more than 1core, dense septate.

The colony growth is very fast, within 3 days

The colony growth is very fast, within 5 days

Slow colony growth, 15 days

Colony growth is slow, 17 days

Colony growth is very slow 20 days

Growth of colony is very slow, 24 days.

Slow growth colony, 16 days.

Colony growth is rather slow, 8 days

Hyphae are branched with non-close septate.

SUFAATI et al. Orchid-mycorrhiza from Spathoglottis plicata

Phylogenetic relationship of orchid-mycorrhiza of

Figure 2. Dendrogram showing the phylogenetic relationship of

B I O D I V E R S IT A S 13 (2): 59-64, April 2012

Fundamentals Grant of 2008. To Prof. Zamir K. Punja

Wu J, Ma H, Lu M, Han S, Zhu Y, Jin H, Liang J, Liu L, Xu J. 2010.

Diversity of macrofungal genus Russula and Amanita in Hirpora

around 750 worldwide species of mycorrhizal mushrooms

MATERIALS AND METHODS

Figure 1. Map showing the collection

aand preservatiion for herbarrium purposess. The spore prints

herbarium purpooses, in funngal collectio