Abbr. Field No. UNIMAS. Voucher No. Accession. No. Mt. Kinabalu. 1527 m. a.s.l. 44.50. M. TK4. TK004. UNIMAS 00371. HM067793. 42.18. F. TK20. TK152920.
TROPICAL AGRICULTURAL SCIENCE
Phylogeny and Phylogeography of Aethalops from Sundaland using Mitochondrial 12S rRNA Gene Tingga, R. C. T.1, 2* and Abdullah, M. T.1 2
ABSTRACT Aethalops Aethalops is generally known as Aethalops alecto in most previous A. alecto and A. using partial A. alecto were and minimum spanning network, there were two major clusters within the genus, with Aethalops A. alecto from the islands were unresolved at the
populations of ancestral populations of
Aethalops
INTRODUCTION Article history:
Aethalops is among of the
Aethalops et
al
et al
et al brown to reddish brown, and thick and long and pointed and forearm length is between et al distinctive characteristics that differentiate this genus from its sister genus, Balionycteris et al Aethalops are tailless, spotless on the wings and have a et al Aethalops are found throughout Peninsular
sites for A. alecto
486
genus, namely A. alecto and , and both are endemic to the mountainous there is a distribution boundary between et al Maharadatunkamsi et al some authors still consider Aethalops in A. alecto rather than et al Aethalops A.
Phylogeny and Phylogeography of Aethalops
of A. alecto
In this paper, the phylogenetic Aethalops were Aethalops
Mt
to construct the phylogenetic relationship of A. alecto and and determine the patterns of gene flow of
M M
-
M
-
M
-
1446 m
-
Madi Mt Murud
M
M M
487
Mt Murud
M M
M
Mrd004
-
Mrd007
-
Mrd008
-
Mrd009
-
Mrd015
-
Mt Mulu
M
-
M
-
M
-
44 Mt Penrissen
746-1000 m
M M M
488
MP03 MP3
MP06
Phylogeny and Phylogeography of Aethalops
Mt Penrissen
746-1000 m -
MP4
MP001
-
MP5
MP016
-
MP6
-
M M Mt Pueh
-
1046
MATERIALS AND METHODS et al
polymerase forest trail, near streams and on the forest measured following Payne et al
et al either as wet or dry specimens, and the
part of the body and preserved either in lysis was
bromide, run on gel electrophoresis for 30 100 bp
sent to private laboratories for sequencing terminator ®
et al
®
489
and A. alecto
A. alecto
400 - 1803
1400
490
Phylogeny and Phylogeography of Aethalops
et al.,
consensus tree from a parsimony heuristic between the populations was performed in
was performed with 100th generations implementing Metropolis-
each with four independent incrementally heated Markov chains, sampling every 100th generation and burn-in of 1000 for convergence of the two runs was assumed when the average standard deviation of the split frequencies has reached less than
et al
phylogenetic relationship study for discrete for MP analysis were performed with 10
was assessed using 1000 bootstrap iterations
et al number of haplotype, segregating sites and total number of mutations were estimated et al st st and m et al
491
values was determined by a permutating test st , which is the population subdivision index, was calculated to describe the reduction in heterozigosity relative to the total population st
is the most common measurement used to describe the genetic differentiation of the populations and was developed by Wright st is the value of probability of two random gametes which were drawn from two populations that are identical by descent, and relative to gametes taken from the entire st values ranging from st
statistical approach, was used to compare the geographical distance and genetic differentiation among the populations; in other words, to test for the isolation by
m migrants per generation, was also et al st
m is more than 1, the populations are
estimated following Mindell et al
having little genetic differentiation, whereas
RESULTS et al st is used to estimate the degree of populations’ subdivision at the nucleotide
et al
et al
method that uses permutations to test were independent of each other and a
and A. alecto A. alecto was unsuccessfully to be captured individuals, including two outroups, 69 were successfully sequenced and aligned for
of genus Aethalops
670
636
901
Pueh
865
591
868
Penrissen
0.32 (0.0-1.4)
Mulu
0.38 (0.0-0.7)
830
0 (0.0)
Murud
Mulu
Murud
0.68 (0.0-3.6)
676
0.0 0.35 (0.0-0.7)
Penrissen
0.0 (0.0)
Pueh
1.61 (0.0-2.1)
Phylogeny and Phylogeography of Aethalops
493
494
1 1 1
9
6
A. alecto.
4
17 1 1
1
Murud
1
1 5
Mulu
1
4
Penrissen
1
Pueh
1 1 1
1 1 1
1
1 1 3 1
Phylogeny and Phylogeography of Aethalops
the genetic distance among the species A. A. alecto, the divergence values
Aethalops
495
496
populations
Phylogeny and Phylogeography of Aethalops
gene successfully extracted eight haplotypes of A. alecto A. A. alecto1 to A. alecto8 were haplotypes of A. alecto from 1 to unique haplotypes of
similar results by grouping the Aethalops
4 were from the
haplotypes of
of A. alecto
A.
In particular, had three shared haplotypes and the most common haplotype was 8 which was shared by
A.
pa component ct sc
Within populations
st
a
index than the observed by chance alone after 1000 permutations
st
the populations of
*
*
*
*
*
497
st
m populations of
st
m
st st
et al
Aethalops, based
498
Phylogeny and Phylogeography of Aethalops
Aethalops
with Aethalops produced slightly different topologies from the Aethalops was revealed by clustering
from supported by high bootstrap value in all
499
Aethalops
A. 8, 6 and 13 were shared by a few individuals from
was deviated from
3 of the
haplotype for each species were denoted by the proportional size of their haplo11 from 8 by
steps before reaching the
sharing haplotypes with other populations of
gene intrapopulation was also low
500
of the Bornean Pigmy Fruit Bats
Phylogeny and Phylogeography of Aethalops
from
node represents a unique haplotype and the node sizes are proportional to the haplotype frequencies of
may due to high frequency of haplotype in the populations of both suggesting that these two populations used in this present analysis were small in et al the small sample size may underestimate the actual haplotype distribution among the had a low level of genetic differences which 501
observed between the geographical distance and net percent nucleotide divergence, the populations of between the populations was not a factor that contributed to the divergence of the sequences in
estimated
st
values among the grouped
differentiation matrix of the populations st
were grouped into three which consisted
st
m A.
that the among group has the highest variation
st
st
, with high
Phylogeny and Phylogeography of Aethalops
m st
from A. alecto st, and the highest level
et al
m
et al
gene flow between these populations is appeared to have the lowest gene flow, suggesting that this population was isolated m comparisons of genetic differentiation A.
b
of
have had high genetic A.
shared haplotypes among the individuals,
genetically similar, they are considered as a single morphotype based on the same DISCUSSION
Aethalops into two major monophyletic groups corresponding to A. alecto and A.
interpopulation relationships of however, the separation between Malaysian
clustered in between two clades consisting
, and should no longer be referred to as A. alecto, and A.
slight morphological difference between populations of these bats is possibly due to the adaptation to food resources to survive the skull of the bats may have evolved to adapt into optimised form to meet the demand of holding and masticating of different food sources, depending on what
common haplotype may be the oldest, with the expectation that the haplotype should is predicted as the ancestral
503
rooting, it is suggested that this may not be the absolute oldest haplotype, but it can relatively be considered as one of the ancestral haplotype as compared to other observed haplotypes of relatively similar case was also observed
for the speciation of were not consistent with the Pleistocene speciation this species was apparently due to dispersal rather than vicariance, changes of sea level et al
were observed to be widely distributed but not placed at the basal clade to be considered also showed that the common haplotypes were not rooted at the basal; however, A. and were rooted at the basal of the monophyletic group of A.
of the observed islands in Indonesia was the earliest population of A. alecto after its separation from earlier, the divergence time between A. alecto and
to be the ancestor population among all the A. alecto populations that had been
still retain its ancestral haplotype which a divergent individual from this species is regarded to be associated with distinct
the first colonised island of A. alecto after this particular species had diverged from undetermined whether the two forms of Aethalops It is possible that A. alecto
largely independent of glaciations events et al et al et al., and A. alecto was estimated
504
et al
by Mindell et al
Phylogeny and Phylogeography of Aethalops
Historical Population of the Bornean Pigmy Fruit Bats time for et al who found no evidence indicating mammals
et al., A. was found to have diverged from its sister species A. alecto approximately
that time, intermittent land bridges allowed et al widespread distribution of that the presence of the fauna characteristic of open woodlands found in the vertebrate et al
et al have likely caused the island to retain a group representative from the populations In the analysis of the current data, all the phylogenetic trees showed a close genetic relationship between the populations in
facilitated the dispersal of A. alecto condition has also been observed in other species of bats, such as Myotis muricola and
colonisation event that occurred before the Pleistocene and was not caused by et al species is endemic to the island and this has not undergone repeated extinction and recolonisation, and it is more likely to have persevered at a particular island since et al
predicted to be the location of the original population of among the populations of in found to be the ancestor towards the other
M. muricola was not clustered accordingly to the population groups based
M. muricola
and thus limits the gene flow between
505
as there were no secondary data available in
refugium for the lowland rainforest species
polymorphism has been used as an inference on the historical patterns of population
et al Pleistocene facilitated the dispersal and genetic exchange of populations
that the pattern of movements of this
from A. alecto of
of a montane bat is similar to a montane bird, where dispersal to another mountain or range occurs along a spinal chain that
chain, with an elevation of more than 1500 recently diverged from the one in southeast
away to form a long stretch of lowland
that are supported by very close genetic
of this ridge could be one of the reasons that had led to genetic divergence between the population at Mt Penrissen and those in the
CONCLUSIONS suggests that the haplotypes from Mt Mulu, of and A. alecto findings conclude that
is a
In fact, this could be the reasons why these
was predicted to be the ancestral group
506
and thus support the previous studies that is no longer known as a subspecies of A. alecto
Phylogeny and Phylogeography of Aethalops
of
genetic distance between
from
high and this is supported by the high value of population and nucleotide subdivision, which produced a new hypothesis on this particular species, with the possibility of two
postulated as the possible ancestral for A.
ACKNOWLEDGEMENTS relationships of among the populations of
in
examine the specimens and tissue samples
similarities, whereby the dispersal was
507
Biological Journal of the Linnean Society,
,
Aethalops alecto Mammalia,
to work at the protected areas, using the and also the permit to enter park number Biochemical Genetics,
for permitting us to collect the samples
Hemigbagrus nemurus Molecular Ecology,
valenciennes
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