TROPICAL AGRICULTURAL SCIENCE Phylogeny

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


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


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)


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


the genetic distance among the species A. A. alecto, the divergence values

Aethalops

495

496

populations


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


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


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


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


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


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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