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Genetic variation and phylogeny assessment of Aciagrion occidentale (Odonata: Coenagrionidae) using mitochondrila cytochrome oxidase subunit I gene

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E. K. Jisha Krishnan
E. K. Jisha Krishnan
E. K. Jisha Krishnan
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Sebastian C. D. at University of Calicut
Sebastian C. D.
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Abstract and Figures

Aciagrion occidentale is a migratory species widely distributed in montane and submontane areas in open grass besides weedy ponds and herbage. Here we used the COI barcode locus (522bp) to clarify the specific taxonomic status of Aciagrion occidentale. Molecular data suggests that this species is having 99% sequence similarity to Aciagrion borneense found in Netherland and confirmed that both are sharing same genus. The N-J tree constructed with BLASTn result depicted that Aciagrion occidentale is phylogenetically very close to damselflies. Anisopterans and Zygopterans are sharing some common characters and the results confirm that Zygoptera is a paraphyletic group derived from the monophyletic groups of Anisoptera and Anisozygoptera.
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International Journal of Science and Research (IJSR)
ISSN (Online): 2319-7064
Index Copernicus Value (2013): 6.14 | Impact Factor (2013): 4.438
Volume 4 Issue 4, April 2015
www.ijsr.net
Licensed Under Creative Commons Attribution CC BY
Genetic Variation and Phylogeny Assessment of
Aciagrion Occidentale (Odonata: Coenagrionidae)
Using Mitochondrial Cytochrome Oxidase Subunit I
Gene
Jisha Krishnan E. K., Sebastian C. D*
Molecular Biology Laboratory, Department of Zoology,
University of Calicut, Kerala 673 635 India
*Corresponding Author: drcdsebastian@gmail.com
Abstract: Aciagrion occidentale is a migratory species widely distributed in montane and submontane areas in open grass besides
weedy ponds and herbage. Here we used the COI barcode locus (522bp) to clarify the specific taxonomic status of Aciagrion occidentale.
Molecular data suggests that this species is having 99% sequence similarity to Aciagrion borneense found in Netherland and confirmed
that both are sharing same genus. The N-J tree constructed with BLASTn result depicted that Aciagrion occidentale is phylogenetically
very close to damselflies. Anisopterans and Zygopterans are sharing some common characters and the results confirm that Zygoptera is
a paraphyletic group derived from the monophyletic groups of Anisoptera and Anisozygoptera.
Keywords: Odonates, Aciagrion occidentale, Mitochondrial DNA, Cytochrome oxidase subunit I gene
1. Introduction
Odonates are often addressed as the ‘guardians of the
watershed’, includes dragonflies and damselflies.
Damselflies tend to be less robust, appearing weak in
flight. Most species hold their wings folded back over the
abdomen at rest. Damselflies eyes occupy much of the
animal's head, nearly touching each other across the face
with typically a gap between the eyes. In nature they
appear as the subject of conservation strategy in the case
of endangered species and indicators of wetland habitat
quality. The habitat selection of adult Odonate species
strongly depends on vegetation structure and their larvae
develop only in clean water. Thus the different stages of
their life cycle are very much sensitive to the ecological
disturbances. Therefore, along with birds and amphibians,
dragonflies can serve as one of the key bio-indicator
groups, whose high species diversity clearly mirrors
favourable conservation state of wider wetland ecosystem
[1]. About 137 species and subspecies of Odonata
spreading over 79 genera, 12 families and 31 subfamilies
are reported from Kerala [2]. Aciagrion occidentale is one
of the important odonate species coming under the family
Coenagrionidae, commonly known as ‘Slims’. They are
widely distributed across Central and South India, Sri
Lanka, Thailand, Cambodia and Vietnam. No major threat
to the species has been described and the species is quite
common across its range. Suitable habitat is shrub
dominated wetlands although it can also be found in bogs,
marshes, swamps, fens, peat lands, small streams and
permanent freshwater lakes. Males are very small and
slender, having 8 abdominal segments with a black
elongate dorsal triangular mark. Females are similar to
male but much more robust and with a stouter abdomen.
Taxonomy is the identification of species, based on the
morphological characters and it requires vast knowledge
about varied organisms and their characters. Identification
using traditional taxonomy is difficult due to the external
changes in the organisms caused by seasonal and
geographical variations. Many organisms alter themselves
physiologically and morphologically due to the
unfavourable conditions in the environment. Thus
adoption of manual taxonomy, often leads to wrong
identification of the species. This problem has thus
influenced the emergence of the molecular taxonomic
frame work studies for the conformation and the
betterment in the identification of species.
Mitochondrial DNA acts as an excellent genetic marker
with the gene flow in its matrilineal inheritance. It
represents maternal haploid genome and accumulates
nucleotide substitution 5-10 times more rapidly than
nuclear DNA, which makes it suitable for examining
population and sub population structures among related
taxa [3]. Cytochrome oxidase subunit I (COI) gene is one
of the most important protein encoding genes of mt DNA
and has been utilized in the studies of molecular evolution
due to lack of introns, simple alignment, limited exposure
to recombination and availability of robust primer sites.
Phylogenetic analysis using COI gene sequences were
extensively carried out by several workers in different
group of organisms like southern house mosquito Culex
quinquefasciatus [4], Armigeres subalbatus mosquito [5]
green bottle fly Lucilia sericata [6], Herpetogramma
stultalis [7], white backed plant hopper Sogatella furcifera
[8], Asian honeybee Apis cerana [9], and lepidopteran
species [10]. Here we amplified cytochrome oxidase I gene
of Aciagrion occidentale found in Kerala for its
phylogenetic analysis.
Paper ID: SUB153250
1121
International Journal of Science and Research (IJSR)
ISSN (Online): 2319-7064
Index Copernicus Value (2013): 6.14 | Impact Factor (2013): 4.438
Volume 4 Issue 4, April 2015
www.ijsr.net
Licensed Under Creative Commons Attribution CC BY
2. Methodology
Genomic DNA was extracted from one of the thoracic legs
of the experimental insect, Aciagrion occidentale (Figure
1). The tissue was homogenized and genomic DNA in the
homogenate was isolated using Ultrapure Mammalian
Genomic DNA Pre Kit. About 2 ng of genomic DNA was
amplified for mitochondrial cytochrome oxidase subunit Ι
(COΙ) gene using forward Primer, 5'-
CATTGGAGATGACCAAATTTA-3'and reverse primer,
5'ATTGGATCTCCACCACCTGC-3'. The The PCR
products were resolved on a 1% TAE- agarose gel, for
confirmation of the target gene amplification. The PCR
product was column purified using UltraClean PCR Clean-
up Kit (Mo Bio Laboratories, Inc. California). The purified
PCR product was sequenced from both ends using
Sanger’s sequencing method[11]. Sequences obtained
were assembled by using ClustalW and the consensus was
taken for the analysis. The final sequence was searched for
its similarity using BLAST of NCBI
(www.ncbi.nlm.nih.gov/) and submitted in the GenBank
for worldwide accession. The phylogenetic tree was
plotted in Neighbor Joining method using by MEGA6
software [12].
3. Results and Discussion
The partial mitochondrial cytochrome oxidase I (COI)
region of Aciagrion occidentale yielded a 522bp long
fragment (GenBank Accession number: KM 096996)
showing 99% sequence similarity to Aciagrion borneense
(GenBank Accession number: KF 369275) found in
Netherland.
Figure 1: Aciagrion occidentale
Odonates are the indicators of environmental quality,
evolutionary developmental biology and conservation
biology [1]. They generally belongs to the base of winged
insect (Pterygota), making them unique and exciting. This
phylogenetic position shows that they can provide most
useful information on the evolution of the morphological
diversification and mechanics of insect wings and the
general body plan of the winged insects. The effect of
changing environmental condition related to climate
change, environmental pollution, and water quality, habitat
fragmentation on life history strategies, population
dynamics and adaptability of animals in the field rather
than in the laboratory makes them an excellent model
system for study purpose [2]. Aciagrion occidentale is a
member of Coenagrionidae family of Odonates. This
damselfly occurs in a wide variety of habitats chiefly along
streams, ponds and swamps.
Figure 2: Phylogenetic tree of Aciagrion occidentale
(JO5CU) using neighbor joining method
GenBank data analysis showed that Aciagrion occidentale
(KM 096996) is having 99% sequence similarity to
Aciagrion borneense COI gene (KF 369275) reported from
Netherland [1]. This indicates that both of them are
sharing a common ancestor. The N-J tree constructed with
BLASTn result depicted that Aciagrion occidentale is
phylogenetically very close to the other species of
damselflies reported from different geographical locations.
Acknowledgement
The financial assistance from Kerala State Council for
Science Technology and Environment,
Thiruvananthapuram under Science Research Scheme is
gratefully acknowledged.
References
[1] [1] K. Dijkstra, KJ Kalkman, RA Dow, FR Stokvis,
Tol Van. Redefining the damselfly families - A
Comprehensive molecular phylogeny of Zygopterans.
Systematic Entomology, 3: 235-256, 2013.
[2] [2] KG Emiliyamma, C Radhakrishnan, Palot
Muhamed Jafer. Odonata (Insecta) of Kerala.
Zoological Survey of India. Kolkota, pp 195, 2007.
[3] [3] AR Hoelzel. Analysis of regional mitochondrial
DNA variation in the killer whale. Genetic ecology of
whales and dolphins. Report to the International
Whaling Commission. Cambridge, UK. 76: 255-233,
1991.
[4] [4] K Rukhsana, KS Ashitha, K Mashoor, CD
Sebastian. Molecular phylogeny analysis of Southern
KC912320.1 Ischnura senegalensis
KC912324.1 Ischnura senegalensis
JN419694.1 Enallagma sp.
KF369414.1 Ischnura aurora
JN419852.1 Ischnura posita
KF369526.1 Pseudagrion pilidorsum
FJ812830.1 Nesobasis sp
FJ812853.1 Nesobasis rufostigma
FJ812839.1 Nesobasis erythrops
FJ812855.1 Nesobasis selysi
FJ812847.1 Nesobasis selysi
KC135957.1 Ischnura asiatica
AF429295.1 Aeshna subpupillata
AF429293.1 Aeshna subpupillata
JO5CU
KF369275.1 Aciagrion borneense
0.2
Paper ID: SUB153250
1122
International Journal of Science and Research (IJSR)
ISSN (Online): 2319-7064
Index Copernicus Value (2013): 6.14 | Impact Factor (2013): 4.438
Volume 4 Issue 4, April 2015
www.ijsr.net
Licensed Under Creative Commons Attribution CC BY
house mosquito, Culex quinquefasciatus (Diptera:
Culicidae) derived from mitochondrial DNA
sequences, International Journal of Research, 1(5):
844-853, 2014.
[5] [5] PU Bindu, CD Sebastian. Genetic structure of
mitochondrial cytochrome oxidase subunit Igene of
the mosquito, Armigeres subalbatus. International
Journal of Research, 1(10): 49-56, 2014.
[6] [6] KP Priya Bhaskaran, CD Sebastian. Molecular
barcoding of green bottle fly, Lucilia sericata
(Diptera: Calliphoridae) using COI gene sequences.
Journal of Entomology and Zoology Studies, 3(1):10-
12, 2014.
[7] [7] VP Akhilesh, CD Sebastian. Molecular barcoding
and phylogeny analysis of Herpetogramma stultalis
(Lepidoptera:Crambidae) using cytochrome oxidase
subunit I gene sequence. International Journal of
Advanced Life Sciences, 7: 463-466, 2014.
[8] [8] K Sreejith, CD Sebastian. Phylogenetic analysis
and sequencing of the mitochondrial cytochrome
oxidase subunit I (COI) of white backed plant hopper,
Sogatella furcifera (Horvath). International Research
Journal of Pharmacy, 5 (12): 887-890, 2014.
[9] [9] K Rukhsana, VP Akhilesh, CD Sebastian.
Deciphering the molecular phylogenetics of the Asian
honey bee, Apis cerana (Hymenoptera: Apidae) and
inferring the phylogeographical relationships using
DNA barcoding, Journal of Entomology and Zoology
Studies, 2(4): 218-220, 2014.
[10] [10] E Pavana, CD Sebastian. Genetic diversity and
phylogenetic analysis of lepidopteran species by
molecular barcoding using CO I gene sequences.
International Journal of Science and Research, 3(5):
450-452, 2014.
[11] [11] Sanger, F. and Coulson, A. R. A rapid method for
determining sequences in DNA by primed synthesis
with DNA polymerase. Journal of Molecular Biology,
94 (3): 441–448, 1975.
[12] [12] K Tamura, G Stecher, D Peterson, A Filipski, S
Kumar. MEGA6: Molecular Evolutionary Genetics
Analysis Version 6.0. Molecular Biology and
Evolution, 30(12): 2725-2729, 2013
Paper ID: SUB153250
1123
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  • Aug 2013
  • SYST ENTOMOL
An extensive molecular phylogenetic reconstruction of the suborder Zygoptera of the Odonata is presented, based on mitochondrial (16S, COI) and nuclear (28S) data of 59% of the 310 genera recognized and all (suspected) families except the monotypic Hemiphlebiidae. A partial reclassification is proposed, incorporating morphological characters. Many traditional families are recovered as monophyletic, but reorganization of the superfamily Coenagrionoidea into three families is proposed: Isostictidae, Platycnemididae and Coenagrionidae. Archboldargia Lieftinck, Hylaeargia Lieftinck, Palaiargia Förster, Papuargia Lieftinck and Onychargia Selys are transferred from Coenagrionidae to Platycnemididae, and Leptocnemis Selys, Oreocnemis Pinhey and Thaumatagrion Lieftinck from Platycnemididae to Coenagrionidae. Each geographically well-defined clade of Platycnemididae is recognized as a subfamily, and thus Disparoneurinae (i.e. Old World ‘Protoneuridae’) is incorporated, Calicnemiinae is restricted, and Allocnemidinae (type genus: Allocnemis Selys) subfam.n., Idiocnemidinae (type genus: Idiocnemis Selys) subfam.n. and Onychargiinae (type genus: Onychargia Selys) subfam.n. and Coperini trib.n. (type genus: Copera Kirby) are described. Half of Coenagrionidae belongs to a well-supported clade incorporating Coenagrion Kirby and the potential subfamilies Agriocnemidinae, Ischnurinae and Pseudagrioninae. The remainder is less well defined, but includes the Pseudostigmatidae and New World Protoneuridae that, with Argiinae and Teinobasinae, may prove valid subfamilies with further evidence. Ninety-two per cent of the genera formerly included in the polyphyletic Amphipterygidae and Megapodagrionidae were studied. Pentaphlebiidae, Rimanellidae and Devadattidae fam.n. (type genus: Devadatta Kirby) are separated from Amphipterygidae, and Argiolestidae, Heteragrionidae, Hypolestidae, Philogeniidae, Philosinidae and Thaumatoneuridae from Megapodagrionidae. Eight further groups formerly placed in the latter are identified, but are retained as incertae sedis; the validity of Lestoideidae, Philogangidae and Pseudolestidae is confirmed. For some families (e.g. Calopterygidae, Chlorocyphidae) a further subdivision is possible; Protostictinae subfam.n. (type genus: Protosticta Selys) is introduced in Platystictidae. Numerous new combinations are proposed in the Supporting Information. Many long-established families lack strong morphological apomorphies. In particular, venation is incongruent with molecular results, stressing the need to review fossil Odonata taxonomy: once defined by the reduction of the anal vein, Protoneuridae dissolves completely into six clades from five families.
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MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0
Article
Full-text available
  • Oct 2013
  • MOL BIOL EVOL
We announce the release of an advanced version of the Molecular Evolutionary Genetics Analysis (MEGA) software, which currently contains facilities for building sequence alignments, inferring phylogenetic histories, and conducting molecular evolutionary analysis. In version 6.0, MEGA now enables the inference of timetrees, as it implements our RelTime method for estimating divergence times for all branching points in a phylogeny. A new Timetree Wizard in MEGA6 facilitates this timetree inference by providing a graphical user interface (GUI) to specify the phylogeny and calibration constraints step-by-step. This version also contains enhanced algorithms to search for the optimal trees under evolutionary criteria and implements a more advanced memory management that can double the size of sequence data sets to which MEGA can be applied. Both GUI and command-line versions of MEGA6 can be downloaded from www.megasoftware.net free of charge.
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A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase
Article
  • Jun 1975
A simple and rapid method for determining nucleotide sequences in single-stranded DNA by primed synthesis with DNA polymerase is described. It depends on the use of Escherichia coli DNA polymerase I and DNA polymerase from bacteriophage T4 under conditions of different limiting nucleoside triphosphates and concurrent fractionation of the products according to size by ionophoresis on acrylamide gels. The method was used to determine two sequences in bacteriophage φX174 DNA using the synthetic decanucleotide A-G-A-A-A-T-A-A-A-A and a restriction enzyme digestion product as primers.
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