Journal of Threatened Taxa |
www.threatenedtaxa.org | 26 June 2023 | 15(6):
23425–23430
ISSN 0974-7907
(Online) | ISSN 0974-7893 (Print)
https://doi.org/10.11609/jott.8439.15.6. 23425-23430
#8439 | Received 17
March 2023 | Final received 23 May 2023 | Finally accepted 01 June 2023
First record and DNA barcode of a
scarab beetle, Adoretus kanarensis
Arrow, 1917 (Coleoptera: Scarabaeidae:
Rutelinae), from Maharashtra, India
Pranil Jagdale
1, Sujata Magdum 2, Aparna Sureshchandra Kalawate 3,
Swapnil Kajale 4
&
Yogesh Shouche
5
1,2 PG Department and Research Centre
in Zoology, MVP Samaj’s KRT Arts, BH Commerce &
AM Science (KTHM) College, Nashik, Maharashtra 422002, India
3 Zoological Survey of India,
Western Regional Centre, Vidya Nagar, Sector 29, P.C.N.T. (PO), Rawet Road, Akurdi, Pune, Maharashtra
411044, India
4,5 National
Center for Microbial Resource, National Centre for Cell Science, NCCS Complex, Ganeshkhind, Pune, Maharashtra 411007, India
1 pranil.jagdale@gmail.com, 2 sujata_magdum@yahoo.com,
3 aparna_ent@yahoo.co.in (corresponding author), 4 swapnilckajale@gmail.com,
5 yogesh@nccs.res.in
Editor: Mandar Paingankar, Government Science College Gadchiroli,
Maharashtra, India. Date of
publication: 26 June 2023 (online & print)
Citation: Jagdale, P., S. Magdum, A.S. Kalawate, S. Kajale & Y. Shouche (2023). First record and DNA barcode of a
scarab beetle, Adoretus kanarensis
Arrow, 1917 (Coleoptera: Scarabaeidae:
Rutelinae), from Maharashtra, India. Journal of Threatened Taxa 15(6): 23425–23430. https://doi.org/10.11609/jott.8439.15.6. 23425-23430
Copyright: © Jagdale et al. 2023. Creative Commons Attribution 4.0
International License. JoTT allows unrestricted use, reproduction, and
distribution of this article in any medium by providing adequate credit to the
author(s) and the source of publication.
Funding: Council of Scientific & Industrial Research for providing fellowship under file no. 08/541(0008)/2018-EMR-I.
Competing interests: The author declares no competing interests.
Acknowledgements: The authors are grateful to Dr. V.B. Gaikwad (principal, KTHM College, Nashik), Dr. Dhriti Banerjee (director,
Zoological Survey of India, Kolkata), and Dr. Basudev Tripathy
(officer-in-charge, Zoological Survey of India, Western Regional Centre, Pune)
for providing necessary facilities and constant encouragement. The authors are
grateful to the subject editor and anonymous reviewers for the constructive
comments for the improvement of the manuscript.
Abstract: Adoretus
kanarensis Arrow, 1917, composed within the
subfamily Rutelinae of family Scarabaeidae,
was described from India about 107 years ago. These conspicuous, attractive,
and tiny leaf chafers are recorded as predators of several agricultural crops.
The mt DNA Barcodes of many Adoretus
Dejean, 1833 species are available in global
databases. In this work, first DNA barcode of A. kanarensis
has been reported. Morphological details with photographs of the species
along with a checklist of the genus Adoretus
from Maharashtra is provided. The mt COI DNA
sequences of Adoretus species recorded until
now for the genus that are listed in GenBank are analyzed.
Keywords: Chafers, dung beetle, Nashik, new
record, polyphagous pest, survey.
The genus Adoretus
Dejean, 1833 is distributed in Africa, Asia, and
Europe (Anonymous 2023). It is a polyphagous and economically important genus,
due to its potential damage to a wide variety of host plants including
important agricultural cash crops like sugarcane, tobacco and ornamental plants
(McQuate & Jameson 2011).
There are about 460 species (Krajcik 2007) reported
in the world, of which 47 are reported in India (Arrow 1917), few literature is
available on this genus from India (Chandra 2009; Chandra et al. 2012; Ghosh et
al. 2020). Some of the species of this genus are of biosecurity concerns (McQuate & Jameson 2011). The accurate identification
based on the morphological characters are important for undertaking proper
control measures. Beetles of the subfamily Rutelinae
are not dung beetles in the true sense. They are phytophagous and commonly
known as May or June beetles or shining leaf chafers (Sreedevi
et al. 2017). The pioneering work on this group in India was undertaken by
Arrow (1917) and Balthasar (1963, 1974).
Since the fauna has not been revised, few scattered publications are
available on this particular group from India, without detailed distribution
records (Kumar & Pandey 2023).
The present study was undertaken
to document the first record of A. kanarensis
from Maharashtra since Arrow (1917) noted the distribution as Bombay: North
Kanara, indicating that North Kanara was included at that time when Bombay was
the Bombay presidency during British India. Considering this, the present
report forms the first report of A. kanarensis
from Maharashtra, India based on voucher specimen with first DNA barcode for
the country.
Though Adoretus
species are widely distributed, only 12–13 mitochondrial cytochrome oxidase
subunit (COI) sequences are currently available in global database. Hence,
during one of our experiments to generate mt DNA
barcodes for the coleopteran species, we report the first mt
DNA barcode of A. kanarensis from Maharashtra,
India.
Material and Methods
Sampling of dung beetles
Specimen for the present study
was collected at night using light trap. The map of the collection locality was
prepared using open free QGIS software. The details of collection locality are
given under material examined and also shown in Figure 1.
Preservation and Identification
The collected specimen was
euthanized in the vapours of ethyl acetate and
brought to the laboratory for further studies. For morphological
identification, the specimen was studied under Leica EZ4E stereomicroscope. The
identification was done following the keys of Arrow (1917). Further, the
voucher specimen was deposited in the National Repository of Zoological Survey
of India, Western Regional Centre, Pune, Maharashtra (India).
DNA isolation, PCR and Sequencing
The ethanol preserved tissue was
used for DNA isolation. DNA from the tissues of the beetle was extracted from
metathoracic leg using DNeasy kit (Qiagen), according
to the manufacturer’s protocol. The obtained DNA was amplified using polymerase
chain reaction (PCR) using ABI thermocycler. Following primers (Meyer et al.
2005) were used for amplification of COI gene: dgLCO
F1 5’GGTCAACAAATCATAAAGAYATYGG 3’ and dgHCO R1
5’TAAACTTCAGGGTGACCAAARAAYCA 3’. PCR reaction was carried out in total volume
of 25 µl containing 2 µl DNA template, 10 pmol of
each primer and 2 µl of dNTP and 0.2 µl of Taq
polymerase (Bangalore GeNei). Thermo-cycling
conditions were as follows:
One initial cycle of 1 min at
95˚C followed by five cycles of 95˚C for 1 min, then denaturation 35 cycles of
95˚C for 1 min, annealing at 52°C for 40 s, extension at 72˚C for 1 min 15 s,
with final extension of 72˚C for 5 min.
From each PCR reaction, 2 μL of the PCR product was visualized on a 2% agarose gel
stained with ethidium bromide, together with a GeneRuler
100 bp Plus DNA Ladder (Thermo
Scientific). The obtained PCR products were sequenced with both, the forward
and reverse, primers using an automated sequencer (3730 DNA analyzer, ABI,
Hitachi).
Data analysis
Sequence was edited to remove
ambiguous base calls and the forward and the reverse sequences were assembled
using Chromas Pro version 1.34 (Technelysium Pty
Ltd., Tewantin, Queensland, Australia). FASTA format
of Adoretus kanarensis
sequences was used for performed BLAST search at NCBI and species identification
tool at Barcode of Life Data System (BOLD). All the obtained sequences were
aligned and manually edited using BioEdit version
7.2.6. The Maximum Likelihood method and General Time Reversible model (GTR)
model of base substitution was used to calculate pairwise genetic distance in
MEGA X version 10.0.5. Additionally, to check the performance of DNA barcoding,
sequences were downloaded from NCBI and BOLD (Table 2, Supplementary data) for
some species of same genus submitted from different geographical areas. Only
sequences which formed monophyletic clades with the sequences studied here were
selected, to avoid use of sequences from wrongly identified species. These
sequences along with our data were used to generate trees using MEGA X (Nei & Kumar 2000; Kumar et al. 2018).
Results and
Discussions
Morphologically, the collected
sample was identified as Adoretus kanarensis Arrow, 1917 (Figure 3).
Systematic account
Order Coleoptera
Linnaeus, 1758
Suborder Polyphaga
Emery, 1886
Super family Scarabaeoidea
Latreille, 1802
Family Scarabaeidae
Latreille, 1802
Subfamily Rutelinae
MacLeay, 1819
Genus Adoretus Dejean, 1833
Adoretus kanarensis
Arrow, 1917
Material examined: ZSI-WRC-ENT-1/2856,
27.i.2017, 01 Female, Sinner, Nashik district (19.8543 N, 73.9922 E),
Maharashtra, coll. Pranil Jagdale.
Diagnosis
Female (Image 1): Length, 10 mm;
width, 5 mm. Bright brownish-yellow, moderately shining. The lateral margins of
head, pronotum, broad sutural line reaching anteriorly till the humeral callus
and posteriorly not reaching the margins and extremities of tibia and complete
tarsus are dark reddish-brown. Head transversely rugose, small, with the
clypeus broadly rounded. The pygidium has a bare apical area. Legs are slender,
the front tibia is armed with three strong teeth, the larger claw of the front
and middle feet is cleft, and the shorter hind claw is more than half the
length of the longer.
DNA Barcode diagnosis
In this research study, A. kanarensis was identified using available literature
and sequence of COI gene isolated from an adult female. No matches were found
among the already-existing entries in the BOLD database after analysis with the
BOLD Identification tool. The genetic difference between the two samples was
over 10%, indicating that the examined species has not yet been recorded in
BOLD. GenBank’s BLAST analysis yielded the same outcome.
The preliminary molecular
analysis was carried out using available material from NCBI GenBank (Table 2).
As expected, A. kanarensis nested
within the genus Adoretus with the
outgroup taxon Anomla ruficapilla
Burmeister, 1855 using Maximum Likelihood method with General Time Reversible
model. In the current study, A. kanarensis
formed a monophyletic clade with A. duvauceli
Blanchard, 1851 species.
Since the species A. kanarensis is of economic significance, the present mt DNA barcode data generated is expected to be helpful in
building a reliable DNA barcode library for the country intimated with a
voucher specimen and helpful in addressing the taxonomic problems as the
morphological characters are cryptic.
Table 1. Checklist of the genus Adoretus Laporte, 1840 from
Maharashtra, India.
|
|
Species |
Distribution |
References |
|
1 |
Adoretus caliginosus Burmeister, 1844 |
India (Karnataka, Haryana,
Himachal Pradesh, Maharashtra, West Bengal, Punjab, Sikkim, Tamil Nadu);
Myanmar; Pakistan. |
Arrow 1917; Mittal 1981;
Chandra 1986; Ghosh et al. 2020; Schoolmeester 2023 |
|
2 |
Adoretus excisus Ohaus, 1914 |
India (Maharashtra, Madhya
Pradesh, West Bengal). |
Arrow 1917; Schoolmeester
2023 |
|
3 |
Adoretus incurvatus Ohaus, 1914 |
India (Bihar, Haryana, Himachal
Pradesh, Maharashtra, Tamil Nadu). |
Arrow 1917; Mittal 1981;
Chandra 1986; Ghosh et al. 2020; Schoolmeester 2023 |
|
4 |
Adoretus kanarensis Arrow, 1917 |
India (Karnataka, Maharashtra
(present study), Madhya Pradesh). |
Arrow 1917 |
|
5 |
Adoretus lasiopygus Burmeister, 1855 |
India (Assam, Haryana, Bihar,
Chhattisgarh, Karnataka, Kerala, Madhya Pradesh, Maharashtra, Odisha, Punjab,
Sikkim, Tamil Nadu, Uttar Pradesh, West Bengal); Bangladesh; Bhutan; Nepal;
Sri Lanka. |
Arrow 1917; Mittal 1981;
Chandra 2000; Ghosh et al. 2020 |
|
6 |
Adoretus limbatus Blanchard, 1850 |
India (Bihar, Chhattisgarh,
Haryana, Jammu & Kashmir, Madhya Pradesh, Maharashtra, Odisha, Uttar
Pradesh, West Bengal); Bangladesh; Myanmar; Thailand. |
Arrow 1917; Mittal 1981;
Chandra 1986; Ghosh et al. 2020 |
|
7 |
Adoretus lobiceps Arrow, 1917 |
India (Gujarat, Madhya Pradesh,
Maharashtra). |
Arrow 1917; Chandra 2000. |
|
8 |
Adoretus pusillus Arrow, 1917 |
India (Maharashtra) |
Arrow 1917; Schoolmeester
2023 |
|
9 |
Adoretus stoliczkae Ohaus, 1914 |
India (Maharashtra, Madhya
Pradesh); Pakistan. |
Arrow 1917; Chandra 2000 |
|
10 |
Adoretus sorex Arrow, 1917 |
India (Maharashtra & Tamil
Nadu), Sri Lanka |
Arrow 1917; Schoolmeester
2023 |
|
11 |
Adoretus versutus Harold, 1869 |
India (Andaman & Nicobar
Islands, Bihar, Haryana, Karnataka, Kerala, Madhya Pradesh, Maharashtra,
Tamil Nadu, Uttarakhand, West Bengal);
Bangladesh; FijI; Indonesia; Malaysia; Mauritius;
Sri Lanka. |
Arrow 1917; Mittal 1981;
Chandra 1986; Chandra 2000; Ghosh et al. 2020 |
Table 2. Details for the mt DNA COI sequences utilized in the construction of the
phylogenetic tree.
|
|
GenBank Accession No./ BOLD ID |
Locality |
Species name as per NCBI |
Publication details as per NCBI |
|
1 |
JN087425 |
Japan |
Adoretus tenuimaculatus Waterhouse, 1875 |
Unpublished |
|
2 |
OQ428819 |
Iraq |
Adoretus hirsutus Ohaus, 1914 |
Unpublished |
|
3 |
KC510117 |
South Korea |
Adoretus tenuimaculatus Waterhouse, 1875 |
Unpublished |
|
4 |
KF939031 |
South Korea |
Adoretus hirsutus Ohaus, 1914 |
Unpublished |
|
5 |
MT129331 |
Republic of Korea |
Adoretus hirsutus Ohaus, 1914 |
Kang et al. 2021 |
|
6 |
KU737493 |
India |
Adoretus duvauceli Blanchard, 1851 |
Unpublished |
|
7 |
AGIRI101-17 |
India |
Adoretus duvauceli Blanchard, 1851 |
Unpublished |
|
8 |
GMPQC103-19 |
Pakistan |
Adoretus flavus Arrow, 1917 |
Unpublished |
|
9 |
KF939033 |
India |
Adoretus flavus Arrow, 1917 |
Unpublished |
|
10 |
MZ836004 |
India |
Adoretus ovalis Blanchard, 1851 |
Unpublished |
|
11 |
KF939027 |
India |
Adoretus duvauceli Blanchard, 1851 |
Unpublished |
|
12 |
KU939028 |
India |
Adoretus duvauceli Blanchard, 1851 |
Unpublished |
|
13 |
OR026029 |
India |
Adoretus kanarensis Arrow, 1917 |
Present study |
For
figures & image - - click here for full PDF
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