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4 Issues


ISSN: 2320-964X (Online) 

ISSN: 2320-7817  (Print)



Dr. Santosh Pawar 




Int. Journal of Life Sciences, 2017; 5(4): 517-526             |        Available online, November 21, 2017

DNA Barcoding of Six Species of Family Rhopalidae (Insecta: Hemiptera: Heteroptera) from India

Kaur Harbhajan* and Sekhon Navpreet Kaur



Department of Zoology and Environmental Sciences, Punjabi University, Patiala.



Received: 01.08.2017   |   Accepted: 09.11.2017    |      Published : 21.11.2017

Rhopalidae is a small family comprising 18 genera and 209 recognised species all over the world. The Oriental Realm embraces approximately 30 species of about 10 genera. In this study, first ever comprehensive DNA barcode analysis of 17 specimens of six species belonging to five genera of family Rhopalidae from northern parts of India has been presented and compared with worldwide available data. Sequences for all the species but Leptocoris augur (Fabricius, 1974) are first submissions from India while six sequences of Liorhyssus rubicundus (Signoret, 1859) are the pioneer submissions for this particular species to the GenBank and BOLD.


Both the species of genus Liorhyssus  viz L. hyalinus (Fabricius, 1794)  and L. rubicundus (Signoret, 1859) show striking phenotypic plasticity but can be easily delineated by distinct barcodes showing minimum interspecific K2P distance of 7.6 % and much lower maximum intraspecific K2P distances of  2.1% and 0.5% respectively. Our two sequences of Stictopleurus pictus (Fieber, 1861) show K2P distance of 0.0% but 2.3% with sequences from Germany. The Rhopalus (Aeschyntelus) sp. nr. maculatus shows average K2P distance of 3.2%  with Rhopalus maculatus (Fieber,1837) and appears to be a cryptic species. Corizus sp. shows distinct barcode which delineates it from its nearest neighbour Corizus hyocyami (Linnaeus, 1758).


Keywords: COI gene, Coreoidea, K2P distance, Evolutionary affinities


Editor: Dr.R.G. Jadhao


Cite this article as:

Kaur Harbhajan and  Sekhon Navpreet Kaur (2017) DNA Barcoding of Six Species of Family Rhopalidae (Insecta: Hemiptera: Heteroptera) from India; International J. of Life Sciences, 5 (4): 517-526. 



We thank the Department of Zoology and Environmental Sciences, Punjabi University, Patiala and Sophisticated Instrumentation Centre, Punjabi University, Patiala for providing all the lab facilities. The funding for the research was provided by University Grants Commission (UGC) under the Basic Scientific Research Scheme -2013 (BSR-2013). The help and assistance provided by Professor Emeritus C.A Viraktamath and his team Dr. Yeshwanth HM and Dr. Nirmala P of Department of Entomology, University of Agricultural Sciences, Bangalore for identification of bugs is highly appreciated


Conflicts of interest: The authors stated that no conflicts of interest.



Copyright: © 2017 | Author(s), This is an open access article under the terms of the Creative Commons Attribution-Non-Commercial - No Derivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.



Ahmad I (1978) Final technical report of systematics and biology of Pentatomorphous subfamilies Coreoidea  and Pentatomoidea of Pakistan. USDA, 1-625

Ahmad I, Shadab MU, Abrar I and Khan AA (1979) Generic and supergeneric keys with reference to a checklist of Rhopalid fauna of Pakistan (Heteroptera : Coreoidea) with notes on their distribution and food plants. Entomological  Society of  Kararachi Pakistan, 3: 1-14.

BioLib Biological Library.1999- till date. Available from:

British Bugs- An online identification guide to UK Hemiptera. 2013-  Till date.  Available from: (21 June 2017)

Chopra NP (1967) The higher classification of family Rhopalidae (Hemiptera). Transactions of Royal Entomological Society of London, 119: 363-399

Coreoidea Species File Online. Version 5.0/5.0. [2017]. (21 June 2017)

Damgaard J (2008) MtDNA diversity and species phylogeny of western Palaearctic members of the Gerris lacustris group (Hemiptera:Heteroptera:Gerridae) with implications for DNA barcoding of water striders. Insect Systematics & Evolution,  39:107–120.

Distant WL (1904) The Fauna of British India including Ceylon and Burma. Rhynchota (Heteroptera) I. Edited by W.T Blanford. Taylor and Francis, London, 416 -420.

Dusoulier F, Streito  JC, Matocq  A,  Minssieux, E. The Heteroptera of the National Park of Mercantour (Alpes-Maritimes, France): first inventory and elements about biogeography, ecology and molecular data, Unpublished Manuscript, CBGP, INRA, campus, Montferrie, France.

 Folmer O, Black M, Hoeh W, Lutz R and Vrijenhoek R (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology, 3: 294–299.

Gwiazdowski RA, Foottit RG, Maw HEL and  Hebert PDN (2015) The Hemiptera (Insecta) of Canada: Constructing a Reference Library of DNA Barcodes. PLoS ONE, 10(4): e0125635.http://dx.doi:10.1371/journal.pone.0125635

Habeeb SKM and Sanjayan KP (2011) Sequencing and phylogenetic analysis of the mitochondrial Cytochrome C Oxidase subunit 1 of Oxycarenus laetus (Hemiptera:  Lygaeidae). International Journal of Plant, Animal and Environmental Sciences, 1: 85–91.

Hebert PDN, Cywinska A, Ball SL and deWaard JR (2003)  Biological identifications through DNA barcodes. Proceedings of Royal Society of London B, 270:313–322.

(DOI 10.1098/rspb.2002.2218.)

Hebert PDN, Stoeckle MY, Zemlak TS and Francis CM (2004) Identification of birds through DNA barcodes. Plos Biology, 2: 1657–1663.

Hua J, Li M, Dong P, Cui Y, Xie Q and  Bu W (2008) Comparative and phylogenomic studies on the mitochondrial genomes of Pentatomomorpha (Insecta: Hemiptera: Heteroptera). BMC Genomics, 9: 610–624.

Jung S, Duwal RK and Lee, S (2011) COI barcoding of true bugs (Insecta, Heteroptera). Molecular Ecology Resources, 11: 266–270.

Kambhampati S and  Rai KS (1991) Mitochondria1 DNA variation within and among populations of the mosquito Aedes albopictus. Genome,  34: 288-292.

Kerr KCR, Stoeckle MY, Dove CJ, Weigt LA, Francis CM and Hebert PDN (2007)  Comprehensive DNA barcode coverage of North American birds. Molecular Ecology Notes, 7: 535–543.

Kaur H and Sharma K (2016) COI based DNA barcoding of some species of Pentatomidae from North India ( Hemiptera: Heteroptera)  [online]. Mitochondrial DNA Part A, DOI:10.1080/24701394.2016.1180513

Kumar S, Stecher G and Tamura K (2015) MEGA 7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33: 1870–1874.

Li M, Liu Q, Xi L, Liu Y, Zhu G, Zhao Yand Bu W (2014) Testing the potential of proposed DNA Barcoding markers in Nezara virudula and Nezara antennata when Geographic variation and closely Related Species Were Considered. Journal of Insect Science, 14: 1–11. http://dx.doi:10.1093/jis/14.1.1 PMID: 25373148

Lis JA, Olchowik J and Bulin´ska-Balas M (2012) Preliminary studies on the usefulness of DNA mini-barcodes for determining phylogenetic relationships within shieldbugs (Hemiptera: Heteroptera: Pentatomidae). Heteroptera Poloniae -Acta Faunistica, 4:13–25.

Memon N, Meier R, Manan A and Su K (2006) On the use of DNA sequences for determining the species limits of a polymorphic new species in the stink bug genus Halys (Heteroptera: Pentatomidae) from Pakistan. Systematic Entomology, 31:703–710.

Park DS, Footit R, Maw E and  Hebert PDN (2011) Barcoding bugs: DNA-based identification of the true bugs (Insecta: Hemiptera:Heteroptera). PLoS ONE, 6: e18749.

Park DS and Oh HW. DNA barcode for agricultural field animals in Korea. Unpublished manuscript. Microbiological Research Center, Korea Research Institute of Bioscience and Biotechnology, South Korea

Ratnasingham S and Hebert PDN (2013) A DNA-Based Registry for All Animal Species: The Barcode Index Number (BIN) System. PLoS ONE, 8(8): e66213.

Ratnasingham S and Hebert PDN (2007) BOLD: The Barcode of Life Data System. Available from: (15 May 2017) 

Raupach MJ, Hendrich L, Küchler SM, Deister F, Morinière J and Gossner MM (2014) Building-Up of a DNA Barcode Library for True Bugs (Insecta: Hemiptera: Heteroptera) of Germany Reveals Taxonomic Uncertainties and Surprises. PLoS ONE, 9: e106940.

http://dx.doi:10.1371/journal.pone.0106940PMID: 25203616

Rebijith KB, Asokan R, Krishna NK, Srikumar KK, Ramamurthy VV and Bhat P (2012) DNA barcoding and development of species specific markers for the identification of tea mosquito bugs (Miridae: Heteroptera) in India. Environmental Entomology, 41:1239–1245.

Remigio EA and Hebert PDN (2003) Testing the utility of partial COI sequences for phylogenetic estimates of Gastropod relationships. Molecular Phylogenetics and Evolution, 29: 641–647.

Rizvi SA (2000) A revision of Rhopalidae Amylot and Serville (Hemiptera :Coreoidea) of Indo- Pakistan subcontinent and their cladistic analysis. PhD Thesis.Department of Zoology, University of Karachi, Pakistan.

Schuh RT and Slater JA (1995) True Bugs of the World (Hemiptera: Heteroptera): Classification and Natural History. Cornell University Press, New York. 281–282.

Steill J and Meyer J (2003) The Rhopalidae of Florida “Scentless Plant Bugs”. Insect Classification Project. 4(30):1–23.

Tembe S, Shouche Y and Ghate HV (2014) DNA barcoding of Pentatomomorpha bugs (Hemiptera: Heteroptera) from Western Ghats of India. Meta Gene, 2:737–745. http://dx.doi:10.1016/j.mgene.2014.09.006PMID: 25606457

Ward RD, Zemlak TS, Innes BH, Last PR and Hebert PDN (2005) DNA barcoding Australia’s fish species. Philosophical transactions of the Royal Society of London B, 360:1847–1857.

Zhou C, Kandemir I, Walsh DB, Zalom FG and Lavine LC (2012) Identification of Lygus hesperus by DNA Barcoding Reveals Insignificant Levels of Genetic Structure among Distant and Habitat Diverse Populations. PLoS ONE,7: e34528.  http://dx.doi: 10.1371/journal.pone.0034528 PMID: 22479640. 







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