Identification of two cryptic species in the Anopheles (Cellia) annularis complex using ribosomal DNA PCR-RFLP (original) (raw)

Abstract

Anopheles (Cellia) annularis Van der Wulp is a complex of two sibling species provisionally designated as species A and B and can only be differentiated on the basis of the paracentric inversion in the ovarian polytene chromosomes. To analyze the distribution of these two species and to develop a molecular method for the identification of these two cryptic species, we sequenced the ribosomal DNA internal transcribed spacer 2 (ITS2) and domain 3 (D3) of A. annularis specimens collected from Sonapur (Assam), Jabalpur (Madhya Pradesh), Ranchi (Jharkhand), and Ghaziabad (Uttar Pradesh). We did not find any sequence variation among the specimens collected from Assam, Madhya Pradesh, and Jharkhand states, whereas two types of sequences were obtained from the specimens collected from the state of Uttar Pradesh, which correspond to species A and B of the A. annularis complex. Species A was more prevalent among the all four regions studied. The ITS2 sequence of species A showed unique restriction sites for _Mva_I and _Eco_24I, while species B displayed _Hin_fI and _Nru_I sites. Similarly, the D3 sequence of species A showed unique restriction site for _Alw_26I, while species B showed a unique _Kpn_I site. In this study, we report for the first time the development of ribosomal DNA polymerase chain reaction-restriction fragment length polymorphism methods for identifying these two cryptic species of the Annularis complex.

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Acknowledgments

The authors would like to thank Prof. A. P. Dash [Director, National Institute of Malaria Research (ICMR), Delhi] for his support and Ms. Sumiti Vinayak for the discussion and for proofreading this manuscript. MTA acknowledges the Council of Scientific and Industrial Research for the Senior Research Fellowship. Financial support for the work granted by the Indian Council of Medical Research and the Department of Biotechnology, Government of India, is acknowledged. We are also grateful for the bioinformatics facilities of the Biotechnology Information System (BTIS). The authors also declare that the experiments performed in this manuscript comply with the current laws of India.

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Authors and Affiliations

1. Department of Biotechnology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029, India
   Mohammad Tauqeer Alam & Yagya D. Sharma
2. National Institute of Malaria Research (Field Station), Car Nicobar, Andaman and Nicobar Islands, India
   Manoj K. Das
3. National Institute of Malaria Research (Field Station), Sonapur, Kamrup, Assam, 782402, India
   Vas Dev
4. Regional Medical Research Centre for Tribes, Jabalpur, Madhya Pradesh, India
   Musharraf A. Ansari

Authors

1. Mohammad Tauqeer Alam
2. Manoj K. Das
3. Vas Dev
4. Musharraf A. Ansari
5. Yagya D. Sharma

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Correspondence toYagya D. Sharma.

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Alam, M.T., Das, M.K., Dev, V. et al. Identification of two cryptic species in the Anopheles (Cellia) annularis complex using ribosomal DNA PCR-RFLP.Parasitol Res 100, 943–948 (2007). https://doi.org/10.1007/s00436-006-0375-9

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• Received: 22 May 2006
• Accepted: 02 November 2006
• Published: 06 December 2006
• Issue date: April 2007
• DOI: https://doi.org/10.1007/s00436-006-0375-9

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