Abstract
Cyst nematodes of the species Globodera rostochiensis and G. pallida are devastating parasites of the potato crop. Early detection of cyst nematodes in the field is critical for adopting an appropriate management strategy. A specific and sensitive loop-mediated isothermal amplification (LAMP) assay using four oligonucleotide primers has been developed to amplify the internal transcribed spacer region (ITS) of ribosomal DNA of potato cyst nematode G. rostochiensis. The PCN-LAMP reaction could be completed within 75 min at 68 °C followed by termination at 85 °C for 7 min. The primers exhibited specificity for G. rostochiensis and did not detect any other tested genera of plant parasitic or entomopathogenic nematodes. LAMP reaction was highly sensitive, suitable for crude genomic DNA and could successfully detect G. rostochiensis DNA up to femtogram quantity. This assay is rapid, cost effective and requires minimal instrumentation. It will facilitate the detection of G. rostochiensis at field and point-of-care labs and help in the interception of infested plant material/soil samples at quarantine stations independent of a professional nematologist.
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Acknowledgements
Senior Research Fellowship to Amit Ahuja from the PG School, ICAR-Indian Agricultural Research Institute (IARI), New Delhi is acknowledged. We thank Dr. M. R. Khan, Pricipal Scientist and Professor, Division of Nematology, ICAR-IARI for discussions and critical suggestions regarding the research. We thank the Director and the Joint Director (Research), ICAR- IARI for internal funding and extending all the support and facilities to complete the study.
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VS and AA conceptualized the study; VJ and SK collected the biological material for the study; AA, GS, AK and CGB performed the experiments; AA and VSS wrote the first draft and revised the manuscript; UR, AA and VSS edited the final draft of the manuscript.
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Ahuja, A., Joshi, V., Singh, G. et al. Rapid and sensitive detection of potato cyst nematode Globodera rostochiensis by loop-mediated isothermal amplification assay. 3 Biotech 11, 294 (2021). https://doi.org/10.1007/s13205-021-02830-8
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DOI: https://doi.org/10.1007/s13205-021-02830-8