Microscopic and Molecular Identification of Bombyx mori Nuclear Polyhedrosis virus (BmNPV) using lef-8 and polhe Primers

Author: Mohd Shahid Raja, Shilpa Patial, Shabir Ahmad Bhat, Romisa Amin, Seerat Showkat and Firdose Ahmad Malik

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Abstract

Silkworms infected with grasserie were collected at the 4th and 5th instars of their rearing. Sample was collected by a needle puncture limbs of thorax, white fluid containing virus was collected. Collected sample was subjected to centrifuge at 2500 rpm for 2 minutes for isolation of virus from debris and process was repeated till clear white color pallet was obtained. The pallet was examined under compound microscope and FESEM for the presence of Bombyx mori Nuclear Polyhedrosis Virus (BmNPV). After positive microscopic results DNA was extracted and examined for the presence of two specific genes viz., late expression factor or lef-8 (conserved in NPV) and polyhedrin or polhe (conserved in BmNPV). PCR products were resolved by electrophoresis on 0.8 % and clear bands was seen at 650 – 700bp for lef-8 gene and at 160 – 170bp for polhe gene. Observed bands clearly confirm the presence of BmNPV virus responsible for grasserie disease.

Keywords

Silkworm, Bm NPV, Polymerase chain reaction, lef-8, polhe, Virus etc.

Conclusion

Based on the result from present study, we can conclude that if we have to go for the fast and early detection grasserie disease. We need fast and effective fast method without any confusion to cope the virus and start precautions. Early detection can help to take necessary measures and help to save the silkworm cocoon crop from futher damage

References

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How to cite this article

Mohd Shahid Raja, Shilpa Patial, Shabir Ahmad Bhat, Romisa Amin, Seerat Showkat and Firdose Ahmad Malik (2024). Microscopic and Molecular Identification of Bombyx mori Nuclear Polyhedrosis virus (BmNPV) using lef-8 and polhe Primers. Biological Forum – An International Journal, 16(12): 106-109.