Molecular Identification and Pathological Characteristics of NPV Isolated from Spodoptera litura (Fabricius) in Pakistan
Jam Nazeer Ahmad1,2,*, Rashid Mushtaq1, Samina Jam Nazeer Ahmad1,2, Sumaira Maqsood3, Ishita Ahuja4 and Atle M. Bones4
NPV infected S. litura: A, typical NPV symptom of NPV ruptured infected larvae, usually die 4–9 days after infection, disintegrate and release a virus-laden fluid; B, slow move NPV infected swollen malformed pale brown larva; C, slow move NPV infected swollen malformed dark larva from 5th instar; D and E, malformed pre-pupae; F, laboratory propagative NPV infected larva; G, healthy larva.
A, NPV infected S. litura viral occlusion body’s polyhedra under light microscopy; B, degenerating (ruptured) infected S. litura cell.
PCR detection of NPV from S. litura infected samples from Pakistan by using NPV LEF8- specific primers: Lane 1-3, non-infected S. litura larva; Lane 4-8, NPV infected S. litura larva (Pak-15 Faisalabad strain); Lane M, 1 kb DNA marker (Invitrogen).
Molecular phylogenetic analysis by maximum likelihood method of Pakistani isolate SltNPV-FSD15 strain based on nucleotide sequences with the corresponding partial late expression factor-8 (Lef-8) gene of 12 nucleopolyhydroviruses associated with S. litura (NPVs). S. litura associated NPV isolates origin and GenBank accession numbers used in the sequence analysis and phylogenetic tree construction; AY706581.1 S37 (Germany), JF276035.1 Banglore (India), AB451187.1 (AB326103.1, AB451187.1, AB583682.1 (Japan). The numbers represent bootstrap percentage values based on 1,000 replicates.