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. 2008 Apr 16;82(13):6697–6710. doi: 10.1128/JVI.00212-08

FIG. 8.

FIG. 8.

Genotyping of BSGfV viral particles in infected hybrids. (A) IC-multiplex PCR allows specific detection of BSGfV particles (DifGfF/R, 670-bp product) and a monitoring of plant DNA contaminations (Actin1F/R, 420-bp product). Lane M, 1-kb ladder (Invitrogen); lanes 1 to 6, coated plant extracts; lanes 7 to 10, plant total DNA. Lanes 1 to 3 show results for F1 AAB hybrids; lanes 4 to 6 show results for the IC control (lane 4, M. balbisiana cv. PKW; lane 5, M. acuminata cv. Grande Naine infected by BSGfV; lane 6, M. acuminata cv. Grande Naine BSGfV-free). Lanes 7 to 10 show results for the PCR control (lane 7, M. balbisiana cv. PKW; lane 8, M. acuminata cv. Grande Naine infected by BSGfV; lane 9, M. acuminata cv. Grande Naine BSGfV-free; and lane 10, water control). (B) Nested PCR using the internal primers VV1F/GfM2 (642-bp product) and increasing PCR product quantity from diluted DifGfF/R PCR product of infected hybrids. Lanes 1 to 10 show the results for AAB F1 hybrids infected with BSGfV; lane M, 1-kb ladder (Invitrogen). (C) TaaI RFLP test (described in Fig. 7A) to genotype the molecular EPRV signature of the viral BSGfV particle. Lanes 1 to 10 show the results for AAB F1 hybrids infected with BSGfV; lane M shows the results for the 50-bp ladder (NEB).

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