FIG. 5.
Slot blot hybridization analysis of accumulation of SMV RNA, soybean PR-1 protein gene transcript up regulation (PR1), and soybean 18S rRNA in soybean trifoliate leaves. Primary leaves of PI 96983 (Rsv1) were mechanically inoculated with buffer (mock) or infectious sap containing progenies of pSMV-G7 (G7), pSMV-G7d (G7d), or their derivative point mutants generated by site-directed mutagenesis. Following inoculation, plants were maintained in a growth chamber (20°C) until a leaflet from trifoliolates 3 and 4 of infected plants was collected about 6 weeks postinoculation. Samples from corresponding trifoliolate leaflets of four independent replicate plants were combined, total RNA was isolated and denatured, and 10 μg was slot blotted onto a membrane and hybridized with 32P-labeled cDNA probes.