Fig. 2.

Infectivity analysis of the full-length cDNA citrus leprosis virus C clone. (A) Necrotic symptoms (necrotic leaf areas and necrotic spots) are visualized in infiltrated N. benthamiana leaves at 8- and 13-days post infiltration, co-infiltrated with pJL89/35S-cDNA1 + pJL89/35S-cDNA2 (pJL89/35S-CiLV-C) and pLXAS/35S-cDNA1 + pLXAS/35S-cDNA2 (pLXAS/35S-CiLV-C) constructs. Mock corresponds to plants infiltrated with the pJL89 or pLXAS constructs carrying the CiLV-C RNA1 mutant, containing a premature stop codon in the CiLV-C polymerase, co-infiltrated with the respective CiLV-C RNA2 construct. (B) Cell-to-cell spread of CiLV-C infectious clone. Small regions of the leaf were infiltrated to evaluate the necrotic symptoms outside the infiltrated region. The red arrow indicates a small leaf area infiltrated and the purple arrows indicate necrotic lesions outside the infiltrated area. (C) Northern blot showing the profile of acid nucleic detection from N. benthamina leaves agroinfiltrated with the CiLV-C pJL89 or pLXAS constructs at 8 days post-infiltration, using a DIG-riboprobe complementary to the CiLV-C p29 and p24 genes. (-) corresponds to a non-infiltrated plant. Each lane corresponds to a different N. benthamiana plant. p1, p2, and p3 = plant 1, plant 2, and plant 3. rRNA stained with ethidium bromide indicates equal loading of samples. The localization of CiLV-C gRNA1 and 2 and sgRNA1, 2, 3, and 4 are indicated. The graphs represent the relative accumulation (a.u) of total CiLV-C RNAs.