Molecular cloning of the double-stranded RNA of beet cryptic viruses

Three of the four dsRNA components of purified beet cryptic virus (BCV) were copied into cDNA and cloned into pUC9. Clones corresponding to RNAs 1, 3 and 4 did not hybridize to each other or to RNA 2, suggesting that there is no significant sequence homology between the four dsRNA components. RNA extracted from 15 BCV-infected beet plants was analysed by Northern blotting using the cDNA clones as probes. Nine plants were found to contain RNAs 1, 3 and 4 whereas in six plants only RNAs 3 and 4 were detectable. The results are compatible with the occurrence of two different viruses. The sensitivity and specificity of the cDNA hybridization assay was greater than that of immunosorbent electron microscopy in the detection of BCVs. Beet cryptic virus (BCV) has isometric particles about 30 nm in diameter and is widespread in different cultivars of Beta vulgaris but induces no apparent symptoms (Kassanis et al., 1977). Furthermore, it is transmitted only through seed and pollen, and cannot be transmitted by mechanical inoculation (Kassanis et al., 1977, 1978). BCV purified from seedlings of beet cv. Sharpes Klein E contains two proteins (mol. wt. 52 500 and 54 500) and four dsRNA components (mol. wt. 1.36 × 10 6, 1.15 x 10 6, 0-94 x 106 and 0.87 x 10 6) (Accotto & Boccardo, 1986). Bee