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Rice plants infected with the rice dwarf virus (RDV) are stunted with white chlorotic specks on leaves4). The virus consists of icosahedral double-shelled particles approximately 70 nm in diameter, containing 12 segments of double-stranded (ds) RNA and several proteins. A severe virus strain (RDV-S) has been reported4) which causes a more pronounced stunting of plants than does the ordinary strain (RDV-O). Major differences in the virus components between the strains are as follows: 1) the molecular size of the constituent protein of the outer capsid, encoded by genome segment S88), is larger (47K) in the RDV-S than that (46K) in RDV-O, when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and 2) the fourth largest genome segment differs in their migration position in SDS-PAGE. As a first step towards the understanding of the mechanisms controlling the differences in symptom severity of the RDV strains, we analyzed the nucleotide sequence of genome segment S8 of RDV-S and compared the deduced gene product with that of RDV-O. RDV-O and RDV-S were purified according to the method reported earlier9). Complementary DNAs (cDNA) to RDV-S genome segments were cloned into pBR322 as described by Koganezawa et al5) Of the 8 clones that hybridized specifically with genome segment S8 dsRNA labelled with [ƒ¿-32P]ATP, two corresponding to the full length of the original dsRNA were selected for sequencing. The nucleotide sequence was determined as described by Noda et al.7) with an automated DNA sequencer (370A, Applied Biosystems, U. S. A.), using fluorescent primers (Applied Biosystems) and Taq DNA polymerase (Promega) for the dideoxy chain termination reaction10). All the parts of one cDNA were sequenced at least twice in each direction and the sequence was confirmed for the other one. As shown in Fig. 1, 26 substitutions in the nucleotide sequence of the genome segment S8 of RDV-S were detected compared with RDV-O8). The result was the same for the two clones analyzed. All the substitutions were located in the open reading frame (residue 24-1283), as in the case of the WTV variants2). The substitutions in the first, the second and the third positions of the triplet numbered 2, 1 and 23, respectively. There was one amino acid substitution from isoleucine to threonine at the residue 235 in the amino acid sequence of RDV-S, upon comparison with RDV-O. No difference was observed between the two strains in the secondary structures of 46K protein by the substitution analyzed using DNASIS (Hitachi, Japan) based on the theory of Chou and Fasman1). To confirm the position of the amino acid substitution, the 46K protein of RDV-O and the 47K

Nucleotide Sequence of the Outer Capsid Protein Gene of a Severe Strain of Rice Dwarf Virus.