Use of Spring beauty latent virus to identify compatible interactions between bromovirus components required for virus infection

Abstract

Spring beauty latent virus (SBLV) is a member of the genus Bromovirus, and is closely related to Brome mosaic virus (BMV) and Cowpea chlorotic mottle virus (CCMV). Compatible interactions between viral components are required for successful infection of plants by BMV and CCMV. To further our understanding of interactions between bromovirus components, we used SBLV to produce reassortants among the three bromoviruses. We found that SBLV RNA 2 functioned with heterologous bromovirus RNA 1 in infections of whole plants and protoplasts of Nicotiana benthamiana, although SBLV RNA 1 did not function with heterologous bromovirus RNA 2. A DNA-based transient assay for 1a and 2a proteins, which are encoded by RNAs 1 and 2, respectively further suggested that SBLV 2a protein may function in combination with heterologous bromovirus 1a protein. Moreover, analysis of the ability of reassortants to spread locally revealed that an RNA 2-mediated interaction between viral components may be required for efficient cell-tocell movement of bromoviruses.

Cowpea chlorotic mottle bromovirus replication proteins support template-selective RNA replication in Saccharomyces cerevisiae

“…In such plant cells, BMV 1a + 2a pol and CCMV 1a + 2a pol support replication and subgenomic mRNA transcription of either BMV or CCMV RNA3 to similar levels, although heterologous 1a/2a pol pairings do not support full RNA replication [8]. Many studies of CCMV and BMV in plants have productively utilized component mixing or hybrid viral proteins and RNAs to study essential mechanisms of RNA replication and viral spread [19–22].…”

Identification and Characterization of theSSB1Locus Involved in Symptom Development bySpring beauty latent virusInfection inArabidopsis thaliana

“…The press blot assay for detecting the distribution of SBLV CP in A. thaliana was performed as previously described (Fujisaki et al 2003b;Takahashi et al 2001). To analyze the accumulation of SBLV CP, infected leaves were ground in Laemmli's sample buffer (Laemmli 1970) and were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (samples from 0.4 mg of fresh weight tissues were loaded into each lane), followed by immunoblot analysis as described previously (Damayanti et al 1999).…”