Reference Report for IND80101671
| Title: | Infection of protoplasts from soybean cell culture with southern bean mosaic and cowpea mosaic viruses. |
| Authors: | Jarvis, N.P., Murakishi, H.H. |
| Source: | J. Gen. Virol. 1980, 48(2):365-376 |
| Abstract: | Soybean protoplasts, isolated from liquid suspension culture, were sucessfully infected with cowpea moasic virus (CPMV) and with southern bean mosaic virus (SBMV). Poly-L-ornithine (PLO) was required for infection with either virus. As detected by fluorescent antibody, 70-90% of the protoplasts were infected by CPMV when the inoculation medium contained 0.4 M sorbitol, 0.5 .mu.g virus/ml, 1.5 .mu.g PLO/ml, 10 mM-potassium phosphate buffer, pH 6.3, and 0.5 mM CaCl2, and inoculation was performed at 23.degree. C. This maximum level of infection was decreased 6-fold when CaCl2 was omitted. Inoculation at temperatures below 10.degree. C also decreased infection substantially. With SBMV a maximum of 30-35% of the protoplasts were infected with 0.4 M sorbitol, 2-2.5 .mu.g virus/ml, 2 .mu.g PLO/ml, 10 mM tris-HCl buffer, pH 8.0, 1 mM MgSO4 and 0.5 M CaCl2 were present in the inoculum. Less than 5% of the protoplasts were infected when Mg and Ca salts were omitted. The major advantage of the use of soybean cell suspension culture for plant virus studies is that it can provide a reliable and continuous source of cells for protoplast isolation. These soybean protoplasts allow synchronous infection and replication under sterile conditions without antibiotics and a high degree of reproducibility. |
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