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It is barely possible to distinguish one from the other in terms of the fine structure of caudal setae (Moritsu, 1983). The 2-D patterns of proteins from both species were almost indistinguishable (Ishikawa and Yamaji, 1985d). In contrast, when endosymbionts from the two aphids were incubated in vifro, it was shown that they produce many different proteins. This suggests that the two endosymbionts have genomes apparently distinct from each other (lshikawa and Yamaji, 198561). That virtually all aphid ENDOSYMBIOSIS IN INSECTS 37 species harbor endosymbionts implies that a common ancestor of these aphids was once associated with one species of bacterium.
H. Howell, 1976). , 1974). It has also been demonstrated that the primary endosymbiont from A . pisum is capable of synthesizing R N A in insect saline. While neither actinomycin D nor a-amanitin at moderate concentrations had any effect, rifampicin at 20 pglml exerted an 80% inhibition on R N A synthesis by the isolated endosymbiont (Ishikawa, 1982b). Since the rifamycin antibiotics inhibit the prokaryotic RNA synthesis very specifically through binding to a certain unique structural component of the prokaryotic RNA polymerases (Buss and Kun, 1978), this result was taken to provide a crucial piece of evidence of the endosymbiont's prokaryotic nature.
The endosymbionts were isolated from the [35S]methionine-injected aphids, and the proteins were separated on SDS-polyacrylamide gels. In ENDOSYMBIOSIS IN INSECTS 31 contrast with the in vivo experiments described in the preceding section, radioactivity of nascent proteins coincided with the stained bands of total proteins. When insects received an injection of cycloheximide prior to injection of [''Slmethionine, labeled proteins found with the endosymbiont were almost exclusively symbionin. Injection of chloramphenicol instead of cycloheximide did not inhibit the synthesis of any protein except symbionin.
