SPOROGENOUS MIDGUT BACTERIA OF THE LEAFROLLER, PANDEMIS-PYRUSANA (LEPIDOPTERA, TORTRICIDAE)

The composition of the microbial nora within the midgut of Pandemis py rusana Kearfott was analyzed by using the polymerase chain reaction (P CR) and culturing techniques. Universal 16S ribosomal RNA gene-specifi c eubacterial primers were used in PCR, and the resulting products clo ned and sequenced. High homology (86-95%) was obtained between the clo ned products and Clostridium ramosum and Clostridium spiroforme when c ompared with computer database sequence information. Initial culturing steps aimed at endospore-forming bacteria yielded reproducible growth from a bacterium that matches most closely with Bacillus licheniformi s when analyzed by partial 16S ribosomal DNA sequencing and biochemica l/cultural profiling. This isolate was transformed with an Escherichia . coli-Bacillus subtilis shuttle vector and reintroduced into the inse ct, where it remained stable under nonselective conditions for at leas t 9 d after initial ingestion. This confirmed the ability of the genet ically altered bacterium to survive within the leafroller midgut in th e midst of competing microorganisms and perhaps to serve as a suitable vehicle for biological control.