Host range validation,molecular identification and release and establishment of a Chinese biotype of the Asian leaf beetle Lilioceris cheni (Coleoptera: Chrysomelidae: Criocerinae) for control of Dioscorea bulbifera L. in the southern United States

Dioscorea bulbifera, an Asian vine, is invasive in the southeastern USA. It rarely flowers but propagates from potato-like bulbils formed in leaf axils, which persist into the subsequent growing season. Lilioceris cheni Gressitt and Kimoto, a foliage-feeding beetle (Coleoptera: Chrysomelidae: Criocerinae) from Nepal, had been tested, proven to be a specialist and approved for release as a biological control agent. Regulatory delays, however, resulted in the demise of quarantine-held colonies, and acquisition of new Nepalese stock proved untenable. Searches then undertaken in southern China resulted in the collection of over 300 similar beetles. Two Chinese Lilioceris species were identified: one confirmed to be L. cheni and the other identified as Lilioceris egena (Weise). Mitochondrial analysis revealed an exact DNA match between some Chinese and one of the two Nepalese c oxidase subunit I haplotypes and all Chinese L. cheni haplotypes clustered as a single species but the comingling of the two species aroused concerns over possible hybridisation. These concerns were allayed by nuclear D2 analysis showing the absence of dual parental sequences. Nonetheless, diligence was exercised to ensure that the Chinese strains were safe to release. Abridged host testing using critical test species verified specificity. Caged releases during autumn 2011 documented the ability of adult beetles to overwinter in south Florida despite a prolonged lack of foliage. Open releases the following year produced vigorous populations that caused extensive defoliation. Preliminary observations indicate that L. cheni now contributes to the control of D. bulbifera and the bulbil-feeding L. egena should complement these effects if its host range proves appropriate.