# Detection of Ochyromera ligustri (Coleoptera: Curculionidae: Curculioninae) in Ligustrum spp. (Oleaceae) Using Newly Developed PCR Primers

## TL;DR

Researchers developed PCR primers to detect a weevil in privet plants, which could help control the invasive shrubs.

## Contribution

New PCR primers were developed for rapid and accurate detection of Ochyromera ligustri in Ligustrum spp.

## Key findings

- The primers specifically detect O. ligustri DNA at low concentrations (0.01 ng).
- The primers do not amplify DNA from five other weevil species.
- O. ligustri DNA was detected in three Ligustrum species common in the southeastern US.

## Abstract

Privets (Ligustrum spp.) are popular hedge shrubs that escape from gardens and cause negative impacts by invading forests in the United States. The narrow host specificity of the seed-feeding weevil, Ochyromera ligustri, suggests that this weevil could be considered a fortuitous biological control agent of privets. To find out the host range of this weevil, time-consuming experiments involving rearing and dissections are often conducted. Herein, we developed PCR primers to rapidly and accurately detect the presence of the weevil in privet fruits. We confirmed that the novel PCR primers specifically work for O. ligustri even with a small quantity of DNA, which was 0.01 ng. Using the newly developed PCR-based detection saves time and labor in research related to the detection and impact of this weevil.

Ligustrum spp. (Oleaceae) have become invasive species in the US and negatively affect native plant diversity and richness in forests. Ochyromera ligustri (Coleoptera: Curculionidae) is considered a potential biological control agent in the US because adults feed on the foliage and larvae are seed-feeders of Ligustrum spp. To discover the relationships between O. ligustri and Ligustrum spp., fruit dissections or rearing and field observations are required. In the current research project, novel PCR primers were developed to rapidly detect the DNA of O. ligustri in molecular analyses without rearing and observation. The developed PCR primers worked even with 0.01 ng of DNA and did not amplify the DNA of the other five curculionid species tested. When the novel primers were tested with three Ligustrum spp. species common in the southeastern US, the DNA of O. ligustri was detected from all three species. We expect that the novel primers will be utilized to find out the presence and impact of O. ligustri on Ligustrum spp rapidly and accurately.