In general, hemipteran insects transmit either obligate or facultative symbionts to their offspring using various transmission mechanisms. Riptortus pedestris (Hemiptera: Alydidae) also have facultative symbiosis with Burkholderia sp. which is acquired from the soil every generation especially during 2nd instar nymphal period. This environmental symbiont transmission makes R. pedestris face a great risk of missing their symbionts, however, previous studies showed that they have intimate associations with their symbionts. Therefore, we suspected that R. pedestris 2nd instar nymphs may have a preference toward Burkholderia sp. to achieve an efficient symbiont acquisition during the limited time of window. In order to identify the preference of 2nd instar nymph, we first conducted dual-choice experiments varying with the number of 2nd instar nymphs (a nymph and 100 nymphs) in plastic cages. Furthermore, Y-tube olfactometer experiments varying with the presence of soil as medium of Burkholderia sp. were performed to determine if the insects respond to bacterial volatile induced from the symbiont. Based on our results, we observed that R. pedestris 2nd instar nymphs may not show any specific behavioral preference or response to chemical volatile from Burkholderia sp. in laboratory conditions (P >0.24). Therefore, we will further conduct a dual-choice experiment in a semi-field condition to investigate if such patterns can be detected in a larger arena simulating a more natural environment.

Vertical transmission of symbiont is known as the most effective way to deliver their offspring. However, it has beenreported that the bean bug, Riptortus pedestris, acquires its gut symbiont, Burkholderia sp., from its environment (e.g.soil) during the 2nd instar nymphal stage. Nevertheless, it is not unknown how they locate their symbiont. For this reason,dual-choice experiments were conducted in both solitary (a nymph) and group conditions (100 nymphs) to investigatewhether or not R. pedestris has preference on Burkholderia sp. treated or untreated solution in laboratory conditions. Inthese experiments, there was no significant difference in the preference between the two treatments. We are testing inthe Y-tube olfactometer system to evaluate the response of R. pedestris on possible chemical cues from Burkholderia sp..

The bean bug, Riptortus pedestris, is a pest of leguminous crops. Recent studies have revealed an interesting relationship between R. pedestris and a gut symbiont Burkholderia sp.. R. pedestris mainly acquires Burkholderia sp. during 2nd instar period, which leads to enhancement of growth and reproductive rate of the host. Nevertheless, how R. pedestris acquires its symbiont is not yet clearly described. Therefore, we investigated whether 2nd instar nymphs have preference on Burkholderia sp. compared to untreated control in laboratory settings. In the choice test, no significant difference was found in the preference by R. pedestris between the two treatments. To further investigate single individual’s behaviors (its first choice, approaching frequency to each treatment, and residence time on the treatment). We video-recorded single individuals in the choice setting for 12 hours. Again, there was no significant difference in any of the behavioral traits recorded. Further choice tests will be conducted in the Y-tube olfactometer systems and semi-field conditions.

It is not uncommon in the nature for insects to have symbiotic relationships with microorganisms. The bean bug, Riptortus pedestris, which harbors symbiont Burkholderia sp., is a serious agricultural pest in South Korea and Japan. It is reported that the symbiont has positive effects on the fitness of R. pedestris. However, virtually no information is available for how this insect finds the symbiont in the nature and how the behavior of R. pedestris is affected by the symbiont. To investigate the symbiont acquisition mechanism, the frequencies of R. pedestris approaching to symbiont or untreated control treatment were compared in a plastic cage (465×290×260mm) (n=20). In the experiment, there was no detectable difference between the two treatments. Further study using video tracking systems will help us to better understand detailed behaviors for symbiont acquisition.

From the course of screening of useful enzyme producing microorganism from insect guts, we isolated 9 lipase producing strains and their lipase producing activities were tested. 16S rDNA sequence analysis showed that they were Gram negative bacteria grouped on Serratia sp., Pseudomonas sp., and Burkholderia sp.. Among them, an excellent lipase producing strain, Burkholderia sp. HY-10 identified by 16S rDNA analysis and biochemical methods, was further studied its lipase producing characteristics. It was isolated from a longcorm beetle, Prionus insularis and showed cell density dependent lipase producing activity in the culture media that contained olive oil as a carbon source. Maximum lipase production was achieved in the M9 media containing 0.5% yeast extract and 0.5% olive oil when cultured at 30℃ for 36-42 hrs.