Bacillus thuringiensis (Bt) is a gram-positive bacterium that produces parasporal crystal proteins known as endotoxins or Cry proteins. The Cry protoxins are then cleaved by insect midgut proteinases to form active Bt toxins. The activated Cry protein then binds to specific receptors at the midgut epithelium. Cadherin-like and aminopeptidase N (APN) proteins are involved in Bt toxin binding by interacting sequentially with different toxin structures. Aminopeptidase N (APNs) from several insect species have been shown to be putative receptors for these toxins. We have characterized four different midgut APNs(APN1, APN2, APN3, APN4) cDNAs from S. exigua. Forward primers and reverse primers for confirmation of four different midgut APNs were designed based on their sequences cloned from the cDNA libraries. Quantitative RT-PCR procedures includes 42℃ for 20min (cDNA synthesis), 99℃ for 5min, and 35 cycles (94℃ for 1min, and 60℃ for 50 s) for collection. Four aminopeptidase N isoforms were confirmed with qRT-PCR. Sequence analysis was performed by BlastX search the National Center for Biotechnology Information(NCBI) nucleotide. Furthermore, double-stranded RNAs(dsRNAs) were synthesized. DsRNAs were determined for bioassay.

Background : Ginseng widely cultivated as a major medicinal herb in Korea, is economically important crop for farmer. Ginseng root disease caused by soil borne pathogens is main factors restricting the quantity and quality of ginseng. The disease can result in harvest loss of up to 20~70% and limits the replanting of ginseng under same field for long time. The traditional control method of agrochemical use is not recommend to control soil borne disease because of difficulty in use and unstable effect. The objective of this study was to evaluate the efficacy of several antagonistic microbes for developing biological control method of ginseng root rot. Methods and Results : To select biocontrol agents against ginseng soil borne disease, several bacteria were isolated from ginseng root and rhizosphere soil evaluated in vitro screening of antifungal bacterial against ginseng root pathogens. Two antagonistic bacteria, ES17 and CJ4, showed the strongest inhibition effect against ginseng root pathogen. In the pot experiment under greenhouse conditions, ginseng seedling dipped in bacterial suspension at inoculum density of 106 cfu/ml for 1 hour were planted in pot containing inoculum. Control effect was examined depend on disease severity index at 30 days after inoculation. Ginseng root treated with CJ4 and ES17 isolate reduced root rot disease development on the ginseng root with degrees of control efficacy of 85% and 70%, respectively. Conclusion : Two biocontrol agent, Burkholderia ambifaria CJ4 and Paenibacillus strain ES17, had strong antifungal efficacy against ginseng soil borne pathogens. These results obtained from in vitro test and pot experiment suggest the potential applicability of the biocontrol agent to control ginseng root rot caused by various soil borne pathogens.