This study focuses on developing diagnostic compositions, kits, and information provision methods for identifying species-specific genes in domestically residing Reticulitermes speratus and Reticulitermes kanmonensis, as well as the recently introduced Cryptotermes domesticus. The core innovation of this invention lies in the utilization of species-specific genetic markers to facilitate rapid and accurate species identification using a PCR (polymerase chain reaction)-based diagnostic technique. This approach enables swift identification of termites at quarantine stages, contributing to efficient management of imported goods and minimizing ecological and economic damages caused by termites. Through genome analysis of termites, this research has identified candidate species-specific genetic markers, developed diagnostic compositions and kits based on these markers, and proposed a rapid diagnostic method capable of determining termite species within a day, optimally within three hours. This invention provides a groundbreaking tool for termite management and research, significantly contributing to pest control and biodiversity conservation efforts.
신령버섯은 항암작용과 더불어 면역강화작용 등 약용으로서 이용되고 있습니다. 최근에는 신령버섯 재배농가에서 새로운 병징 현상이 보고되어 내생세균에 대한 분자생물학적 방법으로 조사하였다. 병징을 나타내는 수집된 기형버섯을 eubacterial 16S rDNA 영역을 이용하여 PCR한 결과 기형버섯만 증폭되었다. 이 영역을 부분적으로 염기서열을 결정한 결과 CFB bacterium과 유사성이 가장 높았고, 이 염기서열을 이용하여 프라이머를 디자인한 후 nested PCR를 부위별로 확인한 결과 기형을 일으킨 갓의 주름살 부위에서 가장 강하게 증폭되었고 포자수확도 되지 않았으며 배양불능세균 group인 CFB bacterium임을 확인하였다.
In this study, we report that the development of a multiplex PCR method using species-specific primers for the simultaneous detection of Poaceae family members, including adlay, barley, maize, rice and wheat, based on the sequence polymorphism of the DNA-directed RNA polymerase beta'' chain (rpoC2) genes Species-specific primers were constructed with common forward primer and each reverse primers which have differences on the basis of sequences. Each primer pairs could amplify PCR products of 443 bp for rice, 346 bp for barley, 278 bp for adlay, 221 bp for wheat and 96 for maize, respectively, from the five chloroplast DNAs. The series of template DNA concentrations were identified by the sensitivity of multiplex PCR. The band of products were clearly amplified from the DNA concentration range of 0.01 to 10 ng/μL. In addition, the species-specific primers were examined for the detection of seven commercial flour mixed products. The combination of the sensitivity of a multiplex PCR with the specificity of the primers for the detection of species would allow to be applied in analyses of processed foods.