중금속은 다양한 경로를 통해 환경 중 배출되어 서식 생물에 노출되며 체내 다양한 생리학적 불균형을 유도한다. 본 연구에서는 수서생물지표종으로 이용되는 깔따구 (Chironomus riparius)를 이용하여 야외 중금속 (Al, Aluminum; Cr, Chromium; Cu, copper; Mn, Manganese; Zn, Zinc) 농도 노출에 따른 다양한 분자발현 반응과 상관성을 분석하였다. 생물 체내 분자 반응을 관찰하기 위해 heat shock protein 40, 70, 90 (HSP40, 70, 90), cytochrome 450 (CYP450), Glutathione S-transferase (GST) and Serine-type endopeptidase (SP)를 이용하였다. 그 결과, 스트레스 분자마 커로 이용되는 HSPs 유전자들은 중금속 노출된 개체들에서 대조군보다 높은 경향을 보였으며 Cu 노출 시 가장 높은 발현을 나타냈다. 해독에 관여하는 CYP450과 GST 유전자 발현 결과, Cr과 Cu에서는 다른 노출군에 비해 높은 발현 경향을 나타냈다. SP 유전자 발현 결과 Al을 제외한 모든 노출군이 대조군과 유사한 발현 패턴을 보였다. 이와 같은 연구 결과는 실내에서 환경 중 존재하는 실제 농도를 반영한 독성실험을 통해 노출물질과 농도에 따라 특이적으로 발현하는 분자마커 패턴을 보고하였다. 또한, 수생태계로 유입되는 중금속이 하천에 서식하는 생물에 주는 유해 영향에 대한 정보와 분자 지표 유전자들의 현장 적용 가능성을 보여준다.

Chironomous is aquatic insect belonging to order Diptera, family Chironomidae. Their larval stage can be found mainly in aquatic benthic environment, hence good model organism to study environmental toxicology assessments and consider as useful bio indicators of contamination of the aquatic environment. In this study, Chironomus Heat Shock Proteins, Cytochrome 450, Glutathione S-transferase, Serine-type endopeptidase gene expressions were compared between polluted field areas (Chironomus plumosus) and under laboratory conditions (Chironomus riparious) to investigate molecular indicators for environmental contaminant stress assessment. Heavy metal (Al, Fe, Mn, Cu, Cr, Zn, Se, Pb, As, Cd) concentrations in sediments collected from three study areas exceeded the reference values. Moreover, HSPs, CYP450 and GST gene expression except SP for C. plumosus showed higher expression than C. riparious gene expression. Similar gene expression pattern was observed in C. riparious that exposed environment waters up to 96 h when compared to C. plumosus exposed to waters that grown in lab conditions. In summary, this comparative gene expression analysis in Chironomous between field and laboratory condition gave useful information to select candidate molecular indicators in heavy metal contaminations in the environment.

Local and seasonal populations of the oriental fruit moth, Grapholita molesta , were monitored with sex pheromone trapping and RAPD (random amplified polymorphic DNA) molecular marker to analyze their movement in apple orchards. To detect their movements among farms, pheromone traps were placed at regions between apple farms (‘outside-farms’) as well as within-farms (‘inside-farms’). Four seasonal adult peaks were evident in apple-cultivating fields from April to October in both trappings of inside- or outside-farms. After overwintering generation, populations of inside-farms were significantly reduced with frequent insecticide applications, compared to populations of outside-farms. Within apple farms, G. molesta tended to be unevenly distributed because of significant sublocal preference. Active movements of local and seasonal populations of G. molesta were supported by gene flow analysis using RAPD marker. Monitoring data using sex pheromone and seasonal reduction in initial genetic differentiation detected in the overwintering populations suggest that there must be significant movement of G. molesta among different orchards in apple-cultivating areas.

An unidentified moth was captured in sex pheromone traps of the oriental fruit moth, Grapholita molesta, especially at spring season in apple orchards and their vicinity. Though the captured males were similar in appearance to G. molesta males, they were easily distinguished by a marked difference in body size. Their occurrence pattern was also similar to that of overwintering G. molesta population from April to May, at which more males were captured in the pheromone traps installed in the vicinity of apple orchards than within apple orchards. After May, they were no longer captured in the pheromone traps. To investigate any larval damage due to this unidentified moth, molecular markers needed to be developed. Four PCR-RFLP markers originated from cytochrome b region of mitochondrial DNA could distinguish this unidentified moth from G. molesta.

사과 해충으로서 복숭아순나방(Grapholita molesta)은 과실에 피해를 주는 일차해충이다. 이에 따라 과실 내부에서 피해를 주는 유충이 방제 대상이나, 살충제와 접촉이 어려워 방제가 어렵다. 이를 극복하기 위한 방제기술이 성페로몬을 이용한 교미교란제 처리이다. 그러나 소규모로 흩어져 있는 국내 사과원 구도는 단일 과원의 교미교란제 처리는 교미된 암컷의 이주에 따라 유효성을 얻기가 힘들다. 이에 따라 광범위한 교미교란제 처리가 불가피하나, 현실적으로 어느 정도의 규모가 처리 대상인지 결정하기 어렵고 불필요한 광역화는 비용면에서 현실성이 없다. 대안으로 복숭아순나방 성충의 교미 반경을 측정할 수 있다면 교미교란제 처리 규모를 합리적으로 결정할 수 있다. 이러한 복숭아순나방 교미 반경 결정을 위해서는 집단간 차이를 보이는 분자지표의 개발이 우선적이다. 본 연구는 이를 위해 세 가지 전략으로 접근하였다. 첫째로, 복숭아순나방 미토콘드리아 전체 염기서열 분석으로 이를 통해 집단간 변이를 보이는 부위를 결정하려 했다. 둘째로 PCR-RFLP 기술로서 미토콘드리아 DNA의 제한효소 다형부위 결정기술이다. 끝으로 PCR-RAPD로서 전체 게놈에 대한 임의적 증폭물에 대한 다형 유전좌위 개발 전략이다.

Oriental fruit moth, Grapholita molesta, is a serious pest on apples. To control this pest in an environmentally friendly method, mating disruption strategy using sex pheromone has been developed. Area-wide application of mating disruption has been needed to be effective, with little understanding on how much size of apple cultivating area should be treated in one time application of the mating disruption technique. On this matter, we needed to determine a minimal mating active zone of G. molesta that should be applied with mating disrupters to be effective. Molecular markers to discriminate a specific population should be developed to trace population migration for reproductive behaviors. Here we developed two effective molecular markers using random amplified polymorphic DNA (RAPD) technique. Different field populations of G. molesta, based on locations and seasons, were analyzed with these markers. In a specific location, G. molesta populations varied in genetic composition with different seasons. Different local populations showed differential variation according to their relative distances among apple orchards. In overall, genetic variation among different populations became lessen with progression of seasons.

Two fruit moths of the oriental fruit moth, Grapholita molesta (Busck), and the peach fruit moth, Carposina sasakii (Matsumura), infest apples in Korea by internally feeding behavior. C. sasakii is a quarantine insect pest from some other countries importing Korean apples. G. molesta is not a quarantine insect pest, but can be incorrectly identified as C. sasakii especially when it is found inside apple fruits at its larval stages because it is not easy to identify the two species by morphological characters alone. This incomplete identification results in massive economical loss by fruits needlessly destroyed or turned away at border inspection stations of the importing nations. This difficulty can be overcome by molecular DNA markers. Several polymorphic regions of mitochondrial DNA of both species were sequenced and used for developing specific restriction sites and polymerase chain reaction (PCR) primers. Based on these sequences, three diagnostic PCR-restriction fragment length polymorphism (RFLP) sites were detected and validated for their practical uses. Also, species-specific PCR primers were devised to develop diagnostic PCR method for identifying the internal feeders.

The development of random amplified polymorphic DNA (RAPD) and expressed sequence tag-derivedsimple sequence repeats (EST-SSRs) provided a useful tool for investigating Korean ginseng genetic diversity. In this study,18 polymorphic markers (7 RAPD and 11 EST-SSR) selected to assess the genetic diversity in 31 ginseng accessions (11Korean ginseng cultivars and 20 breeding lines). In RAPD analysis, a total of 53 unique polymorphic bands were obtainedfrom ginseng accessions and number of amplicons ranged from 4 to 11 with a mean of 7.5 bands. Pair-wise genetic similaritycoefficient (Nei) among all pairs of ginseng accessions varied from 0.01 to 0.32, with a mean of 0.11. On the basis of theresulting data, the 31 ginseng accessions were grouped into six clusters. As a result of EST-SSR analysis, 11 EST-SSR mark-ers detected polymorphisms among the 31 ginseng accessions and revealed 49 alleles with a mean of 4.45 alleles per primer.The polymorphism information content (PIC) value ranged from 0.06 to 0.31, with an average of 0.198. The 31 ginsengaccessions were classified into five groups by cluster analysis based on Nei’s genetic distances. Consequently, the results ofginseng-specific RAPD and EST-SSR markers may prove useful for the evaluation of genetic diversity and discrimination ofKorean ginseng cultivars and breeding lines.