우리나라는 벼 재배를 위한 관개 시설이 비교적 잘 확보되어 염에 의한 피해가 적으나, 간척지를 비롯한 염 토양에서는 복합적 요인에 의해 벼 생육이 저해되므로 내염성 벼의 개발이 필요하다. 본 연구는 자포니카 벼로의 내염성 유전자 도입시 동반되는 열악형질을 최소화하기 위한 초정밀 분자표지 개발을 위해 기존에 보고된, Saltol QTL 분자표지의 이용 가능성을 검토하였다. 본 연구의 결과를 통해 1. 유묘 내염성 반복실험 및 정밀 검정을 통해 내염성이 안정적인 계통을 선발하였다. 2. HR28867/Pokkali와 조평/Saltol STL의 조합에서 SKC1 등 4 개(SKC1, RM10748, RM10793, RM7075)의 분자표지가 표현형과 유전자형이 일치하였으며, 분자표지 Pect4는 HR28867/ Pokkali의 조합에서만 일치하는 것으로 확인되었다. 본 연구의 결과는 자포니카 벼로의 내염성 유전자 도입에 필요한 초정밀 분자표지 개발의 기초 자료로 이용될 것이며, 향후 내염성 벼 육성을 통한 간척지 토지 이용 효율을 높이는 데 기여할 것으로 기대된다.

One of major mechanisms of insecticide resistance is the reduced susceptibility caused by mutations on the main target sites, such as sodium channel and acetylcholinesterase. Bioassay is a useful method to diagnose insecticide resistance of mosquitoes; however, it is hard to establish regional/annual resistance database based on bioassay. Recently, various markers of mutation and copy number variation have been identified through insecticide mechanism studies. Thus, molecular detection of resistance based on the resistance marker is now feasible, which can be readily implemented as a novel resistance monitoring tool to complement or replace the conventional bioassay method. In Korea, the density of vector mosquitoes native to the subtropical areas has increased due to climate change. Therefore, it is required to establish an efficient resistance monitoring system based on molecular markers to facilitate the construction of a nation-wide resistance map for more effective control of mosquitos. In addition, alternative insecticides should be introduced to the areas where mosquitoes develop high levels of resistance to maximize control efficacy against resistant populations

Origin of Red imported fire ant (RIFA : Solenopsis invicta) is Central America, a tropical climate region. It has settled in the invasive area, causing various problems. In recent, Solenopsis invicta were discovered in Busan in 2017 and then in 2018 at a construction site in Busan, Pyongtaek, Incheon and in Daegu. This study aims to confirm the origin of invasive colonies of Solenopsis invicta. We tried to test previously developed microsatellite markers so that we establish the tracing protocol for molecular epidemiology of RIFA. We justified 66 microsatellite markers already developed using DNA from the RIFA found in Incheon Harbor. As results, 30 markers were selected to facilitate amplification and fragment analysis.

1. 전세계에서 식량문제 해결을 위해 다양한 식물 신품종들이 개발되고 있다. 이에 따라 식물 유전자원과 관련된 국내외 분쟁 또한 증가하고 있다. 2. 품종 분쟁 판결에 있어 재배시험, 물질분석 및 분자표지들이 근거로 제시되어 왔다. 그 중에서 유전적 분석 기법과 기능성 분자표지에 대한 연구가 신속성, 재현성, 기술 발달 등의 이유로 활발하게 적용되고 있다. 3. 국내외 7개의 분쟁 사례를 분석한 결과, 결정적인 증거자료로써 분자표지를 이용한 판별기술이 채택되고 있지는 않았다. 반면 목표 형질의 표현형 차이에 대한 재배시험의 통계적 자료가 더욱 결정적인 증거자료로 채택되고 있었다. 분자표지가 충분한 증거로써 채택되지 못하는 원인은 분자표지 제공자의 객관성과 증거로 제시된 표지가 품종의 결정적 차이를 보이는 목적 형질과 연결되지 않았기 때문이었다. 4. 형질과 연결되지 않는 분자표지 대신 유전자 발현표지나 기능성 분자표지를 개발하여 적용하는 것이 더 효율적이고 합리적이라 판단한다. 5. 분쟁에서 근거로써 분자표지의 단점을 보완하여 분쟁에 적용한다면 품종 분쟁에 있어 보다 효율적이고 객관적인 판정이 이루어질 수 있을 것이다.

The mushroom species Cordyceps militaris has been studied and cultivated as a medicinal mushroom due to its multiple valuable biological and pharmaceutical activities. For breeding new strains of C. militaris, multiplex PCR assays were performed using primers specific for its mating type genes, MAT1-1 and MAT1-2. Mating types and mating status were confirmed, as evidenced by DNA bands at 233-bp and 191-bp for MAT1-1 and MAT1-2 respectively. The novel 'Dowonhongcho 2ho’ was developed through mating; they were found to possess high-quality fruiting bodies when grown in artificial media. The stromata of the new strain were club-shaped, with a bright orange-red color, and measured 7.1 cm in length. They had an average cordycepin content of 0.33%. Compared to 'Dowonhongcho,' the new strain had a 7% higher yield, as well as firm fruiting bodies. The optimum temperature for mycelial growth was 20~25°C, and the optimum temperature for stroma development was 18~22 °C. The fruiting bodies developed after 49.1 days from inoculation. The use of mating type molecular markers improved the breeding efficiency of the new strain 'Dowonhongcho 2ho.’ Thus, they may be valuable for artificial cultivation and industrial-scale production of C. militaris with excellent characteristics.

Differentiation of Pleurotus eryngii is laborious and time-consuming tasks especially in mycelial status. For development of a method for differentiation of P. eryngii cultivars, simple sequence repeats (SSR) from whole genomic DNA sequence analysis was used for genotyping and two multiplex-SSR primer sets were developed. These SSR primer sets were employed to distinguish 12 cultivars and strains. Five polymorphic markers were selected based on the genotypes. PCR with the each primer produced one to four distinct bands ranging in size from 200 to 300 bp. Polymorphism information content (PIC) values of the five markers were in range of 0.6627 to 0.6848 with an average of 0.6775. Unweighted pair-group method with arithmetic mean clustering analysis based on genetic distances using five SSR markers classified 12 cultivars into 2 clusters. Cluster I and II comprised of 4 and 8 cultivars, respectively. Two multiplex sets, Multi-1 (SSR312 and SSR366) and Multi-2 (SSR178 and SSR277) completely discriminated 12 cultivar and strains with 21 allele with a PIC value of 0.9090. These results might be useful to provide an efficient method for the identification of P. eryngii cultivars with separate PCR reactions. (This work was supported by a grant from the Golded Seed Project (213003-04-3-SBY20), MIFAFF, Republic of Korea.]

최근 가축의 유전적 다양성 유지 및 식량 안보에 있어서 재래품종의 중요성은 점차 증대되고 있다. 제주흑우는 멸종위험에 처한 품종이며, 2013년 7월 문화재청에 의해 천연기념물 제546호로 지정되었다. 본 연구의 목적은 제주흑우(124두)의 유전적 다양성, 유연관계 및 유전적 구조의 평가이며, 그 비교 대상으로 한우(128두) 및 외래품종 홀스타인(73두)을 공시하였다. 분자유전학적 특성을 평가하기 위해 11개 초위성체 마커(BM1824, BM2113, ETH10, ETH225, ETH3, INRA23, SPS115, TGLA122, TGLA126, TGLA227, TGLA53)의 대립유전자형을 분석하였으며, 그 결과를 토대로 유전적 다양성 지수들을 산출하였다. 품종별 평균 기대이형접합도(HExp)는 0.605-0.738, 관측이형접합도(HOsb)는 0.667-0.747 그리고 다형정보지수(PIC)는 0.644-0.773의 범위를 보였다. 특히, 제주흑우의 유전적 다양성 지수는 가장 낮은 결과를 보였다. STRUCTURE를 이용한 군락 분석 결과 유전적으로 3개의 군락으로 구분되었으며, 주성성분분석(PCA) 결과 또한 3개의 군집으로 분류됨을 확인하였다. 따라서 본 연구의 결과는 제주흑우의 유전적 고유성 및 유전자원으로써 가치 판단을 위한 과학적 근거가 될 것으로 사료된다.

종의장거리산포는집단의확산과집단간개체의흐름에중요한역할을한다. 본연구는독도에서발견된식물을형태학적특징과핵및엽록체DNA의분자마커를이용하여종을식별한결과, 참빗살나무(노박덩굴과)로확인되었다. 얕은토양층과험준한지형등입지적으로열악한대양섬인독도에서목본의분포는의미있는결과이며, 향후외부유입종의지속적인모니터링이요구된다.

Trombiculid mites are known to be the vector of tsutsugamushi disease by transmitting Orientia tsutsugamushi to human. Although the classification of trombiculid mites is necessary for vector surveillance, their classification by morphological observation is only possible at the larval stage and not easy because of similar shapes as well as tiny body sizes. Further the classification need the specimen production process, it takes much time and the accuracy of classification is changed according to the technology of the researcher. The internal transcribed spacers (ITS) regions of 8 trombiculid mite species were analyzed by amplification using tick common ITS primer sets. We designed molecular marker sets for the identification of five Leptotrombidium species, the lengths of marker L. orientale (1078 bp), L. pallidum (820 bp), L. palpale (1202 bp), L. scutellare (447 bp) and for L. zetum (621 bp) and three Neotrombicula species, the lengths of marker N. gardellai (264 bp), N. japonica (460 bp) and N. kwangneungensis (309 bp) based on alignment of ITS sequences. The markers will be helpful for exact classification of trombiculid mites. This study is the first report on molecular marker of ITS regions of trombiculid mites.

The cladistics analysis of subfamily Phylinae, based on molecular data analysis of 3655 base pairs of mitochondrial (16S, COI) and nuclear (18S, 28S) ribosomal DNA for 66 taxa in five tribes were conducted. Partitions of datasets were aligned using different algorithms implemented in MAFFT, further combined through sequence matrix and the combined dataset was analyzed using partitioned maximum likelihood. Clades in analytical method comprise six subfamilies of Miridae as outgroup: Isometopinae, Cylapinae, Bryocorinae, Orthotylinae, Deraeocorinae, and Mirinae, and five tribes of ingroup in subfamily Phylinae: Auricillocorini, Hallodapini, Phylini, Leucophoropterini, and Pilophorini. In analysis, tribe Auricillocorini as sister group to all remaining phylines. Most of the tribes are monophyly, except largest tribe phylini, which is clearly divided as paraphyly.

본 연구에서는 분자마커를 이용한 목표형질분리의 효용성을 검토하였다. Ivory seed(IVS) 유전자는 bHLH 단백질을 암호화하고 있고, 꽃의 안토시아닌과 종자의 프로안토시아닌의 발현에 관여하는 애기장대의 TT8 및 페튜니아의 AN1유전자와 상동성을 가지는 유 전자로 나팔꽃의 꽃과 종자의 착색을 촉진하는 것으로 알려져 있다. 아이보리 종자로부터 화색이 full purple(FP), pale purple(PP), magenta(M) 및 pale red(PR)인 4가지 계통의 식물체가 분리되었다. 분자분 석을 통해 이 식물체 모두 ivs 유전자의 변이형질을 지니는 것으로 확인되었다. 분자마커 선발을 위해 고안 된 프라이머를 사용하여 IVS 유전자의 exon 2와 intron 5 영역을 증폭하여, ivs 대립형질인 ivs-stb 및 ivs-pwt을 야생형 IVS로부터 구분할 수 있었다. PCR 결과로 예측한 IVS 유전자의 대립형질 구성 상태는 Southern blot 분석 결과와 정확하게 일치하였다. PP 와 PR은 ivs-stb에 의한 열성 변이체였으며, M과 FP 계통은 ivs-stb 대립형질에 더하여 각각 야생형 IVS 및 ivs-pwt와의 이형접합체로 확인되었다. 본 연구에서 선발된 분자마커는 IVS 유전자의 대립형질 구분에 매 우 유용하였다.

Three tortricid pests, Grapholta dimorpha (Komai), G. molesta (Busck), and Carposina sasakii (Matsumura) are known as internal apple feeders in Korea. For identify young larvae which occurring serious damage in fruits, the molecular maker was developed from their mitochondrial DNA (mtDNA) sequences. To develop of PCR-RFLP marker, ND4 locus was digested with Swa Ⅰ. ND4-Swa Ⅰ digests showed two bands (396, 292 bp), one band (700 bp), and three bands (408, 178, and 103 bp) of G. dimorpha, G. molesta, and C. sasakii, respectively. Species-specific diagnostic PCR primers were developed in the ND4 locus and gave species-specific PCR products. Finally, these markers were applied to diagnose larvae infesting apples and showed species-specific fruit damage patterns, in which most feeders of G. dimorpha, G. molesta, and C. sasakii showed major feedings in apex, surface, and core of apple fruits, respectively.

Three insect pests internally feed pome fruits in Korea. These include oriental fruit moth (Grapholita molesta), plum fruit moth (Grapholita dimorpha) and peach fruit moth (Carposina sasakii). Molecular markers discriminating these three species were developed using PCR-RFLP technique. Mitochondrial (mt) genomes were analyzed to locate polymorphic loci. Six mtDNA regions (CO-Ⅰ, CO-Ⅱ, CB, 16SrRNA-12SrRNA, ND3, ND4) of G. dimorpha were cloned and sequenced. These six sequences of G. dimorpha were aligned with those of C. sasakii and G. molesta to determine polymorphic restriction sites. Predicted PCR-RFLP markers were confirmed with known insect samples. With the validated PCR-RFLP markers, field male adults collected in traps baited with rubber sept lures impregnated with different ratios of major sex pheromone components of G. molesta were analyzed.

Acquisition of a reference Tetranychusstrains that are completely susceptible to acaricides and retain identical genetic backgrounds to acaricide-resistant strains is an essential step in elucidating mechanisms of resistance. To establish both completely susceptible and acaricide-resistant strains for this purpose, I collected Tetranychus mite populations from various regions in South Korea including both heavily cultivated and remote regions. Suitability as a susceptible or resistant reference strain was tested by determining species identity as Tetranychus urticae along with baseline susceptibility to common acaricides. The majority of mite populations collected from cultivated areas belonged to a monophyletic group of the previously reported green-type T. urticae as determined by mtCOI- and ITS2-based phylogenetic analysis. These strains showed relatively reduced levels of susceptibility to the acaricides tested, suggestive of the development of resistance. Among them, the AbaR strain was classified as a minor group in the T. urticae complex. The UD strain, originally collected from a remote island region, was found to be susceptible to the acaricides tested and generated an independent phylogenetic branch. The UD strain was also considered a minor group in the T. urticae complex. Phenotypic analysis based on morphological characters confirmed that both the AbaR and UD strains were statistically undistinguishable from the major green-type T. urticae. Taken together, I propose that the UD strain be used as a susceptible reference strain as it provides baseline susceptibility to acaricides and a wild-type genetic background for the resistance studies.