Stoneflies (Plecoptera) are known for being sensitive to water pollution and are used as bioindicators for evaluating water quality. Among them, Nemouridae, especially the genus Nemoura, which are commonly referred to as winter stoneflies, can be found around streams even during the cold winter months. Nemoura geei Wu, 1929, among them, was originally described from Beijing and is widely distributed in Korea, China, Japan, and the Russian Far East. Here, we report the development and characterization of new functional microsatellite markers of N. geei using high-throughput sequencing technology. A total of 80,661 microsatellite loci were identified with a total length of 1,801,591 bp. The average length was 22.34 bp, and microsatellites occupied 0.42% of the entire sequence. The novel 20 microsatellite markers developed in this study can be usefully applied to the population genetics analyses as important genetic resources for understanding the ecological and evolutionary characteristics of a stonefly species at the population level in Korea.

The pear pest, Cacopsylla jukyungi (Hemiptera: Psyllidae), is one of the most damaging insect to commercial pears in South Korea. In this study, we developed eight microsatellite markers specific to C. jukyungi and genotyped 132 individuals collected from 11 localities throughout South Korea. Populations showed lower observed heterozygosity than expected heterozygosity and slightly or highly positive values of inbreeding coefficients, suggesting that C. jukyungi is subjected to inbreeding. The nationwide expansion of pear orchards and the replacement with a popular new cultivar during the last 50 years, which may have accompanied the spread of C. jukyungi-bearing pear grafts and scions, are likely sources of such facilitated dispersal. Thus, a management strategy against unintended anthropogenic dispersal of the pear psyllid will be required for better control of C. jukyungi.

A number of Korean Chicken breeds were registered in Domestic Animal Diversity Information System (DAD-IS, http://dad.fao.org/) of the Food and Agriculture Organization (FAO). Evaluation of genetic diversity and relationship of local breeds is an important factor towards the identification of unique and valuable genetic resources. Therefore, this study aimed to analysis the genetic diversity and relationship of 22 Korean Chicken breeds using 12 microsatellite (MS) markers. The mean number of alleles for each variety was 5.52, ranging from a 3.75 (Leghorn F; NF) to a 7.0 (Ross). The most diverse breed was the Hanhyup3 (HCC), which had the highest expected heterozygosity (HExp) (0.754) and polymorphic information content (PIC) (0.711). The NF was the least diverse population, having the lowest HExp (0.467) and PIC (0.413). As a result of the principal coordinates analysis (PCoA) and factorial correspondence analysis (FCA) confirmed that Hy-line Brown (HL) and Lohmann Brown (LO) are very close to each other and that Leghorn and Rhode Island Red (RIR) are clearly distinguished from other groups. Thus, the reliability and power of identification using 12 types of MS markers were improved, and the genetic diversity and probability of individual discrimination were confirmed through statistical analysis. This study is expected to be used as basic data for the identification of Korean chicken breeds, and our results indicated that these multiplex PCR marker sets will have considerable applications in population genetic structure analysis.

본 연구는 한우 개체식별 및 친자감별에 있어 기존의 di-nucleotide repeat microsatellite marker 사용 시 발생했던 stutter로 인한 대립유전자 판별 오류 등의 문제들을 극복하고, 분석결과의 신뢰도와 정확도를 높이기 위해 tri-, tetra-, penta-, hexa-nucleotide repeat microsatellite 좌위들로 이루어진 새로운 개체식별 마커 13 종(BTRC6_01, BTRC19_02, BTRC11_03, BTRC16_05, BTRC9_07, BPC19_08, BTEC17_09, BPC21_10, BTEC4_11, BPC7_12, BPC1_13, BHXC29_14, BPC1_15)을 개발하였다. 선발된 13개의 좌위를 가지고 소 1,530두에 microsatellite typing을 실시한 결과, 총 61개에 대립유전자가 발견되었으며, 좌위별로 평균 4.69개의 대립유전자를 가지는 것으로 확인되었다. 마커의 다형성과 정보력의 척도인 PIC (Polymorphism Information Contents)값은 0.25(BTRC9_07)~0.59(BTEC17_09)로 나타났으며 BHXC29_14, BPC1_13, BTEC17_09, BTRC16_05, BTRC19_02, BTRC6_01 좌위들은 PIC 0.5 이상 그리고 나머지 좌위들 모두 PIC 0.25 이상의 값을 가지는 것으로 확인되어 마커로서 다형성이 있음이 검증되었다. 개발한 마커를 활용하여 한우(영암, 장흥)와 유럽우 7종 (Brown Swiss, Limousin, Angus, Simmental, Hereford, Charolais, Holstein)의 유전적 특성을 분석하였으며, 총 9개 집단의 Heterozygosity와 FIS (inbreeding coefficient) 값을 측정하였다. 기대이형접합율은 0.451(BS)~0.605(AG) 범위 내로, 한우는 0.532(영암), 0.545(장흥)의 값을 가지는 것으로 나타났다. 한우(영암, 한우)와 7종의 유럽우들 간의 유연관계 분석은 특정 대립유전자 빈도를 근거로 한 유전적 거리의 추정으로 이루어졌다. 한우집단과 Simmental 간의 유전적 거리(0.1848)가 가장 가깝고 비교적으로 Brown Swiss와의 유전적 분포(0.3352)가 가장 먼 것으로 나타났으며, 계통발생학적으로 유전적 분화 양상을 확인함으로써 본 마커의 한우 유전적 다양성 및 유연관계 분석에 활용 가능성을 제시하였다.

The aim of this study was to assess the levels of genetic diversity and relationships of Korean native black goat (n = 58), compared with the exotic breed, Boer (n = 97). For the analysis of genetic characterization 11 microsatellite markers (MAF065, INRA063, CSRD247, OarFCB20, SRCRSP5, INRA006, ILSTS008, ILSTS011, INRA005, ILSTS087, SRCRSP8) were genotyped. The number of alleles was observed 3 (INRA005) to 10 (SRCRSP8) each markers. The mean expected and observed heterozygosity (Hexp and Hobs) and polymorphism information content (PIC) for the Korean native black goat breed varied from 0.551 to 0.860, 0.517 to 0.948 and 0.464 to 0.835, respectively. Principal Components Analysis (PCoA) and FCA results showed that Korean native black goat breed was confirmed to be clearly separated from bore breed. These results were scientific evidence that Korean native black goat represents a unique and valuable animal genetic resource.

In this study, we developed ten microsatellite markers specific to L. angelina using the Illumina NextSeq 500 platform. Forty-three individuals of L. angelina collected from three localities in South Korea were genotyped to validate these markers and to preliminarily assess population genetic characteristics. The observed number of alleles, observed heterozygosity (HO), and expected heterozygosity (HE) at a locus ranged from 4–13, 0.211–0.950, and 0.659–0.871 in the population with the largest sample size (20 individuals), respectively, thereby validating the suitability of the markers for population analyses. Our preliminarily assessment of the population genetic characteristics indicates the presence of inbreeding in all populations, an isolation of the most geographically distant population (Seocheon), and lower HO than HE. The microsatellite markers developed in this study will be useful for studying the population genetics of L. angelina collected from additional sites in South Korea and from other regions. †These authors contributed equally to this paper.

In this study, we developed 11 microsatellite markers specific to A. crataegi using NGS to investigate the genetic relationships of A. crataegi populations from South Korea to circumferential Asian countries (China, Russia, Mongolia, and Japan). Further, two mitochondrial DNA (mtDNA) gene segments (COI and CytB) were sequenced from the samples. The population- and individual-based Principal Coordinates and STRUCTURE analyses collectively suggested that the South Korean population of A. crataegi is most differentiated from the Japanese population, whereas it was closer to Mongolian and Chinese populations. These results collectively suggest that northern populations, in particular, Mongolian populations can be considered as the most genetically compatible one as donee population, when the reintroduction program is launched. †These authors contributed equally to this paper.

유전자 친자확인시험법은 동일 종 내에서도 개체를 식별 할 수 있는 강력한 분자진단체계이다. 반달가슴곰 우수리아 종(Ursus thibetanus ussuricus)은 한반도를 비롯한 중국, 러시아 등지에 분포하나, 우리나라에서는 지역적으로 절멸된 것으로 추정되고 있으며, 현재는 멸종위기야생생물Ⅰ급으 로 지정되어 있고, 북한, 러시아, 중국 등에서 재도입되어 종복원이 진행 중에 있다. 본 연구는 우리나라에 재도입된 반달가슴곰집단의 친자확인, 가계도 작성, 미확인개체 추적 등 방사개체의 관리에 요구되는 분자진단체계의 구축을 위하여 차세대염기서열분석법을 통해 결정한 반달가슴곰 전 장유전체 서열(whole genome sequence, WGS)에서 미세 부수체(microsatellite, MS) 마커 개발을 목적으로 하였다. 전장유전체 서열은 총 12개체의 혈액 DNA를 이용하여 결정하였고, 결정된 서열에서 총 1,730,507개의 단순반복서열(simple tandem repeat, STR)의 정보를 발굴하였다. 이 중 반복단위(repeat unit)의 길이가 2-6 nt로 구성된 STR은 총 5,954개였다. WGS 상에서 검출된 STR의 관찰이형접합율(observed heterozygosity, Ho)을 고려하여 총 97종을 1차 선정하였다. 부-모-자 유전자형의 멘델유전(mendelian inheritance)을 만족하는 48종의 마커를 반달가슴곰 전체 집단에 적용하여 시험하였고, 중합효소연쇄반응(polymerase chain reaction, PCR)을 통해 유전자형을 결정하였다. 유전 자형의 다양성지수와다형정보량(polymorphic information content, PIC), 동일개체출현율을 고려하여 최종 15종의 MS 마커 세트를 구성하였다. 분석결과, 전체 집단에서 출현한 평균 대립유전자의 수는 6.267개였으며, 기대이형접합율(expected heterozygosity)는 평균 0.7242을 나타내었다. 평균 PIC는 0.6718, 동일개체 출현율은 1.867×10-14 수준으로 높았으나, 부권부정율은 0.00337로 다소 낮은 수준을 보였다. 재도입된 1세대집단 전체에서는 평균 대립유전자 수 5.867개, 평균 기대이형접합율 0.7217, 평균 PIC 0.6573을 나타내었고, 후손 전체에서는 평균 대립유전자 수 5.933개, 평균 기대이형접합율 0.7140, 평균 PIC 0.6554를 나타내었다. 하지만, 후손집단을 2세대와 3세대로 구분했을 때, 2세대는 평균 대립유전자 수 5.867, 평균 기대이형접합율 0.722, 평균 PIC 0.657, 3세대는 평균 대립유전자 수 4.467, 평균 기대이형접합율 0.694, 평균 PIC 0.601을 나타내어 세대가 진행될수록 대립유전자의 수와 기대이형접합율, PIC 모두 감소하는 경향을 나타내었다. 최종 선정된 15개의 MS 마커체계를 이용하여 반달가슴곰 집단에 대한 미확인 개체 추적, 친자확인 등을 시험하였다. 최근 포획된 개체들 중에서 개체명이 확인되지 않았던 4개체는 자연출생 2세대, 3세대, 전파발신기가 탈락된 방사 1세대로 확인 되었다. 또한 동일성검사 결과 올해 5월 교통사고를 당한 반달가슴곰은 KM-53이며, 올해 출생한 새끼 반달가슴곰들에 대한 검사를 통해 2개체가 인공수정에 의해 출생한 것으로 확인되었다. 이상의 결과들은 본 연구에서 확립한 MS 마커의 조합이 자연방사와 자연출생 개체들로 구성된 반달 가슴곰 집단의 개체관리를 위한 분자마커체계로 유용하게 이용될 수 있음을 보여주고 있다. 또한 본 연구를 통해서 확립된 MS 마커들은 현재 지리산과 수도산에 서식하고 있는 반달가슴곰 집단의 개체관리, 미확인 개체 추적, 친자확인 등 유전자분석에 활용될 수 있을 것으로 기대되며, 아울러 향후 유전적 다양성 증진, 집단간 교류개체 선정 등에도 필요한 유전 정보를 제공할 수 있을 것이다. 결론적으로 반달가슴곰 전장유전체 서열을 토대로 마련된 분자진단체계는 향후 반달가슴곰의 보호와 생태복원을 위한 종복원에 기여할 것으로 기대된다.

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.

The black-veined white, Aporia crataegi (Lepidoptera: Pieridae), which is distributed mainly in Eastern Asia is presumed to be extinct in South Korea, only with some numbers of dried specimens left, whereas the species is found casually in circumferential countries. One of the common conservation practices for such species is to launch introduction program, but prior population genetic analysis between donor and donee populations might be essential for long-term conservation. In this study, we developed 11 microsatellite markers specific to A. crataegi using Illumina paired-end sequencing to investigate the genetic relationships of A. crataegi populations from South Korea and circumferential Asian countries (China, Russia, Mongolia, and Japan). Further, two mitochondrial DNA (mtDNA) gene segments (COI and CytB) were sequenced from the samples. The population- and individual-based Principal Coordinates and STRUCTURE analyses collectively suggested that the South Korean population of A. crataegi is most differentiated from the Japanese population, whereas it was closer to Mongolian and Chinese populations. The STRUCTURE analysis based on two concatenated mtDNA gene sequences also supported different genetic composition of Japanese population from the remaining populations including that of South Korea and rather similar genetic composition between the populations of South Korea and Mongolia. These results collectively suggest that northern populations, in particular, Mongolian populations can be considered as the most genetically compatible one as doner population, when reintroduction program is launched.