GM작물은 세계적으로 재배면적이 지속적인 증가되고 있으며, 이로 인한 GM작물의 잠재적인 환경위해성에 대한 우려도 증가되고 있다. 현재까지 국내에서 GM작물의 상업적 재배는 되고 있진 않지만 GM작물의 안전성 평가를 위한 기술 개발은 크게 요구되고 있는 실정이다. 국내외적으로 다양한 GM작물의 유전자이동성 평가가 수행되어왔으나 화분 공급원인 GM 벼의 개화기 차이에 의한 유전자 이동성 연구는 부족한 실정이다. 본 연구에서는 벼의 개화기 차이에 따른 비타민A 강화 벼(PAC)로부터 모품종인 낙동벼와 중만생종인 일미벼, 조생종인 운광벼, 중생종인 대보벼, 통일형인 세계진미벼로의 화분 매개에 의한 유전자 이동성을 평가하였다. 비타민A 강화벼(PAC)의 도입 유전자를 검출하는 PCR 분석을 통해 유전자 이동성 유무를 최종적으로 검증하였다. 총 파종된 종자수에 대한 교잡율은 개화기가 일치하는 낙동벼에서는 0.0007%, 개화기가 차이가 있는 일미벼, 운광벼, 대보벼, 세계진미벼에서는 0%로 나타났으며, 모든 교잡개체들은 비타민A 강화벼(PAC)에 근접한 2m 내에서 발견되었다. 화분 매개에 의한 비타민A 강화벼(PAC)의 유전자 이동 특성은 기존에 연구된 결과들과 비슷한 결과를 보였으며, 벼 재배품종간의 개화기 차이가 화분에 의한 교잡을 결정하는데 중요한 요인으로 작용하였다. 이에 벼 경작지의 기상 조건과 벼 품종간의 개화시기 중복 여부 등이 GM벼에 의한 일반 재배품종 및 잡초성벼로의 유전자 이동 최소화 기술 개발과 안전관리 기준 작성에서 주요 영향 요소들로 고려해야 한다.
We developed and characterized five polymorphic microsatellites of Nilaparvata lugens from hybridization method using biotin enrichment strategy and two polymorphic microsatellites from Next Generation Sequencing. Also 11 microsatellites that developed by Sun et al. (2011) are employed to carry out genetic analysis of N. lugens in Southeast Asia. The number of alleles per locus ranged from 2 to 12 with an average of 4.63 alleles per locus. The mean observed heterozygosity of the eleven populations ranged from 0.031 to 0.938 and the expected hetetrozygosity ranged from 0.031 to 0.881. Signifiant genetic differentiation was detected among the three N. lugens populations as the FST ranged from 0.028 (Cheong Do and Ha Long) to 0.161 (CH and BN). The results of microsatellite marker suggested that found N. lugens migrated to Korea at least two times in different period or once. Genetic distance of N. lugens between Korea and Hi Pong were mostly closed and genetic distance of Ha Long and HCM were relatively closed. In this study, development of microsatellites should facilitate the study of future population genetics of N. lugens, and eventually elucidate the route of N. lugens migration to Korea. Thus, combining satisfactory microsatellite markers and intensive surveillance methods in paddy field could be easy to understand to the N. lugens migration mechanism.
The pear psylla, Cacopsylla pyricola Foerster (Homoptera: Psyllidae), is a serious insect pest of commercial pear crops. The species, which resides on pear trees throughout its life cycle, is rapidly spreading in some regions of the world. Given the life cycle, it is unclear how such a rapid spread has been facilitated. Presently, the population genetic structure of the species including genetic diversity and gene flow was studied to understand the nature of dispersal and field ecology of the species. Pear psylla was collected from several pear orchards in Korea. The 658-bp region of mitochondrial COI gene and the 716-bp long complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA were sequenced. Unlikely other previously studied insect pests, the COI-based genetic diversity of the pear psylla was extremely low (maximum sequence divergence of 0.15%). This finding allowed us to conclude that the species may have been introduced in Korea relatively recently, possibly with the phenomenon of genetic bottlenecks. ITS2 sequence-based analyses of phylogeny, population differentiation, gene flow, and hierarchical population structure all concordantly suggested that the pear psylla populations in Korea are neither genetically isolated nor hampered for gene flow. These genetic data are concordant with the dispersal of an overwintering winterform morph outside the non-pear habitat in the fall and the possibility of subsequently longer distant dispersal.
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.
국내 4개 지역으로부터 채집된 배추좀나방(Plutella xylostella)의 미토콘드리아 DNA중 COI 유전자 일부 (438 bp)의 염기서열을 결정, 유전적 다양도 및 유전자 이동정도를 파악함으로써 집단 유전적 구조 및 특성에 대하여 연구하였다. 총 21개체로부터 13개의 mtDNA haplotype을 얻었으며 이들의 변이는 0.3~1.4%로 다른 곤충을 대상으로 한 유사연구와 비슷한 크기를 나타내었으며 haplotype 다양도는 매우 높았다(평균 h=0.81). 지리적으로 먼 제주도의 개체군과 경남 김해 두 지역(11km 거리)의 개체군을 비교한 결과, 통계적으로 유의한 정도의 유전적 격리(p<0.05%)는 전혀 관찰되지 않았으며, 대신 상당한 정도의 세대당 암컷 이동률(Nm=2-30)을 보였다. 또한 GenBank에 등록된 하와이의 배추좀나방 haplotype은 본 연구에서 얻은 것들과 유전적으로 흡사하였다. 종합적으로, 국내 배추좀나방은 전체적으로는 많은 haplotype수에 기인한 적절한 크기의 유전적 분화율을 보유하고 있으며 국지적으로는 상당한 이동력에 의한 장거리 이동으로 개체군내 높은 haplotype 다양도를 보이며 동시에 지역간의 유전적 유사성을 나타낸다고 요약되었다.
Genetically modified (GM) crops have been developed worldwide through the recombinant DNA technology and commercialized by global agricultural companies. Until now, GM crops have not been cultivated commercially in Korea. Commercialization of GM crops requires a compulsory assessment of environmental risk associated with the release of GM crops. This study was conducted to evaluate the frequency of pollen mediated gene flow from Bt transgenic rice (Agb0101) to japonica non-GM rice (Nakdongbyeo), indica non-GM rice (IR36), and weedy rice (R55). A total of 729,917, 596,318 and 230,635 seeds were collected from Nakdongbyeo, IR36, and R55, respectively, which were planted around Agb0101. Selection of the hybrids was determined by repeated spraying of herbicide and Cry1Ac1 immunostrip assay. Finally, the hybrids were confirmed by PCR analysis using specific primer. The hybrids were found in all non-GM rice and out-crossing ranged from 0.0005% at IR36 to 0.0027% at Nakdongbyeo. All of hybrids were located within 1.2 m distance from the Agb0101 rice plot. The meteorological elements including rainfall and temperature during rice flowering time were found to be important factors to determine rice out-crossing rate. Consideration should be taken for many factors like the meteorological elements of field and physiological condition of crop to set up the safety management guideline to prevention of GM crops gene flow.
Genetically modified (GM) crops have never been cultivated commercially in Korea, it is necessary for a thorough assessment of the risks associated with their environmental release. We determined the frequency of pollen mediated gene flow from disease resistant GM rice (OsCK1) to non-GM rice (Nagdongbyeo) and weedy rice (R55). A total of 449,711 or 164,604 seeds were collected from non-GM and weedy rice, respectively which were planted around OsCK1. Resistance of the hybrids was determined by repeated spraying of herbicide and DNA analysis using specific primer to confirm hybrids. Though non-GM rice and weedy rice have similar flowering time, the hybrids were found only in non-GM rice and out-crossing ranged from 0.018% at 0.3 m to 0.013% at 0.6 m. All of hybrids were located within 0.6 m distance from the GM rice plot in southerly direction. The meteorological factors including temperature and relative humidity during flowering time were found to be the most important factors for determining rice out-crossing. It should be considered many factors like the local weather condition and flowering time to set up the safety management policy to prevent pollen mediated gene flow between GM and conventional crop.
Genetically modified (GM) crops have never been cultivated commercially in Korea, it is necessary for a thorough assessment of the risks associated with their environmental release. We quantified the amount determined the frequency of pollen mediated gene flow from disease resistant GM rice to weedy rice (R55). A total of 164,604 seeds were collected from weedy rice, which were planted around GM rice. Resistance of the hybrids was determined by repeated spraying of herbicide and DNA analysis using specific primer to confirm hybrids. Though weedy rice has similar flowering time, the hybrids were found only in non-GM rice and out-crossing ranged from 0.018% at 0.3 m to 0.013% at 0.6 m. All of hybrids were located within 0.6 m distance from the GM rice plot in southerly direction. The meteorological factors including temperature and relative humidity during flowering time were found to be the most important factors for determining rice out-crossing. It should be considered many factors like the local weather condition and flowering time to set up the safety management policy to prevent pollen mediated gene flow between GM and conventional crop.
국내에서는 아직까지 유전자변형 작물이 재배되고 있지 않으나 유전자변형 작물의 환경방출을 위해서는 반드시 환경위해성 평가가 수행되어야 한다. 본 연구에서는 병저항성 유전자변형 벼(OsCK1)로부터 비 형질전환 벼(낙동벼)와 잡초성벼(R55)로의 화분 매개에 의한 유전자 이동성을 평가하였다. 비 형질전환 벼로부터 449,711립의 종자를 얻었으며 잡초성벼로부터는 164,604립의 종자를 수확하였다. 교잡개체는 2회의 제초제 살포와 병저항성 벼에 특이적인 프라이머를 이용한 분자생물학적 방법을 통해 저항성 여부를 검증하였다. 개화기간이 서로 일치하였음에도 불구하고 비 형질전환 벼에서만 교잡개체를 확인할 수 있었으며, 교잡율은 교잡개체가 발생한 거리에 따라 0.013~0.018%로 나타났다. 모든 교잡개체들은 병저항성 유전자변형 벼에 근접한 0.6 m내에서 발견되었다. 화분 매개에 의한 병저항성 유전자변형 벼의 유전자 이동성은 앞서 연구된 결과와 유사한 것으로 나타나 농업환경에 방출되더라도 환경에 미치는 영향은 다른 벼들과 비슷할 것으로 추정되었다. 개화기간 중 온도와 습도 등 기상요인이 벼의 교잡율을 결정하는데 중요한 요소이었다. 따라서 유전자변형 벼와 주변 재배종 및 잡초성벼로의 유전자 이동에 의한 안전관리 대책을 마련하기 위해서 지역의 기상요인과 개화기 중복 등의 요인들이 충분히 고려되어야 할 것이다.
CMVP1 (cucumber mosaic virus pathotype 1) has been frequently occurring virus causing damage in pepper farms, and it is hard to control the outbreak due to lack of the genetic source resistant to this specific pathotype. Therefore, we have developed transgenic peppers tolerant of CMVP1 using a CP gene of CMVP0 pathogen. In order to fulfill the requirement of the biosafety assessment criteria, we have studied the horizontal gene flow from GM pepper to non-GM pepper by monitoring the transgene movement. If the pepper farms are located closely each other and the pollen moves from GM pepper to non-GM pepper, it would cause unintended fertilization. Therefore, a buffer zone to separate the cultivation regions is required to avoid the contamination of transgene. Previously, several data regarding the movement distance of pepper pollen were reported by judging the phenotypic change. However, no tool as a trace marker was available. The objective of this study was to assess the frequencies of gene flow from GM peppers to non-GM peppers in neighboring farms using the transgene of CP as a trace marker. The GM and non-GM peppers were cultivated in the isolated farm of Nongwoo Bio Co. (NW GM pepper field) and pepper fruits were collected from the NW GM pepper field as well as the neighboring pepper farms. The pepper seeds collected from the farms were planted and the massive PCR analysis was performed to answer the question how far the pollen of GM pepper migrates. The conclusive data based on the consecutive experiments for 6 years is that the gene flow by pollen movement did not occur in peppers that were separated each other over 30 m.
본 연구에서 실행한 LM 벼의 화분비산 연구는 non-LM벼와 잡초성 벼의 의도치 않은 교잡은 경작지의 생태계에 문제를 야기할 기능성이 커지고 있다. 본 연구는 이를 예방하기 위한 벼의 이격거리 설정을 위하여 수행하였다. 1. 벼의 개화기간의 개화시간(10:00~14:00) 의 주풍은 남풍이었으며, 시간대별 풍속은 0.94~1.77 m/s이었다. 2. 개화기는 LM벼와 Wild벼가 일치하는 기간은 8일이였으며, LM벼와 Wild벼의 최성기는 4일의 차이가 있다. 3. LM벼의 화분이 non-LM벼보다 유의하게 작게 조사되었다. 4. 화분채집량은 위가 노출된 슬라이드 글라스가 위가 덮인 슬라이드 글래스 보다 많았으며 LM벼가 Wild벼보다 비산량이 많았다. 5. 벼 화분의 비산은 오전 10시부터 오후 2시까지 대부분 이루어졌다. 6. 거리에 따른 화분비산량은 1 m까지는 급격히 감소하였으며 2 m이상에서는 서서히 감소하는 전형적인 지수함수를 나타냈으며, 3 m이상에서는 거의 발견되지 않았다.
A gene flow has been tested from transgenic herbicide-resistant Chinese cabbage to major crops in Cruciferae. Hybridizations were made between transgenic Basta-resistant Chinese cabbage and non-transgenic Chinese cabbage, B. napus, B. juncea and Brassicoraphanus(B. campestris × Raphanus sativus, 2n=4x=38) with honey-bee mediated fertilization. Progeny populations were selected by Basta herbicide. The F1 hybrids with pat gene were again self-pollinated or cross-pollinated with nontransgenic Chinese cabbage and rape naturally or artificially. Pod setting ratio were compared as a tentative parameter for potential gene flow.
Key agronomic characters were compared among the F1 hybrid populations obtained. In the days to flowering after cold treatment, the F1 hybrids of Brassicoraphanus x B. campestris and B. napus x B. campestris(GM) were the shortest (75 days), and the GM Chinese cabbage and the F1 hybrid of B. juncea x B. campestris(GM) were 87 and 93 days, respectively. In the stem length, the F1 hybrid of Brassicoraphanus x B. campestris was the longest(175 cm), and followed by the F1 hybrid of B. napus x B. campestris(GM), the F1 hybrid of B. juncea x B. campestris(GM) in order. The naturally occurring pod setting ratios were fluctuated seriously among individual and segregation lines. Natural pod setting of the transgenic Chinese cabbage was 10 to 19 %. The F1 hybrid of B. napus x B. campestris(GM) showed the ratio of 4-11%. However, the F1 hybrids of B. juncea x B. campestris and Brassicoraphanus x B. campestris(GM) did not produced seeds.
The F1 hybrids with pat gene were hybridized with nontransgenic Chinese cabbage and rape by artificial bud pollination to test potential gene flow. All cross combinations except Brassicoraphanus produced seeds and the pod setting ratios ranged from 35 to 100 percentages.
Thus, the potentials of the gene flow from Chinese cabbage with pat gene to the order specise in the same gene interspecies or intergeneric species and from the F1 hybrids with pat gene to nontransgenic Chinese cabbage and rape were detected. Herbicide resistant test to the acquired seeds by natural and artificial pollinations including honey-bee mediated fertilization using the same materials is going on.