The two-spotted spider mite, Tetranychus urticae, is a worldwide agricultural pest that invades a wide range of host crops and rapidly develops resistance to pesticides. T. urticae can be resistant to one acaricide or exhibit multiple resistances or cross resistance to various other acaricides. Acequinocyl inhibits respiration in mitochondria at the ubiquinol oxidation site (Q0) of Complex III of the electron transfer chain. Pyridaben is a METI acaricide that inhibits mitochondrial electron transport at complex I. In this study, we investigated the cross resistance to seven acaricides in acequinocyl- and pyridaben-resistant strain. Furthermore, the frequencies of the I256V and N321S mutations in mitochondrial cytochrome b (cytb) of pyridaben-resistant and fieldcollected strain was analyzed.
The melon and cotton aphid Aphis gossypii Glover (Hemiptera; Aphididae) is one of the most serious pests worldwide. We surveyed insecticide susceptibility in A. gossypii field populations to 12 insecticides to examine resistance ratios. The levels of insecticide resistance were extremely high, especially to neonicotinoids. One point mutation was found in the beta1 subunit loop D region of the nicotinic acetylcholine receptor (nAChR) of the imidacloprid-resistant strain. Feeding behavior analysis using an electrical penetration graph showed that sublethal doses of imidacloprid had significant effects on the duration of phloem ingestion. In addition, higher doses of imidacloprid induced contact toxicity rather than inhibition of feeding behavior. Temperature and insecticide are two important factors that affect survival, reproduction and other physiological processes of insects. To determine interactions of temperature and insecticide treatment on susceptible and imidacloprid-resistant strains of A. gossypii, adults were exposed to three temperatures (17, 22, and 28℃) or combinations of three temperatures and imidacloprid (LC20), and the expression of several genes (heat shock protein 70, cuticle protein, cytochrome P450, and elongation factor) were analyzed. Additionally, the effect of electron beam irradiation on development, reproduction, and several gene expression of imidacloprid-resistant and -susceptible A. gossypii were compared.
Temperature and insecticide are two important factors that affect survival, reproduction and other physiological processes of insects. To determine interactions of temperature and insecticide treatment on susceptible (S) and imidacloprid-resistant strains (IMI-R) of Aphis gossypii, adults were exposed to three temperatures (17, 22, and 28℃) or combinations of three temperatures (17, 22 and 28℃) and imidacloprid (LC20), and the expression of several genes (heat shock protein 70, cuticle protein, cytochrome P450, and elongation factor) were analyzed. The expression level at 17℃ of heat shock protein 70, cuticle protein, and elongation factor in S strain were up-regulated with increased time and higher than those of IMI-R strain. However, expression of cytochrome P450 was not affected by elevated temperature both S and IMI-R strain. Combined treatment of elevated temperature and imidacloprid were significantly up-regulated only cuticle protein in S strain and higher than those of IMI-R strain.
벼멸구에 감수성이며 고품질인 ‘황금누리’와 벼멸구 저항성 유전자 Bph18을 가지고 있는 SR30071-3-7-23-6-1-1-1계통을 교배하여 전통육종법과 분자육종법을 활용하여 새로운 벼멸구 저항성 계통을 선발하여 병해충 저항성과 농업형질을 조사 수행하였다. 1. 20개체의 F1을 양성하여 F2 1,200개체를 포장에 전개하 였다. 포장에서 선발된 개체에 대해 실내미질선발을 수행하여 23개체의 F3를 공시하였으며 벼멸구 유묘검정, 도열병 밭못자 리검정, 흰잎마름병 K1 균계 접종검정을 실시한 후 F4 세대 를 거쳐 100계통을 F5 세대로 공시하였다. 2. F5 세대에서 DNA 마커 검정으로 100계통 중 Bph18은 92계통, Xa3는 96계통, Stv-bi는 98계통에서 저항성 유전자가 확인되었다. 생물검정에서는 100계통 중 벼멸구 유묘검정은 94계통이 저항성을, 흰잎마름병 K3 균계검정은 27계통 및 73 계통이 각각 저항성 및 중도저항성 반응을 보였다. 3. F5 세대 100계통 중 주요 농업적 형질이 우수한 11계통 을 생산력검정시험에 공시하였고, 보통기 보비재배에서 농업 형질이 양호하고, 수량이 높으며 병해충 저항성을 보이는 우 량한 4계통을 선발하여 이삭모양과 초형이 우수하며 출수기는 중만생종이며, 간장이 작고 수량이 높은 HR28011-1-2-3을 ‘익 산562호’로 계통명을 부여하였다. 본 실험에서 육성한 ‘익산562호’는 지역적응성 평가를 통한 품종화를 추진 중이며 벼멸구 및 기타 주요 병해충에 복합저 항성 벼품종육성을 위한 유전자원으로 활용되기를 기대한다.
To determine differential gene expression profiles in a cypermethrin-resistant strain (CR) of diamondback moth, Plutella xylostella Linnaeus (1758), a subtractive cDNA library was constructed by suppression subtractive hybridization. A total of 1196 expressed sequence tags (EST) were clustered and assembled into 579 contigs (100 multiplets and 479 singletons). About 46% (267) of 579 contigs had the matched BLASTx hits (E ≤ 10-5). Among these, 143 contigs had similarity to proteins with assigned molecular function in the Functional Catalogue database, and most of them (86%) were homologous to the genes from insects, particularly to Lepidoptera (56%). The contigs encoding carboxylesterase and cytochrome P450 known to be involved in the insecticide resistance were found in the library. They were identified as pxest3, pxest4, and CYP9G2 gene by 5' and 3' RACE. Among these, pxest4 was determined to be 2-fold over-transcribed in the CR strain by qrtPCR. Several contigs encoding enzymes including cytochrome oxidase subunit I that are likely involved in the insecticide resistance were also identified from the library. Discovery of the genes specific to cypermethrin resistance should promote further studies on the molecular mechanisms of insecticide resistance in P. xylostella.
Cytokinin은 식물의 성장과 발달에 중요한 역할을 하는 필수 호르몬이다. mRNA differential display 방법으로 애기장대 amp1 돌연변이체로부터 cytokinin에 의하여 발현이 유도되는 PR4 유전자를 분리하였다. AtPR4로 명명한 애기장대 PR4 유전자는 212개의 아미노산으로 구성되어 있었으며 분자량은 22,900이고 등전점은 7.89로 추정되었다. Genomic DNA 분석결과, AtPR4는 single copy 유전자인
To assess the environmental risks of transgenic chili pepper with PepEST gene on non-target organisms before it exposes to the agro-ecosystem environments, we conducted the three sets of green peach aphids (Myzus persicae S.) life table experiment under laboratory conditions (Temp. 25℃, R.H. 50-70%, Photoperiod L16 : D8) in series during 2005-2006. We measured the net reproductive rate (R₀)*, the intrinsic rate of increase (r<SUB>m</SUB>), the mean generation time (T<SUB>c</SUB>), fecundity*, life span, and reproduction period between non-transgenic chili peppers and transgenic chili peppers, respectively. The life span of green peach aphids from three sets was 31, 27, 25 days, and the period of life span was similar to the general average length of green peach aphids, 25-29 days. Although the first reproduction of transgenic pepper was similar to the non transgenic pepper (P>0.05), the fecundity and the net reproductive rate (R<SUB>o</SUB>) by using Jackknife method of transgenic pepper were lower than those of non transgenic pepper (P<0.05). Conclusively, we observed the adverse effect from our results but we should execute further experiments to confirm the results at the fields with the similar way.
대부분의 식중독은 단체 급식으로부터 발생하는 경우가 많으며, 특히 위생상태와 연관되어 식중독을 야기 시키는 병인 물질 중 포도상 구균은 많은 부분을 차지 하고 있다. 따라서 본 연구에서는 서부경남지역의 5개 초등학교 급식시설에서 총 98개의 샘플 중 A급식소의 식수, D급식소의 손, E급식소의 냉장고와 앞치마에서 4개의 포도상 구균을 분리하였다. 분리된 균주들은 1개의 메티실린 저항성 혈장응고 효소 음성 황색포도상구균(Methicilline Resistant Coagulase Negative Staphylococcus aureus: MRCNS땃 깨의 메치실린 민감성 혈장응고효소 양성 황색포도상구균 (Methicilline sensitive Coagulase positive Staphylococcus aureus· MSCPS)으로 구분되었다. 한편 포도상 구균은 내열성 내독소로서, 이 중 가장 문제가 되는 내독소 B(enterotoxin B)를 검색하기 위한 PCR을 실시한 결과, A장소의 식수, D장소의 손, E 장소의 냉장고와 앞치마에서 분리된 균주로부터 477b의 생성물을 갖는 sob gene을 확인할 수 있었다. 이들 분리된 황색포도상구균에 대한 항생제 민감성 실험에서는 ampicillin과 penicillin에 대하여 전체적으로 저항성을 가졌으며, 특히 A 식수에서 분리된 균주는 옥사실린 저항성(oxacilline resistant)균주로 나타나 MRSA(methicilline resistant staphylococcus aureus)균주로 확인되었다.
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.
시들음병은 진균성 병원균인 Fusarium oxysporum f. sp. conglutinans에 의해 양배추(Brassica oleracea var. capitata)에 발병하는 심각한 병해이다. 최근, 양배추의 시들음병 저항성 유전자, FocBo1, 과 연관된 분자 마커 (MTK-C)에 대한 보고가 있었다. 본 연구에서는, MTK-C를 이용하여, 국내 시들음병 저항성 육종을 위한 양배추 육종 계통이 FocBo1 유전자와 연관된 것인지 판별하고자 하였다. 상업적으로 판매되고 있는 ‘대박나’를 대조군으로 하여 6가지의 양배추 육종계통을 통제된 실험실 조건에서, 시들음병을 처리하고, 생물검정에 따른 발병도와 식물체내 병원균의 생체량을 확인하여 육종계통의 시들음병에 대한 반응성을 정확히 평가 하였다. 그결과, KR-518, OK-517은 반복 실험을 통해 병징 뿐만 아니라 병원균의 바이오 매스도 거의 검출되지 않는 강한 저항성을 나타냈고, RK-P6-1, MT-624는 실험실내 병 처리 환경에 따라 그 저항성과 감수성의 정도가 달라지는 각각 중도 저항성, 감수성으로 분류 할 수 있으며, JK-2, HY-164는 병징과 바이오매스 모두에서 병에 대한 감수성을 보임을 확인 하였다. FocBo1 간의 상관관계를 알기 위하여, 6개 양배추 계통의 genomic DNA를 추출하고, MTK-C 마커를 이용하여 PCR을 수행한 결과, 흥미롭게도 감수성 유전자원인 HY-164 식물체에서도 FocBo1 밴드가 관찰 되었다. 염기서열의 차이를 확인하고자 증폭된 밴드를 클로닝하여 sequencing 한 결과 감수성 HY-164와 저항성 KR-518 간의 차이는 없었다. 하지만, HY-164의 식물체에서는 FocBo1 유전자의 발현이 없는 것을 cDNA를 이용한 RT-PCR 결과로 확인 하였으며, 더하여, FocBo1 유전자의 발현 정도가 완전 저항성, 중도 저항성을 결정하는 중요 요인이라는 것을 실험을 통해 확인 할 수 있었다. 더하여, 확인된 MTK-C 밴드의 염기서열을 이용하여 단일밴드로 정확히 FocBo1 발현을 확인할 수 있는 프라이머, FocBo1-C를 만들 수 있었다. 본 실험을 통하여, 국내에서 보유한 6개 시들음병 관련 양배추 육종계통의 정확한 병 반응성을 정리 하였으며, 또한 이들의 시들음병 저항성이 FocBo1 유전자 및 발현과 밀접한 관련이 있다는 것을 확인할 수 있었다. 따라서, FocBo1을 이용한 분자육종에서 저항성과 감수성 판별을 위하여 genome 상의 FocBo1 존재뿐만 아니라 그 발현의 정도도 같이 확인 하여야 한다는 것을 확인하였다.
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.
It is necessary to carry out a risk assessment to determine the consequences of releasing a particular plant species containing specific transgenes before transgenic plants can be grown under filed conditions. Gene flow from transgenic plants to wild closely related species has raised concern recently. Since transgenic crops were released in 1996, the global area of transgenic crops has been increasing rapidly. The transgene introgression from transgenic crops to their wild relatives is unavoidable in some species. Transgene introgression is of concern because the crop–wild plant hybrids might be conferred with a selection advantage to increase their performance, which could result in negative ecological consequences to natural ecosystems. The genus Brassica has 159 species, including a number of wild species that are of great importance to the economy. Most transgenic Brassica gene flow research has focused on the most successful cross between transgenic oilseed rape Brassica napus and its wild relatives Brassica rapa, a widely distributed weed in the farming system in Europe and America, since the hybridization can spontaneously happen and the generations can backcross to B. rapa easily in the wild conditions. In this study, we aimed to characterize transgene introgression, segregation, and expression in backcrossed generations between tramsgenic B. napus and B. rapa. These results will contribute to the environmental risk assessment and assist in biosafety management.
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.
Recently whole genome SNP genotyping has been used to do association analysis and to map a gene of interest. Here we report application of bulked segregant analysis(BSA) using Infinium HD assay with ‘BARC Bean6K_3’, a SNP genotyping beadchip containing 5,399 SNPs for common bean to locate a target gene. We used BSA using Infinium HD assay was performed to find the candidate region of a single dominant rust resistant gene in PI310762, a common bean cultivar. And SSR markers were identified and mapped on the candidate region using F2 population derived from the cross of susceptible Pinto114 x resistant PI310762. BSA revealed the candidate region of the resistant gene is on chromosome 4 where we developed nine SSR markers. Three SSR markers (beanssr1170, beanssr1168, and beanssr1167) of them appeared closely linked to the resistant gene which is located between beanssr1167 at 0.1cM and beanssr1170 at 0.5cM on chromosome 4. This study showed BSA using high-throughput whole genome SNP genotyping is a very fast and efficient method to locate a gene of interest on chromosome.
본 연구는 최근 콩 재배에서 심각한 병으로 대두된 콩 불마름병에 대한 저항성 유전자인 rxp 근접분자표지를 개발하고자 수행하였다. 콩 불마름병은 국내에서 전국적으로 발생하는 심각한 세균병으로 이에 관련하여 세균병 접종을 이용한 저항성 품종과 감수성 품종에 대한 연구가 많이 진행되고 있지만 정확한 유전자의 염기서열이 밝혀져 있지 않고 있다. 본 연구는 콩 불마름병에 저항성 품종 8개체와 감수성 품종 8개체를 이용하여 rxp 유전자 근접분자표지를 확인하기