• Journal of Life Science
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• 제9권1호
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• pp.14-16
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• 1999

Genetic variability was monitored by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) in dicofol-susceptible (S), dicofol-resistant (R) and its reverse-selected (RS) strains of two-spotted spider mite, of Tetranychus urticae. Before the reverse-selection, RS strain, selected reversely from R strain, was 23-fold resistance ratio at {TEX}$LC_{50}${/TEX} to S strain. The resistance was started to in incline slowly to the resistance level of S strain after one year, and the resistance ratio was 4-fold in the 7 years after then. PCR-amplification of T. urticae DNA showed polymorphism in the amplifications with 12 primers in 100 kinds of arbitrary DNA sequences. RAPD amplification with primer OPR-12 (5`-ACAGGTGCGT-3`) showed amplified bands at 1,000 base pair in the S-and RS-strain, and at 350 base pair in R-strain. The results of polymorphism are genetic variabilities derived from development and selection of resistance in each strain. The peculiarly amplified fragments were guessed to participate in dicofol resistance. From the analysis of genetic similarity, it is inferred the gene composition of S-and RS-strain is much closer than that of R-strain.

• 환경위생공학
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• 제13권2호
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• pp.34-39
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• 1998

For more convenient and rapidly isolation of Bacillus thuringiensis subsp. israelensis(Bti), 1) heat treatment spore forming bacteria, 2) growth in enrichment culture media for Bacillus sp. and 3) selection of bacteria producing a lecithinase for Bacillus thuringiensis subsp. israelensis, were performed. Spore forming bacteria were counted 4.8 $\times $ 10$^{8}$cells/g soil on NAPGCY media, 9.2 $\times $ 10$^{7}$cells/g soil on NA media, and 3.6 $\times $ 10$^{8}$cells/g soil on NAAC media, respectively. Bacteria producing only a lecithinase were reached at 25.2% on medium contained egg york, bacteria only producing a delta-endotoxin were reached at 23.2% by phase contrast microscope, and bacteria producing a lecithinase & a delta-endotoxin simultaneously were reached at 13.7%. Bacillus thuringiensis which producing a lecithinase and a delta-endotoxin simultaneously among bacteria producing a lecithinase, were reached at 56.5%; A half of Bacillus thuringiensis was produced a delta-endotoxin, but not produced a lecithinase. Among 8 isolates of Bacillus thuringiensis, two strain of Bti which has a mosquito-cidal toxin, were detected by PCR using a specific primer of $\delta $-endotoxin gene from Bti.

• 아시안잔디학회지
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• 제18권4호
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• pp.211-219
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• 2004

Transgenic zoysiagrass (Zoysia japonica cv. Zenith) plants have been obtained by particle bombardment of embryogenic callus with the plasmid pSMABuba, which contains hygromycin resistance (hpt) and bialaphos resistance (bar) genes. Parameters on DNA delivery efficiency of the particle bombardment were partially optimized using transient expression assay of a chimeric $\beta-glucuronidase$(gusA) gene driven by the CaMV 35S promoter. Stably transfarmed zoysiagrass plants were recovered with a selection scheme using hygromycin. Transgenic zoysiagrass plants were confirmed by PCR analysis with specific primer for bar gene. Expression of the transgene in transformed zoysiagrass plants was demonstrated by Reverse transcriptase (RT)-PCR analysis. All the tested transgenic plants showed herbicide BastaR resistance at the field application rate of $0.1\%-0.3\%$.

• 한국산림과학회지
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• 제96권2호
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• pp.125-130
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• 2007

The principal morphological characters used for identification of hydrangea cultivars are often dependent on agroclimatic conditions. Furthermore, information on the selection or the genetic background of the hydrangea breeding is so rare that a molecular marker system for cultivar identification is needed. Amplified fragment length polymorphism (AFLP) markers were employed for fingerprinting Hydrangea macrophylla cultivars and candidate cultivars of H. macrophylla selected in Korea. One AFLP primer combination was sufficient to distinguish 17 H. macrophylla cultivars and 4 candidate cultivars. The profile of 19 loci that can minimize the error of amplification peak detection was constructed. AFLP markers were efficient for identification, estimation of genetic distances between cultivars, and cultivar discrimination. Based on the observed AFLP markers, genetic relationship was reconstructed by the UPGMA method. Seventeen H. macrophylla cultivars and H. macrophylla for. normalis formed a major cluster, and candidate cultivars selected in Korea formed another cluster.

• Mycobiology
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• 제51권6호
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• pp.463-467
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• 2023

Soybean is one of the world's most widely cultivated food crops, and soybean seeds are supplied from national seed resources in Korea. However, the transmission of seed-borne diseases through infected soybean seeds is problematic. Among these diseases, soybean seed decay is caused by Diaporthe spp. Infecting the pods, and the infected seeds show rotting symptoms. Most diseased seeds are removed during the selection process; however, it is difficult to distinguish infected seeds that do not display symptoms. Hence, a sequencebased method was devised to screen Diaporthe-infected seeds. Based on the nuclear ribosomal internal transcribe spacer (ITS) region of the pathogen, a primer was designed to distinguish the infection from other soybean seed pathogens. As a result of the comparison between healthy and Diaporthe-diseased seeds by using the primers, Diaporthe was detected only in the diseased seeds. Therefore, it is possible to distribute healthy soybean seeds by detecting Diaporthe-diseased seeds at the genetic level using the Diaporthe-specific primers.

• 식물조직배양학회지
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• 제24권2호
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• pp.113-117
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• 1997

Simple sequence repeats (SSR)는 진핵생물체에 널리 산재되어 있으며, 큰 다형현상을 나타내고, polymerase chain reaction (PCR)으로 쉽게 분석된다. 이 연구의 목적은 서로 다른 Chlamydomonas reinhardtii계통간의 다형현상과 Chlamydomonas의 SSR 좌위에서의 유전양상을 결정하는데 있었다. C. reinhardtii의 genomic DNA library를 만들어 $^{32}$P로 라벨링한 (AC)$_{11}$ probe를 이용하여 (CA/GT)n 반복서열을 가지는 clone을 선택하기위해 screen하였다. 선택된 clone을 sequencing하여 (CA/GT)n sequence에 인접한 PCR primer set를 제조하였다. PCR은 여러 C. reinhardtii 계통의 SSR 좌위를 증폭하기 위하여 사용하였다. 그 좌위는 및몇 C. reinhardtii 계통에서 다형현상을 보였다. 그러나 그 좌위에서 C. reinhardtii의 6계통중 4계통만 DNA가 PCR 증폭을 하였고 2계통은 증폭을 하지 않았다. C. reinhardtii와 C. smithii의 교배로 생긴 4배체에서 2:2의 분리비를 보여주었는데, 이는 단순한 멘델의 유전양상을 나타낸다. 이러한 결과로 미루어 SSR 다형현상은 Chlamydomonas의 개체 식별, 개체군 연구, 연쇄 분석, 그리고 유전자 지도 작성을 하는데 유용할 것이다.다.

• 원예과학기술지
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• 제16권2호
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• pp.213-214
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• 1998

본 연구는 Agrobacterium tumefaciens를 이용한 형질전환의 효율성을 높이기 위한 기초자료를 제공하고자 실시되었다. 선발배지에 사용되는 kanamycin에 대한 토마토 자엽전편체의 감응성 검정결과 50mg/L가 적합한 것으로 제시되었다. 토마토 절편체에는 부정적인 영향을 주지 않고 배지내 Agrobacterium의 제거에 적합한 cefotaxime의 농도로는 200mg/L가 선정되었다. Agrobacterium과의 공동배양에 의해 자엽절편체로부터의 callus형성과 신초 재분화가 현저히 억제되었으며 공시된 3품종의 토마토의 경우에는 3일간의 공동배양기간이 callus형성과 신초 재분화에 적합한 것으로 판단되었다. 재분화 신초에 대한 형질전환여부검정은 GUS염색법과 NPTII primer를 이용한 PCR검정을 실시하였으며 공시된 3품종에서 모두 형질전환 신초를 획득하였다.

• 한국육종학회지
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• 제43권2호
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• pp.133-138
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• 2011

The groundnut or cultivated peanut (Arachis hypogaea L.) in Korea consists of 36 domestic varieties which have been developed and registered as cultivars for the public during last 25 years. To screen and identify of Korean peanut varieties and genetic resources, we present a simple and reliable method. A methodology based on simple sequence repeat (SSR) markers developed and widely used for prominent gene identification and variety discrimination. For identification of those 36 Korean peanut varieties, 238 unique peanut SSR markers were selected from some previously reported results, synthesized and used for polymerase chain reaction (PCR). Data were taken through acryl amide gel electrophoresis and changed into proper formats for application of data mining analysis using Biomine (all-in-one functional genomics data mining program). Consequently, twelve SSR primers were investigated and revealed the differences between those 36 varieties. These primer pairs amplified 27 alleles with an average of 2.3 allele per primer pair. In addition, those results showed genetic relationship by classification method within 36 varieties. The approach described here could be applied to monitoring of our varieties and adapting to peanut breeding program.

• 식물병연구
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• 제17권1호
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• pp.52-58
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• 2011

우리나라에서 주로 재배되는 십자화과 작물(상추, 콜라비, 무, 배추, 양배추)의 종자 전염 병원균 중에서 세균성 병원균 Xanthomonns campestris pv. campestris(Xcc)와 바이러스 병원균 Lettuce Mosaic Virus(LMV)를 종자에서 동시검출하기 위한 One-step multiplex RT-PCR을 개발하였다. 각각의 병원균을 특이적으로 증폭시키는 병원균 검출용 프라이머 2종(Xcc-F/R, LMV-F/R)을 primerblast 프로그램을 이용하여 제작하였고 이들 프라이머 세트는 프라이머간 또는 병원균 cDNA간의 간섭없이 특이적으로 타겟 병원균만을 검출하였다. PCR을 이용한 병원균의 검출 최소 민감도는 1 ng이었다. 십자화과 작물중에서 유통 중인 콜라비 10품종, 상추 50품종, 무 20품종, 배추 20품종 그리고 양배추 20품종에 대한 종자 감염 병원균 검출을 위한 One-step multiplex RT-PCR 수행 결과, LMV는 전체 120품종 중에서 39품종에서 검출되었고, Xcc는 2개 품종에서 검출되었다. 그리고 50품종의 상추 종자 시료 중에 8품종의 시료에서 LMV와 Xcc가 동시 검출되었다.

• 식물병연구
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• 제17권1호
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• pp.44-51
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• 2011

우리나라에서 주로 재배되는 가지과작물의 종자 전염 병원균 중에서 세균성 병원균 Xanthomonns campestris pv. vesicatoria(Xcv), Clavibacter michiganensis subsp. michiganensis (Cmm), Erwinia carotovora subsp. carotovora (Ecc)와 바이러스 병원균 Pepper mild mottle virus (PMMoV) and Tobacco mild green mosaic virus (TMGMV)를 종자로부터 동시검출하기 위한 One-step Multiplex RT-PCR을 개발하였다. 각각의 병원균을 특이적으로 증폭시키는 병원균 검출용 프라이머 15세트를 primer-blast 프로그램을 이용하여 제작하였고 각각의 병원균을 특이적으로 검출할 수 있는 5세트의 프라이머 세트(Cmm-F/R, Ecc-F/R, Xcv-F/R, PMMoV-F/R, TMGMVF/R)를 선발하였다. 이들 프라이머 세트는 프라이머간의 또는 병원균 cDNA간의 간섭없이 타겟 병원균만을 검출하였다. 가지과 작물 중에서 유통 중인 고추종자 20품종과 토마토종자 10품종에 대한 종자 감염 병원균 검출을 위한 PCR을 수행한 결과, 2개 품종의 토마토종자에서 Ecc가 검출되었고 4개 품종의 고추종자에서도 Ecc가 검출되었다. 특히 고추 종자에선 Ecc가 검출된 4개 품종 외에 1개 품종에서 PMMoV, TMGMV가 동시 검출되었다. 이로 인해 개발된 One-step multiplex RT-PCR은 서로 간섭 현상 없이 병원균을 효율적으로 검출할 수 있음을 보여 주었다.