• Journal of Plant Biology
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• 제35권3호
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• pp.253-257
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• 1992

Potyvirus group에 속하는 것으로 알려진 마늘 모자이크 바이러스 (GMV)가 식물에서 병을 유도하는 메카니즘을 이해하기 위하여, 마늘에 존재하는 GMV에 대한 cDNA clone인 clone 29-6을 분리하였다. Northern blot 분석에 의해 이 바이러스의 genome size는 약 9 kb이고, 또한 clone 29-6은 GLV genome과 유사성이 없었으며, 마늘잎으로부터 분리된 $poly(A)^{+}$ RNA와 강하게 hybridization되었다. 이러한 사실은 이 cDNA clone이 마늘에 감염하여 모자이크 병반을 유도하는 GMV에 대한 cDNA clone 중의 하나인 것으로 생각되었다. Clone 29-6을 probe으로 사용하여 마늘 바이러스의 cDNA 은행을 탐색하여 이와 중첩되는 clone을 선별하여 염기서열을 결정한 결과 이들의 염기서열이 중첩부위에서 서로 일치하지 않았는데 이는 마늘은 여러 형태의 GMV에 의해 감염되어 있음을 암시한다.

• 한국식물병리학회지
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• 제12권4호
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• pp.389-394
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• 1996

도열병균, Magnaporthe grisea, 균주간의 유전적 유연관계를 분석하고 그들의 유전에 관한 '기본 정보를 얻고자 DNA polymorphism 분석을 실시하였다. 기주가 다른 도열병 균주들이 공시되었고 cloning에 의해 벼 도열병균 KJ201레이스 균주로부터 생성된 임의 선발 genomic clone들이 공시균주들간의 polymorphism을 밝히기 위해 사용되었던 바 그중 repectitive sequence를 보유한 repeated copy clone 하나가 선발되었다. Clone pMJ6에 의해 밝혀진 repetitive sequence는 Southern hybridization시 벼 분리균주에는 약 30개, 다른 기주 분리균에도 20∼33개의 밴드를 형성하였다. 반면 피 분리균주에는 단지 두 개의 밴드만을 나타내 분리기주가 다른 균주간에 뚜렷한 polymorphism이 존재하였으며 parsimony 분석에서도 역시 아주 먼 cluster를 형성하여 피 분리균은 다른 기주 분리균과 유전적으로 상당히 먼 것으로 추정되었다. 공시균의 genomic DNA를 HindIII로 처리했을 때 pMJ6에 의한 밴드양상은 공시균을 EcoRI으로 처리했을 때의 MGR probe의 밴드 양상과 유사하여 이 repeated copy clone이 도열병균주간의 유전적 유연관계를 분석하는데 MGR 못지않게 유용할 것으로 보인다.

• 식물조직배양학회지
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• 제26권2호
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• pp.77-83
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• 1999

Agrobacterium tumefaciens $A_4$T에 의해 형질전환된 사리풀 (Hyoscyamus niger L.)의 모상근으로부터 tropane alkaloids (scopolamine, hyoscyamine)의 생산성 향상을 위한 최적배지, 배양기간 및 탄수화물의 영향을 조사하였다. Tropane alkaloids의 생산성이 우수한 모상근 (HNl8, HN57)의 최적 배지는 SH배지이며, 모상근의 생장, tropane alkaloids 함량 및 생산성에서 조사한 6가지 배지보다 높게 나타났다. Tropane alkaloids생산을 위한 최적 배양기간은 HNl8 clone이 6주, HN57 clone은 5주로 나타났으며, 배양기간에 따라 scopolamine의 함량은 계속적으로 증가하는데 반하여 hyoscyamine의 함량은 일정 수준을 유지하는 것으로 나타났다. 두 세포주는 sucrose농도가 7%까지 증가할수록 생장률이 증가하는 반면, tropane alkaloids 함량은 감소하였으며, tropane alkaloids생산성은 3% sucrose처리구에서 가장 높게 나타났다. HN18 clone의 경우 최적 sucrose(3%)및 dextrose (2%)에서 유사한 tropane alkaloids 생산성을 나타내었다. 하지만 HN57 clone은 sucrose 처리구에 비하여 dextrose 처리구에서 매우 낮은 tropane alkaloids생산성을 나타내었다.

• Plant Resources
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• 제4권3호
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• pp.111-120
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• 2001

The efficiency of in vitro regeneration of four clones of Populus euramericana, Canada blanc, Eugenii, I-45/51, and Wisconsin #5, was examined. Cytokinins and the combinations with auxins affected the rate of regeneration from the explants of root segments, stem internodes, and leaf discs. Overall, BA and the combination with auxins were effective in root segments and leaf discs of the Canada blanc clone, whereas zeatin and the combination with auxins were important in stem internodes of the Wisconsin #5 clone. The highest number of shoots averaging 17.6 $\pm$ 0.47 from root segments in the Canada blanc clone,18.2 $\pm$ 3.0 from stem internodes in the Wisconsin #5 clone, and 17.8 $\pm$ 1.92 from leaf discs in the Canada blanc clone were obtained with 2.0 mg/1 BA, 2.0 mg/l zeatin combined with 0.2 mg/l IAA, and 0.5 mg/l BA combined with 0.05 mg/l 2,4-D, respectively. In particular, the addition of 2,4-D into cytokinin medium promoted shoot proliferation.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 1994년도 Proceedings of International Symposium on BIOLOGICAL CONTROL OF PLANT DISEASES Korean Society of Plant Pathology
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• pp.86-102
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• 1994

To understand the molecular structure and pathogenesis mechanism of Korean garlic viruses, we have isolate cDNA clones for garlic viruses. The partial nucleotide sequences of 24 cDNA clones were determined and that of six clones containing poly (A) tail were compared with those of other plant viruses. One of those clones, V9 has 81.8% similarity in nucleotide sequence and 93.0% in deduced amino acid sequence, respectively, to the coat protein gene for garlic mosaic virus (GMV). Northern blot analysis with the clone V9 demonstrated that the genome of GMV is 7.8 kb long and has poly (A) tail. The anti-coat protein antibody for GMV recognizes 35 kDa polypeptide which could be the coat protein of GMV from infected garlic leaf extract or virus preparation. Clone G7 has about 62% of deduced amino acid sequence identity with the members of potyvirus group. Northern blot analysis with the clone G7 demonstrated that the genome of the potyvirus I garlic is 9.0 kb long and has poly (A) tail. The third clone, S81, shows 42% amino acid identity to the potexvirus. The other clones are under the characterization. To test the possibility of producing garlic virus resistant plant, we have designed a hairpin type ribozyme to cleave V9 RNA at the middle of the coat protein gene. From the cleavage reactions in vitro with two different sizes of RNA substrates, V9SUB (144 nucleotides) and V9 RNA (1,361 nucleotides), the ribozyme can cleave V9 sequence effectively at the predicted site. To study the activity of the ribozyme in vivo, plant transformation is in progress. Further possibilities to produce garlic virus resistant plant will be discussed.

• Journal of Plant Biology
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• 제33권4호
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• pp.303-307
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• 1990

A genomic clone carrying a proteinase inhibitor I sequence was isolated and characterized. The clone contained a 0.7 kb EcoRI fragment hybridized with tomato inhibitor I cDNA. The nucleotide sequence of the EcoRI fragment revealed presence of a truncated form of a proteinase inhibitor I gene of potato. The truncated gene contained the 5' flanking region and the first exon of a functional proteinase inhibitor I gene. Although the 5' flanking region contained the regulatory sequences TATAAA and CCACT, a deletion of 40 bp occurred between them.

• BMB Reports
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• 제29권1호
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• pp.52-57
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• 1996

A specific DNA fragment from Korean radish (Raphanus sativus L.) was amplified by performing PCR with oligonucleotide primers which correspond to the highly conserved regions of plant peroxidases. The size of the PCR product was ca. 400 bp, as expected from the known plant peroxidase genes. Comparison of the nucleotide and deduced amino acid sequences of the PCR product to those of other plant peroxidase-encoding genes revealed that the amplified fragment corresponded to the highly conserved region I and III of plant peroxidases. By screening a genomic library of Korean radish using the amplified fragment as a probe, two positive clones, named prxK1 and prxK2, were isolated. Restriction mapping studies indicated that the 5.2 kb Sail fragment of the prxK1 clone and the 4.0 kb EcoRI fragment of the prxK2 clone encode separate isoperoxidase genes. Analyses of the promoter region of the prxK1 clone shows that putative CAAT box, CMT box, and TGA1b binding sequence (5' TGACGT) are present 718 bp upstream from the start codon.

• 농업과학연구
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• 제49권4호
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• pp.873-883
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• 2022

American plum line pattern virus (APLPV), a member of the genus Ilarvirus in the family Bromoviridae, is one of the plant quarantine pathogens in Korea. In this study, 15 candidate primer sets were designed and examined to develop a reverse transcription polymerase chain reaction (RT-PCR) assay for plant quarantine inspection of APLPV. Using APLPV-infected and healthy samples, the primer sets were assessed for APLPV detection. To confirm the occurrence of nonspecific reactions, six ilarviruses (Apple mosaic virus, Asparagus virus 2, Blueberry shock virus, Prune dwarf virus, Prunus necrotic ringspot virus, and Tobacco streak virus) and 10 target plants (Prunus mume, P. yedoensis, P. persica, P. armeniaca, P. dulcis, P. tomentosa, P. avium, P. glandulosa, P. salicina, and P. cerasifera) were examined. Finally, two primer sets were selected. These primer sets could generate the expected amplicons even with at least 1 ng of the total RNA template in concentration-dependent amplifications. In addition, a positive clone was developed for use as a positive control in the abovementioned RT-PCR assay.

• The Plant Pathology Journal
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• 제37권2호
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• pp.162-172
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• 2021

Soybean mosaic virus (SMV) is the predominant viral pathogen that affects the yield and quality of soybean. The natural host range for SMV is very narrow, and generally limited to Leguminosae. However, we found that SMV can naturally infect Pinellia ternata and Atractylodes macrocephala. In order to clarify the molecular mechanisms underlying the cross-family infection of SMV, we used double-stranded RNA extraction, rapid amplification of cDNA ends polymerase chain reaction and Gibson assembly techniques to carry out SMV full-length genome amplification from susceptible soybeans and constructed an infectious cDNA clone for SMV. The genome of the SMV Shanxi isolate (SMV-SX) consists of 9,587 nt and encodes a polyprotein consisting of 3,067 aa. SMV-SX and SMV-XFQ008 had the highest nucleotide and amino acid sequence identities of 97.03% and 98.50%, respectively. A phylogenetic tree indicated that SMV-SX and SMV-XFQ018 were clustered together, sharing the closest relationship. We then constructed a pSMV-SX infectious cDNA clone by Gibson assembly technology and used this clone to inoculate soybean and Ailanthus altissima; the symptoms of these hosts were similar to those caused by the virus isolated from natural infected plant tissue. This method of construction not only makes up for the time-consuming and laborious defect of traditional methods used to construct infectious cDNA clones, but also avoids the toxicity of the Potyvirus special sequence to Escherichia coli, thus providing a useful cloning strategy for the construction of infectious cDNA clones for other viruses and laying down a foundation for the further investigation of SMV cross-family infection mechanisms.

• Journal of Plant Biology
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• 제39권3호
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• pp.197-202
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• 1996

A pollen-specific cDNA clone, LMP50, was isolated from the mature pollen cDNA library of the Easter lily. The LMP50 transcript was highly abundant in mture pollen grains but not detectable in other organs. The LMP50 cDNA clone contains 1383 nucleotides and two open reading frames. The first codes for a peptide of 15 amino acid residues. The role of this peptide is nuclear. The second encodes a protein containing 329 amino acid residues. This protein exhibited a significant homology to human tartrate-resistant acid phosphatase and porcine uteroferrin. Both of these enzymes have been suggested to play a role in iron transport. Therefore, LMP50 may act as an iron carrier protein in mature pollen grains.