• BMB Reports
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• 제41권5호
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• pp.376-381
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• 2008

The discovery of RNA silencing inhibition by virus encoded suppressors or low temperature leads to concerns about the stability of transgenic resistance. RNA-dependent RNA polymerase (RdRp) has been previously characterized to be essential for transgene-mediated RNA silencing. Here we showed that low temperature led to the inhibition of RNA silencing, the loss of viral resistance and the reduced expression of host RdRp homolog (NtRdRP1) in transgenic T4 progeny with untranslatable potato virus Y coat protein (PVY-CP) gene. Moreover, RNA silencing and the associated resistance were differently inhibited by potato virus X (PVX) and tobacco mosaic virus (TMV) infections. The increased expression of NtRdRP1 in both PVX and TMV infected plants indicated its general role in response to viral pathogens. Collectively, we propose that biotic and abiotic stress factors affect RNA silencing-mediated resistance in transgenic tobacco plants and that their effects target different steps of RNA silencing.

• Journal of Plant Biology
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• 제5권3호
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• pp.30-36
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• 1962

The mechanism of synthesis of the toacco mosaic virus(TMV) and the potato virus X(PVX) was investigated using the methods of ultraviolet light irradiation and serological analysis. In vitro irradiation of UV on the infected tobacco juice for 10 minutes caused the infectivity of TMV and PVX to decrease markedly on their respective local lesion indicator hosts, Nicotiana glutinosa L. and Gomphrena globosa L., indicating that UV destroys directly the infectivity of the virus particles. Ten minutes after the UV was irradiated on the leaves of the two indicator hosts before inoculation, the infectivity of TMV decreased as it was irradiated in vitro, whereas that of PVX increased by 26% as compared with the unirradiated control. When the two viruses were mix-inoculated in the common host of tobacco and the synthetic products were analyzed by serological methods for a two week infection period, it was found that both viruses were multiplying more rapidly and abundantly than they were singly inoculated into the same host species. Titers from mixed series were often two times as high as those of singly inoculated series. A mechanism of competition in the synthesis between the mixed viruses in the common host is postulated.

• 한국응용곤충학회지
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• 제18권2호
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• pp.89-100
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• 1979

[ $\lceil$ ]한국산 명나방과(Pyralidae)에 관하여는 이미 필자(1976)에 의하여 언급된바와 같이 1889년 영국인 곤충학자 J.H. Leech가 처음으로 9종의 신종을 발표한데 이어 현재까지 300여종이 기록되어 왔다. 그러나 이들의 기록이 여러종류의 발표문에 산재해 있는 상태로 체계적인 정리가 이루어진바 없으며 학명뿐아니라 우리말 이름조차 갖가지로 쓰여지고 있어 곤충명 사용에 혼란이 야기되고 있는 실정이다. 이에 필자는 현재까지 발표되었던 종들을 조사하여 최근의 분류체계에 따라 학명 및 그들의 동의어(Synonym)를 정리함과 아울러 우리말 이름도 먼저 명명된 이름이나 또는 현재 많이 사용되고 있는 이름을 택하여 통일화하고 ( )안에 그 이명을 표시하였다. Evergestiinae(새들명나방아과, 신칭)은 현재까지는 주로 Pyraustinae(들명나방아과)에 포함되는 족(Tribe)으로 취급되어 왔으나 최근의 분류체계에 따라 독립된 아과로 취급하였다. 이들에 대해서는 Leech에 이어 일본인 Shibuya(1927년에 의하여 13종, 박세욱등 (1958, 1969)에 의하여 27종이 새로 기록되는등 1968년에 출판된 한국 동물명집 곤충편에는 94종이 수록되었다. 최근 필자(1976)에 의하여 우리나라 미기록종으로 발표된 24종과 기타 국내외의 문헌을 통한 기록의 근거를 기초로 작성된 이 목록에는 Evergestiinae에 1속 3종, Pyraustinae에 53속 123종이 포함되었으며 학명이 불분명하거나 채집의 근거가 불확실한 4종은 후미에 별도로 기재하였다. 분포란의 괄호안에 명시된 채집지는 필자에 의하여 채집 확인된 종들로서 그들의 표본은 농업기술연구소 곤충연구담당관실 표본실에 소장되어 있다. 다른 아과에 관하여는 정리가 완료되는대로 추후 발표할 예정이다.nemella hengsungica, Aphelenchoides besseyi 등으로 2아목 7과 11종이었다.는 내연가공한 옷생산에 사용되지 않는 것이 타당하겠다. 주아의 즙액에 의해서도 C. amaranticolor에 기계적 전염이 되었다. 7. C. amaranticolor 상에 계통분리된 마늘 모자이크 바이러스의 내열성은 $65^{\circ}-70^{\circ}C$, 희석한계는 $10^{-}2-10^{-3}$, 그리고 보존한계는 2 일이었다. 8. 마늘 모자이크 바이러스의 순화는 2회의 분획원심과 Sephadex gel filtration에 의해서 가능했다. 9. 전자현미경하에서 관찰한 마늘 모자이크 바이러스는 길이 1200-1225mu 폭 10-12mu의 사상이었다. 10. 혈청학적 미량침강 반응법에 의해 마늘잎에서뿐만 아니라 인편과 주아에서도 마늘 모자이크 바이러스의 검정이 가능했다. 11. 우리나라 5개 지방에서 수집한 마늘 종구 150개와 주아 30개에 대해 혈청학적방법으로 마늘 모자이크 바이러스의 감염률을 조사한 결과 $100\%$의 감염률을 보였다. 12. 마늘 모자이크 바이러스와 크기가 근사한 Potato Virus X. Potato virus Y, Potato virus S, Potato virus M 등과의 혈청학적 유연관계를 조사한 바, 마늘 모자이크 바이러스는 이들과 구별되는 다른 바이러스라고 생각된다. 13. 마늘의 모자이크 감염주에서 단일계통으로 분리하여 본 실험에 사용한 바이러스는 마늘의 바이러스 무감염주를 얻을 수가 없기 때문에 직접 마늘잎에 접종해서 모자이크톤의 병원이라는 것을 확인할 수 없었지만, 검정식물상의 반응, 혈청학적반응, 전자현미경적 관찰등의 간접적인 조사 결과로 미루어 미인록의 마늘모자이크 바이러스라고 생각된다

• The Plant Pathology Journal
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• 제22권2호
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• pp.139-146
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• 2006

The 3' region of Potato virus X (PVX) has the 74 nt 3'-nontranslated region (NTR) that is conserved among all potexviruses and contains several cis-acting elements for minus-strand and plus-strand RNA accumulation. Three stem-loop structures (SL1-SL3), especially formation of SL3 and U-rich sequence of SL2, and near upstream elements in the 3' NTR were previously demonstrated as important cis-acting elements. To Investigate the binding of these cis-acting elements within 3' end with host protein, we used the electrophoretic mobility shift assays (EMSA) and UV-cross linking analysis. The EMSA with cellular extracts from tobacco and RNA transcripts corresponding to the 150 nt of the 3' end of PVX RNA showed that the 3' end of PVX formed complexes with cellular proteins. The specificity of protein binding was confirmed through competition assay by using with 50-fold excess of specific and non-specific probes. We also conducted EMSA with RNAs containing various mutants on those cis-acting elements (${\Delta}10$10, SL3B, SL2A and ${\Delta}21$; J Mol Biol 326, 701-720) required for efficient PVX RNA accumulation. These analyses supported that these cis-acting elements are required for interaction with host protein(s). UV-cross linking analysis revealed that at least three major host proteins of about 28, 32, and 42 kDa in mass bound to these cis-elements. These results indicate that cis-acting elements from 3' end which are important for minus and plus-strand RNA accumulation are also required for host protein binding.

• 농업과학연구
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• 제44권2호
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• pp.145-180
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• 2017

Alternanthera mosaic virus (AltMV) is a member of the genus Potexvirus which has been known for less than twenty years, and has been detected in Australasia, Europe, North and South America, and Asia. The natural host range to date includes species in at least twenty-four taxonomically diverse plant families, with species in at least four other families known to be infected experimentally. AltMV has been shown to differ from Potato virus X (PVX), the type member of the genus Potexvirus, in a number of ways, including the subcellular localization of the Triple Gene Block 3 (TGB3) protein and apparent absence of interactions between TGB3 and TGB2. Differences between AltMV variants have allowed identification of viral determinants of pathogenicity, and identification of residues involved in interactions with host proteins. Infectious clones of AltMV differing significantly in symptom severity and efficiency of RNA silencing suppression have been produced, suitable either for high level protein expression (with efficient RNA silencing suppression) or for Virus-Induced Gene Silencing (VIGS; with weaker RNA silencing suppression), demonstrating a range of utility not available with most other plant viral vectors. The difference in silencing suppression efficiency was shown to be due to a single amino acid residue substitution in TGB1, and to differences in subcellular localization of TGB1 to the nucleus and nucleolus. The current state of knowledge of AltMV biology, including host range, strain differentiation, host interactions, and utility as a plant viral vector for both protein expression and VIGS are summarized.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2015년도 추계학술대회 및 정기총회
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• pp.26-26
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• 2015

Phytophthora infestans is the causal agent of potato and tomato late blight, one of the most devastating plant diseases. P. infestans secretes effector proteins that are both modulators and targets of host plant immunity. Among these are the so-called RXLR effectors that function inside plant cells and are characterized by a conserved motif following the N-terminal signal peptide. In contrast, the effector activity is encoded by the C terminal region that follows the RXLR domain. Recently, I performed in planta functional profiling of different RXLR effector alleles. These genes were amplified from a variety of P. infestans isolates and cloned into a Potato virus X (PVX) vector for transient in planta expression. I assayed for R-gene specific induction of hypersensitive cell death. The findings included the discovery of new effector with avirulence activity towards the Solanum bulbocastanum Rpi-blb2 resistance gene. The Rpi-blb2 encodes a protein with a putative CC-NBS-LRR (a coiled-coil-nucleotide binding site and leucine-rich repeat) motif that confers Phytophthora late blight disease resistance. We examined the components required for Rpi-blb2-mediated resistance to P. infestans in Nicotiana benthamiana. Virus-induced gene silencing was used to repress candidate genes in N. benthamiana and to assay against P. infestans infections. NbSGT1 was required for disease resistance to P. infestans and hypersensitive responses (HRs) triggered by co-expression of AVRblb2 and Rpi-blb2 in N. benthamiana. RAR1 and HSP90 did not affect disease resistance or HRs in Rpi-blb2-transgenic plants. To elucidate the role of salicylic acid (SA) in Rpi-blb2-mediated resistance, we analyzed the response of NahG-transgenic plants following P. infestans infection. The increased susceptibility of Rpi-blb2-transgenic plants in the NahG background correlated with reduced SA and SA glucoside levels. Furthermore, Rpi-blb2-mediated HR cell death was associated with $H_2O_2$, but not SA, accumulation. SA affects basal defense and Rpi-blb2-mediated resistance against P. infestans. These findings provide evidence about the roles of SGT1 and SA signaling in Rpi-blb2-mediated resistance against P. infestans.

• Plant Biotechnology Reports
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• 제5권4호
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• pp.331-344
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• 2011

The well-conserved NBS domain of resistance (R) genes cloned from many plants allows the use of a PCR-based approach to isolate resistance gene analogs (RGAs). In this study, we isolated an RGA (CapRGC) from Capsicum annuum "CM334" using a PCR-based approach. This sequence encodes a protein with very high similarity to Rx genes, the Potato Virus X (PVX) R genes from potato. An evolutionary analysis of the CapRGC gene and its homologs retrieved by an extensive search of a Solanaceae database provided evidence that Rx-like genes (eight ESTs or genes that show very high similarity to Rx) appear to have diverged from R1 [an NBS-LRR R gene against late blight (Phytophthora infestans) from potato]-like genes. Structural comparison of the NBS domains of all the homologs in Solanaceae revealed that one novel motif, 14, is specific to the Rx-like genes, and also indicated that several other novel motifs are characteristic of the R1-like genes. Our results suggest that Rx-like genes are ancient but conserved. Furthermore, the novel conserved motifs can provide a basis for biochemical structural. function analysis and be used for degenerate primer design for the isolation of Rx-like sequences in other plant species. Comparative mapping study revealed that the position of CapRGC is syntenic to the locations of Rx and its homolog genes in the potato and tomato, but cosegregation analysis showed that CapRGC may not be the R gene against PVX in pepper. Our results confirm previous observations that the specificity of R genes is not conserved, while the structure and function of R genes are conserved. It appears that CapRGC may function as a resistance gene to another pathogen, such as the nematode to which the structure of CapRGC is most similar.

• 식물병연구
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• 제28권4호
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• pp.237-244
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• 2022

국화(Dendranthema morifolium)는 한국에서 경제적으로 중요한 화훼식물에 포함되고, 바이러스와 바이로이드 병들에 의해 경제적 피해를 받고 있다. 본 연구에서는 충청남도 예산군에서 재배된 국화 350개체를 대상으로 7종의 바이러스들과 2종의 바이로이드들(Chrysanthemum chlorotic mottle viroid, CChMVd; Chrysanthemum stunt viroid, CSVd; Cucumber mosaic virus, CMV; Chrysanthemum virus B, CVB; Chrysanthemum stem necrosis orthotospovirus, CSNV; Impatiens necrotic spot orthotospovirus, INSV; Potato virus X, PVX; Tomato aspermy virus, TAV; Tomato spotted wilt orthotospovirus, TSWV)을 검정하였다. 그 결과 2종의 바이러스들(CVB-CN-Y, TAV-CN-Y)과 2종의 바이로이드들(CChMVd-CN-Y, CSVd-CN-Y)이 검정되었다. CVBCN-Y는 6개의 국화 식물체들에서 검정되었고, TAV-CN-Y는 한개가 국화 식물체에서만 검정되었다. CChMVd-CN-Y는 97개의 국화 식물체들에서 검정되었으며, CSVd-CN-Y는 21개의 국화 식물체들에서 검정되었다. CVB-CN-Y는 CVB-GS1과 86.9% 염기서열 상동성을 보였으며, TAV-CN-Y는 3개의 분리주들(TAV-Chj, TAV-P, TAV-V)과 100% 염기서열이 일치하였다. CChMVd-CN-Y는 CChMVd-Horst와 99.5% 염기서열 상동성을 보였으며, CSVd-CN-Y는 4개의 분리주들(Au1.1, K4pop, Sagae, Tochigi)과 99.7% 염기서열 상동성을 보였다. 본 연구는 2021년 예산군에서 재배된 국화들을 대상으로 바이러스 및 바이로이드들을 검정하고 그들의 감염률에 관한 보고서이다.

• The Plant Pathology Journal
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• 제40권1호
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• pp.59-72
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• 2024

A comprehensive survey of mungbean-growing areas was conducted to observe leaf spot disease caused by Alternaria alternata. Alternaria leaf spot symptoms were observed on the leaves. Diversity of 50 genotypes of mungbean was assessed against A. alternata and data on pathological traits was subjected to cluster analysis. The results showed that genotypes of mungbean were grouped into four clusters based on resistance parameters under the influence of disease. The principal component biplot demonstrated that all the disease-related parameters (% disease incidence, % disease intensity, lesion area, and % of infection) were strongly correlated with each other. Alt a 1 gene that is precisely found in Alternaria species and is responsible for virulence and pathogenicity. Alt a 1 gene was amplified using gene specific primers. The isolated pathogen produced similar symptoms when inoculated on mungbean and tobacco. The sequence analysis of the internal transcribed spacer (ITS) region, a 600 bp fragment amplified using specific primers, ITS1 and ITS2 showed 100% identity with A. alternata. Potato virus X (PVX) -based silencing vector expressing Alt a 1 gene was constructed to control this pathogen through RNA interference in tobacco. Out of 50 inoculated plants, 9 showed delayed onset of disease. Furthermore, to confirm our findings at molecular level semi-quantitative reverse transcriptase polymerase chain reaction was used. Both phenotypic and molecular investigation indicated that RNAi induced through the VIGS vector was efficacious in resisting the pathogen in the model host, Tobacco (Nicotiana tabacum). To the best of our knowledge, this study has been reported for the first time.

• 한국산림과학회지
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• 제84권1호
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• pp.77-86
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• 1995

임목을 대상으로 삽입된 외래 유전자의 공간적, 시기별 발현 특성을 이해하기 위한 기초연구로서 온실에서 생육 중인 형질전환된 2년생 잡종 포플러 (Populus alba X P. grandidentata) Hansen 클론을 대상으로 삽입된 외래 유전자의 발현정도를 각 기관별로 조사하였다. Agrobacterium binary vector pRT45, pRT102 및 pRT104에 의해서 형질전환된 3계통의 형질전환체 Tr15, Tr345, Tr665 모두는 선발가능한 표식 유전자로서 nos promoter-NPT II 유전자가 대상 식물체의 genome에 삽입되어 있으며, 그외에, pin2 promoter-CAT 유전자(pRT45), nos promoter-PIN2 유전자(pRT102), Cauliflower Mosaic Virus 35s promoter-PIN2 유전자(pRT104)가 3계통의 형질전환체에 제각각 삽입되어 있는 잡종 포플러이다. 이들 3계통의 형질전환 포플러 식물체의 DNA를 PCR 검정 기법을 이용하여 분석해 본 결과 선발 가능한 표식 유전자인 NPT-II가 삽입되어 있음이 입증되었다 발현 정도를 비교 분석하기 위해서 NPT-ELISA 검정을 실시하였다. 삽입된 NPT II 유전자는 형질전환된 포플러의 잎, 엽병, 형성층 조직, 줄기의 목질부, 뿌리에서 발현되었으며, 발현 정도는 형질전환된 식물체의 계통에 따라서, 그리고 형진전환된 식물체의 부위에 따라서 다양하게 나타났다. pRT45에 의해서 형질전환된 Tr15 형질전환체의 경우, 늙은 잎과 엽병에서 NPT II 유전자가 가장 높은 수준으로 발현되었으며, 어린 잎과 뿌리 조직에서 가장 낮게 발현되었다. 삽입된 외래 유전자가 각 식물체간에, 각 기관에 따라서 각각 상이한 발현 정도를 나타내는 이와 같은 결과는 형질전환된 식물체에 대한 효과적인 선발과정이 요구됨을 의미함은 물론이고, 형질전환 식물체의 발달 과정에 따라서 삽입된 외래 유전자가 공간적, 시기적으로 각각 다르게 발현할 수 있다는 것을 나타낸다.