• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.146.1-146
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• 2003

Transgenic Nicotiana benthmiana plants harboring and expressing coat protein (CP) gene of Zucchini green mottle mosaic virus (ZGMMV) were generated for both virus-resistant screening and complementation analysis of related viruses and environmental safety assessment (SA) of living modified organism (LMO) purposes. Transformation of leaf disc of N. benthamiana was performed using Agrobacterium-mediated method and the pZGCPPGA748 containing the ZGMMV CP and NPTII genes. Two kinds of transgenic homozygous groups, virus-resistant and -susceptible lines, were obtained by screening of challenging homologous virus for T1 generations. Complementation of CP-deficient related virus was analyzed using the susceptible line of ZGMMV. These two pathologically different lines can be useful for host-virus interactions and LMO environmental SA.

• The Plant Pathology Journal
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• v.20 no.3
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• pp.206-211
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• 2004

Transgenic Nicotiana benthamiana plants harboring coat protein (CP) gene of Zucchini green mottle mosaic virus (ZGMMV) were generated for virus-resistant screening and complementation analysis of related viruses for environmental safety assessment (SA) of living modified organism (LMO) purposes. Transformation of leaf disc of N.benthamiana was performed by using Agrobacterium-mediated method and the pZGC-PPGA748 containing the ZGMMV CP and NPTII genes. Two kinds of transgenic homozygous groups, virus-resistant and virus-susceptible N.benthamiana lines, were obtained by screening of challenging homologous virus for Tl generations. These two pathologically different lines can be useful for host-virus interactions and LMO environmental SA.

• Journal of the Korean Society of Tobacco Science
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• v.20 no.1
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• pp.50-56
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• 1998

The complementary DNA (cDNA) of potato virus Y- vein necrosis strain (PVY-VN) replicase gene (Nlb) was transformed into tobacco (Nicotiana tabacum cv. Burley 21) plants. Out of 25 putative transformants regenerated, 3 were resistant to PVY-VN, one highly resistant plant with no symptom until seed harvest time and the other two with mild chlorotic spot symptoms at late stages after infection. No symptom was observed in the highly resistant plant, while mild vein necrotic symptoms were developed on suckers of the moderately resistant plants after seed harvest time, In the first generation (T1) via self fertilization, resistance to susceptibility frequency in transgenic plants from the highly resistant transformant was about 3 : 1, while it was lowered much (about 1:2 and 1:19) in T1 of the moderately resistant transformants. In the second generation (T2) of the highly resistant plant, resistance frequencies were similar to T1, but resistance levels varied greatly and appeared to be decreased. Key words : potato virus Y, viral replicate gene, transgenic tobacco plants, resistance.

• Journal of Plant Biotechnology
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• v.37 no.4
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• pp.370-378
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• 2010

This review describes the stratagies of development of virus-resistant Alstroemeria plants using the genetic modification system. Despite of increasing of its importance in cut flower market, improvements of some horticultuirally important traits such as fragrance, long vase-life, virus resistance and tolerance against abiotic stresses are lack of the breeding program in Alstroemeria. Of these traits, virus-resistance is quite difficult to develop in Alstroemeria plants due to the limitations of genetic variation in the existed germplasm. To extend the genetic variation, plant biotechnological techniques such as genetic transformation and tissue culture should be combined to develop virus-resistant line in Alstroemeria. In this review, several strategies for the generation of virus-resistance by using natural resistance genes, pathogen-derived genes and other sources including pathogen-derived proteins, virus-specific antibodies and ribosome-inactivating proteins are presented. Also, brief histories of breeding, tissue culture, and transformation system in Alstroemeria plants are described to inderstand of the application of transgenic approach for the development of virus-resistance in Alstroemeria species.

• Journal of the Korean Society of Tobacco Science
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• v.6 no.2
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• pp.185-189
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• 1984

To classify the inheritance of resistance to potato virus Y, crosses between susceptible flue-cured tobacco variety NC 95 and resistant variety McNair 30 were conducted. The parents, $F_1$ plants, $F_2$ populations, and haploid plants derived from anthers of $F_1$ plants were screened for a resistance of two potato virus Y strains (PVY-VB and PVY-VN) isolated in Korea. The Chi-square values for the $F_3$ populations and haploids of $F_1$ fitted 1 :3 and 1 :1 ratios of resistant to susceptible for two strains, respectively. Therefore, it was found that the resistance of McNair 30 for the potato virus Y was controlled by a single recessive gene. Moreover the resistance to two strains screened was inherited dependently.

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.153-160
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• 1997

Cucumber mosaic virus (CMV) leads to a cause of poor crop productivity and quality. To solve this problem, we attempted to develop a virus-resistance tobacco plants by using viral coat protein (CP) gene. Transgenic tobacco plants expressing CMV CP gene were analysed by the resistance upon CMV infection. The virus-resistance was measured in $\textrm{T}_{1}$, generation by the inhibition of plant growth and the expression of the mosaic symptoms infected with CMV. The transgenic lines were divided into four groups: highly resistant, resistant, moderate and susceptible based on their growth and symptom severity. Out of 39 transgenic lines, 16 lines showed significant virus-resistance. And of resistant lines, 2 lines were designated highly resistant based on the facts that they achieved similar plant height to that of non-infected tobacco plants and showed lower disease symptom than that of other lines. The steady state level of CP RNA and coat protein level were measured by northern blot and immunoblot analysis. The CP RNA was highly accumulated in most resistant and moderate lines but barely detected in susceptible lines. The coat protein was detected in most lines regardless of their resistance to CMV. from this result, virus-resistance appeared to correlate more with CP RNA level than the level of coat protein. However, in two highly resistant lines, CP RNA level was unexpectedly low. This unexpected phenomenon need to be further investigated.

• The Plant Pathology Journal
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• v.19 no.3
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• pp.159-161
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• 2003

The coat protein (CP) gene of Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was selected for the design of virus-specific primers for amplification and molecular detection of the virus in cultivated apple. A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) was performed with a single pair of ApMV-specific primers and crude nucleic acid extracts from virus-infected apple for rapid detection of the virus. The PCR product was verified by restriction mapping analysis and by sequence determination. The lowest concentration of template viral RNA required for detection was 100 fg. This indicates that the RT-PCR for detection of the virus is a 10$^3$times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. The specificity of the primers was verified using other unrelated viral RNAs. No PCR product was observed when Cucumber mosaic virus (Cucumovirus) or a crude extract of healthy apple was used as a template in RT-PCR with the same primers. The PCR product (669 bp) of the CP gene of the virus was cloned into the plasmid vector and result-ant recombinant (pAPCP1) was selected for molecule of apple transformation to breed virus-resistant transgenic apple plants as the next step. This method can be useful for early stage screening of in vitro plantlet and genetic resources of resistant cultivar of apple plants.

• Journal of Plant Biotechnology
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• v.47 no.2
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• pp.164-171
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• 2020

Transgenic Alstroemeria plants resistant to Alstroemeria mosaic virus (AlMV) were generated through RNA-mediated resistance. To this end, the friable embryogenic callus (FEC) of Alstroemeria was induced from the leaf axil tissue and transformed with a DNA fragment containing the coat protein gene and 3'-nontranslated region of AlMV through an improved particle bombardment system. The bar gene was used as a selection marker. More than 300 independent transgenic FEC lines were obtained. Among these, 155 lines resistant to phosphinothricin (PPT) were selected under low stringent conditions. After increasing the stringency of PPT selection, 44 transgenic lines remained, and 710 somatic embryos from these lines germinated and developed into shoots. These transgenic shoots were then transferred to the greenhouse and challenged with AlMV. In total, 25 of the 44 lines showed some degree of resistance. PCR analysis confirmed the presence of the viral sequence. Virus resistance was observed at various levels. Establishment of an efficient transformation system for Alstroemeria will allow inserting transgenes into this plant to confer resistance to viral and fungal pathogens. Accordingly, this is the first report on the production of a transgenic virus-resistant Alstroemeria and lays the foundation for alternative management of viral diseases in this plant.

• The Plant Pathology Journal
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• v.22 no.3
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• pp.265-270
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• 2006

Cultivated hot pepper crops showing severe mosaic symptom were found in Korea in 2004 and their causal agent was identified as Cucumber mosaic virus (CMV). These pepper crops was resistant to the virus in the filled, and they belonged to pathotype 0 (P0) resistant pepper. Resistance screening of selected pepper plants showed that a pepper isolate of CMV was the P0 resistance-breaking virus. This P0 resistance-breaking isolate of CMV, named as Ca-P1, was isolated from leaves of the virus-infected Capsicum annuum cv. Manidda that showed systemic severe mosaic symptom. Ca-P1-CMV could induce systemic mosaic symptoms on P0-susceptible (P0-S) and P0-resistant (P0-R) cultivars whereas an ordinary strain (Fny-CMV) could not infect P0-R. This result suggests that Ca-P1-CMV can overcome P0 resistant pepper cultivars. To analyze its genome sequence, the complete nucleotide sequence of RNA3 of Ca-P1-CMV was determined from the infectious full-length cDNA clone of the virus. RNA3 of Ca-P1-CMV consisted of 2,219 nucleotides. Overall sequence homology of RNA3-encoded two viral proteins (movement protein and coat protein) revealed high similarity (75.2-97.2%) with the known CMV strains. By sequence analysis with known representative strains of CMV, Ca-P1-CMV belongs to a typical member of CMV subgroup IB. The resistance and resistance-breaking mechanisms of pepper and counterpart CMV, respectively, remain to be investigated, which will enrich the genetic resources and accelerate CMV-resistant pepper breeding programs.

• Biomedical Science Letters
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• v.16 no.4
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• pp.229-238
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• 2010

Chicken Mx protein (cMx) induced interferon (IFN) is an antiviral protein to inhibit replication of RNA virus, particularly negative stranded RNA virus, through blockage of transfortation of viral RNA and proteins. In order to determine antiviral effects of cMx from different MHC haplotype chicken, we characterized cMx gene by studying on nucleotide sequencing, antiviral effects to Newcastle disease virus, VSV and MDV, and transcription activities. Three types of eMx genes (2,118 bp) were detected from the different MHC haplotype chickens [B19 (N), B15(F) and B21 (GSP)] chickens, which have showed different susceptible to Marek's disease (MD). Several amino acid substitutions were showed in the cMx. The amino acid 548 and 631 in the cMxs from N and F, chickens susceptible to MD, was Val and Asn which was important on antiviral effects, and showed in resistant cMx. Those in the cMx from GSP, chicken resistant to MD, were same that showed in susceptible cMx. Though every cMx transactivated the expression of the reporter gene, the transcription activation by resistant cMx from N and F was lower compared to that by susceptible cMx from GSP. The decease of the cell growth in the resistant cMx cloned cells was seen in comparison with another cMx clone cells. Replication of NDV and VSV was suppressed in the clones with resistant cMx from N and F. NMx258-transducted cells lack of antiviral effects, and NMx437 or NMx646-transducted cells was showed 60% of antiviral effects compared to NMx705. Mean death time (MDT) and hemaggutination (HA) titer to NDV was long and low in the eggs of N and F lines, but short and high in the egg of GSP line. Interestingly, strong suppression to NDV was observed in the clone with N-Mx and in the eggs of N line. However, the effects of Mx for replication of vvMDV1 have not been. Thus, resistant types of cMx, N- and F-Mx, have showed the anti-viral effects to only RNA virus including NDV and VSV, but not to DNA virus. Antiviral effects of cMx were required whole length of amino acid including Val and Asn in amino acid 548 and 631.