Seung-Kook Choi

The Effect of Cucumber mosaic virus 2b Protein to Transient Expression and Transgene Silencing Mediated by Agro-infiltration

The transient and rapid expression system of a foreign protein in planta is a very useful technique in biotechnology application. We have investigated optimum condition of Agrobacterium-infiltration technique in which expression level of foreign proteins were maximized without detrimental effects on plants using GFP and Cucumber mosaic virus 2b protein, which is known as an enhancer of gene expression and a suppressor of post-transcriptional gene silencing(PTGS). The optimum expression level of both RNA and protein of GFP with minimum leaf impairment was obtained at

Variation in the Pathogenicity of Lily Isolates of Cucumber mosaic virus

Two isolates of Cucumber mosaic virus (CMV) originated from lily plants, named Ly2-CMV and Ly8-CMV, were compared with their pathological features in several host plants. Ly2-CMV and Ly8-CMV could induce systemic mosaic symptom in Nicotiana benthamiana, but Ly2-CMV could not systemically infect tomato and cucumber plants that have been used for CMV-propagative hosts. While Fny-CMV used as a control infected systemically the same host plants, producing typical CMV symptoms. Ly8-CMV could infect systemically two species of tobacco (N. tabacum cv. Xanthi-nc and N. glutinosa) and zucchini squash (Curcubita pepo), but Ly2 failed systemic infection on these plants. As resulted from tissue-print immunoblot assay, different kinetics of systemic movement between Ly2-CMV and Ly8-CMV were crucial for systemic infection in tobacco (cv. Xanthi-nc). Sequence analysis of full-length genome of two lily isolates showed Ly2 and Ly8 belonged to subgroup IA of CMV. The lily isolates shared overall 98 % sequence identity in their genomes. Coat protein, 3a protein, and 2b protein involved in virus movement was highly conserved in genomes of the isolates Ly2 and Ly8. Although there is the low frequency of recombinants and reassortants in natural CMV population, phylogenetic analysis of each viral protein among a number of CMV isolates suggested that genetic variation in a defined population of CMV lily isolates was stochastically produced.

First Report of Myrothecium roridum Causing Leaf and Stem Rot Disease on Peperomia quadrangularis in Korea

Abstract In 2010, symptoms of leaf and stem rot were observed on potted plants (Peperomia quadrangularis) in a greenhouse in Yongin, Korea. The causative pathogen was identified as Myrothecium roridum based on morphological data, internal transcribed spacer sequence analysis, and pathogenicity test. To our knowledge, this is the first report of M. roridum causing leaf and stem rot disease on P. quadrangularis in Korea and elsewhere worldwide.

Genome analysis and characterization of a tobacco mosaic virus isolate infecting balsam (Impatiens balsamina)

The complete RNA genomic sequence of a tobacco mosaic virus isolate infecting Impatiens balsamina, designated as TMV-IM, has been determined. The genomic sequence and the predicted gene products of TMV-IM were similar to those of other members of the genus Tobamovirus. The CP ORF of TMV-IM showed sequence identities of 95.0–99.5% with the corresponding ORFs of other TMV strains. Full-length cDNA of TMV-IM was amplified by RT-PCR with a 5′-end primer harboring a T7 promoter sequence and a 3′-end TMV-specific primer. Subsequently, the full-length cDNA was cloned into plasmid vectors. Capped transcripts synthesized from the cDNA clone were highly infectious and caused characteristic symptoms in balsam plants, similar to wild-type TMV-IM and TMV-U1. These results provide definitive evidence for the natural occurrence of TMV in balsam.

High sequence conservation among Odotoglossum ringspot virus isolates from orchids

The variability in the nucleotide (nt) and amino acid (aa) sequences of the coat protein (CP) of Odontoglossum ringspot virus (ORSV), which naturally infects orchids worldwide, was investigated. The CP genes of 48 ORSV isolates originating from different locations in Korea were amplified using RT-PCR and sequenced. The encoded CP consists of 158 aa. The CP sequences of the Korean isolates were compared at the nt and aa levels with those of the previously published ORSV isolates originating from different countries. The Korean isolates share 94.8–100% and 92.4–100% CP identity to ORSV isolates from other countries at the nt and aa levels, respectively. No particular region of variability could be found in either sequence of the viruses. In the deduced aa sequence, the N-terminal region was more conserved than the C-terminal region in ORSV. The phylogenetic tree analysis and recombination analysis revealed that there was no distinct grouping between geographic locations and sequence identity, and nor distinct intra-specific recombination events among ORSV isolates.

Manipulation of induced resistance to viruses

Induced resistance against plant viruses has been studied for many years. However, with the exception of RNA silencing, induced resistance to viruses remains mechanistically less well understood than for other plant pathogens. In contrast, the induction processes involved in induced resistance, comprising basal resistance signaling, effector-triggered immunity, and phytohormone pathways, have been increasingly well characterized in recent years. This has allowed induced resistance to viruses to be placed in a broader conceptual framework linking it to other defense systems, which we discuss in this review. We also discuss the range of agents, including chemicals and beneficial microorganisms and application methods that can be used to induce resistance to viruses.

Characterisation of a satellite RNA of Cucumber mosaic virus that induces chlorosis in Capsicum annuum

The presence of Cucumber mosaic virus (CMV) satellite RNA dramatically changes symptoms on some hosts. A satellite RNA present in a strain of CMV (PepY-CMV) that induced chlorosis in pepper (Capsicumannuum) was shown to induce chlorosis in pepper in combination with another strain (Fny-CMV) that by itself induced a green mosaic symptom. The location of sequences within the PepY satellite RNA (PepY-satRNA) of CMV that conferred the ability to induce chlorosis on pepper plants were analyzed by exchanging sequence domains between cDNA clones of PepY-satRNA and an attenuated mosaic satellite RNA (Paf-satRNA), as well as site-directed mutagenesis of various clusters of the 22-nt sequence differences between the two satellite RNAs in the delimited central domain. The symptoms induced by site-directed mutants of PepY-satRNA and Paf-satRNA in the presence of Fny-CMV demonstrated an insertion within PepY-satRNA of 11 nt at positions 86–96 relative to Paf-satRNA determined the chlorosis-inducing phenotype. Within the chlorosis-inducing domain, deletion of nucleotides did not affect the satRNA replication but abolished the ability of PepY-satRNA to elicit chlorosis symptom. Conversely, a mutant satellite RNA derived from Paf-satRNA in which eleven nucleotides were inserted indicated that sequences of 11 nucleotides were found to be sufficient for chlorosis induction in pepper.

Analysis of Symptom Determinant of Cucumber mosaic virus RNA3 via Pseudorecombinant Virus in Zucchini Squash

Isolates of Cucumber mosaic virus (CMV) collected in Korea, were compared with their pathological features in tobacco and zucchini squash. Full-length cDNA clone of RNA3 was generated by using long-distance RT-PCR. Transcript RNA3 from the cDNA clone was inoculated onto host plants with transcripts RNA1 and RNA2 of Fny strain, generating RNA3-pseudorecombinant CMV. Timing and severity of systemic symptom was not significantly different among the pseudorecombinant CMVs in tobacco, compared with strains Fny-CMV and Pf-CMV. However, the pseudorecombinant CMVs induced two different systemic symptoms (mosaic vs. chlorotic spot) in zucchini squash. Based on symptom induction, the pseudorecombinant CMVs were categorized into two classes. The severity and timing of symptoms were correlated with viral RNA accumulations in systemic leaves of zucchini squash, suggesting that different kinetics of virus movement associated with CMV proteins are crucial for systemic infection and symptom development in zucchini squash. The analysis of movement proteins (MP) of CMV strains showed high sequence homology, but the differences of several amino acids were found in the C-terminal region between Class-I-CMV and Class-II-CMV. The analysis of coat proteins (CP) showed that the CMV isolates tested belonged to CMV subgroup I and the viruses shared overall 87-99% sequence identity in their genomes. Phylogenetic analysis of MP and CP suggested that biological properties of Korean CMV isolates have relationships associated with host species.

A Simple and Reliable Molecular Detection Method for Tomato yellow leaf curl virus in Solanum lycopersicum without DNA Extraction

This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. A Simple and Reliable Molecular Detection Method for Tomato yellow leaf curl virus in Solanum lycopersicum without DNA Extraction

Biological Characterization and Sequence Analysis of Cucumber mosaic virus isolated from Capsicum annuum

Whereas most of isolates of Cucumber mosaic virus(CMV) can induce green mosaic systemic symptoms on zucchini squash, foliar symptoms of a pepper isolate of CMV (Pf-CMV)-infected zucchini squash revealed systemic chlorotic spots. To assess this biological property, infectious full-length cDNA clones of Pf-CMV were constructed using long-template RT-PCR. The complete nucleotide sequences of RNA2 and RNA3 of Pf-CMV were determined from the infectious fulllength cDNA clones, respectively. RNA 2 and RNA3 of Pf-CMV contain 3,070 nucleotides and 2,213 nucleotides, respectively. Overall sequence homology of two RNAs revealed high similarity (90%) between CMV strains, and 60% similarity to those of Tomato aspermy virus and Peanut stunt virus strains. By sequence analysis with known representative strains of CMV, Pf- CMV belongs to a typical member of CMV subgroup IA. The virus has high evolutionary relationship with Fny-CMV, but the pathology of Pf-CMV in zucchini squash was quite different from that of Fny-CMV. The pesudorecombinant virus, F1P2P3, induced chlorotic spot leaf symptom and timing of systemic symptom in squash plants, similar to the plants infected by Pf-CMV. No systemic symptoms were observed when Pf-CMVinoculated cotyledons were removed at 5 days postinoculation (dpi) while Fny-CMV showed systemic symptom at 2 dpi. These results suggest that the pepper isolate of CMV possesses unique pathological properties distinguishable to other isolates of CMVs in zucchini squash.