• Research in Plant Disease
• /
• v.29 no.2
• /
• pp.193-199
• /
• 2023

The apple stem grooving virus (ASGV) is one of the most harmful latent viruses infecting pear orchards worldwide. To examine the genetic diversity of ASGV in Korean pear orchards, the complete coat protein (CP) gene of five ASGV isolates collected from various regions were identified. The five Korean ASGV isolates showed 88-96% nucleotide identity with the 11 isolates worldwide occurring elsewhere in the world. Phylogenetic analysis of five isolates, as well as the previously sequenced isolates, indicated that the ASGV clusters had no correlation with the host or geographical regions of origin. Recombination analysis showed that one of the five Korean isolates is a recombinant, with a recombination site in the CP gene region (nt 532-708). This study is the first report of natural recombination within the CP gene of ASGV isolates from pears grown in Korea.

• Proceedings of the Zoological Society Korea Conference
• /
• 1994.10a
• /
• pp.243.2-243
• /
• 1994

No Abstract, See Full Text

• The Plant Pathology Journal
• /
• v.33 no.3
• /
• pp.296-306
• /
• 2017

Sugarcane mosaic virus (SCMV) is one of the most damaging viruses infecting sugarcane, maize and some other graminaceous species around the world. To investigate the genetic diversity of SCMV in Iran, the coat protein (CP) gene sequences of 23 SCMV isolates from different hosts were determined. The nucleotide sequence identity among Iranian isolates was more than 96%. They shared nucleotide identities of 75.5-99.9% with those of other SCMV isolates available in GenBank, the highest with the Egyptian isolate EGY7-1 (97.5-99.9%). The results of phylogenetic analysis suggested five divergent evolutionary lineages that did not completely reflect the geographical origin or host plant of the isolates. Population genetic analysis revealed greater between-group than within-group evolutionary divergence values, further supporting the results of the phylogenetic analysis. Our results indicated that natural selection might have contributed to the evolution of isolates belonging to the five identified SCMV groups, with infrequent genetic exchanges occurring between them. Phylogenetic analyses and the estimation of genetic distance indicated that Iranian isolates have low genetic diversity. No recombination was found in the CP cistron of Iranian isolates and the CP gene was under negative selection. These findings provide a comprehensive analysis of the population structure and driving forces for the evolution of SCMV with implications for global exchange of sugarcane germplasm. Gene flow, selection and somehow homologous recombination were found to be the important evolutionary factors shaping the genetic structure of SCMV populations.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2003.10a
• /
• pp.114.2-115
• /
• 2003

Rice stripe virus causes severe damage to rice in Korea, Japan and China. RSV is a type member of the tenuivirus group and transmitted by the small brown planthopper, Laodeiphax striatellus, in a persistant manner. Until now, occurrence of RSV is limited in of southern part of Korea. But recently occurrence of RSV is increasing and spreading in central part of Korea including Chungcheong and Kyonggi province. So we analyzed recent occurrence trend of RSV which is increased and cloned and sequenced coat protein gene for isolating of RSV strain. Infected rice of each species(Ilpumbyeo, Sindongjinbyeo, Keumobyeo-2, Dongjinbyeo, Jongnambyeo, Samcheonbyeo, etc.)is collected, we extracted total RNA from infected leaves and detected RSV viral RNA by reverse transcription(RT)-PCR using specific primer of coat protein gene. The result of RT-PCR, we observed specific band. We already cloned cDNA from the band, is analyzing sequence variety and homology of each species.

• The Plant Pathology Journal
• /
• v.31 no.1
• /
• pp.41-49
• /
• 2015

Sweet cherry is an important fruit crop with increasing economical value in Turkey and the world. A number of viruses cause diseases and economical losses in sweet cherry. Prune dwarf virus (PDV), is one of the most common viruses of stone fruits including sweet cherry in the world. In this study, PDV was detected from 316 of 521 sweet cherry samples collected from 142 orchards in 10 districts of Isparta province of Turkey by double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA). The presence of PDV in ELISA positive samples was confirmed in 37 isolates by reverse transcription- polymerase chain reaction (RT-PCR) method. A genomic region of 862 bp containing the coat protein (CP) gene of PDV was re-amplified from 21 selected isolates by RT-PCR. Amplified DNA fragments of these isolates were purified and sequenced for molecular characterization and determining genetic diversity of PDV. Sequence comparisons showed 84-99% to 81-100% sequence identity at nucleotide and amino acid level, respectively, of the CP genes of PDV isolates from Isparta and other parts of the world. Phylogenetic analyses of the CP genes of PDV isolates from different geographical origins and diverse hosts revealed that PDV isolates formed different phylogenetic groups. While isolates were not grouped solely based on their geographical origins or hosts, some association between phylogenetic groups and geographical origins or hosts were observed.

• Research in Plant Disease
• /
• v.18 no.3
• /
• pp.231-235
• /
• 2012

Indian citrus ring spot virus (ICRSV) is known to cause a serious disease to citrus, especially to Kinnow mandarin, the popular cultivated citrus species in India. In this study, we developed diagnostic systems based on enzyme-linked immunosorbent assay (ELISA). In order to generate antibodies against ICRSV coat protein, we overexpressed the coat protein in Escherichia coli using the pET15b expression vector containing an optimized ICRSV coat protein gene. The recombinant ICRSV coat protein was overexpressed as soluble form at $37^{\circ}C$ upon IPTG induction. The protein was purified to 95% in purity by Ni-NTA column chromatography. The purified protein was immunized to rabbit for the generation of polyclonal antibody (PAb). The PAb showed a specific immunoreaction to recombinant ICRSV coat protein in western blot analysis and ELISA. Diluted rabbit antisera (10,000 fold) could detect less than 10 ng and 5 ng of the target protein in western blot and ELISA analysis, respectively.

• The Plant Pathology Journal
• /
• v.29 no.3
• /
• pp.260-273
• /
• 2013

The incidence and distribution of Tobacco mosaic virus (TMV) and related tobamoviruses was determined using an enzyme-linked immunosorbent assay on 1,926 symptomatic horticultural crops and 107 asymptomatic weed samples collected from 78 highly infected fields in the major horticultural crop-producing areas in 17 provinces throughout Iran. The results were confirmed by host range studies and reverse transcription-polymerase chain reaction. The overall incidence of infection by these viruses in symptomatic plants was 11.3%. The coat protein (CP) gene sequences of a number of isolates were determined and disclosed to be a high identity (up to 100%) among the Iranian isolates. Phylogenetic analysis of all known TMV CP genes showed three clades on the basis of nucleotide sequences with all Iranian isolates distinctly clustered in clade II. Analysis using the complete CP amino acid sequence showed one clade with two subgroups, IA and IB, with Iranian isolates in both subgroups. The nucleotide diversity within each subgroup was very low, but higher between the two clades. No correlation was found between genetic distance and geographical origin or host species of isolation. Statistical analyses suggested a negative selection and demonstrated the occurrence of gene flow from the isolates in other clades to the Iranian population.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 1994.06a
• /
• pp.86-102
• /
• 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.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2003.10a
• /
• pp.75.1-75
• /
• 2003

Barely yellow mosaic(BaYMV) and barely mild mosaic (BaMMV) bymoviruses are both transmitted by the soil-inhabiting fungus Polymyxa gramnis, and are responsible for economic losses in barley crops in Asia and Europe. Because chemical control of the vector is ineffective, the losses can only be prevented by growing resistant barley cultivars. The objective of this study is to produce resistant barley plants by transformation with viral coat protein(cp) genes. Resistance tests of T1 plants transformed with the BaYMV CP gene showed that at least four independent lines had clear resistance to BaYMV but two other lines were highly susceptible with severe symptoms. The CP gene was detected in all resistant T1 plants by genomic PCR. Most of T2 progenies derived from the resistant T1 lines also showed resistance. In contrast, only one out of 21 independent T2 lines transformed with the BAMMV CP gene tested showed clear resistance to BaMMV, and others were very susceptible. Further analyses of resistance and CP gene expression are in progress.

• The Plant Pathology Journal
• /
• v.19 no.1
• /
• pp.19-23
• /
• 2003

Turnip crinkle vims (TCV) inoculation onto resistant Arabidopsis ecotype Dijon（Di-17) leads to a hypersensitive response (HR) on the inoculated leaves. A dominant gene, HRT, which confers an HR to TCV, has been cloned from Di-17 plants by map-based cloning. HRT is a LZ-NBS-LRR class resistance gene and it belongs to a small gene family that includes RPP8, which confers resistance to Peronospora parasitica Emco5. Outside of the LRR region, HRT and RPP8 proteins share 98% amino acid identity while their LRR regions are less conserved (87% identity). HRT-transformed Arabidopsis plants developed an HR but generally remained susceptible to TCV due to a dominant RRT allele, which is not compatible with resistance. However, several transgenic plants that over-expressed HRT much higher than Di-l7 showed micro-HR or no HR when inoculated with TCV and were resistant to infection. Both the HR and resistance are dependent on salicylic acid but independent of NPRI, ethylene, or jasmonic acid. Arabidopsis plants containing both TCV coat protein gene and HRT developed massive necrosis and death in seedlings, indicating that the TCV coat protein is an avirulence factor detected by the HRT.