• The Korean Journal of Pesticide Science
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• v.15 no.4
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• pp.545-572
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• 2011

The genetic diagnosis methods by RT-PCR and Virion capture (VC)/RT-PCR against Rice stripe virus (RSV) were developed. Three diagnosis methods of seedling test, ELISA and RT-PCR were compared in virus detection sensitivity (VDS) for RSV. The VDS of ELISA for RSV viruliferous small brown plant hopper (SBPH) was higher with 40.5% than that of seedling test. The VDS of RT-PCR was higher with 21% than that of ELISA. The VDS of ELISA and VC/RT-PCR was same with 9.2% in average on the SBPH collected from fields at the areas of Gimpo, Pyungtaeg and Sihueng, Gyeonggi province in 2009. The specific primers of RSV for SBPH and rice plant were developed for the diagnosis by Real time PCR. The RQ value of Real time PCR for the viruliferous and non viruliferous SBPH was 1 for 50 heads of non viruliferous SBPH, 96.5 for 50 heads of viruliferous SBPH, 23.1 for 10 heads of viruliferous SBPH + 40 heads of non viruliferous SBPH, and 75.6 for 30 heads of viruliferous SBPH + 20 heads of non viruliferous SBPH. The RQ value was increased positively by the ratio of viruliferous SBPH. Full sequences of 4 genomes of RSV RNA1, RNA2, RNA3 and RNA4 were analysed for the 13 RSV isolates from rice plants collected from different areas. Genetic relationships among the RSV isolates of Korea, Japan and China were classified as China + Korea, and China + Korea + Japan by phylogenetic analysis for RSV RNA1 and RNA2. In case of RNA3 involved in pathogenicity, genetic relationship of RSV among the three countries was grouped into 3 as China, China + Korea, and Korea + Japan. According to the genetic relationships in RSV RNA4, RSV isolates were grouped into 4 as China, Korea, China + Korea + Japan, and Korea + Japan. Viruliferous insect rate (VIR) of RSV in average increased in each year from 2008 to 2010, and the rates were 4.3%, 6.1%, and 7.2%, respectively, at the 28 major rice production areas in 7 provinces including Gyeonggido. The highest VIR in each year was 11.3% of Gyeonggido in 2008, 20.1% of Jellanamdo in 2009 and 14.2% of Chungcheongbukdo in 2010. The highest VIR depending upon the investigated areas was 22.1% at Buan of Jellabukdo in 2008, 36% at Wando and Jindo of Jellanamdo in 2009, and 30.0% at Boeun of Chungcheongbukdo in 2010. Average population density (APD) of overwintered SBPH was 13.1 heads in 2008, 13.9 heads in 2009 and 5.6 heads in 2010. The highest APD was 39.1 and 60.4 heads at Buan of Jellabukdo in 2008 and 2009, respectively, and 14.0 heads at Pyungtaeg of Gyeonggido. The acreage of RSV occurred fields was 869 ha in the western and southern parts, mainly at Jindo and Wando areas, of Jellanamdo in 2008. In 2009, RSV occurred in the acreage of 21,541 ha covered whole country, especially, partial and whole plant death were occurred with infection rate of 55.2% at 3,025 plots in 53 Li, 39 Eup/Myun, 19 Si/Gun of Gyeonggido, Incheonsi, Chungcheongnamdo, Jeollabukdo and Jeollanamdo. Seasonal development of overwintered SBPH was investigated at Buan, Jeollabukdo, and Jindo, Jeollanamdo for 3 years from 2008. Most SBPH developed to the 3rd and 4th instar on the periods of May 20 to June 10, and they developed to the adult stage for the 1st generation on Mid and Late June. In 2009, all SBPH trapped by sky net trap were adult on May 31 to June 1 at Mid-western aeas of Taean, Seosan and Buan, and South-western areas of Sinan and Jindo. The population density of adult SBPH was 963 heads at Taean, 919 at Seocheon and 819 at Sinan area. The origin of these higher population of adult SBPH were verified from the population of non-overwintered SBPH but immigrant SBPH. From Mid May to Mid June in 2010, adult SBPH could not be counted as immigrant insects by sky net trap. The variation of RSV VIR was high with 2.1% to 9.5% for immigrant adult SBPH trapped by sky net trap at Hongsung of Chungcheongbukdo, Buan of Jeollabukdo and so forth in 2009. The highest VIR for the immigrant adult SBPH was 9.5% at Boryung of Chungcheongnamdo, followed by 7.9% at Hongsung of Chungcheongnamdo, 6.5% at Younggwang of Jeollanamdo, and 6.4% at Taean of Cheongcheongnamdo. The infection rate of RSV on rice plants induced by the immigrant adult SBPH cultivated near sky net trap after about 10 days from immigration on June 12 in 2009 was 84.6% at Taean, 65.4% at Buan and 92.9% at Jindo, and 81% in average through genetic diagnosis of RT-PCR. Barley known as a overwintering host plant of RSV had very low infection rate of 0.2% from 530 specimens collected at 10 areas covering whole country including Pyungtaeg of Gyeonggido. Twenty nine plant species were newly recorded as natural hosts of RSV. In winter annual plant species, 11 plants including Vulpia myuros showed RSV infection rate of 24.9%. The plant species in summer annual ecotype were 13 including Digitaria ciliaris with 44.9%, Echinochloa crusgalli var. echinata with 95.2% and Setaria faberi with 65.5% in infection rate of RSV. Five perennial plants including Miscanths sacchariflorus with infection rate of 33.3% were recorded as hosts of RSV. Rice cultivars, 8 susceptible cultivars including Donggin1 and 17 resistant ones including Samgwang, were screened in field conditions at 3 different areas of Buan, Iksan and Ginje in 2009. All the susceptible cultivars were showed typical symptom of mosaic and wilt. In 17 genetic resistant cultivar, 12 cultivars were susceptible, however, 5 cultivars were field-resistant plus genetic resistant to RSV as non symptom expression. When RSV was artificially inoculated at seedling stage to 4 cultivars known as genetic resistant and 3 cultivars known as genetic susceptible, the symptom expression in resistant cultivars was lower as 19.3% in average than that of 53.3% in susceptible ones. In comparison of symptom expression rate and viral infection rate using resistant Nampyung and susceptible Heugnam cultivars by artificial inoculation of RSV at seedling stage, the symptom expression of Heugnam was higher as 28% than 12% of Nampyung. However, virion infection of resistant Nampyung cultivar was higher as 12% reversely than 85% of susceptible Heugnam. Yield loss of rice was investigated by the artificial inoculation of RSV at the seedling stage of resistant cultivars of Nampyung and Onnuri, and susceptible cultivars of Donggin1 and Ungwang for 3 years from 2008. The average yield per plant was 7.8 g, 8.5 g and 13.8 g on rice plants inoculated at seedling stage, tillering stage and maximum tillering stage, respectively. The yield loss rate was increased by earlier infection of RSV with 51% at seedling stage, 46% at tillering stage and 13% at maximum tillering stage. In resistant rice cultivars, there was no statistically significant relation between infection time and yield loss. In natural fields on susceptible rice cultivar of Ungwang at Taean and Jindo areas in 2009, the yield loss rate was increased with same tendency to the infection hill rate having the corelation coefficient of 0.94 when the viral infection was over 23.4%.

• Pediatric Infection and Vaccine
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• v.6 no.2
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• pp.219-233
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• 1999

Purpose : Respiratory syncytial virus(RSV) is the major cause of lower respiratory tract infection in infants and young children. This study was performed to analyze antigenic and genetic variation of G protein of subgroup A RSV. Methods : One hundred seventy-nine strains isolated at the Seoul National University Children's Hospital over 8 years-period from 1990 through 1998 were analysed for antigenic and genetic variability. Analysis was made by reactivity with monoclonal antibodies raised against RSV, and by restriction mapping and, for selected strains, nucleotide sequencing following amplification of full sequence of G gene by reverse transcription-polymerase chain reaction. Results : Restriction fragment analysis of the amplified G protein gene revealed 23 restriction patterns, 12 of which included more than 2 isolate, and the most frequent genetic type comprised 30% of the strains. Indirect immunofluorescent staining with monoclonal antibodies revealed 6 antigenic types with one predominant pattern accounting for 91% of the total strains. The most frequent antigenic type had 21 restriction patterns, and some viruses with same restiction pattern had different monoclonal antibody reaction pattern. Nucleotide sequence homology of subgroup A was 91~93% between reference(A2, Long) and Korean isolates, 93~99% among Korean isolates. Maximum-parsimony analysis demonstrated that Korean isolates were distinct from reference strains and subgroup A strains were clustered in 4 groups. Conclusion : The restriction analysis pattern of G protein gene identified greater diversity within subgroup A than was seen with the monoclonal analysis and a variety of antigenic and genetic types of RSV are circulating in Korea which are different from reference strains or strains isolated from other countries.

• Journal of Life Science
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• v.26 no.8
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• pp.911-920
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• 2016

Norovirus is the most common causative agent of acute gastroenteritis. This study was carried out to investigate molecular epidemiology of norovirus infections from outbreaks in Busan from 2012 to 2015. Total of 581 stool specimens were collected from diarrhea patients in outbreaks in Busan, 71 samples were resulted in positive to norovirus. The data were analyzed according to seasonality, patient, age and gender. Noroviruses were detected most frequently during the winter season from November (25.4%) to February (28.2%). The age group from teens was the most susceptible to norovirus infections. To obtain the molecular genetic information of norovirus, we performed sequencing analyses of the strains detected. Norovirus genotypes have been reported to show high genetic diversity. Four kinds of GI genotypes (GI-1, GI-2, GI-3, GI-5) and five kinds of GII genotypes (GII-1, GII-4, GII-5, GII-6, GII-17) were indentified in outbreaks in Busan. Other previous studies have shown that GII-4 is the most predominant circulating in Korea and worldwide. The most prevalent norovirus genotypes of each year were GII-6 in 2012, GII-6 in 2013, GII-4 in 2014 and GII-6 in 2015. Except for 2014, GII-6 genotype was the most prevalent and predominant in Busan. We described the epidemiological analysis of the noroviruses in outbreaks in Busan. The result of this study will contribute to update the epidemiological data and improve hygiene and public health via sustainable surveillance.

• Korean Journal of Breeding Science
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• v.49 no.3
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• pp.235-244
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• 2017

'Manbaek' is a bacterial blight resistant mid-late maturing rice cultivar with high grain quality. 'Manbeak' was derived from anther culture using the backcross combination, $Hopum^*2/SR30075$. 'Hopum' is a mid-late maturing rice cultivar with high grain quality and 'SR30075' is a pyramid line carrying three bacterial blight resistance genes. 'Manbaek' was selected through the pedigree method, yield trials, and local adaptability tests. 'Manbeak' carrying two bacterial blight resistance gene Xa3 and xa5 showed high-level and broad-spectrum resistance against bacterial blight. 'Manbaek' was resistant to K3a, mostly virulent race in Korea, and exhibited resistance reaction against 16 Korean bacterial blight isolates. 'Manbaek' was a mid-late maturing rice. The heading date of 'Manbaek' was August 19th, which was 5 days later than that of 'Nampyeong'. Manbaek' was a lodging-tolerant rice with short culm and dark green leaf. Due to the low viviparous germination, 'Manbaek' could be a useful material to prevent pre-harvest sprouting. 'Mabeak' was resistant to bacterial blight and rice stripe virus, but susceptible to other virus diseases and insect pests. The yield of 'Manbaek' was similar to 'Nampyeong'. 'Manbaek' showed excellent grain appearance and good tastes of cooked rice, so that it could contribute to improving the quality of bacterial blight resistant cultivars. 'Manbaek', bacterial blight resistant cultivar with high grain quality, is suitable for the cultivation at bacterial blight prone area and has been utilized in the breeding programs for enhancing the resistance against bacterial blight (Registration No. 6069).

• Research in Plant Disease
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• v.19 no.2
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• pp.90-94
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• 2013

Potato cv. 'Haryeong' was bred with high solids, resistance to late blight and good culinary quality. It has been registered as new potato variety in 2005. Tuber necrosis symptoms such as severe superficial necrosis, raised surface lesions and ringed necrotic areas were found in tubers of 'Haryeong' during storage of seed potatoes in 2010. Potato virus Y (PVY) was detected from these symptomatic tubers by the analysis of RT-PCR using a primer set specific to coat protein gene of PVY. The nucleotide sequence of RT-PCR product ($PVY^{Hkr}$) was determined and compared to those of other strains, such as $PVY^{Kor}$, $PVY^N$, $PVY^{NTN}$, $PVY^O$, and $PVY^C$ registered in GeneBank. The result showed that $PVY^{Hkr}$ was exactly the same as $PVY^{Kor}$, Korean isolate reported in 2005, except two nucleotides. To verify the PVY was responsible for the tuber necrosis symptoms shown in the tubers of 'Haryeong', a bioassay was done using two viruses (PVY and Potato leafroll virus) and five potato cultivars ('Haryeong', 'Superior', 'Atlantic', 'Dejima', and 'Chubaek'). As expected, the same necrosis symptom appeared in tubers of 'Haryeong' infected with PVY only during the storage period.

• Korean Journal of Medicinal Crop Science
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• v.9 no.2
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• pp.156-165
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• 2001

Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

• Research in Plant Disease
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• v.20 no.4
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• pp.299-302
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• 2014

The Paprika plant infected with Pepper mild mottle virus (PMMoV) do not produce commercial fruit as causing necrotic spots symptom on the fruit. Ten cultivars of paprika were analyzed to select the resistance cultivars against PMMoV pathotypes, $P_{1.2}$ and $P_{1.2.3}$, using bioassay and genetic markers. $L^1$, $L^3$, and $L^4$ genotypes expressing resistance to the pathotypes existed in those cultivars but $L^2$ genotype did not. $L^4L^4$ in cvs. Easy and Magnifico, $L^4L^3$ in cvs. Scirocco and Orange glory F1, $L^4L^1$ in cv. Special F1, $L^3L^3$ in cvs. Fiesta, Piero and Derby, and $L^3L^1$ in Cupra and Mazzona F1 were identified with SCAR and CAPS markers. The resistant cvs. to the 2 pathotypes were Magnipico, Easy, Scirocco F1, Orange glory and Special F1 and the susceptible cvs. were Fiesta, Piero, Derby, Cupra and Mazzona F1. The susceptible cvs. of the absence of $L^4$ genotype showed systemic infection when inoculated with PMMoV-$P_{1.2.3}$. However, those cvs. despite the presence of $L^3$ genotype showed vein necrosis on the inoculated leaf and hypersensitive necrosis symptom on the upper parts when inoculated with PMMoV-$P_{1.2}$.