• Journal of Sericultural and Entomological Science
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• v.25 no.2
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• pp.1-31
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• 1984

Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.

• Journal of Korean Society of Forest Science
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• v.54 no.1
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• pp.1-24
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• 1981

Kangweon-Do is rich in sightseeing resources. There are three sightseeing areas;first, mountain area including Seolak and Ohdae National Parks, and chiak Provincial Park; second eastern coastal area; third lake area including the watersheds of North Han River. In this paper, several methods of forestation were studied for landscaping the North Han River watersheds centering around Chounchon. In Chunchon lake complex, there are four lakes; Uiam, Chunchon, Soyang and Paro from down to upper stream. The total surface area of the above four lakes is $14.4km^2$ the total pondage of them 4,155 million $m^3$, the total generation of electric power of them 410 thousand Kw, and the total forest area bordering on them $1,208km^2$. The bordering forest consists of planned management forest ($745km^2$) and non-planned management forest ($463km^2$). The latter is divided into green belt zone, natural conservation area, and protection forest. The forest in green belt amounts to $177km^2$ and centers around the 10km radios from Chunchon. The forest in natural conservation area amounts to $165km^2$, which is established within 2km sight range from the Soyang-lake sides. Protection forest surrounding the lakes is $121km^2$ There are many scenic places, recreation gardens, cultural goods and ruins in this lake complex, which are the same good tourist resources as lakes and forest. The forest encirelng the lakes has the poor average growing stock of $15m^3/ha$, because 70% of the forest consists of the young plantation of 1 to 2 age class. The ration of the needle-leaved forest, the broad-leaved forest and the mixed forest in 35:37:28. From the standpoint of ownership, the forest consists of national forest (36%), provincial forest (14%), Gun forest (5%) and private forest(45%). The greater part of the forest soil, originated from granite and gneiss, is much liable to weathering. Because the surface soil is mostly sterile, the fertilization for improving the soil quality is strongly urged. Considering the above-mentioned, the forestation methods for improving landscape of the North Han River Watersheds are suggested as follows: 1) The mature-stage forest should be induced by means of fertilizing and tendering, as the forest in this area is the young plantation with poor soil. 2) The bare land should be afforested by planting the rapid growing species, such as rigida pine, alder, and etc. 3) The bare land in the canyon with moderate moist and comparatively rich soil should be planted with Korean-pine, larch, ro fir. 4) Japaness-pine stand should be changed into Korean-pine, fir, spruce or hemlock stand from ravine to top gradually, because the Japanese-pine has poor capacity of water conservation and great liability to pine gall midge. 5) Present hard-wood forest, consisting of miscellaneous trees comparatively less valuable from the point of wood quality and scenerity, should be change into oak, maple, fraxinus-rhynchophylla, birch or juglan stand which is comparatively more valuable. 6) In the mountain foot within the sight-range, stands should be established with such species as cherry, weeping willow, white poplar, machilus, maiden-hair tree, juniper, chestnut or apricot. 7) The regeneration of some broad-leaved forests should be induced to the middle forest type, leading to the harmonious arrangement of the two storied forest and the coppice. 8) For the preservation of scenery, the reproduction of the soft-wood forest should be done under the selection method or the shelter-wood system. 9) Mixed forest should be regenerated under the middle forest system with upper needle-leaved forest and lower broad-leaved forest. In brief, the nature's mysteriousness should be conserved by combining the womanly elegance of the lakes and the manly grandeur of the forest.

• 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%.