• 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%.

• Applied Biological Chemistry
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• v.11
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• pp.1-27
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• 1969

More than thirty kinds of sea food pickles have been eaten in Korea. Out of these salted yellow tail pickle, salted clam pickle, salted oyster pickle, and salted cuttlefish pickle were employed for the analysis of their components, identification of main fermenting microbes, and determination of enzyme characteristics concerned. Also studied was the effect of enzymic action of microbes, which are concerned with the fermenting of pickles, on the production of flavorous 5'-mononucleotides and amino acids. The results are summarized as follows: 1. Microflora observed in the pickles are: (a) Total count of viable cells after 1-2 months of pickling was found to be $10^7$ and that after 6 months decreased to $10^4$. (b) Microbial occurence in the early stage of pickling was observed to be 10-20% Micrococcus spp., 10-20% Brevibacterium spp., 0-30% Sarcina spp., 20-30% Leuconostoc spp., ca 30% Bacillus spp., 0-10% Pseudomonas spp., 0-10% Flavobacterium spp., and 0-20% yeast. (c) Following the early stage of pickling, mainly halophilic bacteria such as Bacillus subtilis, Leuconostoc mesenteroides, Pediococcus halophilus and Sarcina litoralis, were found to exhibit an effect on the fermentation of pickle and their enzyme activities were in direct concern in fermentation of pickles. (d) Among the bacteria participating in the fermentation, Sarcina litoralis 8-14 and 8-16 strains were in need of high nutritional requirement and the former was grown only in the presence of purine, pyrimidine and cystine and the latter purine, pyrimidine and glutamic acid. 2. Enzyme characteristics studied in relation to the raw materials and the concerned microbes isolated are as follows: (a) A small amount of protease was found in the raw materials and 30-60% decrease in protease activity was demonstrated at 7% salt concentration. (b) Protease activity of halophilic bacteria, Bacillus subtilis 7-6, 11-1, 3-6 and 9-4 strains, in the complete media decreased by 10-30% at the 7% salt concentration and that of Sarcina litoralis 8-14 and 8-16 strains decreased by 10-20%. (c) Proteins in the raw materials were found to be hydrolyzed to yield free amino acids by protease in the fermenting microbes. (d) No accumulation of flavorous 5'-mononucleotides was demonstrated because RNA-depolymerase in the raw materials and the pickles tended to decompose RNA into nucleoside and phosphoric acid. (e) The enzyme produced in Bacillus subtilis 3-6 strain isolated from the salted clam pickles, was ascertained to be 5'-phosphodiesterase because of its ability to decompose RNA and thus accumulating 5'-mononucleotide. (f) It was demonstrated that the activity of phosphodiesterase in Bacillus subtilis 3-6 strain was enhanced by some components in the corn steep liquor and salted clam pickle. The enzyme activity was found to decrease by 10-30% and 40-60% at the salt concentration of 10% and 20%, respectively. 3. Quantitative data for free amino acids in the pickles are as follows: (a) Amounts of acidic amino acids such as glutamic and aspartic acids in salted clam pickle, were observed to be 2-10 times other pickles and it is considered that the abundance in these amino acids may contribute significantly to the specific flavor of this food. (b) Large amounts of basic amino acids such as arginine and histidine were found to occur in salted yellow tail pickle. (c) It is much interesting that in the salted cuttlefish pickle the contents of sulfur-containing amino acids were exceedingly high compared with those of others: cystine was found to be 17-130 times and methionine, 7-19 times. (d) In the salted oyster pickle a high content of some essential amino acids such as lysine, threonine, isoleucine and leucine, was demonstrated and a specific flavor of the pickle was ascribed to the sweet amino acids. Contents of alanine and glycine in the salted oyster pickle were 4 and 3-14 times as much as those of the others respectively. 4. Analytical data for 5'-mononucleotides in the pickles are as follows: (a) 5'-Adenylic acid and 3'-adenylic acid were found in large amounts in the salted yellow tail pickle and 5'-inosinic acid in lesser amount. (b) 5'-Adenylic acid, especially 3'-adenylic acid predominated in amount in the salted oyster pickle over that in the other pickles. (c) The salted cuttlefish pickle was found to contain only 5'-adenylic acid and 3'-adenylic acid. It has become evident from the above fact that clam and the invertebrate lack of adenylic deaminase and contain high content of adenylic acid. Thus, they were demonstrated to be the AMP-type. (d) 5'-Inosinic acid was contained in the salted yellow tail pickle in a significant concentration, and it might be considered to be IMP-type. 5. Comparative data for flavor with regard to the flavorous amino acids and the contents of 5'-mononucleotides are: (a) A specific flavor of salted yellow tail pickle was ascribed to the abundance in glutamic acid and aspartic acid, and to the existence of a small amount of flavorous 5'-inosinic acid. The combined effect of these components was belived to exhibit a synergistic action in producing a specific fiavor to the pickle. (b) A specific flavor of salted clam pickle has been demonstrated to be attributable to the richness in glutamic acid and aspartic acid rather than to that of 5'-mononucleotides.