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Construction of a Transgenic Tobacco Expressing a Polydnaviral Cystatin (폴리드나바이러스 유래 시스타틴 유전자 발현 형질전환 담배 제작)

  • Kim, Yeongtae;Kim, Eunsung;Park, Youngjin;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.54 no.1
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    • pp.7-15
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    • 2015
  • CpBV (Cotesia plutellae bracovirus) is a polydnavirus and encodes a cystatin (CpBV-CST1) gene. Its overexpression suppresses insect immunity and alters insect developmental processes. This study aimed to construct a genetically modified (GM) tobacco to further explore the physiological function of the viral cystatin and to apply to control insect pests. To this end, the transgenic tobacco lines were screened in expression of the target gene and assessed in insecticidal activity. A recombinant vector (pBI121-CST) was prepared and used to transform a bacterium, Agrobacterium tumefasciens. The transformed bacteria were used to generate transgenic tobacco lines, which were induced to grow callus and resulted in about 92% of shoot regeneration. The regenerated plants were screened by PCR analysis to confirm the insertion of the target gene in the plant genome. In addition, the expression of the target gene was assessed in the regenerated plants by quantitative real-time PCR (qRT-PCR). The qRT-PCR analysis showed that the transgenic line plant expressed the target gene about 17 times more than the control tobacco, indicating a stable insertion and expression of the target gene in the transgenic tobacco line. The insecticidal activity was then analyzed using the screened transgenic tobacco lines against the teneral 1st instar larvae of the oriental tobacco budworm, Helicoverpa assulta. Though there was a variation in the insecticidal efficacy among transgenic lines, T9 and T12 lines exhibited more than 95% mortality at 7 days after feeding treatment. These results suggest that CpBV-CST1 is a useful genetic resource to be used to generate GM crop against insect pests.

Analysis and verification of vitamin B12 in animal foods for update of national standard food composition table (국가표준식품성분표 개정을 위한 동물성 식품 비타민 B12 분석 및 검증)

  • Jeong, Yon Na;Park, Su-Jin;Lee, Sang Hoon;Choi, Youngmin;Choi, Kap Seong;Chun, Jiyeon
    • Korean Journal of Food Science and Technology
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    • v.52 no.4
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    • pp.317-324
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    • 2020
  • In order to create the national food nutrient database, a total of 41 animal foods (ham, seafood, edible insects and eggs) were analyzed for their vitamin B12 content and the applied immunoaffinity-HPLC was verified. Ham vitamin B12 contents were 0.30-0.65 ㎍/100 g. Seafood showed relatively high vitamin B12 level, where the values of fermented clam were the highest (26.80 ㎍/100 g) followed by fermented pollack roe. Vitamin B12 was not detected in silkworm pupae and beetles, while relatively high levels were found in the two-spotted cricket imago (6.70 ㎍/100 g). Chicken and quail egg yolk had roughly 100- and 30-times higher vitamin B12 levels as compared to their egg white. Vitamin B12 contents in quail and chicken eggs were significantly enhanced by boiling (p<0.05). Results based on accuracy (97-102% recovery) and precision (<5% RSD) indicate that this study provides reliable vitamin B12 information on animal foods consumed in Korea.

Host Dependent Oviposition and Development of Azuki Bean Weevil (Callosobruchus chinensis L.) in Different Leguminous Seeds (기주에 따른 팥바구미(Callosobruchus chinensis L.)의 산란 선호성 및 성장)

  • Kim, Seul Gi;Lee, Su Mi;Park, Jun Hong;Song, Kuk;Shin, Byung Sik
    • Korean Journal of Environmental Biology
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    • v.34 no.4
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    • pp.346-352
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    • 2016
  • To investigate the oviposition preference and development of azuki bean weevil (Callosobruchus chinensis L.), the following six different leguminous seeds were used in this study: red bean (Vigna angularis (Willd.) Ohwi & Ohashi), black soybean (Glycine max (L.) Merr.), soybean (Glycine max (L.) Merr.), seoritae (Glycine max (L.) Merr.), small black bean (Rhynchosia nulubilis) and kidney bean (Phaseolus vulgaris var. humilis Alef.). In the study of oviposition preference, the numbers of eggs per leguminous seed on red bean, black soybean, soybean, seoritae and small black bean were 1.23, 0.61, 0.69, 1.05 and 1.13, respectively. The maximum daily number of eggs was observed at 48 hours and the minimum was at 96 hours. According to each host leguminous seed, developmental time for each host seed was different. The shortest adult emergence time was on red bean (25.27 days). The other five leguminous seeds increased or doubled the adult emergence time. Adult emergence rates feeding on red bean, seoritae, black soybean, soybean, small black bean were 83.33%, 28.23%, 27.87%, 20.44%, and 11.59%, respectively. Emergence rate on red bean was four times higher than the rate on other seeds. The longevity of emerged female adults was almost all longer than that of males. The male adults weighed the lowest of feeding on small black bean. Female adults weighed the lowest of feeding on soybean. Adult weights were the heaviest for both males and females feeding on red bean. As a result, hosts of azuki bean weevil could decrease oviposition rate, emergence rate, adult longevity, and adult weight but increase emergence time. Especially in kidney bean, adult was not completely emerged. No eggs were laid. These results suggest that there might be emergence inhibitors in kidney bean. These imformation might be used to control damages caused by azuki bean weevils.

Effects of Electrolyzed Alkaline Reduced Water on Echinostoma hortense Infection and Immune Response in C57BL/6 Mice (C57BL/6 생쥐에서 전해알칼리환원수가 호르텐스극구흡충 감염과 면역에 미치는 영향)

  • Kim, Dong-Heui;Deung, Young-Kun;Jin, Dan;Huang, Xue Zhu;Qi, Xu Feng;Kim, Kwang-Yong;Lee, Kyu-Jae
    • Applied Microscopy
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    • v.38 no.1
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    • pp.11-19
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    • 2008
  • To examine the effect of the electrolyzed alkaline reduced water (ERW) on animal immunity, by employing Echinostoma hortense that is a parasite in the small intestine, the immune response of C57BL/6 was examined. To C57BL/6 mice, Echinostoma hortense metacercaria 15 per animal was in oculate dorsally, the worm was collected after 2 weeks, and the change of goblet cells and mast cells in the mucosa of small intestine was examined, and by using a protein chip, the change of cytokines in the serum was compared and observed. As a result, average 8.3 worms were collected from the C57BL/6 mice infected with E. hortense, and in the group fed with the ERW, average 10 worms were collected. In regard to the examination of the change of goblet cells, in the experimental group infected with E. hortense and fed with the ERW, average 4.3 worms per villus were detected, hence, it was found that the expression of goblet cells was low (p<0.001). Regarding the examination of the change of mast cells, similarly, in the group infected with E. hortense and fed with the ERW, average 11 worms per villus were detected, and it appears to be less than control group (p<0.001). Regarding the expression of cytokines in mouse serum, in comparison of the experiment group infected with E. hortense and control group, in the expression of the Th1 cytokines IL-6, IL-$1{\beta}$, IFN-${\gamma}$, TNF-${\alpha}$, and IL-2, and the Th2 cytokines IL-4, IL-5, IL-10, and IL-13, a significant difference was not detected. In our study, it was found that in the infection of E. hortense, the ERW mediates its effect on the number of goblet cells and mast cells in the intestinal mucosa, and simultaneously, the worm expulsion was delayed, and thus the conclusion that the ERW mediated its effect on the intestinal immunity of mice was obtained.

A Numerical Study of Hydraulic Fractures Propagation with Rock Bridges (Rock bridges를 고려한 수치 해석적 수압파쇄 균열거동 연구)

  • 최성웅
    • Proceedings of the Korean Society for Rock Mechanics Conference
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    • 2000.09a
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    • pp.201-210
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    • 2000
  • Rock bridge in rock masses can be considered as one of several types of opening-mode fractures, and also it has been known to have a great influence on the stability of structures in rock mats. In the beginning of researching a rock bridge it used to be studied only in characteristics of its behavior, as considering resistance of material itself. However the distribution pattern of rock bridges, which can affect the stability of rock structures, is currently researched with a fracture mechanical approach in numerical studies. For investigating the effect of rock bridges on the development pattern of hydraulic fractures, the author analyzed numerically the stress state transition in rock bridges and their phenomena with a different pattern of the rock bridge distributons. From the numerical studies, a two-crack configuration could be defined to be representative of the most critical conditions for rock bridges, only when cracks are systematic and same in their length and angle. Moreover, coalescence stresses and onset of propagation stresses could be known to increase with decreasing s/L ratio or increasing d/L ratio. The effect of pre-existing crack on hydraulic fracturing was studied also in numerical models. Different to the simple hydraulic fracturing modeling in which the fractures propagated exactly parallel to the maximum remote stress, the hydraulic fractures with pre-existing cracks dial not propagate parallel to the maximum remote stress direction. These are representative of the tendency to change the hydraulic fractures direction because of the existence of pre-existing crack. Therefore s/L, d/L ratios will be identical as a function effective on hydraulic fractures propagation, that is, the $K_{I}$ vague increase with decreasing s/L ratio or increasing d/L ratio and its magnification from onset to propagation increases with decreasing s/L ratio. The scanline is a commonly used method to estimate the fracture distribution on outcrops. The data obtained from the scanline method can be applied to the evaluation of stress field in rock mass.s.

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A Numerical Study of Hydraulic Fractures Propagation with Rock Bridges (Rock bridges를 고려한 수치 해석적 수압파쇄 균열거동 연구)

  • 최성웅
    • Tunnel and Underground Space
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    • v.10 no.3
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    • pp.447-456
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    • 2000
  • Rock bridge in rock masses can be considered as one of several types of opening-mode fractures, and also it has been known to have a great influence on the stability of structures in rock mass. In the beginning of researching a rock bridge it used to be studied only in characteristics of its behavior, as considering resistance of material itself. However the distribution pattern of rock bridges, which can affect the stability of rock structures, is currently researched with a fracture mechanical approach in numerical studies. For investigating the effect of rock bridges on the development pattern of hydraulic fractures, the author analyzed numerically the stress state transition in rock bridges and their phenomena with a different pattern of the rock bridge distributions. From the numerical studies, a two-crack configuration could be defined to be representative of the most critical conditions for rock bridges, only when cracks are systematic and same in their length and angle. Moreover, coalescence stresses and onset of propagation stresses could be known to increase with decreasing s/L ratio or increasing d/L ratio. The effect of pre-existing crack on hydraulic fracturing was studied also in numerical models. Different to the simple hydraulic fracturing modeling in which the fractures propagated exactly parallel to the maximum remote stress, the hydraulic fractures with pre-existing cracks did not propagate parallel to the maximum remote stress direction. These are representative of the tendency to change the hydraulic fractures direction because of the existence of pre-existing crack. Therefore s/L, d/L ratios will be identical as a function effective on hydraulic fractures propagation, that is, the K$_1$ value increase with decreasing s/L ratio or increasing d/L ratio and its magnification from onset to propagation increases with decreasing s/L ratio. The scanline is a commonly used method to estimate the fracture distribution on outcrops. The data obtained from the scanline method can be applied to the evaluation of stress field in rock mass.

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Studies on the Effects of various Levels of Protein in the Artificial Diet on Nutritional Physiology of the Silkworm, Bombyx mori (인공사료의 단백질수준이 가잠의 영양생리에 미치는 영향에 관한 연구)

  • 설광열
    • Journal of Sericultural and Entomological Science
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    • v.23 no.2
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    • pp.37-49
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    • 1982
  • Larvae of the silkworm (Bombyx mori L.) were reared during the 5th instar on the four kinds of artificial diets on the basis of the different amounts of soybean meal used as the protein source. In this experiment it was shown that the various levels of protein in the diet affected not only the growth and silk production but the digestibility of the diet. haemolymph protein and uric acid excretion. The results obtained are summarized as follows; 1. By an increase of the level of protein in the diet the apparant digestibility was increased. but the protein digestibility was comparatively decreased. 2. Larval body weight increment was not observed by the 3rd day of the 5th instar, but was increased from the 4th day as the level of protein was increased in the diet. 3. After the 3rd day of the 5th instar, protein content in the hemolymph was increased steeply by an increase of the protein content in the diet. However, the percentage of hemolymph protein to the ingested protein was decreased from the 2nd day of the 5th instar and increased more or less after the 4th day. 4. An increase of the uric acid excretion was observed as the content of protein in the diet was increased but the pattern of the uric acid excretion was different between high and low-protein diet. However, the percentage of the uric acid excretion to the ingested protein and to the hemolymph protein were both decreased steeply after the 2nd day of the 5th instar. 5. It was also evident that the high-protein diet increased the cocoon productivity. 6. It showed that the feed efficiency for body weight increment and silk formation was high by an increase of the level of protein in the diet, but the protein efficiency was not.

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Use of Sprinkler System for Control of the Pine Needle Gall Midge, Thecodiplosis japonensis Uchida et Inouye (분무장치(噴霧裝置)를 이용(利用)한 솔잎혹파리 방제(防除)에 관(關)한 연구(硏究))

  • Chung, Sang Bae;Kim, Chul Su
    • Journal of Korean Society of Forest Science
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    • v.83 no.3
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    • pp.311-321
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    • 1994
  • Experiments were conducted to evaluate the efficacy of ground and foliar sprinkling of natural water for control of the pine needle gall midge, Thecodiplosis japonensis, by disturbing the reproductive behavior of adults such as copulation and oviposition. Diurnal ground spray for whole area application and foliar spray for single tree treatment were tested at day time during the period of adult emergence. The results obtained are as follows : 1. The population density of overwintering larvae in soil was not affected by the water spray during the adult emergence period. 2. Spraying the water showed no effect on the whole period and number of adult emergence, but retarded early emergence and facilitated Late emergence. The difference in the emerging time by 50% emergence day was approximately 7 days. 3. Ground spray was significantly different from control in gall formation rates but was not sufficient for effective control. As a single-tree treatment, the foliar spray was found to be highly effective for control of the pine needle gall midge, Average gall formation of spray plots and untreated ones were 2.6% and 38%, respectively ; control value exceeded 90%. 4. Ground spray of water was not effective in reducing larvae in gall of needle. Foliar spray, however, reduced 45% of larval numbers. 5. Foliar spray accelerated shoot growth by 15%-18%. 6. Foliar spray was highly effective for the control of sucking insects, such as mites and aphids on pine trees.

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Soil Chemical Characteristics and Comparison with Infested Status of Nematode(Meloidogyne spp.) in Plastic House Continuously Cultivated Oriental Melon in Songju (성주 지역 시설참외 연작지의 토양특성 및 토양선충 변화)

  • Jun, Han-Sik;Park, Woo-chul
    • Korean Journal of Environmental Agriculture
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    • v.20 no.2
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    • pp.127-132
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    • 2001
  • This study was conducted for ten years to evaluate the effective soil management for preventing the infection of root-knot nematode in the field of continuous cultivation with oriental melon under plastic house in Songju area of kyongbuk province. The content of available phosphate, total nitrogen, organic matter, CEC, and exchangeable base in the soil increased with the increase of continuous cultivation year. Especially salt content in the soil increased form 1.2 to 4.55 mS/cm and the yield of oriental melon dramatically decreased with the continuous cultivation year. The number of root-knot nematode was 91 per $300\;cm^3$ of soil in the field of continuous cultivation for 3 years and showed slight damage on the oriental melon, but it was 518 in the field of $4{\sim}6$ years continuous cultivation and showed that 50% of plants died in August, and the yield of late season was less than 50% compared to normal plant. For the seasonal changes in infection rate of root-knot nematode on oriental melon plant, 15% of the normal plant was infected by nematode in February and increased gradually by $10{\sim}20%$ per month, 60% of plants was infected in July. The density of root-knot nematode nymph was 167 in February and increased to 1,625 in August. The infection rate of nematode was 35%, and the number of nematode was about 54 in nursery soil originated from paddy soil, upland soil, and river sand. There were no relationship between the number of nematode and available phosphate or exchangeable base in the soil of plastichouse where oriental melon plants were grown.

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Effect of rice seed dressing with imidacloprid WS on early occurring rice insect pests (본답 초기해충의 생력방제를 위한 imidacloprid의 종자분의 효과)

  • Choi, Byung-Ryul;Yoo, Jae-Ki;Lee, Sang-Guei;Lee, Jeong-Oon
    • The Korean Journal of Pesticide Science
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    • v.2 no.1
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    • pp.85-90
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    • 1998
  • A series of experiments were carried out to determine the effect of seed dressing application of chemicals on the early occurring rice insect pests. The number of small brown planthopper adults per 3hills in seed-dressing plot of Imidacloprid WS (Im WS) was 1.6 and its control efficacy was over 90%, compared with that of untreated plot. Rice water weevil was observed in the density of 8.8 larvae per 5 hills seed-dressed Im WS plots(showing 95.5% of control efficacy). Control efficacy of Im WS and Im GR against adult weevils lasted for 26 days after treatment and was higher than that of Carbofuran GR. Rates of injured stems by rice stem maggot and injured leaves by rice leaf miner were 3% and 3.7% in Im WS treatment plot respectively. Seed germination rate after seed dressing with recommended dosage (3 g/seed kg) of Im WS was 71 % on the 1st day and increased on 5th day up to that of untreated seed. The residual amount of Im in seed dressing plot was 0.11 ppm in rice roots and 0.05 ppm in leaves on the 40th day after treatment. Residual effect of Im WSI sustained for 50 days with over 95% insecticidal effect for the rice water weevil and over 90% for the brown planthopper. Quantity of the chemical applied in the field was calculated as 0.084 kg a.i./ha in seed dressing and 0.3 kg a.i./ha in seed box treatment, respectively It took 1 hour to treat insecticide by seed dressing, 2.5 hours by seed box applying, and 3.6 hours by water surface releasing per 990 $m^2$.

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