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Generation of Transgenic Rice without Antibiotic Selection Marker through Agrobacterium-mediated Co-transformation System (아그로박테리움 동시 형질전환 시스템을 통한 항생제 선발 마커가 없는 형질전환벼의 생산)

  • Park, Soo-Kwon;Kwon, Tack-Min;Lee, Jong-Hee;Shin, Dong-Jin;Hwang, Woon-Ha;Song, You-Chun;Cho, Jun-Hyun;Nam, Min-Hee;Jeon, Seung-Ho;Lee, Sang-Yeol;Park, Dong-Soo
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1152-1158
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    • 2012
  • Development of transgenic plant increasing crop yield or disease resistance is good way to solve the world food shortage. However, the persistence of marker genes in crops leads to serious public concerns about the safety of transgenic crops. In the present paper, we developed marker-free transgenic rice inserted high molecular-weight glutenin subunit (HMW-GS) gene ($D{\times}5$) from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation method. Two expression cassettes comprised of separate DNA fragments containing only the $D{\times}5$ and hygromycin resistance (HPTII) genes were introduced separately into Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring $D{\times}5$ or HPTII was infected into rice calli at a 3: 1 ratio of EHA105 with $D{\times}5$ gene and EHA105 with HPTII gene expressing cassette. Then, among 66 hygromycin-resistant transformants, we obtained two transgenic lines inserted with both the $D{\times}5$ and HPTII genes into the rice genome. We reconfirmed integration of the $D{\times}5$ and HPTII genes into the rice genome by Southern blot analysis. Wheat $D{\times}5$ transcripts in $T_1$ rice seeds were examined with semi-quantitative RT-PCR. Finally, the marker-free plants containing only the $D{\times}5$ gene were successfully screened at the $T_1$ generation. These results show that a co-infection system with two expression cassettes could be an efficient strategy to generate marker-free transgenic rice plants.

Vibrio Vulnificus Induces the Inflammation of Mouse Ileal Epithelium: Involvement of Protein Kinase C and Nuclear Factor-Kappa B (회장 상피세포에서 비브리오균(Vibrio vulnificus)의 염증 유도 기작 연구: protein kinase C와 nuclear factor kappa-B의 관련성)

  • Han, Gi Yeon;Jung, Young Hyun;Jang, Kyung Ku;Choi, Sang Ho;Lee, Sei-Jung
    • Journal of Life Science
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    • v.24 no.6
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    • pp.664-670
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    • 2014
  • In the present study, we investigate the role of V. vulnificus in promoting the inflammation of mouse ileal ephitelium and its related signaling pathways. ICR mice were infected orally with V. vulnificus ($1{\times}10^9CFU$) for 16 h as a representative model of food-borne infection. To find the major portal of entry of V. vulnificus in mouse intestine, we have measured the levels of bacterial colonization in small intestine, colon, spleen, and liver. V. vulnificus appeared to colonize in intestine and colon in the order of ileum >> jejunum> colon, but lack in the duodenum, spleen, and liver. V. vulnificus in ileum caused severe necrotizing enteritis and showed shortened villi heights accompanied by an expanded width and inflammation, compared with the control mice. V. vulnificus induced ileal epithelium inflammation by activating phosphorylation of PKC and membrane translocation of $PKC{\alpha}$. V. vulnificus induced the phosphorylation of ERK and JNK, but did not affect p38 MAPK phosphorylation. Notably, V. vulnificus stimulated the I-${\kappa}B$-dependent phosphorylation of NF-${\kappa}B$ in mouse ileal epithelium. Finally, the ileal infection of V. vulnificus resulted in a significant increase in expression of proinflammatory cytokines and Toll-like receptors, respectively, compared to the control. Collectively, our results indicate that V. vulnificus induces ileal epithelium inflammation by increasing NF-${\kappa}B$ phosphorylation via activation of PKC, ERK, and JNK, which is critical for host defense mechanism in food-borne infection by V. vulnificus.

Development of Marker-free Transgenic Rice Expressing the Wheat Storage Protein, Glu-1Dy10, for Increasing Quality Processing of Bread and Noodles (빵과 면의 가공적성 증진을 위한 밀 저장단백질 Glu-1Dy10을 발현하는 마커프리 형질전환 벼 개발)

  • Park, Soo-Kwon;Shin, DongJin;Hwang, Woon-Ha;Hur, Yeon-Jae;Kim, Tae-Heon;Oh, Se-Yun;Cho, Jun-Hyun;Han, Sang-Ik;Lee, Seung-Sik;Nam, Min-Hee;Park, Dong-Soo
    • Journal of Life Science
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    • v.24 no.6
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    • pp.618-625
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    • 2014
  • Rice flour is used in many food products. However, dough made from rice lacks extensibility and elasticity, making it less suitable than wheat for many food products such as bread and noodles. The high-molecular weight glutenin subunits (HMW-GS) of wheat play a crucial role in determining the processing properties of the wheat grain. This paper describes the development of marker-free transgenic rice plants expressing a wheat Glu-Dy10 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation. Two expression cassettes, consisting of separate DNA fragments containing Glu-1Dy10 and hygromycin phosphotransferase II (HPTII) resistance genes, were introduced separately into Agrobacterium tumefaciens EHA105 for co-infection. Each EHA105 strain harboring Glu-1Dy10 or HPTII was infected into rice calli at a 3: 1 ratio of Glu-1Bx7 and HPTII. Among 290 hygromycin-resistant $T_0$ plants, we obtained 29 transgenic lines with both the Glu-1Dy10 and HPTII genes inserted into the rice genome. We reconfirmed the integration of the Glu-1Dy10 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the Glu-1Dy10 in transgenic rice seeds were examined by semi-quantitative RT-PCR and Western blot analysis. The marker-free plants containing only the Glu-1Dy10 gene were successfully screened in the $T_1$ generation.

Correlation between the Serum Leptin Level and the Expression of Leptin in Stomach Cancer Patients (위암에서 혈청 렙틴 레벨과 조직 내 렙틴 발현과의 상관관계)

  • Kim, Ji-Hyun;Jung, Hun;Jun, Kyong-Hwa;Kim, Sung-Keun;Chin, Hyung-Min;Jung, Ji-Han;Kim, Wook;Jeon, Hae-Myung;Park, Cho-Hyun;Park, Seung-Man;Park, Woo-Bae;Lim, Keun-Woo;Kim, Seung-Nam
    • Journal of Gastric Cancer
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    • v.8 no.4
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    • pp.176-181
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    • 2008
  • Purpose: The adipocyte-derived cytokine leptin plays a major role in the control of stable body weight by suppressing food intake and increasing energy metabolism. Leptin regulates the cell proliferation of various epithelial cells and it may be involved in the promotion of cancer. Leptin and its receptor are highly expressed in gastric adenocarcinoma, but the association between the serum leptin level and the tissue expression of leptin is uncertain. We evaluated the serum leptin level and the expressions of leptin and leptin receptor in gastric cancer, and we explore the possible mechanism and role of leptin in the carcinogenesis of gastric cancer. Materials and Methods: 72 carcinomas that were curatively resected at our hospital from October 2005 to March 2007 were included in this study. By immunoassay and immunohistochemical staining, we evaluated the serum leptin level and the expressions of leptin and its receptor, and we analyzed their relationship together with the clinicopathological variables. Results: The serum leptin level was increased as the patient's BMI increased and it was decreased in H. pylori infected patients. The expression of leptin was increased as the TNM stage increased (P=0.014), and the expression of leptin receptor in the intestinal type gastric adenocarcinoma was higher than that in the diffuse type gastric adenocarcinoma (71.4% vs 28.6%, respectively, P=0.033). Conclusion: There was no significant correlation between the serum leptin level and expression of leptin in gastric cancer patients. The expression of leptin was associated with the TNM stage, but its role in the pathogenesis of gastric cancer has to be elucidated.

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An Analysis of Growth Conditions of old Trees in Yangdong Villages (양동마을의 노거수 생육실태 분석)

  • Kim, Young-Hun;Deng, Bei-Jia;You, Ju-Han
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.38 no.2
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    • pp.95-107
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    • 2020
  • The purpose of this study is to provide the basic data by analyzing and analysing the Growth Conditions of old Trees in Yangdong village. This study investigated about the conducted on tree information, soil information, and tree health. The result are as follows. The trees information in Yangdong Village consists of Juniperus chinensis, Salix chaenomeloides, Salix pseudolasiogyne, Celtis sinensis, Zelkova serrata, Gleditsia japonica, and Gleditsia sinensis trees, The range of height was 4.0~17.0m, and the diameter was 0.51~1.34m, Juniperus chinensis trees of No.17 was most large. In the results of soil analysis, there showed that acidity was pH4.1~6.3, hardness of 5~48mm, organic matter content of 21.2~29.1g/kg, electrical conductivity(EC) of 0.34~1.76dS/m, available P2O5 of 79.8~451.6mg/kg, exchangeable K of 0.22~1.71cmol+/kg, exchangeable Ca of 4.98~7.44cmol+/kg, exchangeable Mg of 0.67~2.19cmol+/kg, exchangeable Na of 0.19~1.04cmol+/kg and cation exchange capacity(C.E.C) of 7.23~13.02cmol+/kg. As a result, the highest number of tree health levels is 8 of 11trees of Celtis sinensis, 2 of 7trees of Zelkova serrata, and 3 of Gleditsia sinensis, and 13 of 30 trees of health levels, The Older trees with high infection, spoil and hollowed part were the remaining trees except for the healthy part. Relatively, more than half of the number of targets is infected, decay, and the hollowed site, and it is necessary to perform surgery on the damaged area. In addition, preservation and protection measures should be implemented by supplying root nutrients for trees, controlling nutrients in the body to prevent secondary and tertiary damages that cause the infection site to metastasize to the health site, In order to continually monitor the trees, measures to improve the location environment and management of the trees should be sought.

New Technologies for the Removal of Bacteriophages Contaminating Whey and Whey Products as Cheese by-Products: A Review (치즈 부산물인 유청과 유청 제품에 감염된 박테리오파지 제거를 위해 새롭게 개발된 기술: 총설)

  • Kim, Dong-Hyeon;Chon, Jung-Whan;Kim, Hyun-Sook;Kim, Hong-Seok;Song, Kwang-Young;Hwang, Dae-Geun;Yim, Jin-Hyuk;Kang, Il-Byung;Lee, Soo-Kyung;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.32 no.2
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    • pp.93-100
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    • 2014
  • In general, whey obtained from various cheese batches is being reused, so as to improve the texture and to increase the yield and the nutrient value of the various final milk-based products. In fact, re-usage of whey proteins, including whey cream, is a common and routine procedure. Unfortunately, most bacteriophages can survive heat treatments such as pasteurization. Hence, there is a high risk of an increase in the bacteriophage population during the cheese-making process. Whey samples contaminated with bacteriophages can cause serious problems in the cheese industry. In particular, the process of whey separation frequently leads to aerosol-borne bacteriophages and thus to a contaminated environment in the dairy production plant. In addition, whey proteins and whey cream reused in a cheese matrix can be infected by bacteriophages with thermal resistance. Therefore, to completely abolish the various risks of fermentation failure during re-usage of whey, a whey treatment that effectively decreases the bacteriophage population is urgently needed and indispensable. Hence, the purpose of this review is to introduce various newly developed methods and state-of-the-art technologies for removing bacteriophages from contaminated whey and whey products.

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Investigational Studies on Reproductive Failures of Slaughtered Cows (도살빈우의 번식장애사례 조사연구)

  • 이용빈;임경순
    • Korean Journal of Animal Reproduction
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    • v.6 no.1
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    • pp.19-30
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    • 1982
  • 1. The cows slaughtered at age of 3, 4, 6, 7, 8, and 9 years old were 1.5, 1.5, 15.0, 62.5 and 4.4% respectively. 2. The cows slaughtered at 351-450kg and more than 500kg were 60 and 28% respectively. 3. Best, very good, good and bad cows in nutritional condition were 1.6, 25.8, 62.9, and 9.7% respectively. Among the six cows which were bad nutrition, the two were with severe endometritis, the three were normal in genital function and one was on 70 days of pregnancy. 4. Holstein cows(55.2%) showed higher reproductive failure than the Korean cows(33.3%). 5. The slaughted ratio of the Korean cattle and Holstein cows was 36 and 64% respectively. 6. Pregnant cows were about 16% among the slaughtered one. 7. Reproductive failures were composed of 46% in uterus, 32% in ovaries, 8% in udder, 6% in oviduct, 4% in cervix of uterine, 2% in vagina and 2% inmummified fetus. 8. Forty six percentages of uterine diseases were as follows; horn, 13%, body of uterus, 32% and ovary diseases were 32%, that is, 12% of ovary atrophy, 8% of ovarycyst and 6% of lutealcyst. 9. The cows of reproductive failures were commonly infected with 1.6 kinds of diseases. 10. According to classification, six type of ovaries were as follows; normal, 58%, ovary-cyst, 11%, luteum cyst, 4%, coexistence of follicles and corpus luteum, 16%, weak function of ovaries, 10% and ovarian atrophy, 1%. 11. Major axis, minor axis and thickness of right ovary were larger than those of left one both in Korean cattle and Holstein cows. Holstein cow had generally larger size of ovary than these of the Korean cattle.. 12. The left and right oviducts showed no difference in length, but Holstein had longer oviduct than Korean cow. 13. There was no difference in the length of uterine horn between right and left in the Korean cows, but the right was longer than the left in Holstein cows. 14. Holstein had longer horn and body of uterine than the Korean cows. 15. The weight of right ovary was heavier than that of left in both breeds, but there was no differences in weight of left ovary between two breeds and right ovary of Holstein breed was heavier than that of the Korean cow. 16. The weight of right oviduct and uterine born was heavier than that of the left, and Holstein had heavier oviducts and uterine horns than the Korean cows. 17. Holstein had heavier uterine body and cervix of uterine than the Korean cows. 18. The length of reproductive systems of Korean cow is as follows; Major and minor diameter and thickness ofovary are 3.6${\pm}$0.7, 2.3${\pm}$0.4 and 1.6${\pm}$1.4 cm in left and 3.7${\pm}$0.6, 2.5${\pm}$0.5 and 1.8${\pm}$0.5 cm in right. Oviduct is 28.4${\pm}$3.1 cm in left and 27.8${\pm}$3.3 cm in right. Uterine horn is 27.4${\pm}$4.5 cm in left and 27.7${\pm}$4.9 cm in right. Uterine body and cervix are 3.4${\pm}$1.1 and 6.5${\pm}$1.7 cm. 19. The length of female reproductive systems ofHolstein cow is as follows; Major and minor diameter and thickness of ovary are 3.9${\pm}$1.3, 2.3${\pm}$0.5, and 1.5${\pm}$0.6 cm in left and 4.0${\pm}$0.8, 2.8${\pm}$0.6 and 1.8${\pm}$0.6 cm in right. Oviduct is 29.4${\pm}$4.2 cm in left and 29.3${\pm}$4.1 cm in right. Uterine horn is 30.2${\pm}$7.4 cm in left and 32.6${\pm}$8.4 cm in right. Uterine body and cervix are 4.5${\pm}$2.5 and 7.8${\pm}$2.9 cm. 20. The weight of reproductive systems of Korean cow is as follows; Ovary is 8.4${\pm}$4.1 g in left and 9.3${\pm}$3.6g in right. Oviduct is 1.5${\pm}$0.5 g in left and 1.6${\pm}$0.5 g in right. Uterine horn is 109${\pm}$27 g left and 118${\pm}$32 g in right. Uterine body and cervix are 30.4${\pm}$14.1 and 76.7${\pm}$38.4g. 21. The weight of reproductive systems of Holstein cow is as follows; Ovary is 8.2${\pm}$3.1 g in left and 12.5${\pm}$5.6 g in right. Oviduct is 1.7${\pm}$0.6 g in left and 1.9${\pm}$0.9 g in right. Uterine horn is 199${\pm}$14.2 g in left and 221${\pm}$111.2g in right. Uterine body and cervix are 58.2${\pm}$46.5 and 126.7${\pm}$103.3 g.

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Effects of a Soil-Born Paenibacillus spp. Strain KPB3 on Suppression of Bacterial Wilt Disease Caused by Ralstonia solanacearum (토양에서 분리한 Paenibacillus spp. KPB3의 Ralstonia solanacearum에 의한 세균성 풋마름병 억제 효과)

  • Suk, Jung-Ki;Ipper, Nagesh S.;Lee, Seon-Hwa;Shrestha, Anupama;Park, Duck-Hwan;Cho, Jun-Mo;Hur, Jang-Hyun;Kim, Byung-Sup;Lim, Chun-Keun
    • The Korean Journal of Pesticide Science
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    • v.10 no.4
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    • pp.313-319
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    • 2006
  • Two hundred bacterial strains were isolated from the soil around healthy tomato plants in a polyvinyl house, where most of the other plants showed bacterial wilt symptoms. The strains were screened in vitro for their antibacterial activity. Among them, a strain, KPB3 showed strong bactericidal activity against bacterial wilt pathogen, Ralstonia solanacearum. The strain KPB3 was identified using physiological and biochemical tests, and 16S rRNA analyses. Based on these tests, the strain was found to be closer to genus Paenibacillus. To control the bacterial wilt caused by R. solanacearum, greenhouse experiments were conducted to determine the effectiveness of the Paenibacillus strain KPB3. Drench application of this strain ($4{\times}10^8$ CFU $mL^{-1}$) into the pots containing tomato plants, post-inoculated with the pathogen, R. solanacearum could drastically reduce the disease severity, compared to the non-treated plants. To evaluate effectiveness of this strain under field conditions, experiments were carried out in polyvinyl houses infested with R. solanacearum, during spring and autumn of the year 2006. It was observed that, during spring, bacterial wilt was more prevalent compared to the autumn. During spring, 50.9% disease incidences occurred in non-treated controls, while, Paenibacillus strain KPB3 treated plants showed 24.6% disease incidences. Similarly, during autumn, around 17.2% plants were infected with bacterial wilt in non- treated polyvinyl houses, compared to the Paenibacillus strain KPB3 treated plants, which showed 7.0% disease incidences. These results demonstrated that, Paenibacillus strain KPB3 is a potential biological control agent against bacterial wilt caused by R. solanacearum, effective under greenhouse as well as field conditions. This is the first report showing biocontrol of R. solanacearum using a Paenibacillus spp. under field conditions.

Several causes of non virus-induced mosaic symptom on potato leaves and its induction by herbicides (감자 이상모자이크증상의 몇 가지 발생원인 및 제초제에 의한 증상 유기)

  • Kwon, Min;Hahm, Young-Il;Kim, Hyun-Jun;Yiem, Myoung-Soon
    • The Korean Journal of Pesticide Science
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    • v.5 no.2
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    • pp.45-50
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    • 2001
  • In recent, non virus-induced mosaic symptoms(NVMS) on potato leaves were observed in the seed potato fields, and its incidence rate was $5{\sim}20%$ nationwide. It made difficult to rogue out virus-infected plants, and caused much arguments between seed potato production farmers and seed potato inspectors. The objectives of these experiments were to find out the causes of NVMS, and also to induce mosaic symptom(phytotoxicity) on potato plants by treatment of several herbicides. No significant correlations were found between incidence rates of NVMS and values from soil analyses; soil pH, soil EC, organic matter content, and contents of inorganic constituents($P_2O_5,\;NO_3$, Ca, Mg, K) in the soil around the potato planted. The examinations by ELISA, virus indicator plants, and TEM showed that NVMS on potato leaves was not caused by the viruses infection. But, the use of herbicides could induced the NVMS on potato leaves. The incidence rates of potato treated with pendimethalin linuron of 400 mL/10 a, pendimethalin of 200 mL/10 a, pendimethalin.oxadiazon of 300 mL/10 a, and control were 61.1%, 47.2%, 19.4%, and 1.4%, respectively. Based on these results, we confirmed that the treatment of pendimethalin alone and in mixture with other herbicides were the reason of NVMS on potato leaves. The yields among test plots were similar except dicamba treated plot, which decreased by about 23% compared to control plot. When their progenies harvested in 1999 were planted in the following season, no symptoms of mosaic were observed.

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Identification and Characterization of Three Isolates of Cucumber mosaic virus Isolated from Weed Hosts (잡초에서 분리한 3종 Cucumber mosaic virus의 동정과 특성)

  • Lee, Hyeok-Geun;Kim, Sung-Ryul;Jeon, Yong-Woon;Kwon, Soon-Bae;Ryu, Ki-Hyun;Choi, Jang-Kyung
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.15-20
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    • 2008
  • Three isolates of Cucumber mosaic virus (CMV) were isolated from weed hosts showing typical mosaic symptoms, and some properties of the viruses were investigated. CMV isolates, designated as Is-CMV, Jd-CMV and Pla-CMV from Isodon inflexus, Jeffersonia dubia and Phryma leptostachya var. asiatica, respectively, were identified and characterized by biological reaction in several host plants, serological property, dsRNA analysis, reverse transcription-polymerase chain reaction (RT-PCR), restriction fragment-length polymorphism (RFLP). All isolates systemically infected in Nicotiana benthamiana, Cucurbita pepo cv. Black beauty and Cucumis sativus, and did not reveal any differences in these host plants between the isolates. However, remarkable difference in the symptoms was found between the CMVs in Capsicum annuum. Is-CMV induced an asymptomatic symptoms, while Jd-CMV and Pla-CMV produced severe mosaic symptoms in C. annuum plants. In dsRNA analysis, all isolates revealed four major bands with estimated molecular size of 3.4, 3.2, 2.1 and 1.0 kbp. The cDNAs of coat protein gene of the isolates were amplified by RT-PCR using a genus-specific single pair primers that designed to amplify a DNA fragment of approximately ranging from 938 to 966 bp. By restriction mapping analysis using RFLP of the RT-PCR products as well as by serological properties of gel diffusion test, the CMV isolates belong to a typical members of CMV subgroup IA. This is the first report on the occurrence of CMV in the three weed hosts.