• 한국작물학회지
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• 제23권2호
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• pp.113-117
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• 1978

대두 자반병에 대하여 저항성인 계통 선발을 위한 Mass-screening technique를 확립하고 이 종자잡종법을 이용하여 수집재내종 및 각 품종의 저항성검정을 실시한 결과는 다음과 같다. 1. 조숙성인 수집재내종 50계통의 자반병 자연감염률과 종자접종에 의한 자주빛 색소 형성률간에 나타난 정의 상관관계는 r=0.121로서 고도의 유의성이 인정되었으며 이 종자접종법은 효과약인 Mass-screening technique으로서 방사선을 이용한 돌연변이육종에서 자반병저항성 계통선발에 이용가능성이 높았다. 2. 자반병 자연감염률이 극히 낮은 만숙성 수집재내종 85계통은 종자접종에 의하여 대부분이 이병성 반응을 보였으며 이는 포장에서의 만숙계통의 저항성이 유전적인 특성이기 보다는 병해도피 현상에 기인한 것으로 생각되었다. 3. 대두품종의 저항성검정에서 우리나라 대두 36품종을 비롯하여 일본과 대만의 각 16품종, 베트남 11품종, 필리핀 8품종 및 미국 38품종 등 대부분이 이병성이였고 미국품종인 Sac, Hill, Harosoy등이 비교적 저항성 이었다. 4. 각 지역에서 분리한 대두 자반병균 244 isolates 중에서 배지에서 자주빛 색소를 형성하지 않는 두 균주를 제외하고는 모두가 암조건에서는 감자한천배지에서, 광조건에서는 오트밀한친배지에서 자주빛 색소형성이 잘 되었으며 배지에서의 색소형성 정도는 종자에서의 병원성과 밀접한 상관이 있었다.

• The Plant Pathology Journal
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• 제21권2호
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• pp.158-163
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• 2005

Virus-induced gene silencing (VIGS) is a recently developed gene transcript suppression technique for characterizing the function of plant genes. However, efficient VIGS has only been studied in a few plant species. In order to extend the application of VIGS, we examined whether a VIGS vector based on TRV would produce recognizable phenotypes in soybean. Here, we report that VIGS using the Tobacco rattle virus (TRV) viral vector can be used in several soybean cultivars employing various agro-inoculation methods including leaf infiltration, spray inoculation, and agrodrench. cDNA fragments of the soybean phytoene desaturase(PDS) was inserted into TRV RNA-2 vector. By agrodrench, we successfully silenced the expression of PDS encoding gene in soybean. The silenced phenotype of PDS was invariably obvious 3 weeks after inoculation with the TRV-based vector. Real-time RT-PCR analyses showed that the endogenous level of GmPDS transcripts was dramatically reduced in the silenced leaf tissues. These observations confirm that the silenced phenotype is closely correlated with the pattern of tissue expression. The TRV-based VIGS using agrodrench can be applied to functional genomics in a soybean plants to study genes involved in a wide range of biological processes. To our knowledge, this is the first high frequency VIGS method in soybean plants.

• The Plant Pathology Journal
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• 제22권4호
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• pp.348-352
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• 2006

Barley mild mosaic virus(BaMMV) is a soilborne Bymovirus vectored by root-infecting fungus, Polymyxa graminis. Mechanism of cultivar's resistance to BaMMV in field tests are difficult to assess since resistance could be either due to the virus or to P. graminis, or both. Whereas, available mechanical inoculation methods for BaMMV and other related viruses are labor intensive, give inconsistent results and generally result in low infection rates. Inoculation method using stick with gauze(SWG) was developed for BaMMV. The improved method proved to be simple, efficient, and reliable. The infected leaf tissues were preserved by drying in a frozen state under high vaccum(freeze dried barley infected leaves) to circumvent reduction of virus infectivity during storage. Five Korean barley cultivars were mechanically inoculated with BaMMV-infected sap by the improved method. Infection rates obtained were compared with natural infection. Cultivar Naehanssalbori showed resistance to BaMMV in the field trials but was found highly susceptible in the greenhouse tests by mechanical inoculation, indicating that the field resistance may be possibly due to resistance to P. graminis.

• 대한수의학회지
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• 제13권1호
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• pp.39-46
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• 1973

Cultural method, dark field microscopy & fluorescent antibody technique were compared for their sensitivity of the detection of leptospires from experimentally infected mice. Two groups of mice were infected with L. icterohemorrhagiae (M20) and L. australis (Ballico), and the infected blood, urine and a number of organs were subjected to the bacterial isolation. The results obtained were summarized as follows: 1. L. icterohemorrhagiae (M20) and L. australis (Ballico) in blood, urine and various tissues of experimentally infected mice were detected with a negrigible non specificity, by the fluorescent antibody technique. 2. The fluorescent antibody technique, as applied to detection of leptospires in blood, urine and various infected tissue, proved to be better than cultural method and dark-field microscopy. 3. Early detection of leptospires by fluorescent antibody technique were possible in blood at 2 days after inoculation, whereas detection of organisms in liver, spleen, lung and kidney were observed later. By means of fluorescent antibody technique, the detection of leptospires in kidney and urine was possible up to 34 days postinoculation, whereas those in other parts were impossible. 4. Fluorescent antibody reaction of leptospires were highly specific to homologous antigen rather than to heterologous one. 5. Fluorescent antibody technique may be of value in the application for the demonstration of leptospira from clinical specimens.

• 대한수의학회지
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• 제35권2호
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• pp.317-326
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• 1995

The purpose of this study was to establish a method for in vitro culture of C parvum isolated in Korea by determination of suitable cell model to complete development of this parasite. The result obtained were summerized as follows: 1. To determine the most suitable cell line, six types of cell line were examined by microscopy. All cell lines were infected with C parvum and showed the highest infection score in HmLu cells. 2. The staining methods including DMSO-modified acid-fast(A-F) stain, hematoxylin-eosin(H & E) stain and immunofluorescence antibody(IFA) stain were applied to examine the infection of C parvum in cell culture. These staining methods were possible to examine the infection of C parvum in cell culture. The most sensitive one was IFA staining technique. 3. Developmental stages of C parvum in HmLu cell were observed. After the initial 8 hour incubation period, some trophozoites were observed. The meronts and gametes were appeared at 24-48 hour post inoculation(PI), and oocysts were observed firstly at 48-72 hour PI. 4. In H & E stain, the parasite appeared as basophilic within parasitophorous vacuole membrane(PVM) and lying in cytoplasm at near the nucleus of the host cells. It was able to distinguish the type I, type II meronts and gametes. 5. In DMSO-modified acid-fast stain, specific stained parasites were appeared firstly after 48 hour PI. The parasites were showed with different degrees of staining bright red color within PVM. 6. The endogenous stages of parasites in HmLu cell recovered at 48, 96, 120 and 144 hour after inoculation were reacted with rabbit immunized serum in immunofluorescence antibody and avidin-biotin complex peroxidase staining technique.

• 한국동물위생학회지
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• 제15권2호
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• pp.184-194
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• 1992

This study was done to identify Newcastle disease virus(NDV) antigens in paraffin sections of various organs from experimentally NDV-infected chicken using peroxidase-antiperoxidase(PAP) technique. Sections were Incubated with rabbit anti-NDV polyclonal as first antibody, followed by incubation with goat anti-rabbit IgG conjugate and peroxidase anti-peroxidase ( PAP ). Positive reactions were often detected in the epithelim of trachea and in the lymphocyte of spleen at 24 hours after virus inoculation. The viral antigen was localized mainly in the cytoplasm of infected cells. The method approved to be highly specific for the indetification of NDV and allowed a precise localization of the viral antigens in infected cells.

• 대한수의학회지
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• 제30권3호
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• pp.309-315
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• 1990

The present experiment was done to identify newcastle disease virus(NDV) antigens in frozen sections of various oragns from experimentally NDV-infected with indirect immunoperoxidase method. Section were incubated with rabbit anti-NDV polyclonal as first antibody, followed by incubation with goat anti-rabbit or protein A peroxidase conjugate. Positive reactions were often detected in the epithelium of trachea and in the lymphocyte of spleen at 24 hours after virus inoculation. the viral antigen was localized mainly in the cytoplasm of infected cells. The method approved to be highly specific for the identification of NDV and allowed a precise localization of the viral antigens in infected cells.

• 대한의생명과학회지
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• 제11권1호
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• pp.71-77
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• 2005

The etiologic agents of haemorragic fever with ranal syndrom (HFRS) in Korea are Hantaan and Seoul virus in the genus Hantavirus, family Bunyaviridae. In order to elucidate the role of maternal immunity to Hantavirus infection in rats, the protective effect of the maternal antibody were studies by using rats experimentally infected with Seoul virus strain HR80-39. Antibody titers of sera and viral antigen against Seoul virus were investigated by indirect immunofluorscence antibody technique (IFA). The dam sera had IFA antibody titers ranging from 1:128 to 1:1,024 after parturition. In fetuses, IFA antibody titers ranged from 1: 16 to 1:64 just after birth, increased to peak titers ranged from 1:256 to 1:1,024 in the 2nd week after birth. Challenged newborn rats had IFA antibody titers ranging from 1:64 to 1:1,024 after inoculation. No viral antigen was detected in lungs or other organs of the newborn rats. The maternal antibody to Seoul virus was transferred prenatally through placenta and postnatally via colostrum from immune dams to their offspring. These results demonstrated that maternal antibody to Seoul virus was quite effective in protecting newborn rats against same virus infection.

• 한국응용곤충학회지
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• 제12권1호
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• pp.23-27
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• 1973

갈색엽고병(Fusarium niwate)의 효과적인 접종방법을 찾아내기 위해 본 시험을 수행하였던 바 다음과 같은 결과를 얻었다. 1) 본 균의 분생포자의 증류수 현탁액을 벼 유묘에 분무접종할 때, 접종 후 습실에 1일 보존했을 경우에는 거의 발병이 되지 않았으나 2일간 보존한 겅우에는 어느 정도 병반이 나타났다. 2) 벼 유묘에 바람이나 또는 나무막대기로 물리적인 상처를 입힌 다음 증류수 포자현탁액을 분무한 결과 상당한 병만이 형성되었다. 3) 포자현탁액에 벼 잎 추출액, glucose, polypeptone, yeast exract 등의 $1\%$ 용액을 첨가 분무접종했을 때 병반형성이 양호하였다. 4) 포자발아상태는 종류수중에서는 불량하였으나 영양분을 첨가한 용액중에서는 발아력도 좋았고 균사의 융합도 양호하게 일어났다. 5) 본 접종시험의 결과 통일품종은 재래 장려품종인 풍광에 비해 본 병에 대해 약했다.

• 한국작물학회지
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• 제45권2호
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• pp.143-149
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• 2000

From the soils of soybean fields in Cotton Branch Station (CBS) and Pine Tree Station (PTS), Arkansas, USA, various single spore isloates of sudden death syndrome (SDS) pathogen were obtained on modified Nash ＆ Snyder's medium (MNSM) with dilution plating technique and transferred to potato dextrose agar (PDA) medium to identify the cultural colony shape. The colony shapes of these isolates resembled F. solani isolate 171 which was white and chalky shaped on MNSM and most of them had unique form of morphology which produced white margin and blue center colony on PDA. Although, some of these isolates had more dark blue or showed slightly different color, all isolates that were selected randomly for green-house inoculation assay produced typical foliar symptoms on leaves of soybean, Hartz 6686. To determine the genetic differences among the isolates, mitochondrial DNA restriction fragment length polymorphism (RFLP) was conducted with fourty isolates from both fields, using mtDNA probes, 2U18 and 4U40, derived from Colletotrichum orbiculare. We obtained distinctive RFLPs in each treatment of restriction enzyme, EcoRI and HaeⅢ. Isolates, 11-2-5 and 14-3-1-1, from CBS and isolates, 104-3-1-2 and 701-1-5-1, from PTS showed different band patterns from 171 in both or in either treatment of restriction enzymes. Even if some of these isolates showed heterogeneous, they were more closer to 171 than PN603. And, also, rest of the thirty-six isolates had exactly same polymorphisms as 171 in each treatment of restriction enzyme. Although, some of the isolates showed the different morphological shape on PDA and slightly different band patterns on RFLPs, all of the isolates selected on MNSM due to their distinctive colony shape from other fungi produced the typical foliar symptoms on soybean leaves in greenhouse inoculation assay. It might be suggested that these isolates were not genetically different from check isolate 171 and they were unique strain of F. solani.