• The Plant Pathology Journal
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• 제31권4호
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• pp.334-342
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• 2015

Two fumonisin-nonproducing strains of Fusarium verticillioides and their fumonisin producing progenitors were tested for aggressiveness toward maize, sorghum, rice, and beetroot seedlings grown under greenhouse conditions. None of the plants showed obvious disease symptoms after root dip inoculation. Fungal biomass was determined by species-specific real-time PCR. No significant (P = 0.05) differences in systemic colonization were detected between the wild type strains and mutants not producing fumonisins. F. verticillioides was not detected in any of the non-inoculated control plants. The fungus grew from roots to the first two internodes/leaves of maize, rice and beet regardless of fumonisin production. The systemic growth of F. verticillioides in sorghum was limited. The results showed that fumonisin production was not required for the infection of roots of maize, rice and beet by F. verticillioides.

• Animal cells and systems
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• 제13권3호
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• pp.297-304
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• 2009

A complete mRNA sequence of transferrin from the wax moth, Galleria mellonella, was obtained, and compared with those of other species. We previously reported that the sequence was most similar to those of Manduca sexta and Bombyx mori. As in other moths, G. mellonella transferrin had only one iron-binding site at its N-terminal region. Semi-qRT PCR was conducted to investigate tissue-specific distribution and transcriptional regulation of the wax moth transferrin mRNA. Larval muscle and fat body contained larger quantity of mRNA than other tested tissues. In this study, it was observed that iron and cadmium regulated transferrin transcription, and this regulation pattern was tissue specific. Iron up-regulated transferrin mRNA level in fat body, while suppressed it in the Malpighian tubules and silk glands. Cadmium decreased the mRNA level in fat body, muscle, and Malpighian tubules, but significantly increased the mRNA level in silk glands. In addition, the mRNA expression was induced by all tested pathogen-associated molecular patterns (PAMPs) including LPS, lipoteichoic acid (LTA), glucan, and even chitin.

• Mycobiology
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• 제41권4호
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• pp.252-255
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• 2013

Fungal pathogens have caused severe damage to the commercial production of Pleurotus eryngii, the king oyster mushroom, by reducing production yield, causing deterioration of commercial value, and shortening shelf-life. Four strains of pathogenic fungi, including Trichoderma koningiopsis DC3, Phomopsis sp. MP4, Mucor circinelloides MP5, and Cladosporium bruhnei MP6, were isolated from the bottle culture of diseased P. eryngii. A species-specific primer set was designed for each fungus from the ITS1-5.8S rDNA-ITS2 sequences. PCR using the ITS primer set yielded a unique DNA band for each fungus without any cross-reaction, proving the validity of our method in detection of mushroom fungal pathogens.

• 한국어류학회지
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• 제18권4호
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• pp.300-310
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• 2006

한국 서해안의 서천 및 고창지역과 남해안의 부산지역으로부터 채취한 전어(Konosirus punctatus) 3개 집단의 개체로부터 genomic DNA를 분리 추출하여 PCR로 반복해서 증폭시켰다. 8개의 decamer와 20-mer를 사용하여 전체적으로 서천의 전어집단에서 713개의 loci, 부산집단에서 791개 및 고창 전어집단으로부터 732개의 100 bp에서 2,800 bp의 크기에 해당되는 total loci를 얻어냈다. 우리는 서천 전어집단에서 독특한 50개의 unique loci, 부산 전어집단으로부터 70개의 unique loci 그리고 고창의 전어집단으로부터 130개의 unique loci를 각각 확인하였고, 또한 3개 전어집단 모두에 대해서 공통적으로 가지고 있는 120개의 shared loci도 확인하였다. 특이한 specific loci를 확인한 결과 서천 전어집단에서는 108개(15.1%), 부산집단에서는 74개(9.4%) 그리고 고창 전어집단에서는 67개(9.2%)를 각각 얻어냈다. 또한 8개의 primer를 통해서 서천 전어집단에서 48개 (6.7%), 부산 전어집단에서는 26개 (3.3%) 그리고 고창 전어집단에서 16개 (2.2%)의 polymorphic loci를 얻어냈다. Similarity matrix를 통해서 볼 때 서천 전어집단에서 0.756에서 0.936까지, 부산집단에서 0.800에서 0.938까지 그리고 고창 전어집단에서 0.731에서 0.959까지의 공유가(bandsharing value)를 확인하였다. 8개의 primer를 이용하여 얻어진 dendrogram을 통해서 볼 때 genetic cluster는 cluster 1 (SEOCHEON 01~SEOCHEON 10), cluster 2 (BUSAN 11~BUSAN 20과 GOCHANG 23~GOCHANG 24) 그리고 cluster 3 (GOCHANG 21, 22, 25, 26, 27, 28, 29 및 30)와 같이 3개의 cluster로 나누어졌다. 위에서와 같이 고창 전어집단의 일부 개체는 부산 전어집단에 속하는 것으로 나타났으며, 따라서 2 전어집단의 일부 개체들은 부분적으로 오고 가는 이주현상을 나타내는 것으로 사려된다. 이러한 결과를 볼 때 RAPD-PCR 분석 방법을 통해서 우리는 지리적으로 떨어져 있는 3개의 전어 집단에 존재하는 유의성이 있는 유전적 거리를 확인할 수 있었다. 여러 가지 decamer와 20-mer를 이용한 RAPD-PCR 분석 방법은 종 및 지리적 집단과 지리적 전어집단에 존재하는 유전적 다양성, 다형성 및 유전적 유사성을 확인하는데 필요로 하는 독특한 specific/polymorphic marker를 확인할 수 있는 이용 가능한 방법이라고 할 수 있다.

• 한국균학회지
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• 제24권2호통권77호
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• pp.155-165
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• 1996

느타리 버섯속 6종 23균주에 대한 rDNA의 ITS II 부위의 DNA 염기서 열을 비교 분석함으로서 종간 및 종내의 근연관계를 조사하였다. rDNA의 ITS II 부위를 증폭하고자 5.8S rDNA의 3'말단 부위와 285 rDNA의 5'말단 부위에 두개 프라이머를 이용하여 PCR증폭을 행하였다. 느타리버섯 6종의 게놈 DNA로 부터 ITS II 부위를 증폭한 결과 종에 따라 밴드 길이에 차이가 있었으며 ITS II 부위의 길이 차이로 느타리 버섯 6종을 구분할 수 있었다. 같은 종내 개체간 구분을 위하여 느타리 버섯 6종 23균주의 PCR 증폭 결과는 느타리버섯(P. ostreatus) ASI 2096, ASI 2025와 노랑 느타리 버섯(P. cornucopiae) ASI 2038 균주외에 동일 종내 균주의 ITS II 길이는 동일한 양상을 보였다. 느타리 버섯 6종 및 ASI 2095, ASI 2025 그리고 ASI 2038에 대한 ITS II 부위의 염기서열을 비교해 보면 염기서열의 변이가 ITS II 부위가 시작되는 부분과 말단 부분에서 주로 존재하였다. ASI 2095와 ASI 2025 균주들은 동일 종내 느타리 버섯(P. ostreatus, ASI 2001) 균주와 ASI 2038와 ITS II 부위의 DNA 염기서열에 차이를 보였다. 이들 염기서열을 기초로 하여 Neighbor program을 이용한 균주간 유연관계는 느타리 버섯(P. ostreatus), 사철 느타리 버섯(P. florida), 여름 느타리 버섯(P. cornucopiae) 그리고 맛 느타리 버섯(P. eryngii)들이, 노랑 느타리 버섯(P. cornucopiae)과 호고 느타리 버섯(P. cystidious)이 각각 서로 가까운 유연관계를 나타내었으나 ASI 2025와 ASI 2038 균주 들은 특이한 계통분지를 나타내었다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권2호
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• pp.184-193
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• 2007

In mammalian ovary, steroidogenic acute regulatory (StAR) protein mediates the true rate-limiting step of transport of cholesterol from outer to inner mitochondrial membrane. Appropriate expression of StAR gene represents an indispensable component of steroidogenesis and its regulation has been found to be species specific. However, limited information is available regarding StAR gene expression during estrous cycle in buffalo ovary. In the present study, expression, localization and hormonal regulation of StAR mRNA were analyzed by semi-quantitative RT-PCR in buffalo ovary and partial cDNA was cloned. Total RNA was isolated from whole follicles of different sizes, granulosa cells from different size follicles and postovulatory structures like corpus luteum and Corpus albicans. Semi-quantitative RT-PCR analyses showed StAR mRNA expression in the postovulatory structure, corpus luteum. No StAR mRNA was detected in total RNA isolated from whole follicles of different size including the preovulatory follicle (>9 mm in diameter). However, granulosa cells isolated from preovulatory follicles showed the moderate expression of StAR mRNA. To assess the hormonal regulation of StAR mRNA, primary culture of buffalo granulosa cells were treated with FSH (100 ng/ml) alone or along with IGF-I (100 ng/ml) for 12 to 18 h. The abundance of StAR mRNA increased in cells treated with FSH alone or FSH with IGF-I. However, effect of FSH with IGF-I on mRNA expression was found highly significant (p<0.01). In conclusion, differential expression of StAR messages was observed during estrous cycle in buffalo ovary. Also, there was a synergistic action of IGF-I on FSH stimulation of StAR gene.

• The Plant Pathology Journal
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• 제22권1호
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• pp.36-45
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• 2006

Twenty isolates of Didymella bryoniae were isolated from infected cucurbit plants in various growing areas of southern Korea in 2001 and 2002. Random Amplified Polymorphic DNA (RAPD) group [RG] I of D. bryoniae was more virulent than RG IV to watermelon. Virulence of the RG I isolate was strong to moderate to cucumber, whereas that of the RG IV varied from strong, moderate to weak. Two hundred seventy-three amplified fragments were produced with 40 primers, and were analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYSPC. At the distance level of 0.7, two major genomic DNA RAPD groups were differentiated among 20 isolates. The RG I included 7 isolates from watermelon and one isolate from melon, whereas the RG IV included 12 isolates from squash, cucumber, watermelon and melon. Amplification of internal transcribed spacer (ITS) region and small subunit rRNA region from the 20 isolates yielded respectively a single fragment. Restriction pattern with 12 restriction enzymes was identical for all isolates tested, suggesting that variation in the ITS and small subunit within the D. bryoniae were low. Amplification of the genomic DNAs of the tested isolates with the sequence characterized amplified regions (SCAR) primer RG IF-RG IR specific for RG I group resulted in a single band of 650bp fragment for 8 isolates out of the 20 isolates. Therefore, these 8 isolates could be assigned into RG I. The same experiments done with RG IIF-RG IIR resulted in no amplified PCR product for the 20 isolates tested. An about 1.4 kb-fragment amplified from the RG IV isolates was specifically hybridized with PCR fragments amplified from genomic DNAs of the RG IV isolates only, suggesting that this PCR product could be used for discriminating the RG IV isolates from the RG I isolates as well other fungal species.

• The Plant Pathology Journal
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• 제33권2호
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• pp.140-147
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• 2017

Fusarium wilts of strawberry, caused by Fusarium oxysporum f. sp. fragariae, is a serious soil-borne disease. Fusarium wilt causes dramatic yield losses in commercial strawberry production and it is a very stubborn disease to control. Reliable chemical control of strawberry Fusarium wilt disease is not yet available. Moreover, other well-known F. oxysporum have different genetic information from F. oxysporum f. sp. fragariae. This analysis investigates the genetic diversity of strawberry Fusairum wilt pathogen. In total, 110 pathogens were isolated from three major strawberry production regions, namely Sukok, Hadong, Sancheong in Gyeongnam province in South Korea. The isolates were confirmed using F. oxysporum f. sp. fragariae species-specific primer sets. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses were executed using the internal transcribed spacer, intergenic spacer, translation elongation factor1-${\alpha}$, and ${\beta}$-tubulin genes of the pathogens and four restriction enzymes: AluI, HhaI, HinP1I and HpyCH4V. Regarding results, there were diverse patterns in the three gene regions except for the ${\beta}$-tubulin gene region. Correlation analysis of strawberry cultivation region, cultivation method, variety, and phenotype of isolated pathogen, confirmed that genetic diversity depended on the classification of the cultivated region.

• 한국동물위생학회지
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• 제33권4호
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• pp.345-351
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• 2010

Toxoplasma gondii is a species of parasitic protozoa in the genus Toxoplasma. The definitive host of T. gondii is the cat, but the parasite can be carried by the vast majority of warm-blooded animals, including humans. It is often found in the tissues of food animals including pigs and sheep. To determine the regional prevalence of infection with T. gondii, bloods (n=300) from domestic pigs and tissues (n=200) from slaughter pigs in Gyeongnam province were tested using an enzyme-linked immunosorbent assay (ELISA) and a polymerase chain reaction (PCR) for detection of antibody and antigen. A total of 115 sero-positive pigs were identified for a prevalence rate of 38.3%. Of the 50 herds from domestic pigs tested, 34 had at least one sero-positive pig for a herd prevalence rate of 68.0%. Sero-positive rates of pigs in fattening farm were higher than that of pigs in breeding company. Sero-positive rates of sows were higher than that of growing pigs. Seasonally, sero-positive rates of pigs were highest in winter (80.0%) and lowest in spring (23.8%). According to farm size, sero-positive rates of pigs were higher in small size farms (${\leq}$2,000) than that of big size farms (>2,000). However, none of the bloods (n=300) from domestic pigs and tissues (n=200) from slaughter pigs were positive for T. gondii specific DNA by PCR.

• 생명과학회지
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• 제13권5호
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• pp.651-657
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• 2003

형태적으로 매우 유사한 적조생물 C. polykrikoides, G. impudicum, G. catenatum을 RAPD 방법을 이용하여 유전적 유연관계를 조사했다. 12개의 primer 중 4종류만 선택되었고 증폭된 밴드수는 59개이며 그 크기는 0.2에서 3.0 kb까지였다. C. polykrikoides, G. impudicum, G. catenatum의 다형화된 밴드수는 16개, 8개, 16개로 각각 나타났다. 반면에, 17개의 밴드만 동일하였다. C. polykrikoides, G. impudicum, G. catenatum의 종 특이적인 밴드수는 26개, 34개, 26개로 각각 보였다. C. polykrikoides와 G. impudicum/G. catenatum의 유전적 유사성은 0.83이며, G. impudicum과 G. catenatum은 0.78로 나타났다. 이러한 결과로 볼 때 형태적으로는 유사하게 보이지만, RAPD 분석에 의하면 C. polykrikoides, G. impudicum, G. catenatum은 현저하게 상이한 적조생물이다. 앞으로 RAPD 기법을 이용하면 이러한 와편모조류의 유전적 변이를 탐색하는데 유용할 것으로 보인다.