• The Plant Pathology Journal
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• 제35권2호
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• pp.100-111
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• 2019

Glomerella leaf spot (GLS) caused by Colletotrichum spp. is a destructive disease of apple restricted to a few regions worldwide. The distribution and evolution of GLS symptoms were observed for two years in Uruguay. The recurrent ascopore production on leaves and the widespread randomized distribution of symptoms throughout trees and orchard, suggest that ascospores play an important role in the disease dispersion. The ability of ascospores to produce typical GLS symptom was demonstrated by artificial inoculation. Colletotrichum strains causing GLS did not result in rot development, despite remaining alive in fruit lesions. Based on phylogenetic analysis of actin, ${\beta}$-tubulin and glyceraldehyde-3-phosphate dehydrogenase gene regions of 46 isolates, 25 from fruits and 21 from leaves, C. karstii was identified for the first time causing GLS in Uruguay and C. fructicola was found to be the most frequent (89%) and aggressive species. The higher aggressiveness of C. fructicola and its ability on to produce abundant fertile perithecia could help to explain the predominance of this species in the field.

• 식물병연구
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• 제26권4호
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• pp.279-282
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• 2020

In 2019, symptoms of Botrytis mold on the peony (Paeonia lactiflora Pall.) 'Sarah Bernhardt' were observed during a survey of the commercial greenhouses of Gangjin County, South Korea. The initial symptoms, small brown spots, were observed mainly at the leaf margins. The lesions extended to the interior of leaves forming irregular spots in which abundant conidia developed. Fungal colonies were obtained from surface-sterilized tissue excised from growing edges of the lesions that were transferred to potato dextrose agar. Melanized irregular sclerotia were formed in these colonies after 40 days at 8℃. Molecular phylogeny based on sequences of genes for glyceraldehyde-3-phosphate dehydrogenase, heat-shock protein 60, and RNA polymerase subunit II were highest for the PBC-2 isolate to the type strains of Botrytis cinerea, rather than other Botrytis species associated with peony diseases. Following Koch's postulates, healthy Sarah Bernhardt plants were inoculated with a foliar application of conidial suspensions of the isolate PBC-2. Following incubation under humidity with a 12 hr photoperiod for 7 days, symptoms developed on the leaf margins that were identical to those observed in the greenhouses. This study is the first report of Botrytis blight caused by B. cinerea on peonies grown in commercial greenhouses in South Korea.

• 식물병연구
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• 제27권2호
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• pp.70-75
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• 2021

The abnormal brown sunken lesions were observed on cv. RubyS apple fruits in an orchard located in Gunwi, Gyeongbuk province, Korea. The primary observed symptoms such as small round sunken lesions and small black dots on the symptomatic area were different from the reported apple diseases. The affected apple fruits were sampled and subjected to isolation of the causal agent. Cultural and morphological characteristics of isolated fungal strain, designated KNUF-20GWA4, were similar to that of Colletotrichum spp. Based on multilocus sequence analyses using internal transcribed spacer regions and partial sequences of β-tubulin, glyceraldehyde-3-phosphate dehydrogenase, chitin synthase, and actin genes, strain KNUF-20GWA4 showed 99.2-100% similarities with C. aenigma ICMP 18608 and the isolate clustered together with several other strains of this species in the phylogenetic tree. To our knowledge, this is the first report of bitter rot on apple fruits caused by C. aenigma.

• Mycobiology
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• 제49권6호
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• pp.599-603
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• 2021

CRISPR/Cas9 genome editing systems have been established in a broad range of eukaryotic species. Herein, we report the first method for genetic engineering in pyogo (shiitake) mushrooms (Lentinula edodes) using CRISPR/Cas9. For in vivo expression of guide RNAs (gRNAs) targeting the mating-type gene HD1 (LeA1), we identified an endogenous LeU6 promoter in the L. edodes genome. We constructed a plasmid containing the LeU6 and glyceraldehyde-3-phosphate dehydrogenase (LeGPD) promoters to express the Cas9 protein. Among the eight gRNAs we tested, three successfully disrupted the LeA1 locus. Although the CRISPR-Cas9-induced alleles did not affect mating with compatible monokaryotic strains, disruption of the transcription levels of the downstream genes of LeHD1 and LeHD2 was detected. Based on this result, we present the first report of a simple and powerful genetic manipulation tool using the CRISPR/Cas9 toolbox for the scientifically and industrially important edible mushroom, L. edodes.

• 한국균학회지
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• 제51권4호
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• pp.277-286
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• 2023

Tan spot, caused by Pyrenophora tritici-repentis, is a major foliar disease in wheat worldwide. In April 2021, tan spot symptoms were observed in a commercial wheat field in Suncheon, Jeonnam Province, Korea, with over 5% of the wheat leaves exhibiting symptoms. These symptoms included oval-shaped tan necrosis surrounded by a bright halo. The three representative isolates exhibited irregular mycelial growth on V8-potato dextrose agar and produced pseudothecia. Based on the concatenated sequence datasets of four multi-genes, including the internal transcribed spacer, large subunit ribosomal RNA, glyceraldehyde-3-phosphate dehydrogenase, and RNA polymerase II second-largest subunit genes, phylogenetic analysis revealed that these three isolates clustered in the same clade as P. tritici-repentis. Results of pathogenicity test indicated that the initial symptoms appeared 5 days post-inoculation (dpi), with typical tan spot symptoms developing at 7 dpi. The pathogen was successfully re-isolated from the symptomatic tissues, thus fulfilling Koch's postulates. Furthermore, we selected three fungicides that effectively inhibited the mycelial growth of P. tritici-repentis by more than 90% in vitro. To the best of our knowledge, this is the first report of tan spot disease in wheat in Korea.

• Asian-Australasian Journal of Animal Sciences
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• 제26권3호
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• pp.423-432
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• 2013

Zi geese (Anser anser domestica) belong to the white geese and are excellent layers with a superior feed-to-egg conversion ratio. Quantitative gene expression analysis, such as Real-time qRT-PCR, will provide a good understanding of ovarian function during egg-laying and consequently improve egg production. However, we still don't know what reference genes in geese, which show stable expression, should be used for such quantitative analysis. In order to reveal such reference genes, the stability of seven genes were tested in five tissues of Zi geese. Methodology/Principal Findings: The relative transcription levels of genes encoding hypoxanthine guanine phosphoribosyl transferase 1 (HPRT1), ${\beta}$-actin (ACTB), ${\beta}$-tubulin (TUB), glyceraldehyde-3-phosphate-dehydrogenase (GADPH), succinate dehydrogenase flavoprotein (SDH), 28S rRNA (28S) and 18S rRNA (18S) have been quantified in heart, liver, kidney, muscle and ovary in Zi geese respectively at different developmental stages (1 d, 2, 4, 6 and 8 months). The expression stability of these genes was analyzed using geNorm, NormFinder and BestKeeper software. Conclusions: The expression of 28S in heart, GAPDH in liver and ovary, ACTB in kidney and HPRT1 in muscle are the most stable genes as identified by the three different analysis methods. Thus, these genes are recommended for use as candidate reference genes to compare mRNA transcription in various developmental stages of geese.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권4호
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• pp.292-296
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• 2004

The recombinant soluble human tumor necrosis factor-alpha (hTNF-$\alpha$) was expressed in a yeast Saccharomyces cerevisiae and its cytotoxicity was evaluated. A cDNA encoding hTNF-$\alpha$ was placed under the control of two different promoters: a glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter and a yeast hybrid ADH2-GPD promoter, consisting of alcohol dehydrogenase II (ADH2) and the GPD promoter. A Northern blot analysis revealed that, although variation in the expression level of hTNF-$\alpha$ existed among transformants, the higher expression was obtained with the GPD promoter. Expressed hTNF-$\alpha$ protein (rhTNF-$\alpha$) was successfully secreted into the culture medium, producing 2.5 mg per liter of culture filtrate, with no changes in cell growth. The bioassay for observing the cytotoxicity to the murine L929 fibroblast cell line, with serial dilution of rhTNF-$\alpha$, indicated that the secreted rhTNF-$\alpha$ was bioactive and its dose-response was improved eight to ten times over that of the E. coli-derived rhTNF-$\alpha$.

• Journal of Microbiology and Biotechnology
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• 제13권4호
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• pp.537-543
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• 2003

The mouse interferon gene (MuIFN-${\gamma}$) was cloned and then used to transform Saccharomyces cerevisiae. Expressed MuIFN-$\{gamma}$ protein (MuIFN-${\gamma}$) was successfully secreted into culture medium due to the presence oi the signal peptide of rice amylase 1A. Two different promoters fused to MuIFN-${\gamma}$ were tested: glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter and a yeast hybrid ADH2-GPD (AG) promoter consisting of alcohol dehydrogenase II (ADH2) and GPD promoter. Using the hybrid promoter, the accumulation of MuIFN-${\gamma}$transcript was the highest after the 24 h cultivation, and then gradually decreased as the cultivation proceeded. However, both cell growth and recombinant MuIFN-${\gamma}$production reached their peaks after the 4-day cultivation. It was possible to produce 6.5 mg/l of MuIFN-${\gamma}$ without any changes in cell growth. Using GPD promoter, the MuIFN-${\gamma}$ transcript accumulation and the recombinant MuIFN-${\gamma}$ production followed the same pattern as the cell growth. However. compared to that of the hybrid promoter, the production of recombinant MuIFN-${\gamma}$ was 0.2 mg/l. The secreted MuIFN-${\gamma}$ had estimated molecular masses of 21 kDa and 23 kDa, which were larger than that of the encoded size due to glycosylation. The protection assay against the viral infection indicated that the recombinant MuIFN-${\gamma}$ was bioactive.

• 한국수산과학회지
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• 제51권1호
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• pp.95-106
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• 2018

To evaluate appropriate reference genes for the normalization of quantitative reverse transcription PCR (RT-qPCR) data with embryonic and larval samples from Russian sturgeon Acipenser gueldenstaedtii, the expression stability of eight candidate housekeeping genes, including beta-actin (ACTB), elongation factor-1A (EF1A), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), histone 2A (H2A), ribosomal protein L5 (RPL5), ribosomal protein L7 (RPL7), succinate dehydrogenase (SDHA), and ubiquitin-conjugating enzyme E2 (UBE2A), were tested using embryonic samples from 12 developmental stages and larval samples from 11 ontogenic stages. Based on the stability rankings from three statistic software packages, geNorm, NormFinder, and BestKeeper, the expression stability of the embryonic subset was ranked as UBE2A>H2A>SDHA>GAPDH>RPL5>EF1A>ACTB>RPL7. On the other hand, the ranking in the larval subset was determined as UBE2A>GAPDH>SDHA>RPL5>RPL7>H2A>EF1A>AC TB. When the two subsets were combined, the overall ranking was UBE2A>SDHA>H2A>RPL5>GAPDH>EF1A>ACTB>RPL7. Taken together, our data suggest that UBE2A and SDHA are recommended as suitable references for developmental and ontogenic samples of this sturgeon species, whereas traditional housekeepers such as ACTB and GAPDH may not be suitable candidates.

• 동의생리병리학회지
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• 제24권6호
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• pp.962-969
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• 2010

The purpose of this investigation was to confirm the effect of Nelumbinis Semen on the recovery of the cardiac muscle activity. We studied the effect of Nelumbinis Semen on the recovery of ischemic SD rat hearts perfused with Nelumbinis Semen, using a model of ex-vivo perfusion (Non-working Langendorff perfusion system) and working heart perfusion system at the same time. To explore the effect of Nelumbinis Semen at the level of proteome, two-dimensional electrophoresis and MALDI-TOF analysis were performed. We found out that the proteins increased after perfusion of Nelumbinis Semen are Mitochondrial aconitase, ATP synthase alpha chain, Lactate dehydrogenase B, Creatine kinase, Glyceraldehyde 3-phosphate dehydrogenase, Alpha B-crystallin, Myosin and Heart fatty acid binding protein. Almost, all of them are concerned with ATP production in the cardiac muscle with glucose metabolism.