• Journal of Korean Society of Forest Science
• /
• v.95 no.1
• /
• pp.139-144
• /
• 2006

Chestnut trees have been severely damaged by the various cankers in Korea. Field surveys for elucidating the relationships between disease occurrence and environmental factors, i.e., tree age, temperature, rainfall precipitation, direction, degree of slope, chemical properties of soils, and so on, were carried out in the chestnut tree plantations located at Gyeongnam, Jeonnam, Chungnam, and Gyeonggi-Do Province. Totally, one thousand and one hundred thirty seven trees from 36 experiment plots were investigated. Trees infected by at least one of the canker fungi were estimated up to 66%. The most severely infested area was Sunchon in Jeonnam Province with the rate of 80%, while the most lightly infested area was Kongju in Chungnam Province with 39%. Disease occurrence was found to be highly correlated with tree age. The disease occurred most frequently on trees ranged from 7 to 12 years old, while it was relatively low on trees below 5 years old or above 25 years old. Canker positions on the tree formed by the pathogenic fungi were most often at 1.6-2.0m above the ground. Over five hundreds bark samples from the canker area on the tree were collected and the pathogenic fungi were isolated. The most commonly isolated fungus was identified as Cryphonectria parasitica, the chestnut blight fungus.

• The Plant Pathology Journal
• /
• v.18 no.1
• /
• pp.23-29
• /
• 2002

A total of 672 Cryphonectria parasitica was isolated from 2,536 blight lesions on chestnut twigs, which were collected from major chestnut plantations all over Korea. Isolation rates of each province ranged from 13.5% in Jeonbuk-ds to 37.4% in Gyeongnam-do, with an average rate of 25.6%. The isolates were classified into six groups according to color and shape of colony on PDA: smooth margin (S), irregular margin (I), yellow to brown (Y), white (W), and white with yellow center (C). Among these groups, IY was the most abundant with an isolation rate of 65%. On the other hand, SW, SC, IW, and SY were quite rare, with isolation rates ranging from 1.5% to 5.8%. When the 672 isolates were inoculated on the chestnut twigs,380 isolates (56.5%) caused lesions larger than the standard virulent isolate EPISS-2, while 158 isolates (23.4%) caused smaller lesions than the standard hypovirulent isolate UEP-1. However, 87.4% of the isolates belonged to the virulent group and only 12.6% belonged to the hypovirulent group based on Bavendamm test. In the provinces of Jeonnam-do, Jeonbuk-do, and Gyeongnamdo, which have high density of chestnut trees, the rates of hypovirulent-like isolates were over 20%.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2002.11a
• /
• pp.83.1-83
• /
• 2002

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2002.11a
• /
• pp.83.2-84
• /
• 2002

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2002.11a
• /
• pp.84.1-84
• /
• 2002

• Journal of Microbiology and Biotechnology
• /
• v.14 no.3
• /
• pp.620-627
• /
• 2004

On the foundation of a database of genome sequences and protein analyses, the ability to clone a gene based on a peptide analysis is becoming more feasible and effective for identifying a specific gene and its protein product of interest. As such, the current study conducted a protein analysis using 2-D PAGE followed by MALDI- TOF and ESI-MS to identify a highly expressed gene product of C. parasitica. A distinctive and highly expressed protein spot with a molecular size of 47.2 kDa was randomly selected and MALDI-TOF MS analysis was conducted. A homology search indicated that the protein appeared to be a fungal enolase (enol). Meanwhile, multiple alignments of fungal enolases revealed a conserved amino acid sequence, from which degenerated primers were designed. A screening of the genomic $\lambda$ library of C. parasitica, using the PCR amplicon as a probe, was conducted to obtain the full-length gene, while RT-PCR was performed for the cDNA. The E. coli-expressed eno 1 exhibited enolase enzymatic activity, indicating that the cloned gene encoded the C. parasitica enolase. Moreover, ESI-MS of two of the separated peptides resolved from the protein spot on 2-D PAGE revealed sequences identical to the deduced sequences, suggesting that the cloned gene indeed encoded the resolved protein spot. Northern blot analysis indicated a consistent accumulation of an eno1 transcript during the cultivation.

• Proceedings of the Microbiological Society of Korea Conference
• /
• 2005.05a
• /
• pp.159-161
• /
• 2005

Hypovirus infection of the chestnut blight fungus Cryphonectria parasitica is a useful model system to study the hypoviral regulation of fungal gene expression. The hypovirus is known to downregulate the fungal laccase1 (lac 1), the modulation of which is tightly governed by the inositol triphosphate ($IP_3$) and calcium second messenger system in a virus-free strain. We cloned the gene cplc1 encoding a phosphatidyl inositol-specific phospholipase C (PLC), in order to better characterize the fungal gene regulation by hypovirus. Sequence analysis of the cplc1 gene indicated that the protein product contained both the X and Y domains, which are the two conserved regions found in all known PLCs, with a 133 amino acid extension between the 2nd ${\beta}$-strand and the ${\alpha}$-helix in the X domain. In addition, the gene organization appeared to be highly similar to that of a ${\delta}$ type PLC. Disruption of the cplc1 gene resulted in slow growth and produced colonies characterized by little aerial mycelia and deep orange in color. In addition, down regulation of lac1 expression was observed. However, temperature sensitivity, osmosensitivity, virulence, and other hypovirulence-associated characteristics did not differ from the wild-type strain. Functional complementation of the cplc1-null mutant with the PLC1 gene from Saccharomyces cerevisiae restored lac1 expression, which suggests that the cloned gene encodes PLC activity. The present study indicates that the cplc1 gene is required for appropriate mycelial growth, and that it regulates the lac1 expression, which is also modulated by the hypovirus. Although several PLC genes have been identified in various simple eukaryotic organisms, the deletion analysis of the cplc1 gene in this study appears to be the first report on the functional analysis of PLC in filamentous fungi.

• Mycobiology
• /
• v.47 no.4
• /
• pp.512-520
• /
• 2019

Statistical experimental methods were used to optimize the medium for mass production of a novel laccase3 (Lac3) by recombinant Saccharomyces cerevisiae TYEGLAC3-1. The basic medium was composed of glucose, casamino acids, yeast nitrogen base without amino acids (YNB w/o AA), tryptophan, and adenine. A one-factor-at-a-time approach followed by the fractional factorial design identified galactose, glutamic acid, and ammonium sulfate, as significant carbon, nitrogen, and mineral sources, respectively. The steepest ascent method and response surface methodology (RSM) determined that the optimal medium was (g/L): galactose, 19.16; glutamic acid, 5.0; and YNB w/o AA, 10.46. In this medium, the Lac3 activity (277.04 mU/mL) was 13.5 times higher than that of the basic medium (20.50 mU/mL). The effect of temperature, pH, agitation (rpm), and aeration (vvm) was further examined in a batch fermenter. The best Lac3 activity was 1176.04 mU/mL at 25 ℃, pH 3.5, 100 rpm, and 1 vvm in batch culture.

• Molecules and Cells
• /
• v.42 no.4
• /
• pp.363-375
• /
• 2019

Fungal sectorization is a complex trait that is still not fully understood. The unique phenotypic changes in sporadic sectorization in mutants of CpBck1, a mitogen-activated protein kinase kinase kinase (MAPKKK) gene, and CpSlt2, a mitogen-activated protein kinase (MAPK) gene, in the cell wall integrity pathway of the chestnut blight fungus Cryphonectria parasitica have been previously studied. Although several environmental and physiological factors cause this sectoring phenotype, genetic variants can also impact this complex morphogenesis. Therefore, RNA sequencing analysis was employed to identify candidate genes associated with sectorization traits and understand the genetic mechanism of this phenotype. Transcriptomic analysis of CpBck1 and CpSlt2 mutants and their sectored progeny strains revealed a number of differentially expressed genes (DEGs) related to various cellular processes. Approximately 70% of DEGs were common between the wild-type and each of CpBck1 and CpSlt2 mutants, indicating that CpBck1 and CpSlt2 are components of the same MAPK pathway, but each component governs specific sets of genes. Functional description of the DEGs between the parental mutants and their sectored progenies revealed several key pathways, including the biosynthesis of secondary metabolites, translation, amino acid metabolism, and carbohydrate metabolism; among these, pathways for secondary metabolism and translation appeared to be the most common pathway. The results of this comparative study provide a better understanding of the genetic regulation of sector formation and suggest that complex several regulatory pathways result in interplays between secondary metabolites and morphogenesis.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2003.10a
• /
• pp.69.2-70
• /
• 2003

For the selection and breeding of chestnut varieties resistant to the chestnut blight fungus Cryphonectria parasitica, disease resistance and susceptibility of 28 varieties widely planted and growing in Korea were evaluated by artificial inoculation of a pathogenic fungus. For this experiment, a typical virulent strain (KCPC-19) was selected. Artificial inoculation was conducted into all varieties by using two different materials and methods, i.e., bark and wood tissue sections in the laboratory and living trees in the field. In the bark and wood tissue section method, the size of necrotic area and canker development on chestnut varieties were examined and compared 4 days after inoculation. There were wide variations of chestnut varieties in disease resistance and susceptibility against chestnut blight fungus, but 3 varieties, Daebo！, Ishizuchi, and Sandae, were shown to be relatively resistant to the disease with the necrotic area of 0.95-1.03 cm2, while Arima was the most susceptible with the size of 2.0 cm2. In the living tree inoculation examined 5 weeks after inoculation, 3 varieties, Daebo, Ishizuchi, and Riheiguri, showed the higher resistance, but Tono 2 did the highest susceptibility among tested varieties.