• Mycobiology
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• 제37권4호
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• pp.247-250
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• 2009

Twenty-five isolates of Fusarium fujikuroi acquired from rice seeds and rice plants evidencing symptoms of Bakanae disease were evaluated to determine their mating types and characterize the formation of their sexual state. The mating types of the isolates were evaluated via multiplex PCR with the diagnostic primers of the mating-type (MAT) region: GFmat1a, GFmat1b, GFmat2c, and GFmat2d. Among the 25 isolates, 11 were identified as MAT-1 (male), and 14 as MAT-2 (female). Four MAT-1 isolates and three MAT-2 isolates were mated and cultured to evaluate the optimal culture conditions for the production of their sexual states. Among four tested media, 10% V8 juice agar proved optimal for the perithecial production of the isolates. The isolates also generated the largest numbers of perithecia when incubated at 23oC in alternating cycles of 12 hr fluorescent light and NUV fluorescent light and 12 hr darkness.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.85.1-85
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• 2003

In most ascomycetes, a single mating type locus, MAT, with two alternate forms (MAT1-1 and MAT1-2) called idiomorphs, controls mating ability. In heterothallic ascomycetes these alternate idiomorphs reside in different nuclei. In contrast, most homothallic ascomycetes carry both MAT1-1 and MAT1-2 in a single nucleus, usually closely linked. An example of the latter is Gibberella zeae, a producer of mycotoxins such as trichothecene and zearalenone that threaten human and animal health. We asked if G. zeae could be made strictly heterothallic by manipulation of MAT. Targeted gene replacement was used to differentially delete MAT1-1 or MAT1-2 from a wild type haploid MAT1-1 MAT1-2 strain, resulting in MAT1-1；mat1-2, mat1-1；MAT1-2 strains that were self-sterile, yet able to cross to wild type testers and more importantly, to each other. These results indicated that differential deletion of MAT idiomorphs eliminates selfing ability of G. zeae, but the ability to outcross is retained. To our knowledge, this is the first report of complete conversion of fungal reproductive strategy from homothallic to heterothallic by targeted manipulation of MAT. Practically, this approach opens the door to simple and efficient procedures for obtaining sexual recombinants of G. zeae that will be useful for genetic analyses of mycotoxin production and other traits, such as ability to cause disease.

• The Plant Pathology Journal
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• 제19권3호
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• pp.143-147
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• 2003

Sensitivity of Phytophthora infestans isolates to fungicides metalaxyl and ethaboxam in Korea was examined with 260 isolates for 3 years (9 isolates in 2000,93 isolates in 2001, and 158 isolates in 2002). Both Al and A2 mating types were found from the isolates collected for 3 years. Al mating type was dominant in the population with 8 isolates (88.9%) in 2000, 84 isolates (89.4%) in 2001, and 138 isolates (87.3%) in 2002. Only some isolates from diseased tomatoes in Buyergun and diseased potatoes in Pyeongchanggun were of the A2 mating type. As for metalaxyl sensitivity, 77.0% of the isolates were moderately resistant with 8 isolates (88.9%) in 2000, 73 isolates (77.7%) in 2001, and 120 isolates (75.9%) in 2002. Meanwhile, those found resistant were 1 isolate (11.1%) in 2000, 16 isolates (17.0%) in 2001, and 33 isolates (20.9%) in 2002. Only 5 isolates (3.2%) were sensitive to metalaxyl in 2002. There was no significant difference in the sensitivity among years. As for ethaboxam, no isolate was able to grow at 5.0 $\mu\textrm{g}$ /ml, and only four isolates (1.5 %) grew at 1.0 $\mu\textrm{g}$ /ml with heavy retardation compared with the untreated control. Based on these 3-year results, the minimum inhibitory concentration (MIC) of ethaboxam to p. infestans was determined to be 0.2-1.0 $\mu\textrm{g}$ /ml. Results indicate that resistance development by p. infestans to ethaboxam is not likely to occur in the natural condition. furthermore, there was no indication of cross resistance between metalaxyl and ethaboxam because all the isolates, regardless of classification for their sensitivity to metalaxyl, were not able to grow at 5.0 $\mu\textrm{g}$ /ml of ethaboxam.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1223-1232
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• 2017

Cordyceps militaris, a member of Ascomycota, a mushroom referred to as caterpillar Dong-chung-ha-cho, is commercially valuable because of its high content of bioactive substances, including cordycepin, and its potential for artificial cultivation. Cordycepin (3'-deoxyadenosine) is highly associated with the pharmacological effects of C. militaris. C. militaris is heterothallic in that two mating-type loci, idiomorph MAT1-1 and MAT1-2, exist discretely in two different spores. In this study, nine C. militaris strains were mated with each other to prepare newly bred strains that produced a larger amount of cordycepin than the parent strains. Nine strains of C. militaris were identified by comparing the internal transcribed spacer sequence, and a total of 12 single spores were isolated from the nine strains of C. militaris. After the MAT idiomorph was confirmed by PCR, 36 mating combinations were performed with six single spores with MAT1-1 and the others with MAT1-2. Eight mating combinations were successfully mated, producing stroma with perithecia. Cordycepin content analysis of all strains by high-performance liquid chromatography revealed that the KASP4-bred strain produced the maximum cordycepin among all strains, regardless of the medium and stroma parts. Finally, universal rice primer-PCR was performed to demonstrate that the bred strains were genetically different from the parental strains and new C. militaris strains. These results may be related to the recombination of genes during mating. The newly produced strains can be used to meet the industrial demand for cordycepin. In addition, breeding through mating suggests the possibility of producing numerous cordycepin-producing C. militaris strains.

• Mycobiology
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• 제46권1호
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• pp.64-71
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• 2018

The aim of this study was to investigate the pattern of distribution of mating type (MAT) genes of Tuber indicum in ectomycorhizosphere soils from natural T. indicum-producing areas and cultivated truffle orchards and ascocarp samples from different regions. Quantitative real-time PCR and multiplex PCR were used to weight the copy numbers of MAT1-1-1 and MAT1-2-1 in natural truffle soils and cultivated orchard soils. The effect of limestone on the pattern of truffle MAT genes and the correlation between soil properties and the proportion of MAT genes were also assessed. These results indicated that an uneven and nonrandom distribution of MAT genes was common in truffle-producing areas, cultivated truffle orchards, and ascocarps gleba. The competition between the two mating type genes and the expansion of unbalanced distribution was found to be closely related to truffle fructification. Limestone treatments failed to alter the proportion of the two mating type genes in the soil. The content of available phosphorus in soil was significantly correlated with the value of MAT1-1-1/MAT1-2-1 in cultivated and natural ectomycorhizosphere soils. The application of real-time quantitative PCR can provide reference for monitoring the dynamic changes of mating type genes in soil. This study investigates the distributional pattern of T. indicum MAT genes in the ectomycorhizosphere soil and ascocarp gleba from different regions, which may provide a foundation for the cultivation of T. indicum.

• 한국버섯학회지
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• 제1권1호
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• pp.9-14
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• 2003

새로운 백색 팽이 품종을 육성하고자 수집균주 중 백색자실체를 생성하는 5개의 이핵계통을 선발하고 이들로부터 단핵균주를 분리하였다. 분리된 단핵균주 계통의 교배형을 검정한 결과 A1B1A2B2 이었으며, 이핵균주들에 비하여 균사생장 속도가 느리고 변이폭도 좁다는 것을 알 수 있었다. 분리된 단핵계통의 근연간 교배를 수행한 결과 육성된 모든 이핵균주는 백색이었으며 모본보다 우수한 균주들을 선발할 수 있었다. 특히 Fv 4-1균주는 가장 높은 수량성과 안정성을 나타내었다. 또한 상호교배에 의해 일부교배조합에서 세포질효과를 확인할 수 있었다. 따라서 근연간 교배법은 기존 재배품종들이 가진 백색과 같은 특성을 유지하면서 새로운 품종을 육성하기 위한 방법으로 사용할 수 있을 것으로 생각된다.

• The Plant Pathology Journal
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• 제17권5호
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• pp.281-289
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• 2001

We analyzed 88 strains of Gibberella fujikuroi (Analmorph: Fusarium section Liseola) from maize in Korea for mating population, mating type, fumonisin production vegetative compatibility, and random amplified polymorphic DNA (RAPD) patterns. We found 50 strains that were MATA-2, 22 that were MATA-1, 1 that was MATD-1, and 15 that were not reproducibly fertile with any of the mating type testers. Of the 50 MATA-2, 15 were female fertile, while 10 of the 22 MATA-1 strains were female fertile. A total of 1,138 nitrate non-utilizing (nit) mutants were recovered from a total of 88 strains. These strains were grouped into 39 vegetative compatability groups (VCGs) by demonstrating heterokaryosis between nit mutants. A single maize ear could be infected by more than one VCG of F. moniliforme. RAPD analysis measured genetic diversity among 63 strains of F. moniliforme. Several VCGs were distinguished by RAPD fingerprinting patterns. Most strains produced significant levels of fumonisins. However, 6 MATA-2 strains from a single VCG produced higher levels of fumonisin $\textrm{B}_3$ than that of fumonisin $\textrm{B}_1$ or $\textrm{B}_2$. From these data, we concluded that most Korean strains of F. moniliforme associated with maize belonged to mating population A and produced significant levels of fumonisins. Futhermore, RAPD analysis could differentiate strains associated with different VCGs.

• 한국버섯학회지
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• 제11권1호
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• pp.1-8
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• 2013

본 연구에서 한국에 자생하는 12종의 민자주방망이버섯과 13종의 프랑스, 스위스, 포루투갈, 일본 등지에서 버섯을 수집하였다. 이들 한국, 외국 품종의 민자주방망이버섯은 단핵균사를 얻기 위해 이들 버섯으로부터 단포자를 얻어 YB배지에 배양하였다. 또한 민자주방망이버섯 육종을 위하여 자실체의 형태적 특성과 단핵균주의 특성 등을 고려하여 교배 단핵균주를 선별하였다. 그 결과 선별된 11개의 교배계통($K1{\times}K2$, $K1{\times}K3$, $K1{\times}K4$, $K1{\times}K6$, $K1{\times}K5$, $K2{\times}K4$, $K2{\times}K3$, $K2{\times}K6$, $K3{\times}K4$, $K4{\times}K5$, $K4{\times}K6$)에서 clamp connection을 관찰하였으며, 또한 RAPD 패턴 분석을 통해 교배 여부를 판단하였다. 교배된 2핵균주를 다시 PDA 배지에 배양하여 교배균주의 양상을 관찰한 결과 선별된 11개의 국내 야생 민자주방망이버섯으로 이루어진 교배계통은 야생품종 2핵 균사 또는 외국 민자주방망이버섯 품종 보다 성장속도가 빠른 것을 확인 하였다. 이는 앞으로 민자주방망이버섯의 한국 고유 품종개발 시 우수 교배형 선택에 있어, 기초자료로서 유용할 것으로 판단된다.

• 대한수의학회지
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• 제28권1호
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• pp.115-118
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• 1988

Mating experiments were performed to, elucidate the distribution of perfect state of Microsporum gypseum complex originated from animals and soils in Korea. A total of 30 strains of M. gypseum complex, composed of 16 from animals and 14 from soils, ere mated with the tester strains of Nannizzia incurvata, N. gypsea and N. fulva. Among 30 strains of M. gypseum complex examined, 16 strains(53.3%) were N. incurvata, 13 strains(43.3%) N. gypsea and 1 strain (3.3%) N. fulva. Among 13 strains of dog isolates, 8 strains(61.5%) were N. incurvata and 5 strains(38.5%) N. gypsea. And each strain isolated from Korean native goat, monkey and albino rat was N. gypsea. Among 14 strains of soil isolates, 8 strains(57.2%) were N. incurvata, 5 strains (35.7%) N. gypsea and 1 strain(7.2%) N. fulva. And -mating type was more frequently observed than +mating type in the strains of N. incurvata and N. gypsea. By this study, N. fulva was isolated the first time in Korea.

• Mycobiology
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• 제51권6호
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• pp.452-462
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• 2023

Opportunistic infections due to Cryptococcus neoformans and C. gattii species complexes continue to rise unabated among HIV/AIDS patients, despite improved antifungal therapies. Here, we collected a total of 20 environmental and 25 presumptive clinical cryptococcal isolates from cerebrospinal fluid (CSF) samples of 175 patients enrolled in an ongoing clinical trial Ambition 1 Project (Botswana-Harvard Partnership). Identity confirmation of the isolates was done using MALDI-TOF MS and PCR. We describe the diversity of the isolates by PCR fingerprinting and sequencing (Oxford Nanopore Technology) of the intergenic spacer region. Mating types of the isolates were determined by amplification of the MAT locus. We report an unusual prevalence of 42.1% of C. neoformans × C. deneoformans hybrids Serotype AD (n = 16), followed by 39.5% of C. neoformans Serotype A (n = 15), 5.3% of C. deneoformans, Serotype D (n = 2), 7.9% of C. gattii (n = 3), and 5.3% of C. tetragattii (n = 2) in 38 representative isolates that have been characterized. Mating type-specific PCR performed on 38 representative environmental and clinical isolates revealed that 16 (42.1%) were MATa/MAT𝛼 hybrids, 17 (44.7%) were MAT𝛼, and five (13.2%) possessed MATa mating type. We used conventional and NGS platforms to demonstrate a potential link between environmental and clinical isolates and lay a foundation to further describe mating patterns/history in Botswana.