• Journal of Microbiology and Biotechnology
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• 제16권5호
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• pp.724-730
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• 2006

A sequence characterized amplified region (SCAR) marker, which was tightly linked with the A1 mating type locus in Phytophthora infestans, was developed. During the random amplified polymorphic DNA-based phylogenic studies of 33 isolates of P infestans collected from year 2002 to 2004, we found an A1 mating type-specific DNA fragment. This 573-bp DNA fragment was generated only in the genomic DNA of the A1 mating types, when OPC-5 primer was used. Based on the specific DNA sequence, we designed the primer sets for generating the A1 mating type-specific 569-bp DNA fragment. When 33 genomic DNAs of P. infestans were subjected to PCR amplification using different primer combinations, the A1 mating type-specific DNA was amplified, when LB-1F and LB-2R primers were used. The specific 569-bp DNA fragment was generated only from all 18 A1 strains, but not from 15 A2 mating type strains. These results corresponded to the mating type discriminating bioassay of 33 isolates of P. infestans. Therefore, the primer combination of LB-1F/LB2R was chosen as a SCAR marker. Overall, this study indicates that the SCAR marker could be developed into a useful tool for mating type determination of P. infestans.

• Mycobiology
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• 제49권6호
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• pp.582-588
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• 2021

The interaction of mating pheromone and pheromone receptor from the B mating-type locus is the first step in the activation of the mushroom mating signal transduction pathway. The B mating-type locus of Lentinula edodes is composed of Bα and Bβ subloci, each of which contains genes for mating pheromone and pheromone receptor. Allelic variations in both subloci generate multiple B mating-types through which L. edodes maintains genetic diversity. In addition to the B mating-type locus, our genomic sequence analysis revealed the presence of a novel chromosomal locus 43.3 kb away from the B mating-type locus, containing genes for a pair of mating pheromones (PHBN1 and PHBN2) and a pheromone receptor (RCBN). The new locus (Bα-N) was homologous to the Bα sublocus, but unlike the multiallelic Bα sublocus, it was highly conserved across the wild and cultivated strains. The interactions of RcbN with various mating pheromones from the B and Bα-N mating-type loci were investigated using yeast model that replaced endogenous yeast mating pheromone receptor STE2 with RCBN. The yeast mating signal transduction pathway was only activated in the presence of PHBN1 or PHBN2 in the RcbN producing yeast, indicating that RcbN interacts with self-pheromones (PHBN1 and PHBN2), not with pheromones from the B mating-type locus. The biological function of the Bα-N locus was suggested to control the expression of A mating-type genes, as evidenced by the increased expression of two A-genes HD1 and HD2 upon the treatment of synthetic PHBN1 and PHBN2 peptides to the monokaryotic strain of L. edodes.

• 대한미생물학회지
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• 제21권1호
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• pp.127-131
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• 1986

The mating types and serotypes of 10 clinical isolates of Cryptococcus neoformans have been investigated. Seven isolates were serotype A and three were serotype D and thus they fell in C. neoformans var. neoformans. Mating types of six isolates were found $\alpha$ and two were $\alpha$ but another two isolates were untypable. Enzyme linked immuno-sorbent assay(ELISA) using rabbit hyper-immune serum to cryptococcal polysaccharides was well adapted to the analysis of capsular polysaccharides in sera of the patients with cryptococcal meningitis.

• 한국균학회지
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• 제16권2호
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• pp.60-63
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• 1988

고추역병균(疫病菌) 103개균주(個菌株)를 분리(分離)하여 배우자형(配偶子型)을 분류(分類)하였다. 분리(分離)된 균주중(菌株中) 54개균주(個菌株)가 mating type $A_1$으로, 49개균주(個菌株)가 mating type $A_2$로 분류(分類)되었다. 서로 다른 배우자형(配偶子型)인 mating type $A_1$과 $A_2$균주(菌株)를 각각(各各) 5% V-8 쥬스배지(培地)에 대치배양(對置培養)한 결과(結果) 유성세대(有性世代)인 난포자(卵胞子)를 형성(形成)하였다.

• Mycobiology
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• 제51권1호
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• pp.26-35
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• 2023

The diversity of A mating type in wild strains of Lentinula edodes was extensively analyzed to characterize and utilize them for developing new cultivars. One hundred twenty-three A mating type alleles, including 67 newly discovered alleles, were identified from 106 wild strains collected for the past four decades in Korea. Based on previous studies and current findings, a total of 130 A mating type alleles have been found, 124 of which were discovered from wild strains, indicating the hyper-variability of A mating type alleles of L. edodes. About half of the A mating type alleles in wild strains were found in more than two strains, whereas the other half of the alleles were found in only one strain. About 90% of A mating type combinations in dikaryotic wild strains showed a single occurrence. Geographically, diverse A mating type alleles were intensively located in the central region of the Korean peninsula, whereas only allele A17 was observed throughout Korea. We also found the conservation of the TCCCAC motif in addition to the previously reported motifs, including ATTGT, ACAAT, and GCGGAG, in the intergenic regions of A mating loci. Sequence comparison among some alleles indicated that accumulated mutation and recombination would contribute to the diversification of A mating type alleles in L. edodes. Our data support the rapid evolution of A mating locus in L. edodes, and would help to understand the characteristics of A mating loci of wild strains in Korea and help to utilize them for developing new cultivars.

• 한국균학회지
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• 제48권4호
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• pp.389-396
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• 2020

표고는 한국에서 가장 많이 소비되는 식용버섯 중 하나이다. 표고는 사극성의 교배계를 따르며, 표고의 교배형은 자웅이주성의 다른 담자균류와 마찬가지로 서로 독립적인 두 유전자좌, A와 B에 의해 결정된다. 표고의 A 유전자좌에는 한 쌍의 homeodomain (HD) 전사인자가 암호화되어 있으며 이들의 N말단에서 나타나는 높은 변이가 A 교배형의 다양성에 중요한 것으로 알려져 있다. 본 연구에서는 표고 품종과 야생종에서 많이 발견되는 11종의 A 교배형을 구분할 수 있는 CAPS 마커를 개발하고자 하였다. A 유전자좌에서 변이가 큰 부분을 PCR을 통해 증폭한 뒤 두 가지 제한효소 HaeIII와 EcoRI로 절단하여 DNA 단편의 크기 및 양상을 살펴봄으로써 11종의 A 교배형을 서로 구별할 수 있었다. 또한 해당 방법이 이핵균주의 교배형을 확인하는 데 활용할 수 있는지도 살펴보았다.

• 생명과학회지
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• 제12권2호
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• pp.173-181
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• 2002

북미산 S. commune UVMl-34의 A $\alpha$ 3 mating locus를 지니는 cosmid clone pSC13의 mating activity에 필요한 sequence 함유여부를 확인한 후 sequencing을 행하고 이에 대한 비교분석을 실시하였다. 그 결과 blast program을 사용한 전체 nucleotide 염기서열의 homology비교분석에서 남미산 1-71에 패하여 약 96%의 높은 homology를 나타내었으며, 1-71 A $\alpha$ 3 mating locus에서 모두 7개로 추정되는 exon의 염기서열의 비교실험에서도 거의 96% 이상에 이르는 homology를 나타내었다. 부분적으로는 AR (acidic rich region)에서 약 97%, HD (homeodomain)에서는 약 99%, BR (basic rich region)에서 약 97%, 그리고 Ser (serine rich regon)에서 약 95%의 높은 homology를 나타내어 지역간에 유전자상의 큰 변화는 나타나지 않는 것으로 확인되었다. 그리고 translated polypeptide sequence 를 이용하여 자생지가 다른 남미산 S. commune A $\alpha$ 3 mating locus 내의 Y-region은 비롯하여 다른 $A\alpha$ alleles내의 Y-region과 비교 분석한 결과 남미산에 대해 전체적으로 약 97%의 높은 homology를 나타내고 있지만 그 외 $A\alpha$ mating alllele gene의 Y-region과는 41~49%의 낮은 homology로 mating activity에 관여하고 있는 것으로 나타났다. 특히 mating에 있어 transcription regulator로 알려진 homeodomain에서는 약 98%의 homology가 나타남으로서 Z-region의 74%에 비하여 대륙간에 상당히 높은 유전자 보존상이 확인되었다. 또한 AR에서 97%, BR에서 100% 그리고 Ser에서도 98%의 상당히 높은 homology를 지니는 사실을 확인하였다. 이러한 결과로 보아 남미와 북미에 자생하는 같은 mating allele type간에는 상당히 높은 비율의 염기서열 보존이 이루어지고 있음을 알 수 있었으며, 비록 다른 $A\alpha$ alleles간의 비교 이지만 다른 mating alleles간의 약 50% homology와 비교할 때 보다 상당히 높은 결과로 보인다. 특히 homeodomain motif의 비교에서 Y1을 비롯한 다른 mating allele gene과도 85% 이상의 높은 homology를 나타내었으며 그 외 AR, BR, Ser 부위에서는 10~50%에 이르는 낮은 비율로 나타났다.

• Mycobiology
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• 제47권2호
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• pp.200-206
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• 2019

Allelic differences in A and B mating-type loci are a prerequisite for the progression of mating in the genus Pleurotus eryngii; thus, the crossing is hampered by this biological barrier in inbreeding. Molecular markers linked to mating types of P. eryngii KNR2312 were investigated with randomly amplified polymorphic DNA to enhance crossing efficiency. An A4-linked sequence was identified and used to find the adjacent genomic region with the entire motif of the A locus from a contig sequenced by PacBio. The sequence-characterized amplified region marker $7-2_{299}$ distinguished A4 mating-type monokaryons from KNR2312 and other strains. A BLAST search of flanked sequences revealed that the A4 locus had a general feature consisting of the putative HD1 and HD2 genes. Both putative HD transcription factors contain a homeodomain sequence and a nuclear localization sequence; however, valid dimerization motifs were found only in the HD1 protein. The ACAAT motif, which was reported to have relevance to sex determination, was found in the intergenic region. The SCAR marker could be applicable in the classification of mating types in the P. eryngii breeding program, and the A4 locus could be the basis for a multi-allele detection marker.

• 대한의생명과학회지
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• 제19권3호
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• pp.213-223
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• 2013

The objectives of this study are to develop a molecular diagnosis to differentiate serotypes and mating-types of C. neoformans/C. gattii complex isolates from pigeon droppings in Korea and to elucidate molecular epidemiology of the isolates. Phenotypes and genotypes of C. neoformans/C. gattii complex isolates were identified by biochemical properties and PCR using specific CNLAC1 gene, respectively. To classify serotypes and mating-types of C. neoformans/C. gattii complex isolates, the five reference strains and thirty-three isolates in Korea were investigated by restriction fragment length polymorphism (RFLP) analysis using CNLAC1 gene for varieties, by random amplified polymorphic DNA (RAPD) for serotyping, and by PCR using specific primer sets for mating typing. All isolates in Korea were belonged to C. neoformans var. grubii (serotype A) by RFLP and RAPD patterns which showed high sensitivity and specificity. Therefore, RFLP and RFLP were available to differentiate varieties and serotypes of C. neoformans. Amplification patterns of the five reference strains by specific PCR for mating typing were differentiable, and all isolates were classified into $MAT{\alpha}$. All C. neoformans environmental isolates in Korea were Cr. neoformans serotype A and $MAT{\alpha}$ which is a more virulent pathogen. This study suggests that RFLP and RAPD are rapid and correct molecular diagnosis tools for epidemiology of C. neoformans/C. gattii complex isolates.

• 한국균학회지
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• 제30권2호
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• pp.152-156
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• 2002

1995년부터 1998년 사이 국내에 분포하는 고추역병균 Phytophthora capsici 집단에 대해 A1과 A2 교배형의 발생분포 및 변화를 조사하였다. 전국의 75개 고추포장들로부터 분리된 973개 P. capsici 균주들의 A1 : A2 교배형 발생비는 573 : 400으로 나타났다. A1과 A2 교배형의 P. capsici 균주들은 조사를 실시한 모든 지방에서 발견되었다. 조사포장별로는 A1 또는 A2 교배형의 병원균들만이 분리된 포장들과 A1과 A2 교배형의 병원균들이 동시에 발생하는 포장들이 관찰되었으며, 각 단일 포장내 P. capsici 균주들의 A1 : A2 교배형 발생비율은 해마다 다양하게 나타났다. 한편, 동일 포장에서의 연차적인 교배형 발생비 추이를 조사한 결과 1995년과 1997년에 A2 교배형의 P. capsici만이 발생했던 공주포장에서는 1998년에 A1 교배형의 균주가 새롭게 발견되었다. 따라서 국내 고추 역병균 P. capsici 집단은 병발생 포장단위별로 다양한 A1 : A2 교배형 균주들의 발생비가 나타나고 있어 단일포장 내에서 유성생식이 이루어질 가능성이 높으며, 포장단위별 균주들 간에는 유전적인 차이가 다양하게 존재할 것으로 판단된다.