• The Plant Pathology Journal
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• 제15권3호
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• pp.131-136
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• 1999

In fungi known as ascomycetes, ability to mate is controlled by a single mating type (MAT) locus with two dissimilar sequences called idiomorphs carrying genes encoding transcription factors that are unrelated to each other. Fungi requiring strains with different MAT genes to complete the sexual process are heterothallic (self-sterile); species in which as single strain is able to undergo sexual reproduction are homothallic (self-fertile). Previous analysis of sequences from several heterothallic and homothallic species of the ascomycete genus Cochliobolus showed that homothallics evolve from heterothallics and that each known Cochliobolus homothallic species arose independently, from a different heterothallic ancestral species. Here we report detailed comparative analyses of MAT sequences ad their flanking regions, and show that: (1) The level of MAT gene similarity is not correlated with reproductive life style; (2) MAT proteins from all Cochliobolus species are conserved within the transcription factor signature sequences; they are not conserved in the carboxy terminal half of MAT-1, or third of MAT-2, except in those from very closely related species; (3) A gene (ORF1) of unknown function, consistently found on the MAT flank, is more conserved than are the MAT genes themselves; (4) The intergenic sequences diverge sharply among species.

• 한국균학회지
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• 제20권3호
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• pp.222-228
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• 1992

사철 느타리버섯 영양요구성(營養要求性) 균주(菌株)를 팽이버섯 leu 2 유전자를 지닌 pM 301 백터를 이용(利用)하여 영양요구성(營養要求性)을 보완(補完) 시킴으로써 형질전환(形質轉換) 하였다. 형질전환균주(形質轉換菌株)의 균사생장(菌絲生長)을 버섯 완전배지(完全培地)나 버섯 최소(最小) 배지(培地)에서 비교(比較)하였다. 형질전환주(形質轉換株)는 1 핵(核) 균사(菌絲)와 교배(交配)후 자실체(子實體) 형태(形態)를 비교(比較)하고 포자 분석(分析)에 의하여 유전분석(分析)을 하였다. 자실체(子實體) 발생(發生)과 자실체(子實體) 모양이 형질전환주(形質轉換株)는 모균주와 다른 특성(特性)이 나타났다. 모균주(母菌株)는 교배형(交配型)이 $A_1B_1$으로 $A_2B_1$ 교배형(交配型)인 1핵(核) 균주(菌株)와 교배(交配)에 의하여 자실체(子實體)를 형성(形成)하지 못하나 형질전환주(形質轉換株)는 교배형(交配型)이 $A_1B_1$이고 $A_2B_1$ 1핵(核) 균주(菌株)와 교배(交配) 하였을때 자실체(子實體)를 형성(形成)하였다.

• 한국균학회지
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• 제25권1호통권80호
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• pp.26-29
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• 1997

여름느타리버섯의 담자포자에 자외선을 조사하여 5'-fluoro-orotic acid(FOA) 선발배지에서 pyrimidine 영양요구성 균주를 직접 선발하였다. 24개의 5'-FOA 저항성 균주중 13개의 pyrimidine 영양요구성 균주를 선발할 수 있었으며, 이들의 mating type group과 성장속도를 측정하여 형질전환을 위한 최적의 DNA수용체를 선발할 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권12호
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• pp.1805-1811
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• 2007

A simulation study was conducted to evaluate the effect of reciprocal cross on the detection and characterization of parent-of-origin (POE) QTL in $F_2$ QTL populations. Data were simulated under two different mating designs. In the one-way cross design, six $F_0$ grand sires of one breed and 30 $F_0$ grand dams of another breed generated 10 $F_1$ offspring per dam. Sixteen $F_1$ sires and 64 $F_1$ dams were randomly chosen to produce a total of 640 $F_2$ offspring. In the reciprocal design, three $F_0$ grand sires of A breed and 15 $F_0$ grand dams of B breed were mated to generate 10 $F_1$ offspring per dam. Eight $F_1$ sires and 32 $F_1$ dams were randomly chosen to produce 10 $F_2$ offspring per $F_1$ dam, totaling 320 $F_2$ offspring. Another mating set comprised three $F_0$ grand sires of B breed and 15 $F_0$ grand dams of A breed to produce the same number of $F_1$ and $F_2$ offspring. A chromosome of 100 cM was simulated with large, medium or small QTL with fixed or different allele frequencies in parental breeds. A series of tests between Mendelian and POE models were applied to characterize QTL as Mendelian, paternal, maternal or partial expression QTL. The overall detection powers were similar between the two mating designs. However, the proportions of paternally expressed QTL that were declared as paternal QTL type were greater in the reciprocal cross design than in the one-way cross, and vice versa for Mendelian QTL. When QTL alleles were segregating in parental breeds, a significant proportion of Mendelian QTL were spuriously declared POE QTL, suggesting that care must be taken to characterize imprinting QTL in a QTL mapping population with a small number of $F_1$ parents.

• 한국수정란이식학회지
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• 제28권3호
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• pp.215-222
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• 2013

Bovine coat color is decided by the melanocortin receptor 1 (MC1R) genotype mutation and melanogenesis. Specially, in the various cattle breeds, dominant black coat color is expressed by dominant genotype of $E^D$, red or brown is expressed in the frame shift mutation of recessive homozygous e by base pair deletion and wild type of $E^+$ is expressed in various coat colors. However, not very well known about the effected of MC1R genotype mutation on the coat color through family lines in KBC. Therefore, this study were to investigate effect of MC1R genotype mutation on the coat color, and to suggest mating breed system in accordance with of MC1R genotype for increased on brindle coat color appearance. Parents (sire 2 heads and dam 3 heads) and offspring (total : 54 heads) from crossbreeding in KBC family line with the MC1R genotype and phenotype records were selected as experimental animals. The relationship between melanocortin 1 receptor (MC1R) genotypes expression verified by PCR-RFLP, and brindle coat color appearance to the family line of the cross mating breed from MC1R genotype pattern was determined. As a result, 4MC1R genetic variations, $E^+/E^+$ (sire 1), $E^+/e$ (sire 2 and dam 3), $E^+/e$ with 4 bands of 174, 207 and 328 bp (dam 1) and $E^+/e$ with 3 bands of 174, 207, 328 and 535 bp (dam 2) from parents (sire and dam) of KBC. However, 3 genetic variations, e/e (24%), $E^+/E^+$ (22%) and $E^+/e$ (56%) were identified in offspring. Also, brindle coat color expressrated was the e/e with the 0%, $E^+/E^+$ with 67% and $E^+/e$ with 77% from MC1R genotype in offspring on the cross mating of KBC. Furthermore, when the sire had $E^+/e$ genotype and the dam had $E^+/E^+$ with the 3 bands or $E^+/e$ genotype, and both had whole body-brindle coat color, 62% of the offspring had whole body-brindle coat color. Therefore, the seresults, the mating system from MC1R genotype patterns of the sires ($E^+/e$) and dams ($E^+/E^+$ with the 3 bands or $E^+/e$) with brindle coat color may have the highest whole body-brindle coat color expression in their offspring.

• Journal of Microbiology and Biotechnology
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• 제28권9호
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• pp.1573-1579
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• 2018

Transcriptional gene silencing is regulated by the chromatin structure, which is by various factors including histones. Saccharomyces cerevisiae contains transcriptionally silenced regions such as telomeric regions and hidden mating (HM) loci. The positively-charged amino acids on the histone H4 tail were reported to be critical for the telomeric silencing in yeast, by interacting with Dot1, a specific methyltransferase for the $79^{th}$ lysine on histone H3. However, Dot1 did not affect gene silencing within HM loci, but whether the positively-charged amino acids on the H4 tail affect HM silencing has not been defined. To elucidate the function of the H4 tail on HM silencing, we created several MATa-type yeast strains bearing the substitution of arginine with alanine or lysine on the histone H4 tail and checked the sensitivity of MATa-type yeast to alpha pheromone. The arginine point mutants substituted by alanine (R17A, R19A, and R23A) did not show sensitivity to alpha pheromone, but only two arginine mutants substituted by lysine (R17K and R19K) restored the sensitivity to alpha pheromone-like wild type. These data suggested that the basic property of arginine at $17^{th}$ and $19^{th}$ positions in the histone H4 tail is critical for maintaining HM silencing, but that of the $23^{rd}$ arginine is not. Our data implicated that the positive charge of two arginine residues on the histone H4 tail is required for HM silencing in a manner independent of Dot1.

• Mycobiology
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• 제38권1호
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• pp.26-32
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• 2010

The cyclic AMP (cAMP) pathway plays a major role in growth, sexual differentiation, and virulence factor synthesis of pathogenic fungi. In Cryptococcus neoformans, perturbation of the cAMP pathway, such as a deletion in the gene encoding adenylyl cyclase (CAC1), causes defects in the production of virulence factors, including capsule and melanin production, as well as mating. Previously, we performed a comparative transcriptome analysis of the Ras- and cAMP- pathway mutants, which revealed 163 potential cAMP-regulated genes (38 genes at a 2-fold cutoff). The present study characterized the role of one of the cAMP pathway-dependent genes (serotype A identification number CNAG_ 06576.2). The expression patterns were confirmed by Northern blot analysis and the gene was designated cAMP-regulated gene 1 (CAR1). Interestingly, deletion of CAR1 did not affect biosynthesis of any virulence factors and the mating process, unlike the cAMP-signaling deficient cac1$\Delta$ mutant. Furthermore, the car1$\Delta$ mutant exhibited wild-type levels of the stress-response phenotype against diverse environmental cues, indicating that Car1, albeit regulated by the cAMP-pathway, is not essential to confer a cAMP-dependent phenotype in C. neoformans.

• The Korean Journal of Ecology
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• 제17권3호
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• pp.299-310
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• 1994

종 내의 클론 상호작용은 개체군 구조에 중요한 영향을 미친다. 본 연구는 클론간의 경쟁에 의해 개체군 내에서 진화적 변화가 일어날 수 있는 가를 조사하고, 균일한 실험실 배양 조건에서 어떻게 다양한 클론이 진화하는지를 밝히고자 하였다. 자원소비율과 교배형이 다른 Polysphondylium pallidum의 클론들이 선택되었다. 실험은 T0에서 T4까지 4번의 시간 간격을 두고 시행되었다. 접종 후 자실체를 수확할 때까지의 한 번의 실험기간은 10~14일 정도 소요되었다. T0의 50개의 클론으로부터 일련의 실험기간 T1, T2, T3, T4에서 50 클론이 두 세트씩 만들어졌다. 개체군들의 자원소비율 변동은 T0와 두세트의 T4 클론들간에 비교되었다. 각 클론은 실험 시작 후 48~56세대가 지난 후에도 균일하고 제한된 Cerophyl 배지 환경에서 다양한 자원 소비율 즉, 다양한 생장율을 보였다. 생장율이 다른 다양한 클론은 서식처가 이질적인 토양 미세 환경 뿐만 아니라 한 장소의 균질한 배양 접시 속에서도 공존 할 수 있었다. 높은 클론 다양성은 한 개체군내의 클론에서 다른 클론을 만날 기회를 제공해 주었고 개체군내의 클론 경쟁이 활발히 일어났다. T0에서 T4까지의 총 실험 기간동안 교배형 I이었던 37개의 클론중에서 한 개체군의 31개가 교배형 II로 바뀌어 클론간에 일어난 경쟁배타율은 0.838이었다. 각 교배형의 출현 빈도는 연속적인 다음 세대마다 0.93~1.29% 만큼 바뀌었다. 2.6X108밀도의 박테리아를 먹이로 제공해 준 실험실 환경에서 한 세대가 지날 때 마다 한 클론에서 다른 클론으로 변화하는 진화적 변화확률이 1.26~1.75%라는 것을 의미한다.

• The Plant Pathology Journal
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• 제21권1호
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• pp.33-38
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• 2005

Changes of control efficacy of chemical to potato late blight caused by Phytophthora infestans in potato fields from 2001 to 2004 were examined. Control efficacy of metalaxyl was suddenly decreased from 100% in 2002 to 50% in 2004 and that of dimethomorph also was similar to those of metalaxyl. However, the control efficacy of ethaboxam no great change. Both A1 and A2 mating type isolates were isolated from 2001 to 2004 in several areas in Korea. The majority of the P. infestans isolates were A1 mating type. Total 939 isolates of P. infestans obtained from several areas in Korea from 2001 to 2004 were examined for changes of sensitivity to metalaxyl. Frequencies of metalaxyl resistance isolates were gradually increased from 17% in 2001 to 84.2% in 2004, but isolation frequencies of metalaxyl sensitive and intermediate resistant isolate were decreased. Cause of decreasing control efficacy of metalaxyl was thought by increase of resistance isolates in A1 mating type population according to increasing metalaxyl use. Most isolates were grown at 0.5 ${\mu}g/ml of dimethomorph and isolates grown at 1 ${\mu}g/ml of dimethomorph were approximately 10.2-22.9%. However, no isolate was able to grow at 5.0 ${\mu}g/ml. Based on these results, minimum inhibitory concentrations (MIC) of dimethomorph to P. infestans were determined to be 0.5-1.0 ${\mu}g/ml. Our results indicated that the reason decreasing control efficacy of dimethomorph was not caused by occurrence of resistant isolates. About 5% and 12.1% isolates among the total isolates collected in 2003 and 2004 were grown on V-8 juice rye agar containing 1.0 ${\mu}g/ml ethaboxam. The 2.1 and 25.4% isolates had MICs of 0.2-0.4 ${\mu}g/ml, and MIC values of 87.9% and 74.3% isolates were less than 0.2 ${\mu}g/ml concentrations of ethaboxam. Therefore, resistance development by P. infestans to ethaboxam is not likely to occur in the natural condition.

• 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.