• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.376-383
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• 2005

Alcaligenes sp. JMP228 carrying 2,4­dichlorophenoxyacetic acid (2,4-D) degradative plasmid pJP4 was inoculated into natural soil, and transfer of the plasmid pJP4 to indigenous soil bacteria was investigated with and without 2,4-D amendment. Plasmid pJP4 transfer was enhanced in the soils treated with 2,4-D, compared to the soils not amended with 2,4-D. Several different transconjugants were isolated from the soils treated with 2,4-D, while no indigenous transconjugants were obtained from the unamended soils. Inoculation of the soils with both the donor Alcaligenes sp. JMP228/pJP4 and a recipient Burkholderia cepacia DBO 1 produced less diverse transconjugants than the soils inoculated with the donor alone. Repetitive extragenic palindromic-polymerase chain reaction (REP-PCR) analysis of the transconjugants exhibited seven distinct genomic DNA fingerprints. Analysis of 16S rDNA sequences indicated that the transconjugants were related to members of the genera Burkholderia and Pandoraea. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that inoculation of the donor caused clear changes in the bacterial community structure of the 2,4-D­amended soils. The new 16S rRNA gene bands in the DGGE profile corresponded with the 16S rRNA genes of 2,4-D­degrading transconjugants isolated from the soil. The results indicate that introduction of the 2,4-D degradative plasmid as Alcaligenes sp. JMP228/pJP4 has a substantial impact on the bacterial community structure in the 2,4-D-amended soil.

• Journal of Microbiology and Biotechnology
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• 제31권11호
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• pp.1591-1600
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• 2021

Streptomyces coelicolor is a filamentous soil bacterium producing several kinds of antibiotics. S. coelicolor abs8752 is an abs (antibiotic synthesis deficient)-type mutation at the absR locus; it is characterized by an incapacity to produce any of the four antibiotics synthesized by its parental strain J1501. A chromosomal DNA fragment from S. coelicolor J1501, capable of complementing the abs- phenotype of the abs8752 mutant, was cloned and analyzed. DNA sequencing revealed that two complete ORFs (SCO6992 and SCO6993) were present in opposite directions in the clone. Introduction of SCO6992 in the mutant strain resulted in a remarkable increase in the production of two pigmented antibiotics, actinorhodin and undecylprodigiosin, in S. coelicolor J1501 and abs8752. However, introduction of SCO6993 did not show any significant difference compared to the control, suggesting that SCO6992 is primarily involved in stimulating the biosynthesis of antibiotics in S. coelicolor. In silico analysis of SCO6992 (359 aa, 39.5 kDa) revealed that sequences homologous to SCO6992 were all annotated as hypothetical proteins. Although a metalloprotease domain with a conserved metal-binding motif was found in SCO6992, the recombinant rSCO6992 did not show any protease activity. Instead, it showed very strong β-glucuronidase activity in an API ZYM assay and toward two artificial substrates, p-nitrophenyl-β-D-glucuronide and AS-BI-β-D-glucuronide. The binding between rSCO6992 and Zn²⁺ was confirmed by circular dichroism spectroscopy. We report for the first time that SCO6992 is a novel protein with β-glucuronidase activity, that has a distinct primary structure and physiological role from those of previously reported β-glucuronidases.

• Journal of Microbiology
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• 제38권2호
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• pp.80-87
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• 2000

The nucleotide sequence of the luxC gene coding for lux-specific fatty acyl-CoA reductase and the upstream DNA (325bp)of the structural gene from bioluminescent bacterium, Photobacterium phosphoreum, has been deternubed. An open reading frame extending for more than 20 codons in 325 bp DNA upstream of luxC was not present in both directions. The lux gene can be translated into a polypeptide of 54 kDa and the amino acid sequences of lux specific reductases of P. phosphoreum shares 80, 65, 58, and 62% identity with those of the Photobacterium leiognathi, Vibrio fischeri, Vibrio harveyi, and Xehnorhabdus luminescenens reductases, respectively. Analyses of codon usage, showing that a high frequency (2.3%) of the isoleucine codon, AUA, in the luxC gene compared to that found in Escherichia coli genes (0.2%) and its absence in the luxA and B genes, suggested that the AUA codon may play a modulator role in the expression of lux gene in E. coli. The structural genes (luxC, D, A, B, E) of the P. phosphoreum coding for luciferase (${\alpha}$,${\beta}$) and fatty acid reductase (r, s, t) polypeptides can be expressed exclusively in E. coli under the T7 phage RNA polymerase/promoter system and identificationof the [35S]methionine labelled polypeptide products. The degree of expression of lux genes in analyses of codon usage. High expression of the luxC gene could only be accomplished in a mutant E. coli 43R. Even in crude extracts, the acylated acyl-CoA reductase intermediate as well as acyl-CoA reductrase activities could be readily detected.

• Journal of Microbiology and Biotechnology
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• 제8권6호
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• pp.650-655
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• 1998

The chromosomal DNA fragment from Phaffia rhodozyma CBS 6938 which is able to autonomously replicate in the yeast Saccharomyces cerevisiae was cloned on an integrative URA3 plasmid. Its minimal fragment exhibiting autonomously replicating activiy in the S. cerevisiae gave a higher frequency transformation efficiency than that found for centromere-based plasmid, and enabled extrachromosoma1ly stable transmission of the plasmids in one copy per yeast cell under non-selective culture condition. The 836-bp DNA element lacked an ORF and did not contain any acceptable match to an ARS core consensus. Sequence analysis, however, displayed a cluster of three hairpin-Ioop-sequences with individual $\triangle {G_{25}}^{\circ}C$ free energy value of -10.0, -17.5, and -17.0 kcal. $mor^{-l}$as well as a 9-bp sequence with two base pair mismatches to the S. cerevisiae/E. coli gyrase-binding site. This 836-bp sequence also included one 7-bp sequence analogous to the core consensus of centromeric DNA element III (CDEIII) of S. cerevisiae, but CDEIII-like 7 bp sequence alone did not give a replicative function in this yeast.

• Mycobiology
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• 제43권2호
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• pp.170-173
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• 2015

Herein, we report the first occurrence of web blight of rosemary caused by Rhizoctonia solani AG-1-IB in Gangneung, Gangwon Province, Korea, in August 2014. The leaf tissues of infected rosemary plants were blighted and white mycelial growth was seen on the stems. The fungus was isolated from diseased leaf tissue and cultured on potato dextrose agar for identification. The young hyphae had acute angular branching near the distal septum of the multinucleate cells and mature hyphal branches formed at an approximately $90^{\circ}$ angle. This is morphologically identical to R. solani AG-1-IB, as per previous reports. rDNA-ITS sequences of the fungus were homologous to those of R. solani AG-1-IB isolates in the GenBank database with a similarity percentage of 99%, thereby confirming the identity of the causative agent of the disease. Pathogenicity of the fungus in rosemary plants was also confirmed by Koch's postulates.

• Mycobiology
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• 제38권1호
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• pp.7-12
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• 2010

A green mold species that has not previously been reported in Korea was isolated from oak log beds used for shiitake (Lentinula edodes) cultivation that were infested by mushroom flies. In this study, we identify the mold species as Gliocladium viride (an anamorph of Hypocrea lutea) and describe its mycological properties. The fungus was cottony on both potato dextrose agar (PDA) and Czapek yeast extract agar (CYA), but was colored white on PDA and became yellowish green and brown on CYA. Mycelial growth on PDA attained a diameter of 73 mm at $30^{\circ}C$ after 5 days. The fungus grew faster on malt extract agar (> 80 mm, 5 days at $25^{\circ}C$) compared to CYA and PDA (< 68 mm, 5 days at $25^{\circ}C$). Penicillate conidiophores of the fungus are hyaline, smooth walled, branching above typically in four stages, and $120\sim240\;{\mu}m$ in length. Club-shaped or slender phialides are formed on the metulae. Conidia of the fungus were ovate and elliptic, yellowish brown and green, and $2.5\sim3.0\;{\mu}m\times1.8\sim2.3\;{\mu}m$ in size. Typically, slimy conidia are formed in a mass and colored brown to dark green to almost black. The internal transcribed spacer rDNA and translation elongation factor 1 alpha gene sequences of the fungus isolated here show 99% identity with previously identified G. viride strains.

• 한국버섯학회지
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• 제14권4호
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• pp.168-173
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• 2016

청양과 장흥에 소재한 버섯 재배사에서 새로 육종된 표고 품종의 현장 실증검증 도중 문제를 일으킬 수 있는 잠재적 진균을 파악하고자 재배사내 공기 모니터링을 수행하여 오던 중 국내에 기록이 없는 Mortierella parvispora, Doratomyces purpureofuscus, Periconia byssoides, Periconia pseudobyssoides 등 네 종의 진균을 분리하여 동정하였다. 이중 두 종은 식물병원균으로 알려진 종이었고 다른 두 종은 부생성 균으로 다량의 포자를 생산하고 버섯재배 환경에서 오염균으로 작용할 가능성이 있는 균이었다. 본 연구에서는 이들 동정된 진균에 대한 형태적 특성, internal transcribed spacer (ITS) 와 18S rDNA region 염기서열 분석에 기반한 계통학적 관계, 그리고 알려진 정보 등에 대하여 보고하고자 한다.

• 한국식품조리과학회지
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• 제24권1호
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• pp.11-15
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• 2008

발효차로부터 분리 동정한 JY-1 효모는 18S rDNA 유전자 분석에서는 Rhodosporidium azoricum과의 유의성 99%, 생화학적 분석 VITEK system에서는 Rhodotorula glutinis/Rhodotorula mucilaginosa와 93%의 유의성을 나타내였다. 배양조건에서 최적의 온도는 $25-30^{\circ}C$, pH는 5.0이었으며 염의 농도는 2%, 최적의 에탄올 농도는 4%이었다. 일반적인 실험결과에서도 Rhodosporidium 속의 특이성과 비교하여 보았을 때 형태적, 생리적, 생화학적 유사성을 나타내었다. 이상의 결과에서 앞으로의 연구과제는 많이 음용되고 있는 발효차와 관련한 또 다른 미생물군을 분리 동정하여 우수한 균주를 확보하는데 있으며 이를 이용한 발효차 음료산업 개발에 목적을 두고 있다.

• Applied Biological Chemistry
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• 제50권2호
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• pp.101-106
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• 2007

국내 다양한 토양시료로부터 우수 선충포식곰팡이 9 균주를 선발하였다. 순수분리된 선충포식곰팡이는 포식기관의 형태에 따라 3차원적 점착성 그물구조(3-dimensional adhesive nets)를 나타내는 선충포식곰팡이(A 그룹), 2차원적 점착성 그물구조(2-dimensional adhesive nets)를 나타내는 선충포식곰팡이(B 그룹)와 수축성 고리구조(constricting ring)를 나타내는 선충포식곰팡이(C 그룹)로 크게 3개의 형태그룹으로 분류되었다. 이들 각 그룹에 속하는 선충포식곰팡이의 균사체, 분생포자병, 분생포자의 모양과 크기, 분생포자 형성 개수, 분생포자 마디(node), 분생포자 격막의 수와 격막의 위치, 휴면포자의 형성과 크기 및 색 등 형태학적 특징과 선충포식곰팡이의 rDNA ITS 영역을 PCR로 증폭하여 염기서열을 분석한 결과, 분리된 포식곰팡이는 Monacrosporium thaumasium(Kan-2, Kan-4, Kan-11), Arthrobotrys oligospora(Kan-9, Kan-13, Kan-20, Kan-21), A. musiformis (Kan-12), A. dactyloides(Kan-22)로 동정되었다.

• 한국균학회지
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• 제41권3호
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• pp.137-141
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• 2013

오서산의 소나무와 일본잎갈나무 두 수종의 침엽에서 내생균의 다양성을 분석하였다. 총 13개체의 숙수식물에서 채집한 침엽을 표면살균하여 분리한 균주들은 형태적인 특징과 rDNA 유전자(ITS 부위) 분석을 수행하였다. 그 결과 총 37개의 균주가 분리되었으며 이들은 17개의 분류군으로 묶을 수 있었다. 그 중 59%는 Leotiomycetes에 속하였으며, 30%는 Sordariomyetes에, 8%는 Dothideomycetes에 속하였으며, 3%는 Agaricomycetes에 속하는 균으로 판명 되었다. 이러한 결과들은 선행연구의 결과들과 매우 유사하였으며 소나무에서보다 일본잎갈나무에서 내생균의 종 다양성이 높게 나타났다. 특히 Lophodermium 속에 속하는 분류군들이 내생균의 다양성에서 주요한 균류로 확인되었으며, 한국에 분포하는 Lophodermium 속 내 종들에 대한 심도 있는 연구가 요구된다.