• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1223-1229
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• 2009

To construct a new recombinant strain of Bacillus velezensis that has antifungal and insecticidal activity via the expression of the insecticidal Bacillus thuringiensis crystal protein, a B. thuringiensis expression vector (pHT1K-1Ac) was generated that contained the B. thuringiensis cry1Ac gene under the control of its endogenous promoter in a minimal E. coli-B. thuringiensis shuttle vector (pHT1K). This vector was introduced into a B. velezensis isolate that showed high antifungal activities against several plant diseases, including rice blast (Magnaporthe grisea), rice sheath blight (Rhizotonia solani), tomato gray mold (Botrytis cinerea), tomato late blight (Phytophthora infestans), and wheat leaf rust (Puccinia recondita), by electroporation. The recombinant B. velezensis strain was confirmed by PCR using cry1Ac-specific primers. Additionally, the recombinant strain produced a protein approximately 130 kDa in size and parasporal inclusion bodies similar to B. thuringiensis. The in vivo antifungal activity assay demonstrated that the activity of the recombinant B. velezensis strain was maintained at the same level as that of wild-type B. velezensis. Furthermore, it exhibited high insecticidal activity against a lepidopteran pest, Plutella xylostella, although its activity was lower than that of a recombinant B. thuringiensis strain, whereas wild-type B. velezensis strain did not show any insecticidal activity. These results suggest that this recombinant B. velezensis strain can be used to control harmful insect pests and fungal diseases simultaneously in one crop.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2009년도 추계학술대회 및 정기총회
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• pp.14-16
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• 2009

Poria cocos is an importantant medical macrofungus,the sclerotium of Poria cocos has specific value as the drug material. There are few papers about its breeding and spawn rejuvenation. In this project, the protoplasts of cultivated strain T and wild strain L were prepared and treated separately by ultraviolet and heating, then fused with the PEG6000. The tural fusants were selected and identified by the affinity and ISSR analysis. 71 incompatibility strains between parents and reg regenerations were obtained from 118 regenerations by the affinity analysis. Five incompatibility strains were amplified with different primers, the results were showed that they had specific bands of both parents in the profile amplified with 3 primers, which proved these 5 strains were fusants by means of molecular biology marker. On the other hand, 25 strain were selected from 168 protoplast regenerations of cultivated strain T for cultivation experiment. The fresh sclerotium weight of these protoplast regenerations were better than the original strain.significantil 3 strains (T-1, T-4, T-7) increased respectively 118%, 73% and 73% than original strain. This method could be the effective in the rejuvenation Poria cocos.

• Journal of Species Research
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• 제9권3호
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• pp.204-209
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• 2020

In 2019, as a subset study to discover indigenous yeast species in Korea, a total of 20 yeast species were isolated from soil samples collected in Pochoen-si. Among them, eight strains were unreported species. From the high 26S rRNA gene sequence similarity and formation of a robust phylogenetic clade with the closest species, it was determined that each strain belonged to independent and predefined yeast species. The 20 strains were assigned to the genera Aureobasidium (1 strain) and Meyerozyma (1 strain) of the phylum Ascomycota and Cystofilobasidium (2 strains), Filobasidium (1 strain), Naganishia (2 strains), Bullera (3 strains), Leucosporidium (9 strains) and Sampaiozyma (1 strain) of the phylum Basidiomycota. There is no official report of the following species in Korea: Leucosporidium creatinivorum (4 strains), Leucosporidium escuderoi(2 strains), Leucosporidium golubevii(1 strain) and Leucosporidium intermedium (2 strains). Basic biochemical characteristics, colony and cell morphology are also described in the species description section.

• 한국균학회지
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• 제46권3호
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• pp.307-314
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• 2018

본 연구는 알코올 발효능이 우수한 야생 효모를 막걸리 발효에 응용할 목적으로 충남 예산군 예당저수지 야생화에서 분리한 비병원성 야생 효모들 중 에탄올 생산 우수 효모로 선발한 Aureobasidium pullulans P-1의 균학적 특성과 막걸리 발효 조건을 조사하였다. A. pullulans P-1는 출아에 의해 영양증식을 하였고 자낭포자를 생성하는 유포자 효모로서 내당성과 에탄올 내성이 강한 호염성 야생 효모이었다. A. pullulans P-1의 yeast extract-peptone-dextrose 배양액을 주모로 증자미와 입국과 물이 혼합된 술밑에 5% 첨가한 후 $25^{\circ}C$에서 1~10일간 발효시키면서 발효 중의 이화학적 성질의 변화를 조사한 결과, 에탄올 함량은 $25^{\circ}C$로 10일간 발효시켰을 때 가장 많은 8.45%를 생성하였고 관능 특성이 우수하였으며 항고혈압 활성 안지오텐신 전환효소 저해활성이 71.1%로 높았다.

• Journal of Microbiology and Biotechnology
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• 제17권2호
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• pp.280-286
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• 2007

We evaluated a commercial biopreparation of plant growth-promoting rhizobacteria (PGPR) strains Bacillus subtilis GB03 and B. amyloliquefaciens IN937a formulated with the carrier chitosan (Bio Yield) for its capacity to elicit growth promotion and induced systemic resistance against infection by Cucumber Mosaic Virus (CMV) and Pseudomonas syringae pv. tomato DC3000 in Arabidopsis thaliana. The biopreparation promoted plant growth of Arabidopsis hormonal mutants, which included auxin, gibberellic acid, ethylene, jasmonate, salicylic acid, and brassinosteroid insensitive lines as well as each wild-type. The biopreparation protected plants against CMV based on disease severity in wild-type plants. However, virus titre was not lower in control plants and those treated with biopreparation, suggesting that the biopreparation induced tolerance rather than resistance against CMV. Interestingly, the biopreparation induced resistance against CMV in NahG plants, as evidenced by both reduced disease severity and virus titer. The biopreparation also elicited induced resistance against P. syringae pv. tomato in the wild-type but not in NahG transgenic plants, which degrade endogenous salicylic acid, indicating the involvement of salicylic acid signaling. Our results indicate that some PGPR strains can elicit plant growth promotion by mechanisms that are different from known hormonal signaling pathways. In addition, the mechanism for elicitation of induced resistance by PGPR may be pathogen-dependent. Collectively, the two-Bacilli strain mixture can be utilized as a biological inoculant for both protection of plant against bacterial and viral pathogens and enhancement of plant growth.

• 한국어병학회지
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• 제6권2호
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• pp.93-101
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• 1993

효모의 RNAI유전자는 RNA processing에 관여 하는지 혹은 RNA transport에 관여 하는지 아직까지 유전자의 기능이 정확히 알려져 있지 않은 실정이다. 효모의 RNAI 유전자의 기능을 파악하기 위한 방법으로 본 연구에서는 rna1-1 mutant gene을 cloning하여 이에 대한 DNA sequence를 조사함으로써 RNAI 유전자와 rna1-1 유전자의 차이점을 이해하고자 하였다. rna1-1 marker를 갖는 yeast strain(R49)로 부터 genomic DNA를 추출하여 이를 BglII로 절단하여 genomic southern blotting을 행한 결과 wild type의 경우와 동일하게 3.4 kb에서 hybridization되는 signal을 얻었으며, RNAl 및 rna1-1이 yeast genome내에 single site로 존재함을 보여 주는 결과를 얻었다. mutant strain으로 부터 얻은 3.4 kb의 BglI fragment를 pUC19의 BamHI site에 subcloning하여 transformant들을 얻었고, wild type RNAl 유전자를 probe로 하여 rna1-1 mutant 유전자를 cloning할 수 있었다. pUC19에 cloning된 RNA1유전자 및 rna1-1유전자로부터 다양한 Ba131유도체를 얻어 이들에 대한 염기 서열을 비교한 결과 transcription initation site에서부터 down stream쪽으로 17 아미노산위치에 TCC가 TTC로 대치되어 있었으며 그 결과 serine이 phenylalanine으로 변환되는 결과를 얻었다. Wild type RNAI gene의 5'-region에는 3군데의 TATA-like sequence가 true TATA box인지 확인하기 위하여 Bal3I deletion에 의해 -103nt까지 deletion된 유도체를 얻었으며 ${\Delta}RNAI$, rna1-1, 81-2-6 clone이 rna1-1 allele와 complementation한지 확인하였으나 ${\Delta}RNAI$은 TS-complementation을 하지 못하였다. 따라서 현재까지 TATA-box라고 알려진 부분은 promoter로 작용하지 못함을 확인하였다.

• 한국미생물·생명공학회지
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• 제47권4호
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• pp.581-584
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• 2019

In this study, the carbon monoxide (CO)-fermenting acetogen, Clostridium sp. AWRP was subjected to chemical mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine (NTG) to generate a CO-resistant mutant. Among the 26 colonies obtained, the highest alcohol production was observed in one isolate, named C1. Compared to the wild-type strain, the C1 strain exhibited 1.5- and 3.4-fold higher CO consumption rate and alcohol selectivity, respectively. The total CO consumption of strain C1 could be further enhanced by increasing the content of metal ions, such as nickel and iron. The highest ethanol titer (5.7 g/l) was achieved by 5-fold increase in the iron concentration.

• KSBB Journal
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• 제14권3호
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• pp.351-357
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• 1999

A marine bacterium Bacillus cereus ASK202, agarase producing strain, was treated with some mutagenic agents, ultraviolte(UV), 1-methyl-3-nitro-1-nitrosoguanidine(NTG), and ethyl methane sulfonate(EMS), several times for the increasing of the agarase production After mutagen treatment, we isolated one mutant strain treated with NTG showed the highest stability and agarase productivity and named as Bacillus cereus ASK202-N3. This Bacillus cereus ASK202-N3 strain was well grown in the modified marine medium containing 0.5%(w/v) agar, 0.3%(w/v) yeast extract, and 5.0%(w/v) NaCl, and the optimal initial pH, temperature and culture time were 7.8, $25^{\circ}C$ and 32h, respectively. In the optimal culture conditions, the agarase production was increased to 5.3 fold(850units/L) compared to that of the wild type.

• Journal of Microbiology and Biotechnology
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• 제7권5호
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• pp.287-292
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• 1997

The role of glyoxylate bypass in lysine production by Corynebacterium glutamicum ssp. lactofermentum ATCC21799 was analyzed by using cloned aceA and aceB genes which encode enzymes catalyzing the bypass. Introduction of a plasmid carrying aceA and aceB to the strain increased enzyme activities of the bypass to approximately 5 fold on acetate minimal medium. The strain with amplified glyoxylate bypass excreted 25% more lysine to the growth medium than the parental strain, apparently due to the increased availability of intracellular oxaloacetate. The final cell yield was lower in the strain with amplified glyoxylate bypass. These changes were specific to the lysine-producing C. glutamicum ssp. lactofermentum ATCC21799, since the lysine-nonproducing wild type Corynebacterium glutamicum strain grew faster and achieved higher cell yield when the glyoxylate bypass was amplified. These findings suggest that the lysine producing C. glutamicum ssp. lactofermentum ATCC21799 has the ability to efficiently channel oxaloacetate, the TCA cycle intermediate, to the lysine biosynthesis pathway whereas lysine-nonproducing strains do not. Our results show that amplification of the glyoxylate bypass efficiently increases the intracellular oxaloacetate in lysine producing Corynebacterium species and thus results in increased lysine production.

• 한국미생물·생명공학회지
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• 제23권2호
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• pp.164-169
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• 1995

An endogenous cryptic plasmid, pBL1, which has been used to construct plasmid vectors for coryneform bacteria producing amino acids, was eliminated from Brevibacterium lactofermentum. The pBL1 was partially digested with Sau3AI and the resulting DNA fragments were subcloned into a suicide vector pEM1 which contains a kanamycin-resistant (km$^{r}$) gene. KM$^{r}$ B. lactofermentum transconjugants were obtained by conjugal transfer of the pEM1 derivatives containing pBL1 DNA fragments from Escherichia coli into B. lactofermentum. A km$^{r}$ transconjugant was analyzed to contain a plasmid pEB14, which occurred in vivo by homologous recombination between pBL1 and the conjugal-transferred plasmid. The pEB14 including the pEM1-derived km$^{r}$ gene was found to be lost concomitantly with km$^{r}$ phenotype, resulting in the construction of a pBL1-free strain of B lactofermentum. Based on transformation efficiencies and plasmid stability, the resultant pBL1- free strain is more useful than wild strain as a host cell for genetic manipulation. It could be concluded that foreign plasmid DNAs are efficiently isolated and analyzed from the pBL1-free strain because of the absence of endogenous pBL1 plasmid.