• International Journal of Industrial Entomology and Biomaterials
• /
• 제25권2호
• /
• pp.195-203
• /
• 2012

In reared populations of Allomyrina dichotoma, commercial insects, the skin of last instar larvae was changed softer with opaque white, and infested grubs eventually died. To clarify the cause of the symptom, we collected the larvae of A. dichotoma from five farms and examined their intestinal bacterial florae using pyrosequencing technique. From those results, a member of Paenibacillus was found only in the larvae showing the symptom of disease. Through PCR analysis using a Paenibacillus specific primer set, we obtained the partial 16S rRNA gene sequence and confirmed the microbe as Paenibacillus sp. For clear identification, a whole guts was extracted from each larva showing the sign of the disease and incubated at $70^{\circ}C$ for 15 min to isolate spore forming bacteria. After then, each content of guts was cultured on $MYPGP_{NAL}$ agar medium($12.5{\mu}g/ml$ of nalidixic acid) at $30^{\circ}C$. The 16S rRNA gene sequence analysis for the isolated bacteria showed that they were closely related to P. rigui(97.9% similarity), to P. chinjuensis(96.1% similarity), and to P. soli(95.3% similarity). Additional tests including API test and cellular fatty acid composition analysis were performed, but the strain couldn't be identified at species level, suggesting it may represent novel species of the genus Paenibacillus.

• The Plant Pathology Journal
• /
• 제34권5호
• /
• pp.435-444
• /
• 2018

Receptor-like proteins (RLPs) are involved in plant development and disease resistance. Only some of the RLPs in tomato (Solanum lycopersicum L.) have been functionally characterized though 176 genes encoding RLPs, which have been identified in the tomato genome. To further understand the role of RLPs in tomato, we performed genome-guided classification and transcriptome analysis of these genes. Phylogenic comparisons revealed that the tomato RLP members could be divided into eight subgroups and that the genes evolved independently compared to similar genes in Arabidopsis. Based on location and physical clustering analyses, we conclude that tomato RLPs likely expanded primarily through tandem duplication events. According to tissue specific RNA-seq data, 71 RLPs were expressed in at least one of the following tissues: root, leaf, bud, flower, or fruit. Several genes had expression patterns that were tissue specific. In addition, tomato RLP expression profiles after infection with different pathogens showed distinguish gene regulations according to disease induction and resistance response as well as infection by bacteria and virus. Notably, Some RLPs were highly and/or unique expressed in susceptible tomato to pathogen, suggesting that the RLP could be involved in disease response, possibly as a host-susceptibility factor. Our study could provide an important clues for further investigations into the function of tomato RLPs involved in developmental and response to pathogens.

• 한국작물학회지
• /
• 제49권5호
• /
• pp.434-439
• /
• 2004

Medicago truncatula is a model plant for molecular genetic studies of legumes and plant-microbe interactions. To accelerate finding of genes that play roles in the early stages of nodulation and stress responses, a trans-genic plant was developed that contains a promoter­reporter fusion. The promoter of rip], a Rhizobium-induced peroxidase gene, was fused to the coding region of $\beta-glucuronidase (GUS)$ gene and inserted into a modified plant transformation vector, pSLJ525YN, in which the bar gene was preserved from the original plasmid but the neomycin phosphotransferase gene was replaced by a polylinker. Transformation of M. truncatula was carried out by vacuum infiltration of young seedlings with Agrobacterium. Despite low survival rates of infiltrated seedlings, three independent transformants were obtained from repeated experiments. Southern blot analyses revealed that 7 of 8 transgenic plants of the T 1 generation contained the bar gene whereas 6 $T_1$ plants contained the GUS gene. These results indicate that vacuum infiltration is an effective method for transformation of M. truncatula. The progeny seeds of the transgenic plants will be useful for mutagenesis and identification of genes that are placed upstream and may influence the expression of rip] in cellular signaling processes including nodulation.

• 환경생물
• /
• 제24권4호
• /
• pp.307-313
• /
• 2006

2004년 9월과 11월, 2005년 1월, 5월 및 8월의 총 5회에 걸쳐 대한민국 통영연안의 1개 정점에서 표층해수를 계절별로 채취하여 해양세균 군집을 분석하였다. 선택배지에서 순수분리하여 VITEK Microbe ID system으로 동정한 배양법과 16S rRNA PCR-DGGE로 분석한 결과를 상호비교하였다. 배양법에 의한 각 계절별 해양세균 군집의 종조성은 2004년 9월은 5종, 11월은 5종, 2005년 1월은 4종, 5월은 6종 및 8월은 10종으로 조사되었고, Pseudomonas fluorescens와 Acinetobacter lwoffii는 계절과 관계없이 모두 검출되었으며, 그 외에 Pseudomonas stutzeri, Sphingomonas paucimobilis 및 Burkholderia mallei 등이 동정되었다. 16S rRNA PCR-DGGE에 의한 군집분석 결과는 2004년 9월은 10개체군, 11월은 11개체군, 2005년 1월은 6개체군, 5월은 9개체군 및 8월에는 13개체군이 조사되었고, Acinetobacter lwoffii, Burkholderia mallei, Pseudomonas fluoresence, Actinobacillus ureae, Burkholderia sp., Pseudomonas stutzeri, Roseobacter sp., Vibrio parahaemolyticus, Sphingomonas paucimobilis 및 Rugeria algocolus 등이 동정되었다. 이로부터 해양세균 군집의 분포특성을 파악하는 데는 16S rRNA PCR-DGGE가 배양법보다 더 효율적임이 판명되었다.

• 한국약용작물학회지
• /
• 제22권4호
• /
• pp.306-312
• /
• 2014

Endophytes are microorganisms that live in the internal tissues of plants without harming the host plants. In this symbiotic relationship, the host plants provide nutrients and shelter to the endophytes, in turn, endophytes can promote the growth of host plants and act as a biological control agents against plant pathogens. Plant-microbe interactions like this are noted for natural methods for sustainable agriculture and environmental conservation. However, in spite of the infinite potential, there are only a few reports on the endophytes present in ginseng. In this study, we isolated and identified the endophytes from Panax ginseng seeds and evaluated the biological activities (IAA production ability, nitrogen fixation ability, phosphate solubilization capacity, siderophore production ability, and antifungal activities) of the endophyte isolates. Eight different endophytes were identified by 16S rRNA sequencing. Most of the endophytes have antibiotic and plant growth promoting (PGP) activities. Particularly, PgSEB5-37E have the highest antibiotic activity, both PgSEB5-37B and PgSEB5-37H have high PGP traits such as an abilities to produce IAA, solubilize phosphate and fix nitrogen. These results indicated that the endophytes from P. ginseng seeds may have applicable value to many industries. In order to use the isolated endophytes, quantitative analysis and field tests are needed to be performed.

• 한국산학기술학회논문지
• /
• 제16권1호
• /
• pp.868-876
• /
• 2015

실내 공기 중 부유세균은 실내공기오염을 유발하고 재실자의 건강상 위해를 초래할 수 있다. 본 연구는 다중이용시설을 대상으로 주요목적으로 사용되는 실내에서 부유세균의 농도와 세균의 종류를 동정하고 실내공기오염물질 중 부유세균에 대한 기초정보 제공을 목적으로 한다. 노인시설 7개소 (노인요양시설, 노인전문병원, 복지관 포함), 대규모점포 4개소, 대학병원 4개소, 어린이집 7개소 (유치원, 어린이집, 보육원 포함), 지하역사 4개소 및 버스터미널 4개소 등 총 30개소 120개 지점에서 총부유세균의 농도를 측정 분석하고 세균의 종류를 동정하였다. 모든 시설군에서 측정된 실내 부유세균의 농도는 유지기준 $800CFU/m^3$이하 이었으나, I/O비가 1.09-2.36으로 실내의 총부유세균의 농도가 실외에 비해 높았고, 어린이집, 대학병원, 노인시설에서 동정된 세균의 종류별 높은 검출빈도 보였다. 국내 다중이용시설의 주요목적에 따른 실내 부유세균에 대한 지속적 관찰이 필요하며, 특히 어린이집, 대학병원, 노인시설 등에 대한 관심이 요구되고, 추후 배양 가능한 세균 뿐 아니라 알레르겐 등을 포함한 미생물학적 실내오염물질에 대한 추가 연구가 필요할 것으로 사료된다.

• 한국미생물·생명공학회지
• /
• 제5권4호
• /
• pp.171-175
• /
• 1977

분리된 미생물은 pH 4.5내지 pH 8.0에서 100% 안정성을 나타내며 에너지원이 전혀 공급되지 않은 상태에서도 37$^{\circ}C$의 용액중에서 48시간 이고 경과시도 80% 이상의 높은 생존율을 나타낸다. 생리식염수에서는 4$0^{\circ}C$에서 한 시간 방치하는 경우도 90% 이상의 높은 생존율을 나타낸다는 것을 알수 있었다. 이러한 조건들을 검토해볼 때 생장내 조건에서 상당히 안정하며 경구투여제로 투여하기에 충분히 적당하다고 결론지을 수 있겠다. 특히 포만시에 투여하는 것이 더욱 효과적인 것으로 사료되므로 소화제등과 혼용한 약제로 축발하는 것이 바람직하리라 기대된다.

• Archives of Pharmacal Research
• /
• 제28권6호
• /
• pp.660-666
• /
• 2005

The intestinal microbiota are important to the host with regard to resistance they impart against bacterial infections and their involvement in mediating metabolic functions. Lactic acid producing bacteria such as Lactobacillus play an important physiological role in these matters. The aim of the present study was to isolate Lactobacillus sp. that inhibits enteric pathogens. Initially, 17 isolates from healthy Koreans were collected on Lactobacillus selective medium. Resistance of the isolates to antibiotics including rifampicin, streptomycin, clindamycin and vancomycin was measured. One of the isolate was identified as Lactobacillus ruminus on the basis of bacterial cell morphology, cultural characteristic and biochemical characteristics, 16S rRNA sequence analysis and PCR-RAPD. Antimicrobial activity of the bacterium against Vancomycin Intermediate Resistant Staphylococcus aureus (VISA) and Vancomycin-Resistant Enterococci (VRE) was measured. About $10^4$ cells of VISA or VRE were mixed with 1, 5, and 9 mL of L. ruminus SPM 0211 and the final volume was adjusted to 10 mL with brain heart infusion (BHI) broth. The cell suspension was incubated for 3, 6, 9, and 24 h, serially diluted and then plated on BHI agar plates. As numbers of L. ruminus SPM 0211 were increased, viable cell count of VISA and VRE decreased. The strongest antimicrobial activity of SPM 0211 was observed after 9 h incubation in any mixture, almost completely inhibiting the growth of these two bacteria. The results suggest that the freshly isolated L. ruminus SPM 0211 may be used as a pro-biotic microbe that prevents the colonization of enteric pathogens and can thereby promote good gastrointestinal health.

• 한국토양비료학회지
• /
• 제29권1호
• /
• pp.67-73
• /
• 1996

식물(植物) 생육촉진(生育促進) 유용미생물(有用微生物)을 지리산 일대의 침엽수와 활엽수 밑의 부엽토(腐葉土)에서 선발하고, 고추묘에 접종하여 묘의 생장에 미치는 효과를 조사한 결과는 다음과 같다. 1. 분리된 8종의 균주를 동정한 결과 Micrococcus sp., Bacillus subtilis, Enterobacter agglomerans, Bacillus megaterium, Pseudomonas putida, Pseudomonas fluorescens, Xanthomonas maltophilia, Staphylococcus xylosus등 8종의 균주가 동정되었다. 2. 8종의 혼합균주 처리구의 고추묘의 초장(草長)은 무처리구에 비해 처리 10일 이후, 엽수(葉數)는 44일 이후부터 생육이 크게 진전되었다.

• Journal of Animal Science and Technology
• /
• 제51권5호
• /
• pp.427-432
• /
• 2009

본 연구에서는 xylanase, mannanase를 생산하는 미생물균주를 분리하기 위해 1차로 토양 및 낙엽 등에서 곰팡이 균주를 약 50여주 분리하였으며, 1차 선발된 균주로부터 xylanase, mannanase를 생산하는 미생물 분리를 위해 xylanase 생성균주 선별배지, mannanase 생성균주 선별배지에 single colony를 접종한 후 clear zone이 생기는 15종의 균주를 2차 선발하였다. 2차 선발된 균주를 개별적으로 배양하여, DNS 방법을 활용하여 xylanase, mannanase 효소활성을 측정하여 6종의 균주를 선발하였다. 선별된 균주는 액상배양에서 생산한 xylanase, mannanase 효소활성이 각각 0.9~1.6 unit/mL, 0.2~0.4 unit/mL 범위로 나타났다. 이 중 결과가 좋은 3종을 선정하여 고상배양으로 배양한 균주의 xylanase, mannanase 효소활성은 각각 103.7~220.0 unit/g, 20.1~40.3 unit/g으로 분석되었다. 선별된 3종의 균주중 xylanase, mannanase 효소활성이 각각 197.3 unit/g, 39.9 unit/g으로 가장 높은 E-3 균주를 최종적으로 선발하였다. 최종으로 분리한 E-3 균주는 형태학적 특징과 DNA 염기서열을 비교한 결과 Aspergillus niger와 99% 일치하였다.