• Title/Summary/Keyword: sequencing analysis

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The Antioxidant and Skin-whitening Effects of Saccharomyces cerevisiae FT4-4 Isolated from Berries Grown in Sunchang (화장품 소재로서 순창 베리류 유래 Sacchromyces cerevisiae FT4-4의 항산화 활성 및 미백 효과)

  • Seo, Ji won;Ryu, Myeong Seon;Yang, Hee-Jong;Jeong, Su-Ji;Jeong, Do-Youn
    • Journal of Life Science
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    • v.31 no.2
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    • pp.175-182
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    • 2021
  • Saccharomyces lysate has the well-known function of soothing the skin in various ways: it is an anti-irritant and can treat skin care conditions, such as skin whitening and antioxidative activity. However, data on the safety for use of Saccharomyces lysate in cosmetics and skin care products are still limited. To design a new cosmetic material with antioxidant and skin-whitening effects, 80 yeast strains were isolated from berries grown in Sunchang. Among the isolates, the FT4-4 strain, which exhibited superior biological activities, was selected for further experiments. The FT4-4 strain was identified as Saccharomyces cerevisiae by 18S rRNA gene sequencing analysis. S. cerevisiae FT4-4 showed higher DPPH radical-scavenging (51.41%), superoxide dismutase (62.23%), and tyrosinase inhibition (64.75%) activities. The highest yield of biomass (3.16 g/l) and maximum growth rate of S. cerevisiae FT4-4 were observed within 16 h. Furthermore, the cytotoxicity potential of S. cerevisiae FT4-4 on B16F10 melanoma cells was measured by an MTT assay, and the results indicated that S. cerevisiae FT4-4 had a capacity to inhibit melanin up to 72.02% at an initial 10 mg/ml concentration. These results suggest that S. cerevisiae FT4-4 could be a promising candidate as a multi-functional material for application in the cosmetic industry, especially because of its antioxidant and skin-whitening effects.

Isolation of a Pseudoalteromonas sp. JH-1 Producing Agarase and Characterization of its Agarase (Agarase를 생산하는 Pseudoalteromonas sp. JH-1의 분리·동정 및 agarase의 특성 연구)

  • Lee, Dong-Geun;Kim, Ju-Hui;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.31 no.5
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    • pp.496-501
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    • 2021
  • In this study, the marine agar-degrading bacterium Pseudoalteromonas sp. JH-1 was isolated, and its growth and agarase properties were investigated. Seawater was collected from the offshore of the Yonggung Temple in Busan, and agar-degrading bacteria were isolated and cultured with marine agar medium. The bacterium Pseudoalteromonas sp. JH-1 was isolated through 16S rRNA gene sequencing. The extracellularly secreted enzyme was obtained from the culture broth of Pseudoalteromonas sp. JH-1 and was used to characterize its agarase. The extracellular agarase exhibited a maximum activity of 116.6 U/l at 50℃ and pH 6.0 of 20 mM Tris-HCl buffer. Relative activities were 31, 59, 94, 100, 45, and 31% at 20, 30, 40, 50, 60, and 70℃, respectively. Relative activities were 49, 85, 100, 86, 81, and 67% at pH 4, 5, 6, 7, 8, and 9, respectively. Residual activity was more than 85% after exposure at 20, 30, and 40℃ for 2 hr, and more than 82% after exposure at 50℃ for 2 hr. Zymogram analysis confirmed that Pseudoalteromonas sp. JH-1 produced at least two agarases of 55 and 97 kDa. As the products of α-agarase and β-agarase have antioxidation, antitumor, skin-whitening, macrophage activation, and prebiotic effects, further studies are needed on the agarase of Pseudoalteromonas sp. JH-1.

PCR-based markers for discriminating Solanum demissum were developed by comparison of complete chloroplast genome sequences of Solanum species (가지속 식물의 엽록체 전장유전체 비교를 통한 PCR 기반의 Solanum demissum 특이적 분자마커 개발)

  • Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.48 no.1
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    • pp.18-25
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    • 2021
  • Solanum demissum is one of the wild Solanum species originating from Mexico. It has wildly been used for potato breeding due to its resistance to Phytophthora infestans. S. demissum has an EBN value of four, which is same as that of S. tuberosum, so that it is directly crossable for breeding purposes with the cultivated tetraploid potato (S. tuberosum). In this study, the chloroplast genome sequence of S. demissum obtained by next-generation sequencing technology was described and compared with those of seven other Solanum species to develop S. demissum-specific markers. Thetotal sequence length of the chloroplast genome is 155,558 bp, and its structural organization is similar to those of other Solanum species. Phylogenetic analysis with ten other Solanaceae species revealed that S. demissum is most closely grouped with S. hougasii and S. stoloniferum followed by S. berthaultii and S. tuberosum. Additional comparison of the chloroplast genome sequence with those of seven other Solanum species revealed two InDels specific to S. demissum. Based on these InDels, two PCR-based markers for discriminating S. demissum from other Solanum species were developed. The results obtained in this study will provide an opportunity to investigate more detailed evolutionary and breeding aspects in Solanum species.

Assessment of Rhizosphere Microbial Community Structure in Tomato Plants after Inoculation of Bacillus Species for Inducing Tolerance to Salinity (토마토에 염류 내성을 유도하는 바실러스 균주 처리 후 근권 미생물 군집 구조 연구)

  • Yoo, Sung-Je;Lee, Shin Ae;Weon, Hang-Yeon;Song, Jaekyeong;Sang, Mee Kyung
    • Korean Journal of Environmental Agriculture
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    • v.40 no.1
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    • pp.49-59
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    • 2021
  • BACKGROUND: Soil salinity causes reduction of crop productivity. Rhizosphere microbes have metabolic capabilities and ability to adaptation of plants to biotic and abiotic stresses. Plant growth-promoting bacteria (PGPB) could play a role as elicitors for inducing tolerance to stresses in plants by affecting resident microorganism in soil. This study was conducted to demonstrate the effect of selected strains on rhizosphere microbial community under salinity stress. METHODS AND RESULTS: The experiments were conducted in tomato plants in pots containing field soil. Bacterial suspension was inoculated into three-week-old tomato plants, one week after inoculation, and -1,000 kPa-balanced salinity stress was imposed. The physiological and biochemical attributes of plant under salt stress were monitored by evaluating pigment, malondialdehyde (MDA), proline, soil pH, electrical conductivity (EC) and ion concentrations. To demonstrate the effect of selected Bacillus strains on rhizosphere microbial community, soil microbial diversity and abundance were evaluated with Illumina MiSeq sequencing, and primer sets of 341F/805R and ITS3/ITS4 were used for bacterial and fungal communities, respectively. As a result, when the bacterial strains were inoculated and then salinity stress was imposed, the inoculation decreases the stress susceptibility including reduction in lipid peroxidation, enhanced pigmentation and proline accumulation which subsequently resulted in better plant growth. However, bacterial inoculations did not affect diversity (observed OTUs, ACE, Chao1 and Shannon) and structure (principle coordinate analysis) of microbial communities under salinity stress. Furthermore, relative abundance in microbial communities had no significant difference between bacterial treated- and untreated-soils under salinity stress. CONCLUSION: Inoculation of Bacillus strains could affect plant responses and soil pH of tomato plants under salinity stress, whereas microbial diversity and abundance had no significant difference by the bacterial treatments. These findings demonstrated that Bacillus strains could alleviate plant's salinity damages by regulating pigments, proline, and MDA contents without significant changes of microbial community in tomato plants, and can be used as effective biostimulators against salinity stress for sustainable agriculture.

Effect of ammonia nitrogen and microorganisms on the elevated nitrogenous biochemical oxygen demand (NBOD) levels in the Yeongsan river in Gwangju (광주지역 영산강의 NBOD 발생에 대한 암모니아성 질소 및 미생물 영향 연구)

  • Jang, Dong;Cho, Gwangwoon;Son, Gyeongrok;Kim, Haram;Kang, Yumi;Lee, Seunggi;Hwang, Soonhong;Bae, Seokjin;Kim, Yunhee
    • Journal of Korean Society of Water and Wastewater
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    • v.36 no.2
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    • pp.81-95
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    • 2022
  • The present study was performed to investigate the effects of NH3-N and nitrifying microorganisms on the increased BOD of downstream of the Yeongsan river in Gwangju. Water samples were collected periodically from the 13 sampling sites of rivers from April to October 2021 to monitor water qualities. In addition, the trends of nitrogenous biochemical oxygen demand (NBOD) and microbial clusters were analyzed by adding different NH3-N concentrations to the water samples. The monitoring results showed that NH3-N concentration in the Yeongsan river was 22 times increased after the inflow of discharged water from the Gwangju 1st public sewage treatment plant (G-1-PSTP). Increased NH3-N elevated NBOD levels through the nitrification process in the river, consequently, it would attribute to the increase of BOD in the Yeongsan river. Meanwhile, there was no proportional relation between NBOD and NH3-N concentrations. However, there was a significant difference in NBOD occurrence by sampling sites. Specifically, when 5 mg/L NH3-N was added, NBOD of the river sample showed 2-4 times higher values after the inflow of discharged water from G-1-PSTP. Therefore, it could be thought other factors such as microorganisms influence the elevated NBOD levels. Through next-generation sequencing analysis, nitrifying microorganisms such as Nitrosomonas, Nitroga, and Nitrospira (Genus) were detected in rivers samples, especially, the proportion of them was the highest in river samples after the inflow of discharged water from G-1-PSTP. These results indicated the effects of nitrifying microorganisms and NH3-N concentrations as important limiting factors on the increased NBOD levels in the rivers. Taken together, comprehensive strategies are needed not only to reduce the NH3-N concentration of discharged water but also to control discharged nitrifying microorganisms to effectively reduce the NBOD levels in the downstream of the Yeongsan river where discharged water from G-1-PSTP flows.

Simotang Alleviates the Gastrointestinal Side Effects of Chemotherapy by Altering Gut Microbiota

  • Deng, Lijing;Zhou, Xingyi;Lan, Zhifang;Tang, Kairui;Zhu, Xiaoxu;Mo, Xiaowei;Zhao, Zongyao;Zhao, Zhiqiang;Wu, Mansi
    • Journal of Microbiology and Biotechnology
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    • v.32 no.4
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    • pp.405-418
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    • 2022
  • Simotang oral liquid (SMT) is a traditional Chinese medicine (TCM) consisting of four natural plants and is used to alleviate gastrointestinal side effects after chemotherapy and functional dyspepsia (FD). However, the mechanism by which SMT helps cure these gastrointestinal diseases is still unknown. Here, we discovered that SMT could alleviate gastrointestinal side effects after chemotherapy by altering gut microbiota. C57BL/6J mice were treated with cisplatin (DDP) and SMT, and biological samples were collected. Pathological changes in the small intestine were observed, and the intestinal injury score was assessed. The expression levels of the inflammatory factors IL-1β and IL-6 and the adhesive factors Occludin and ZO-1 in mouse blood or small intestine tissue were also detected. Moreover, the gut microbiota was analyzed by high-throughput sequencing of 16S rRNA amplicons. SMT was found to effectively reduce gastrointestinal mucositis after DDP injection, which lowered inflammation and tightened the intestinal epithelial cells. Gut microbiota analysis showed that the abundance of the anti-inflammatory microbiota was downregulated and that the inflammatory microbiota was upregulated in DDP-treated mice. SMT upregulated anti-inflammatory and anticancer microbiota abundance, while the inflammatory microbiota was downregulated. An antibiotic cocktail (ABX) was also used to delete mice gut microbiota to test the importance of gut microbiota, and we found that SMT could not alleviate gastrointestinal mucositis after DDP injection, showing that gut microbiota might be an important mediator of SMT treatment. Our study provides evidence that SMT might moderate gastrointestinal mucositis after chemotherapy by altering gut microbiota.

Development of PCR-based markers specific to Solanum brevicaule by using the complete chloroplast genome sequences of Solanum species (엽록체 전장유전체 비교를 통한 PCR 기반의 Solanum brevicaule 특이적 분자마커 개발)

  • Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.49 no.1
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    • pp.30-38
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    • 2022
  • Solanum brevicaule is one of the tuber-bearing wild Solanum species. Because of its resistance to several important pathogens infecting potatoes during cultivation, it can be used for potato breeding. However, the fact that S. brevicaule used in this study has an EBN value of two causes the sexual reproduction barriers between the species and cultivated potatoes. In this study, specific markers for discriminating S. brevicaule from other Solanum species were developed on the basis of the results of sequence alignments with the whole chloroplast genomes of S. brevicaule and seven other Solanum species. The chloroplast genome of S. brevicaule was completed by next-generation sequencing technology described in other recent studies. The total sequence length of the chloroplast genome of S. brevicaule is 155,531 bp. Its structure and gene composition are similar to those of other Solanum species. Phylogenetic analysis revealed that S. brevicaule was closely grouped with other Solanum species. BLASTN search showed that its genome sequence had 99.99% and 99.89% identity with those of S. spegazzinii (MH021562) and S. kurtzianum (MH021495), respectively. Sequence alignment identified 27 SNPs that were specific to S. brevicaule. Thus, three PCR-based CAPS markers specific to S. brevicaule were developed on the basis of these SNPs. This study will facilitate in further studies on evolutionary and breeding aspects in Solanum species.

An Analysis of the Current Status and Characteristics of Airborne Fungi in Indoor Air in Multi-Use Facilities Nationwide (전국 다중이용시설의 실내공기 부유 곰팡이의 현황 및 특성 분석)

  • Park, Yongsung;Kwon, Soonhyun;Park, Song-Yi;Kee, Sun-Ho;Yoon, Wonsuck
    • Journal of Environmental Health Sciences
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    • v.48 no.5
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    • pp.282-289
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    • 2022
  • Background: Airborne fungi are ubiquitous in the air and exposure to an airborne fungus can be a significant risk factor. The composition of fungi has been potentially important for human health, especially for respiratory diseases like asthma and atopic dermatitis. Therefore, we attempted to ascertain what kind of airborne fungi affect human health at a nationwide level. Objectives: This study was carried out to provide information on indoor fungi distribution at multi-use facilities throughout South Korea. Methods: We classified our data by region and public facility after collection, cultivation, and identification via the sequencing of the ITS (internal transcribed spacer) region. We investigated whether or not the proliferation of HaCaT cells was affected by the identified airborne fungi. Results: In our data, the most isolated airborne fungi by region were Penicillium spp (Seoul, Daegu), Periconia sp (Gyeonggi-do), Iprex sp (Gangwon-do), Phanerochaete sp (Busan), Bjerkandera sp (Gwangju), and Aspergillus sp (Jeju-do). In the public facilities, the most detected fungi were Cladosporium sp (public transport), Penicillium sp (apartment house, retail market, financial institution, karaoke room), Bjerokandera sp (underground parking lot, public toilet, medical institution), Periconia sp (retail store), and Fusarium sp (general restaurant). Next, we selected twenty airborne fungi to examine their cytotoxicity and proliferation of human skin cells. In this experiment, the proliferation of the cells was influenced by most of the identified fungi. In case of the cytotoxicity test, most genera except for Rhodotorula sp and Moesziomyces sp showed cytotoxicity in HaCaT cells. Conclusions: The distribution of mold in the indoor air in multi-use facilities in South Korea differs from region to region, and this is an indicator that should be considered in future health impact studies. In addition, as a result of culturing about 20 types of bacteria dominant in indoor air, it was found that most (90%) inhibit the growth of skin cells, which can be harmful to health. An in-depth study of the health effects of floating fungi is needed.

PCR-based markers to select plastid genotypes of Solanum acaule (Solanum acaule 색소체 유전자형 선발을 위한 특이적 분자마커 개발)

  • Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.178-186
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    • 2022
  • The tetraploid Solanum acaule is a wild potato species from Bolivia widely used for potato breeding because of its diverse attractive traits, including resistance to frost, late blight, potato virus X, potato virus Y, potato leafroll virus, potato spindle tuber viroid, and cyst nematode. However, the introgression of useful traits into cultivated potatoes via crossing has been limited by differences in endosperm balance number between species. Somatic fusion could be used to overcome sexual reproduction barriers and the development of molecular markers is essential to select proper fusion products. The chloroplast genome of S. acaule was sequenced using next-generation sequencing technology and specific markers for S. acaule were developed by comparing the obtained sequence with those of seven other Solanum species. The total length of the chloroplast genome is 155,570 bp, and 158 genes were annotated. Structure and gene content were very similar to other Solanum species and maximum likelihood phylogenetic analysis with 12 other species belonging to the Solanaceae family revealed that S. acaule is very closely related to other Solanum species. Sequence alignment with the chloroplast genome of seven other Solanum species revealed four InDels and 79 SNPs specific to S. acaule. Based on these InDel and SNP regions, one SCAR marker and one CAPS marker were developed to discriminate S. acaule from other Solanum species. These results will aid in exploring evolutionary aspects of Solanum species and accelerating potato breeding using S. acaule.

Isolation and Identification of Fungi Associated with Decay of Quercus mongolica (신갈나무의 부후에 관여하는 곰팡이 분리 및 동정)

  • HAM, Youngseok;AN, Ji-Eun;LEE, Soo Min;CHUNG, Sang-Hoon;KIM, Sun Hee;PARK, Mi-Jin
    • Journal of the Korean Wood Science and Technology
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    • v.49 no.3
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    • pp.234-253
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    • 2021
  • The Quercus mongolica has a high utilization value in industrial economic sector. The species is distributed throughout Korea, however, the damage caused by deterioration such as discoloration and decay is severe. For this reason, the deterioration of Q. mongolica is an obstacle to its use as wood, but research on deterioration factors is insufficient. In this study, we focused on fungi as a factor influencing the deterioration of Q. mongolica, and isolated and identified the fungi from the deteriorated Q. mongolica. In additions, in order to confirm whether the identified fungi actually affects wood deterioration, enzyme activities of the identified fungi were evaluated and related mass loss of wood treated with the fungi was measured by wood decay test. As a result of sequencing analysis using the ITS region of the genomic DNA of the fungi isolated from Q. mongolica, Mucor circinelloides, Cunninghamella elegans, and Umbelopsis isabellina 3 species belonging to Mucoromycota phylum, and Ophiostoma piceae and Aureobasidium melanogenum 2 species belonging to Ascomycota phylum were identified. These five fungi had enzyme (i.e. cellulase, laccase) activities related to wood decay and reduced the mass of heartwood and sapwood of Q. mongolica in practice. In particular, O. piceae and A. melanogenum, which have both cellulase and laccase activities, showed 6.9% and 1.5% mass loss, respectively. These results indicated that five fungi identified in this study influence the deterioration of Q. mongolica and are wood decaying fungi for Q. mongolica potentially.