• Title/Summary/Keyword: 26S rRNA gene sequencing and phylogenetic analysis

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DNA Sequencing and Phylogenetic Analysis of the 18S rRNA Gene of Atractylodes japonica Koidz and Analysis of Atractylon (삽주의 18S rRNA 유전자의 염기서열 결정, 계통분류학적 분석 및 atractylon 분석)

  • Bae, Young-Min
    • Korean Journal of Medicinal Crop Science
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    • v.17 no.1
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    • pp.26-32
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    • 2009
  • The region containing 18S rRNA gene, ITS 1 and part of the 5.8S rRNA gene of the Atractylodes japonica Koidz was amplified by PCR and the product cloned in a pBluescript SK II plasmid. DNA sequence of the cloned DNA was determined and submitted to the GenBank (accession number EU678363). Phylogenetic analysis of the ITS 1 DNA showed close similarity with the other plant species of the family Compositae. The extract of the plant materials of five different members of the family Compositae was analyzed by HPLC to detect atractylon. Extract of the A. japonica Koidz showed presence of significant amount of atractylon. However, noticeable amount of atractylon was not detected by the same analyses from the extracts of the other plants belonging to the family Compositae including Artemisia capillaris, Chrysantemum zawadskii, Eclipta prostrata or Taraxacum platycarpum.

Detection and Potential Abundances of Anammox Bacteria in the Paddy Soil

  • Khanal, Anamika;Lee, Seul;Lee, Ji-Hoon
    • Korean Journal of Environmental Agriculture
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    • v.39 no.1
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    • pp.26-35
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    • 2020
  • BACKGROUND: Microbes that govern a unique biochemical process of oxidizing ammonia into dinitrogen gas, such as anaerobic ammonium oxidation (anammox) have been reported to play a pivotal role in agricultural soils and in oceanic environments. However, limited information for anammox bacterial abundance and distribution in the terrestrial habitats has been known. METHODS AND RESULTS: Phylogenetic and next-generation sequencing analyses of bacterial 16S rRNA gene were performed to examine potential anammox bacteria in paddy soils. Through clone libraries constructed by using the anammox bacteria-specific primers, some clones showed sequence similarities with Planctomycetes (87% to 99%) and anammox bacteria (94% to 95%). Microbial community analysis for the paddy soils by using Illumina Miseq sequencing of 16S rRNA gene at phylum level was dominated by unclassified Bacteria at 33.2 ± 7.6%, followed by Chloroflexi at 20.4 ± 2.0% and Acidobacteria at 17.0 ± 6.5%. Planctomycetes that anammox bacteria are belonged to was 1.5% (± 0.3) on average from the two paddy soils. CONCLUSION: We suggest evidence of anammox bacteria in the paddy soil. In addition to the relatively well-known microbial processes for nitrogen-cycle, anammox can be a potential contributor on the cycle in terrestrial environments such as paddy soils.

Optimization of Citric Acid Production by Immobilized Cells of Novel Yeast Isolates

  • Hesham, Abd El-Latif;Mostafa, Yasser S.;AlSharqi, Laila Essa Omar
    • Mycobiology
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    • v.48 no.2
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    • pp.122-132
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    • 2020
  • Citric acid is a commercially valuable organic acid widely used in food, pharmaceutical, and beverage industries. In this study, 260 yeast strains were isolated from soil, bread, juices, and fruits wastes and preliminarily screened using bromocresol green agar plates for their ability to produce organic acids. Overall, 251 yeast isolates showed positive results, with yellow halos surrounding the colonies. Citric acid production by 20 promising isolates was evaluated using both free and immobilized cell techniques. Results showed that citric acid production by immobilized cells (30-40 g/L) was greater than that of freely suspended cells (8-19 g/L). Of the 20 isolates, two (KKU-L42 and KKU-L53) were selected for further analysis based on their citric acid production levels. Immobilized KKU-L42 cells had a higher citric acid production rate (62.5%), while immobilized KKU-L53 cells showed an ~52.2% increase in citric acid production compared with free cells. The two isolates were accurately identified by amplification and sequence analysis of the 26S rRNA gene D1/D2 domain, with GenBank-based sequence comparison confirming that isolates KKU-L42 and KKU-L53 were Candida tropicalis and Pichia kluyveri, respectively. Several factors, including fermentation period, pH, temperature, and carbon and nitrogen source, were optimized for enhanced production of citric acid by both isolates. Maximum production was achieved at fermentation period of 5 days at pH 5.0 with glucose as a carbon source by both isolates. The optimum incubation temperature for citric acid production by C. tropicalis was 32 ℃, with NH4Cl the best nitrogen source, while maximum citric acid by P. kluyveri was observed at 27 ℃ with (NH4)2 SO4 as the nitrogen source. Citric acid production was maintained for about four repeated batches over a period of 20 days. Our results suggest that apple and banana wastes are potential sources of novel yeast strains; C. tropicalis and P. kluyveri which could be used for commercial citric acid production.

Isolation and Identification of Chlorella sp. CMS-1 and the Chemical Composition of Its Hot Water Extract (클로렐라 균주의 분리 동정 및 열수 추출물의 이화학적 조성)

  • Cha, Jae-Young;Kim, Jung-Wook;Park, Bo-Kyung;Jin, Hyun-Jin;Kim, Sung-Young;Cho, Young-Su
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1723-1727
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    • 2008
  • Chlorella sp. CMS-1 strain was isolated from the outdoors cultivation pools in Culmansa Co., Ltd. This strain was found to be a rounded type of 3 ${\mu}m$. Phylogenetic analysis by the 18S rRNA sequencing using isolated strain is most similar to Chlorella sp. IFRPD 1018 gene at the level of nucleotide sequence identity at 99%. Accordingly, the isolated Chlorella strain was named as Chlorella sp. CMS-1 based on its morphological and phylogenetic properties. The concentrations of crude protein and fat were 59% and 0.01%, respectively. Major compositional amino acids (mg%) were glutamic acid 6.21, alanine 5.76, aspartic acid 5.44%, glycine 4.29%, and threonine 3.09% and major free amino acids (mg%) were ${\gamma}$-aminobutyric acid (GABA) 7.13%, L-alanine 1.44%, L-glutamic acid 0.90, L-leucine 0.26% and L-glycine 0.20%. The concentrations of major minerals were P 2.25%, K 2.25%, Na 1.09%, Mg 0.63%, and Ca 0.28%.

A Study of the Diversity and Profile for Extracellular Enzyme Production of Aerobically Cultured Bacteria in the Gut of Muraenesox cinereus (갯장어(Muraenesox cinereus) 장으로부터 호기적 조건에서 분리된 미생물의 다양성 및 세포외 효소 생산능 분석에 관한 연구)

  • Lee, Yong-Jik;Oh, Do-Kyoung;Kim, Hye Won;Nam, Gae-Won;Sohn, Jae Hak;Lee, Han-Seung;Shin, Kee-Sun;Lee, Sang-Jae
    • Journal of Life Science
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    • v.29 no.2
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    • pp.248-255
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    • 2019
  • This research confirmed the diversity and characterization of gut microorganisms isolated from the intestinal organs of Muraenesox cinereus, collected on the Samcheonpo Coast and Seocheon Coast in South Korea. To isolate strains, Marine agar medium was basically used and cultivated at $37^{\circ}C$ and pH7 for several days aerobically. After single colony isolation, totally 49 pure single-colonies were isolated and phylogenetic analysis was carried out based on the result of 16S rRNA gene DNA sequencing, indicating that isolated strains were divided into 3 phyla, 13 families, 15 genera, 34 species and 49 strains. Proteobacteria phylum, the main phyletic group, comprised 83.7% with 8 families, 8 genera and 26 species of Aeromonadaceae, Pseudoalteromonadaceae, Shewanellaceae, Enterobacteriaceae, Morganellaceae, Moraxellaceae, Pseudomonadaceae, and Vibrionaceae. To confirm whether isolated strain can produce industrially useful enzyme or not, amylase, lipase, and protease enzyme assays were performed individually, showing that 39 strains possessed at least one enzyme activity. Especially the Aeromonas sp. strains showed all enzyme activity tested. This result indicated that isolated strains have shown the possibility of the industrial application. Therefore, this study has contributed for securing domestic genetic resources and the expansion of scientific knowledge of the gut microbial community in Muraenesox cinereus of South Korea.