• Title/Summary/Keyword: ITS rDNA

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Genetic Diversity of Culturable Endophytic Fungi Isolated from Halophytes Naturally Growing in Muan Salt Marsh (무안 염습지에 자생하는 염생식물(halophyte)의 뿌리로부터 분리된 내생진균(endophytic fungi)의 유전학적 다양성)

  • You, Young-Hyun;Yoon, Hyeok-Jun;Seo, Yeong-Gyo;Kim, Mi-Ae;Kang, Myung-Suk;Kim, Chang-Mu;Ha, Sang-Chul;Cho, Ga-Youn;Kim, Jong-Guk
    • Journal of Life Science
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    • v.22 no.7
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    • pp.970-980
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    • 2012
  • Native halophytes, such as Suaeda maritima, Limonium tetragonum, S. japonica, Zoysia sinica, and Phragmites australis were collected from the Muan salt marsh. Ninety endophytic fungi were isolated from the roots of the collected halophytes. Molecular insights inferred by internal transcribed spacer containing ITS1, 5.8s, and the ITS2 region showed that all the fungal strains belong to ten orders, i.e., Capnodiales (4.44%), Cystofilobasidiales (1.11%), Dothideales (3.33%), Eurotiales (53.33%), Glomerellales (3.33%), Hypocreales (8.89%), Mucorales (1.11%), Pleosporales (15.56%), Sordariales (1.11%), and Trichosphaeriales (1.11%). The rest (6.67%) of all fungal isolates were not identified. Ninety fungal strains were confirmed at the genus level, containing Acremonium, Alternaria, Aspergillus, Aureobasidium, Cephalosporium, Chaetomium, Cladosporium, Colletotrichum, Cryptococcus, Didymella, Dothideomycete, Emericellopsis, Epicoccum, Eupenicillium, Fusarium, Gibberella, Gongronella, Macrophoma, Microsphaeropsis, Nigrospora, Paecilomyces, Paraconiothyrium, Penicillium, Phaeomyces, Phoma, Pleosporales, Purpureocillium, and Talaromyces. Of all the endophytic fungi identified from the various halophytes, Aspergillus and Penicillium of Eurotiales had the highest abundance.

Characterization of a cDNA Encoding Transmembrane Protein 258 from a Two-spotted Cricket Gryllus bimaculatus (쌍별귀뚜라미(Gryllus bimaculatus)의 GbTmem258 cDNA 클로닝과 발현분석)

  • Kisang Kwon;Honggeun Kim;Hyewon Park;O-Yu Kwon
    • Journal of Life Science
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    • v.33 no.10
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    • pp.828-834
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    • 2023
  • The cDNA that encodes transmembrane protein 258 (Tmem258) was cloned from Gryllus bimaculatus and named GbTmem258. This protein comprises 80 amino acids, has no N-glycosylation site, and contains five potential phosphorylation sites at two serines, two threonines, and one tyrosine. The predicted molecular mass of GbTmem258 is 9.06 kDa, and its theoretical isoelectric point is 5.5. The tertiary structure of GbTmem258 was predicted using the available secondary structure information, which suggests the presence of alpha helices (52.5%), random coils (22.5%), extended strands (16.25%), and beta turns (8.75%). Homology analysis revealed that GbTmem258 exhibits high similarity at the amino-acid level to Tmem258 found in other species. The effect of starvation and refeeding on GbTmem258 mRNA expression was also examined in this study. It was found that GbTmem258 mRNA expression in the hindgut progressively increased throughout the starvation period, peaking at almost 1.5 times the control level after six days of starvation. However, refeeding for one to two days after the six-day starvation period restored GbTmem258 mRNA expression to the control level. In fat body, GbTmem258 mRNA expression was almost 3-fold higher during starvation compared to the control level. Refeeding for one to two days after the six-day fast resulted in a decline in the expression to about a 2.5-fold increase over the control level. Throughout the starving and refeeding periods, no other tissues showed any discernible alterations in GbTmem258 mRNA expression.

Characterization of β-agarase from Isolated Simiduia sp. SH-4 (분리된 Simiduia sp. SH-4가 생산하는 β-agarase의 특성조사)

  • Kim, Jae-Deog;Lee, Sol-Ji;Jo, Jeong-Gwon;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.26 no.4
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    • pp.453-459
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    • 2016
  • Agarases are classified into α-agarase and β-agarase that produce agarooligosaccharides and neoagarooligosaccharides, respectively. Neoagarooligosaccharides have whitening effect of skin, delay of starch degradation, and inhibition of bacterial growth etc. Hence, the object of this study was to isolate a novel agarase producing marine bacterium and characterization of its β-agarase. A novel agar-degrading bacterium was isolated from seashore of Namhae at Gyeongnamprovine, Korea and purely cultured with Marine agar 2216 media. The isolated bacterium was identified as Simiduia sp. SH-4 after 16S rRNA gene sequencing. The enzymatic sample was obtained from culture media of Simiduia sp. SH-4. Enzymatic activity was highly increased from 20(30% relative activity) to 30℃ (100%) and decreased from 30 to 40℃(75%) and so more. Relative activity was 100% at pH 6 while those were about 91% and 59% at pH 5.0 and 7.0, respectively, meaning the enzyme possesses narrow optimal pH range. Hence, the enzyme exhibited the maximal activity with 120.4 units/l at pH 6.0 and 30℃ in 20 mM Tris-HCl buffer. Thin layer chromatography (TLC) analysis showed that Simiduia sp. SH-4 produces β-agarase, which hydrolyze agarose to produce biofunctional neoagarooligosaccharides such as neoagarotetraose and neoagarobiose. Hence, broad applications would be possible using Simiduia sp. SH-4 and its enzyme in the food industry, cosmetics and medical fields.

Isolation and Characterization of Three Kinds of Lipopeptides Produced by Bacillus subtilis JKK238 from Jeot-Kal of Korean Traditional Fermented Fishes (한국 전통젓갈에서 분리한 Bacillus subtilis JKK238 균주 유래 세 종류 Lipopeptide의 분리 및 특성)

  • Yoon Sang-Hong;Kim Jung-Bong;Lim Yoong-Ho;Hong Seong-Ryeul;Song Jae-Kyeung;Kim Sam-Sun;Kwon Soon-Wo;Park In-Cheol;Kim Soo-Jin;Yeo Yun-Soo;Koo Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.295-301
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    • 2005
  • About seven hundred bacterial strains were collected from Jeot-Kal, a Korean traditional fermented fishes, in various Korean districts. One of the strains designated JKK238 has its ability to antagonize in vitro the growth of a wide variety of plant pathogenic fungi responsible for diseases of economical importance. The JKK238 strain was isolated from Oh-Jeot, a kind of fermented shrimps, of Kangkyeung in Korea, and was identified as Bacillus subtilis based on its physiological characteristics, fatty acids compositions of cellular wall, and 16S rDNA sequence analysis. We isolated simply antimicrobial lipopeptides (AMLP) by $25\%$ ammonium sulfate precipitation of 3 days-old tryptic soy broth cultures of the JKK238 strain. Further analysis of AMLP revealed that B. subtilis JKK238 produces a wide variety of antifungal lipopeptide isomers from the iturin, fengycin and surfactin families simultaneously. Above results indicate that the JKK238 strain can be added to the limited number B. subtilis strains reported to co-produce the three kinds of lipopeptide families.

Antifungal Activity of Bacillus Subtilis HK2 against Trichothecium Roseum Causing Pink Rot of Melon and White Stain Symptom on Grape (멜론 분홍빛썩음병과 포도 흰얼룩병의 원인균인 Trichothecium Roseum에 대한 Bacillus Subtilis HK2의 항균활성)

  • Oh, Soh-Young;Lee, En-Young;Nam, Ki-Woong;Yoon, Deok-Hoon
    • Korean Journal of Plant Resources
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    • v.29 no.1
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    • pp.39-45
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    • 2016
  • Pink Rot on melon and White Stain Symptom on grape are caused by Trichothecium roseum, one of the most important diseases of grape and melon. These diseases have been occurred in national-wide in Korea and causes irreversible damage on the grape and the melon at harvest season. This research presents the evaluation of the capacity of Bacillus subtillis HK2 to protect both melon and grape against T. reseum and establishes its role as a biocontrol agent. In this study, we isolated a Bacillus strain HK2 from rhizosphere soil, identified it as Bacillus subtillis by 16S rRNA analysis and demonstrated its antifungal activity against T. roseum. Under I-plate assay it was observed that the effect of hyphal growth inhibition was not due to production of volatile compounds. The optimum culture condition of HK2 was found at 30℃ and initial pH of 7.0. Application of HK2 culture suspension reduced 90.2% of white stain symptom on grape as compared to control, resulting in greater protection to grape against T. roseum infestation. Butanol extract of HK2 culture purified using flash column chromatography. The antifungal material was a polar substance as it showed antifungal activity in polar elute. Therefore, our results indicated a clear potential of B. subtilis HK2 to be used for biocontrol of Pink rot in melon and white stain symptom on grape caused by T. roseum.

Parvatrema duboisi (Digenea: Gymnophallidae) Life Cycle Stages in Manila Clams, Ruditapes philippinarum, from Aphae-do (Island), Shinan-gun, Korea

  • Jung, Bong-Kwang;Chang, Taehee;Shin, Hyejoo;Ryoo, Seungwan;Hong, Sooji;Lee, Jeonggyu;Song, Hyemi;Cho, Jaeeun;Kim, Deok-Gyu;Jun, Hojong;Kim, Min-Jae;Won, Eun Jeong;Han, Eun-Taek;Shin, Eun-Hee;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.59 no.1
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    • pp.83-88
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    • 2021
  • Life cycle stages, including daughter sporocysts, cercariae, and metacercariae, of Parvatrema duboisi (Dollfus, 1923) Bartoli, 1974 (Digenea: Gymnophallidae) have been found in the Manila clam Ruditapes philippinarum from Aphae-do (Island), Shinan-gun, Jeollanam-do, Korea. The daughter sporocysts were elongated sac-like and 307-570 (av. 395) ㎛ long and 101-213 (av. 157) ㎛ wide. Most of the daughter sporocysts contained 15-20 furcocercous cercariae each. The cercariae measured 112-146 (av. 134) ㎛ in total length and 35-46 (av. 40) ㎛ in width, with 69-92 (av. 85) ㎛ long body and 39-54 (av. 49) ㎛ long tail. The metacercariae were 210-250 (av. 231) ㎛ in length and 170-195 (av. 185) ㎛ in width, and characterized by having a large oral sucker, genital pore some distance anterior to the ventral sucker, no ventral pit, and 1 compact or slightly lobed vitellarium, strongly suggesting P. duboisi. The metacercariae were experimentally infected to ICR mice, and adults were recovered at day 7 post-infection. The adult flukes were morphologically similar to the metacercariae except in the presence of up to 20 eggs in the uterus. The daughter sporocysts and metacercariae were molecularly (ITS1-5.8S rDNA-ITS2) analyzed to confirm the species, and the results showed 99.8-99.9% identity with P. duboisi reported from Kyushu, Japan and Gochang, Korea. These results confirmed the presence of various life cycle stages of P. duboisi in the Manila clam, R. philippinarum, playing the role of the first as well as the second intermediate host, on Aphae-do (Island), Shinan-gun, Korea.

Development and Validation of Multiplex Polymerase Chain Reaction to Determine Squid Species Based on 16s rRNA Gene (오징어류 종 판별을 위한 다중 유전자 검사법 개발 및 검증)

  • Kim, Hyunsu;Seo, Yong Bae;Choi, Seong-Seok;Kim, Jin-Hee;Shin, Jiyoung;Yang, Ji-Young;Kim, Gun-Do
    • Journal of Food Hygiene and Safety
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    • v.30 no.1
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    • pp.43-50
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    • 2015
  • In this study, single PCR and multiplex PCR tests were examined for identification of four types of squid species (giant squid, cuttlefish, octopus, beka squid) purchased from fish market as well as aquatic processed products in Busan. To design the specific primers against each species, the nucleotide sequences of the mitochondrial 16s rRNA gene of Architeuthis dux, Todarodes pacificus, Enteroctopus dofleini, Enteroctopus megalocyathus, Uroteuthis chinensis, Uroteuthis duvauceli, Uroteuthis edulis groups were analyzed for the identification of each species registered in the GeneBank (www.ncbi.nlm.nih.gov) and have been used for comparative analysis. In order to obtain the size variation of amplified fragments on multiplex PCR, we designed KOJ-F, OJ-F, OCT-F, HAN-F, ALLR primers for each species. The optimal PCR conditions and primers were selected for four types of squid species to determine target base sequences in its PCR products. In the case of single PCR, giant squid was only amplified by KOJ-F/ALLR primer; cuttlefish was only amplified by OJ-F/ALLR primer; octopus was only amplified by OCT-F/ALLR primer; and beka squid was only amplified by HAN-F/ALLR primer. For multiplex PCR, the mixture of four kinds of genomic DNA (giant squid, cuttlefish, octopus, beka squid) been prepared as a template and used together with the mixture of KOJ-F/OJ-F/OCT-F/HAN-F/ALLR primers in the reaction. By the multiplex PCR, it is confirmed that four samples are correspond to multiple simultaneous amplicon. Finally, we validated the established methods of multiplex PCR in the aquatic processed products. Although the mitochondrial 16s rRNA primers used in this study was useful as a marker for detection of each species among them, the study indicated that the established multiplex PCR method can be more useful tool for monitoring the processed products.

Investigation of Fungicides Inhibitory Effect of on Summer Patch Disease, Caused by Magnaporthiopsis poae, in Kentucky bluegrass (여름잎마름병(Summer patch) 병원균에 대한 살균제의 억제효과 조사)

  • Lee, Jung Han;Shim, Gyu Yul;Kim, Jeong Ho;Jeon, Chang Wook;Kwak, Youn-Sig
    • Weed & Turfgrass Science
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    • v.6 no.2
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    • pp.151-156
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    • 2017
  • Summer patch is the most serious disease at turfgrass field or golf course established with Kentucky bluegrass during high temperature season in Korea. Nevertheless, chemicals for the summer patch control are not yet registered in Korea. We isolated the pathogens from the turfgrass showing typical summer patch symptoms and identified as Magnaporthiopsis poae by using the internal transcribed spacer ITS1 and ITS4 sequences of rDNA. The inhibition rates of the pathogen were investigated for 10 fungicides. As results, the pathogen growth was suppressed when chemicals concentration increased and negatively correlated with incubation period with the chemicals. In triazole group, all chemicals (metconazole, myclobutanil, propiconazole and tebuconazole) treated showed the inhibition rates by 100%. Thiophanate-methyl showed the next highest inhibition effect against a summer patch pathogen. In strobilurin group, pyraclostrobin was the highest suppression effect compared with azoxystrobin and trifloxystrobin. Inhibition effect of fludioxonil and fluxapyroxad on pathogen was similar to the trifloxystrobin. Based on the results, triazole and carboxamide groups are strongly recommended due to the highest inhibition effect on the summer patch pathogen, Magnaporthiopsis poae.

Culture characteristics and genetic relationship of morel mushroom (Morchella spp.) isolates from Korea and other countries (곰보버섯 (Morchella spp.) 수집균주의 배양적특성 및 유전적 유연관계)

  • Min, Gyeong-Jin;Park, Hye-sung;Lee, Eun-ji;Lee, Chan-Jung
    • Journal of Mushroom
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    • v.18 no.1
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    • pp.100-106
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    • 2020
  • Eight morel mushroom species were collected from Korea and other countries. The culture characteristics, genetic relationships, and beta-glucan content of the strains were analyzed. The mycelia of Morchella species exhibited optimal growth when cultured in dark at 25 ℃ in media with pH 7. The mycelia had a distinctive mycelial scent and characteristically changed color, being white initially, and then turning dark yellow to dark brown as it grew. The mycelia were classified into five types based on morphology. The isolates were identified as Morchella conica, two M. sextelata, M. importuna, M. esculenta, and three M. crassipes, based on ITS-rDNA sequences. PCR polymorphisms were variably produced within Morchella spp. using Universal Fungal Fingerprinting Primers (UFPF) and classified into four groups at the intra and inter species level. The strains, KMCC04971 and KMCC04407, showed the same banding pattern as M. conica and M. sextelata, respectively; however, these results were different from those of ITS analysis. Glucan content analysis by strain showed that the KMCC 04973 strain of M. importuna had the highest alpha- and beta-glucan content, at 16.4 g and 33.1 g per 100 g, respectively.

Screening for the 3' UTR Polymorphism of the PXR Gene in South Indian Breast Cancer Patients and its Potential role in Pharmacogenomics

  • Revathidevi, Sundaramoorthy;Sudesh, Ravi;Vaishnavi, Varadharajan;Kaliyanasundaram, Muthukrishnan;MaryHelen, Kilyara George;Sukanya, Ganesan;Munirajan, Arasambattu Kannan
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.3971-3977
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    • 2016
  • Background: Breast cancer, the commonest cancer among women in the world, ranks top in India with an incidence rate of 1,45,000 new cases and mortality rate of 70,000 women every year. Chemotherapy outcome for breast cancer is hampered due to poor response and irreversible dose-dependent cardiotoxicity which is determined by genetic variations in drug metabolizing enzymes and transporters. Pregnane X receptor (PXR), a member of the nuclear receptor superfamily, induces expression of drug metabolizing enzymes (DMEs) and transporters leading to regulation of xenobiotic metabolism. Materials and Methods: A genomic region spanning PXR 3' UTR was amplified and sequenced using genomic DNA isolated from 96 South Indian breast cancer patients. Genetic variants observed in our study subjects were queried in miRSNP to establish SNPs that alter miRNA binding sites in PXR 3' UTR. In addition, enrichment analysis was carried out to understand the network of miRNAs and PXR in drug metabolism using DIANA miRpath and miRwalk pathway prediction tools. Results: In this study, we identified SNPs rs3732359, rs3732360, rs1054190, rs1054191 and rs6438550 in the PXR 3; UTR region. The SNPs rs3732360, rs1054190 and rs1054191 were located in the binding site of miR-500a-3p, miR-532-3p and miR-374a-3p resulting in the altered PXR level due to the deregulation of post-transcriptional control and this leads to poor treatment response and toxicity. Conclusions: Genetic variants identified in PXR 3' UTR and their effects on PXR levels through post-transcriptional regulation provide a genetic basis for interindividual variability in treatment response and toxicity associated with chemotherapy.