• Title/Summary/Keyword: sequencing analysis

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Manufacture of Cutting-Gouda Cheese using Lactobacillus rhamnosus_p1 and the Physicochemical Properties of Gouda Cheese during Ripening Periods (Lactobacillus rhamnosus_p1을 이용한 절단형 고다치즈 제조방법 및 숙성 중 품질특성)

  • Park, Jong-Hyuk;Jung, Hoo-Kil;Moon, Hye-Jung;Oh, Jeon-Hui;Lee, Joo-Hee;Kim, Myung-Kon;Na, Sang-Eon;Kim, Youn-Jeong;Hwang, Young-Tae
    • Journal of Dairy Science and Biotechnology
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    • v.32 no.2
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    • pp.131-139
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    • 2014
  • The aim of this study was to manufacture Cutting-Gouda cheese and to investigate the change in physicochemical properties of Cutting-Gouda cheese made with Lactobacillus rhamnosus_p1. Lactic acid bacteria were isolated from Gouda cheese ripened for more than 1 year. They were identified as 2 strains of L. rhamnosus, Lactobacillus casei, Lactobacillus curvatus, and Staphylococcus saprophyticus by 16S rDNA sequencing and named L. rhamnosus_p1, L. casei_p2, L. curvatus_p3, L. rhamnosus_p4 and S. saprophyticus_p5. The proteolytic activities of isolated strains against casein were measured using prepared skim milk agar plates. L. rhamnosus_p1 showed the highest proteolytic activity. Cutting-Gouda cheese was made with L. rhamnosus_p1, and its physicochemical properties (moisture, protein, fat, ash and free amino acid content) were measured during ripening periods. Because of the modified atmosphere packaging ($N_2{^-}$), there was no change in moisture, protein, fat, and ash in the experimental group. The total amount of free amino acids in the control and experimental group gradually increased during ripening periods. The sensory evaluation showed that the experimental group was preferable to the control group. This result suggests that L. rhamnosus_p1 has potential to be developed as a new starter for Gouda cheese.

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Occurrence and eradication of Plum pox virus on Ornamentals in Korea, 2016-2017 (2016-2017년 국내 핵과류에서의 자두곰보병 발생 및 방제)

  • Kim, Mikyeong;Kim, Gi-Su;Kwak, Hae-Ryun;Kim, Jeong-Eun;Seo, Jang-Kyun;Hong, Seong-Jun;Lee, Gyeong-Jae;Kim, Ju-Hui;Choi, Min-Kyeong;Kim, Byeong-Ryeon;Kim, Ji-Gwang;Han, In-Yeong;Lee, Hyeon-Ju;Won, Heon-Seop;Kang, Hyo-Jung;Han, Jong-Woo;Ko, Suk-Ju;Kim, Hyo-Jeong;Kim, Seung-Han;Lee, Jung-Hywan;Choi, Hong-Soo
    • Research in Plant Disease
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    • v.25 no.1
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    • pp.8-15
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    • 2019
  • Plum pox virus (PPV) is a significant viral disease in Prunus spp. worldwide. A nationwide survey was started in Prunus spp. orchards, since PPV was first detected from peach in Korea, 2015. During 2016-2017, samples were collected from 30,333 trees in 1,985 orchards of stone fruits in 8 provinces and 4 cities, Korea and tested by RT-PCR using specific PPV primer set. As a result, 21 trees including peach (9 trees), Japanese apricot (4 trees), plum (1 tree), apricot (7 trees) in 10 orchards were infected and controlled by eradication program. Amplicons of the expected size (547 bp) were obtained from total RNA of seven peach trees in 2016, and directly sequenced. BLAST analysis revealed the highest nucleotide (NT) identity (99%) with a PPV D isolates (LC331298, LT600782) in Genbank. The seven isolates from shared nt sequence identities of 98 to 100% with one another. Phylogenetic analysis showed the isolates in peach clustered closely with the PPV-D isolates from Korea, Japan, USA, and Canada. This is, to our knowledge, the first report of the presence of PPV in Prunus spp. orchards in Korea, 2016-2017, we hope that our results and efforts will contribute to effective measures for eradication of PPV.

Isolation of Agarivorans sp. JS-1 and Characterization of Its β-Agarase (한천분해세균 Agarivorans sp. JS-1의 분리 및 β-아가라제의 특성 규명)

  • Jin Sun Kim;Dong-Geun Lee;Go-Wun Yeo;Min-Joo Park;Sang-Hyeon Lee
    • Journal of Life Science
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    • v.33 no.4
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    • pp.357-362
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    • 2023
  • This report looks at an agar-degrading marine bacterium and characterization of its agarase. Agar-degrading marine bacterium JS-1 was isolated with Marine agar 2216 media from seawater from the seashore of Sojuk-do, Changwon in Gyeongnam Province, Korea. The agar-degrading bacterium was named as Agarivorans sp. JS-1 by phylogenetic analysis based on 16S rRNA gene sequencing. The extracellular agarase was prepared from the culture media of Agarivorans sp. JS-1 and used for characterization. Relative activities at 20℃, 30℃, 35℃, 40℃, 45℃, 50℃, 55℃, and 60℃ were 70%, 74%, 78%, 83%, 87%, 100%, 74%, and 66%, respectively. Relative activities at pH 5, 6, 7, and 8 were 91%, 100%, 90%, and 89%, respectively. Its extracellular agarase showed maximum activity (207 units/l) at pH 6.0 and 50℃ in 20 mM Tris-HCl buffer. The residual activity after heat treatment at 20℃, 30℃, and 50℃ for 30 minutes was 90%, 70%, and 50% or more, respectively. After a 2-hour heat treatment at 20℃, 30℃, 35℃, 40℃, and 45℃, the residual activity was 80%, 68%, 65%, 63%, and 57%, respectively. At 50℃ and above, after heat treatment for 30 minutes, the residual activity was below 60%. Thin layer chromatography analysis suggested that Agarivorans sp. JS-1 produces extracellular β-agarases as they hydrolyze agarose to produce neoagarooligosaccharides such as neoagarohexaose (20.6%), neoagarotetraose (58.5%), and neoagarobiose (20.9%). Agarivorans sp. JS-1 and its thermotolerant β-agarase would be useful in the production of neoagarooligosaccharides, showing functional activity such as inhibition of bacterial growth and delay of starch degradation.

Association Between the Polymorphism in the Promoter Region of Porcine A-FABP Gene and Growth Traits in Duroc Pigs (돼지 Duroc 품종에서 A-FABP promoter의 다형성과 성장형질의 연관성)

  • Han, Sang-Hyeon;Jo, In-Cheol;Lee, Jong-Eon;Kim, Hyo-Seon;Lee, Jeong-Gyu;Jeon, Jin-Tae;O, Mun-Yu;Go, Mun-Seok
    • Journal of Animal Science and Technology
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    • v.48 no.1
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    • pp.21-28
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    • 2006
  • A polymorphism was found in the promoter region of porcine adipocyte fatty acid binding protein gene(A-FABP) gene which plays a key role in the binding and transportation of free fatty acid in adipocyte and deposition of intramuscular fat. Mutation was detected a substitution(T406C) using SSCP analysis and subsequently confirmed by sequencing the fragment in Duroc pigs. This T-406C mutation might change the binding activity for transcription factor nuclear factor 1(NF1). In this population, this mutation was genotyped using HinfⅠRFLP, and found three kinds of genotypes(TT, TC, and CC) showing their frequencies of 42.3, 44.3, and 13.4%, respectively. We statistically analyzed the association between the A-FABP genotypes and growth traits and found that the body weights of the pigs containing 406C/(TC or CC) were heavier for the body weight at the age of 20 weeks than those containing genotype TT(P<0.05), but not for those at the age of 0, 3, and 10 weeks. Pigs containing genotype CC had also a higher value for the average daily gain and lower values for the date for 90kg of body weight and food conversion ratio than those of 406T/- genotype. In addition, without the significant difference of back fat thickness, there was a significant association between the existence of allele CC and lean meat and eye muscle area(P<0.05). As a result of this study, we suggest that the allele T406C in the promoter region of A-FABP gene play an important role in deposition of intramuscular fat and weight in the later growth period. This polymorphism will be an useful molecular marker for breeding of Duroc pigs.

Microbial profile of asymptomatic and symptomatic teeth with primary endodontic infections by pyrosequencing (원발성 치근단 치주염을 갖는 감염근관에서 증상유무에 따른 세균분포의 pyrosequencing 분석)

  • Lim, Sang-Min;Lee, Tae-Kwon;Kim, Eun-Jeong;Park, Jun-Hong;Lee, Yoon;Bae, Kwang-Shik;Kum, Kee-Yeon
    • Restorative Dentistry and Endodontics
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    • v.36 no.6
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    • pp.498-505
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    • 2011
  • Objectives: The purpose of this in vivo study was to investigate the microbial diversity in symptomatic and asymptomatic canals with primary endodontic infections by using GS FLX Titanium pyrosequencing. Materials and Methods: Sequencing was performed on 6 teeth (symptomatic, n = 3; asymptomatic, n = 3) with primary endodontic infections. Amplicons from hypervariable region of the small-subunit ribosomal RNA gene were generated by polymerized chain reaction (PCR), and sequenced by means of the GS FLX Titanium pyrosequencing. Results: On average, 10,639 and 45,455 16S rRNA sequences for asymptomatic and symptomatic teeth were obtained, respectively. Based on Ribosomal Database Project Classifier analysis, pyrosequencing identified the 141 bacterial genera in 13 phyla. The vast majority of sequences belonged to one of the seven phyla: Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, Proteobacteria, Spirochetes, and Synergistetes. In genus level, Pyramidobacter, Streptococcus, and Leptotrichia constituted about 50% of microbial profile in asymptomatic teeth, whereas Neisseria, Propionibacterium, and Tessaracoccus were frequently found in symptomatic teeth (69%). Grouping the sequences in operational taxonomic units (3%) yielded 450 and 1,997 species level phylotypes in asymptomatic and symptomatic teeth, respectively. The total bacteria counts were significantly higher in symptomatic teeth than that of asymptomatic teeth (p < 0.05). Conclusions: GS FLX Titanium pyrosequencing could reveal a previously unidentified high bacterial diversity in primary endodontic infections.

Antifungal Activity of Bacillus sp. AM-651 Against Phytophthora capsici (고추역병 유발병원균 Phytophthora capsici에 대한 Bacillus sp. AM-651의 항진균활성)

  • Lee, Jung-Bok;Shin, Jeong-Hak;Jang, Jong-Ok;Shin, Kee-Sun;Choi, Chung-Sik;Kim, Kun-Woo;Jo, Min-Sub;Jeon, Chun-Pyo;Kim, Yun-Hoi;Kwon, Gi-Seok
    • Microbiology and Biotechnology Letters
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    • v.36 no.3
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    • pp.227-232
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    • 2008
  • Biological antagonists of Phytophthora capsici were isolated from soil in Gyeongbuk, Korea. Among the isolated bacteria, a Bacillus sp. was identified from l6S rDNA sequence analysis and named Bacillus sp. AM-651. Bacillus sp. AM-65l strain which can strongly a antifungal activity against Phytophthora capsici. Culture conditions for the maximum production of the antagonistic substance were optimized. The production of antibiotic were high on modified Davis mineral medium pH 7 at $30^{\circ}C$. The medium for highest production of the agonistic substance optimized. It is composed the best activity on glucose, $(NH_4)_2SO_4$ and $K_2HPO_4$ at 0.5%, 0.1%, and 0.7%, respectively. By time course of culture solution selected Bacillus sp. AM-65l, the culture solution after 48hrs had strongly growth inhibition rate against P. capsici. And culture solution of Bacillus sp. AM-651 was stable within a pH range $5{\sim}11$ and temperature range $4{\sim}70^{\circ}C$. Bacillus sp. AM-651 cultured broth shown fungal growth inhibitory activity against B. sorokiniana, B. cinerea, R. solani avove and beyond P. capsici and comparatively showed a high activity against C. gloeosporioides, B. dothidea, B. cinerea and F. graminearum by agar diffusion method.

Selection of Salt-Tolerant Silage Rice Through in vitro Screening and Saltol QTL Analysis (기내 선발과 Saltol QTL 분석을 통한 내염성 증진 사료용 벼 선발)

  • Cho, Chuloh;Kim, Kyung Hwa;Ahn, Eok-Keun;Park, Hyangmi;Choi, Man-Soo;Chun, Jaebuhm;Seo, Mi-Suk;Jin, Mina;Kim, Dool-Yi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.214-221
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    • 2020
  • Salinity is one of the major abiotic stressors that inhibits the growth, yield, and productivity of crop plants. Therefore, it is necessary to develop crops with increased salt tolerance for cultivation in saline soils such as is found in reclaimed land. The objective of this study was to develop a salt-tolerant silage rice line that grows on reclaimed land. In order to develop this salt-tolerant silage rice, we transferred Saltol, a major QTL associated with salt tolerance, from IR64-Saltol, a salt-tolerant indica variety, into Mogyang, a susceptible elite japonica variety. To determine the effect of salt stress, Mogyang and IR64-Saltol cultivars were grown on a medium containing various concentrations of NaCl in in vitro conditions. Shoot length was found to decrease with increasing salt concentrations, and root growth was almost arrested at NaCl concentrations over 50 mM in the Mogyang cultivar. Based on these preliminary results, we screened five salt-tolerant lines showing superior growth under salt stress conditions. Polymerase chain reaction and sequencing results showed that the introgression types of Saltol QTL were derived from the IR64-Saltol cultivar in almost all selected lines. Based on the observed growth and physiological characteristics, the new Saltol introgression lines showed higher salt tolerance compared to the Mogyang parental cultivar. The salt-tolerant lines identified in this study could be used as a genetic resource to improve rice salt tolerance.

Development and Characterization of EMS-induced Mutants with Enhanced Salt Tolerance in Silage Maize (EMS 유도 내염성 증진 사료용 옥수수 돌연변이체 선발 및 특성 분석)

  • Cho, Chuloh;Kim, Kyung Hwa;Seo, Mi-Suk;Choi, Man-Soo;Chun, Jaebuhm;Jin, Mina;Kim, Dool-Yi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.4
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    • pp.406-415
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    • 2020
  • Maize (Zea mays L.) is one of the most valuable agricultural crops and is grown under a wide spectrum of environmental conditions. However, maize is moderately sensitive to salt stress, and soil salinity is a serious threat to its production worldwide. In this study, we used ethyl methane sulfonate (EMS) to generate salt-tolerant silage maize mutants. We screened salt-tolerant lines from 203 M3 mutant populations by evaluating the morphological phenotype after salt stress treatment and selected the 140ES91 line. The 140ES91 mutant showed improved plant growth as well as higher proline content and leaf photosynthetic capacity compared with those of wild-type plants under salt stress conditions. Using whole-genome re-sequencing analysis, 1,103 single nucleotide polymorphisms and 71 insertions or deletions were identified as common variants between KS140 and 140ES91 in comparison with the reference genome B73. Furthermore, the expression patterns of three genes, which are involved in salt stress responses, were increased in the 140ES91 mutant under salt stress. Taken together, the mutant line identified in our study could be used as an improved breeding material for transferring salt tolerance traits in maize varieties.

Physiological Characteristics of Melon Plants Showing Leaf Yellowing Symptoms Caused by CABYV Infection (CABYV 감염 멜론의 황화증상에 따른 생리적인 특성)

  • Lee, Hee Ju;Kim, Mi-Kyeong;Lee, Sang Gyu;Choi, Chang Sun;Choi, Hong-Soo;Kwak, Hae Ryun;Choi, Gug Seoun;Chun, Changhoo
    • Horticultural Science & Technology
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    • v.33 no.2
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    • pp.210-218
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    • 2015
  • Melon leaves showing yellowing symptoms were analyzed using electron microscopy and RT-PCR for major cucurbit-infecting-viruses (CMV, MNSV, CGMMV, SqMV, WMV, KGMMV, PRSV and ZYMV) reported in Korea, but these viruses were not detected. As the result of further analysis by next-generation sequencing (NGS), the virus was identified as Cucurbit aphid-borne yellows virus (CABYV), and then confirmed by RT-PCR using CABYV-specific primers. When photosynthetic capacity was measured based on chlorophyll fluorescence yield (ChlFY), the leaves of the diseased plants showed $4.09{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, which was one-third of the readings observed for unaffected normal plants ($12.36{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$). The root functions of plants affected by leaf yellowing symptoms (LYS) was $0.28mg{\cdot}g^{-1}$, about half that measured for the normal unaffected plants ($0.48mg{\cdot}g^{-1}$). Cytological observations revealed that there were no morphological differences in the palisade parenchyma and mesophyll spongy cells of the leaves between the diseased and the normal plants. However, the same leaf cells of the affected plants contained more starch granules compared to those of the normal, unaffected plants. We conclude that the LYS of muskmelon is not merely a physiological disorder but a viral disease caused by CABYV and spread by aphids.

Breeding of the Scab-Resistant Pear Cultivar 'Greensis' (배 검은별무늬병 저항성 품종 '그린시스' 육성)

  • Kim, Yoon-Kyeong;Kang, Sam-Seok;Won, Kyung-Ho;Shin, Il-Sheob;Cho, Kwang-Sik;Ma, Kyeong-Bok;Kim, Myung-Su;Choi, Jang-Jeon;Choi, Jin-Ho
    • Horticultural Science & Technology
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    • v.34 no.4
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    • pp.655-661
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    • 2016
  • To develop scab-resistant pear (Pyrus spp.) varieties with fruits that are as crisp and juicy as Asian pears, a cross was made between 'Whangkeumbae' and 'Bartlett' varieties (P. pyrifolia ${\times}$ P. communis) at the Pear Research Institute of the National Institute of Horticultural & Herbal Science, Rural Development Administration, in 1994. Among the 285 seedlings, 'Greensis' was first selected in 2006 for its good eating quality and named in 2012 after regional adaptation tests in nine regions and ten experimental plots from 2007 to 2012. The tree showed a vigorous growth habit and semi-spreading characteristics, like 'Whangkeumbae'. The optimum fruit harvest date was also around Sept. 26 and fruit was round in shape and green in skin color at maturity. Average fruit weight was 470g, and the soluble solids content was $12.4^{\circ}Brix$. The flesh was very crisp and juicy, and had good eating quality. Its' leaf size was similar with 'Bartlett' and smaller than 'Whangkeumbae'. The average of full bloom date of 'Greensis' was determined as Apr. 26, which was six days later than 'Whangkeumbae' and similar with 'Bartlett'. S genotypes of 'Greensis' were identified as $S_4S_e$ by S-allele PCR product sequencing analysis. It seems that the $S_4$ allele was inherited from 'Whangkeumbae' and the Se allele from 'Bartlett'. 'Greensis' displayed strong resistance to scab disease caused by Venturia nashicola, similar to European pear cultivars like 'Beurre Hardy' and, 'Conference'. 'Greensis' was also highly resistant to black leaf spot (Alternaria kikuchiana) in the field