• Title/Summary/Keyword: Whole-genome sequence

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Flanking Sequence and Copy-Number Analysis of Transformation Events by Integrating Next-Generation Sequencing Technology with Southern Blot Hybridization

  • Qin, Yang;Woo, Hee-Jong;Shin, Kong-Sik;Lim, Myung-Ho;Cho, Hyun-Suk;Lee, Seong-Kon
    • Plant Breeding and Biotechnology
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    • v.5 no.4
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    • pp.269-281
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    • 2017
  • With the continual development of genetically modified (GM) crops, it has become necessary to develop detailed and effective molecular characterization methods to select candidate events from a large pool of transformation events. Relative to traditional molecular analysis methods such as the polymerase chain reaction (PCR) and Southern blot hybridization, next generation sequencing (NGS) technology for whole-genome sequencing of complex crop genomes had proven comparatively useful for in-depth molecular characterization. In this study, four transformation events, including one in Bacillus thuringiensis (Bt)-resistant rice, one in resveratrol-producing rice, and two in beta-carotene-enhanced soybeans, were selected for molecular characterization. To merge NGS analysis and Southern blot-hybridization results, we confirmed the transgene insertion sites, insertion construction, and insertion numbers of these four transformation events. In addition, the read-coverage depth assessed by NGS analysis for inserted genes might provide consistent results in terms of inserted T-DNA numbers in case of complex insertion structures and highly duplicated donor genomes; however, PCR-based methods can produce incorrect conclusions. Our combined method provides an effective and complete analytical approach for whole-genome visual inspection of transformation events that require biosafety assessment.

Application of genotyping-by-sequencing (GBS) in plant genome using bioinformatics pipeline

  • Lee, Yun Gyeong;Kang, Chon-Sik;Kim, Changsoo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.58-58
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    • 2017
  • The advent of next generation sequencing technology has elicited plenty of sequencing data available in agriculturally relevant plant species. For most crop species, it is too expensive to obtain the whole genome sequence data with sufficient coverage. Thus, many approaches have been developed to bring down the cost of NGS. Genotyping-by-sequencing (GBS) is a cost-effective genotyping method for complex genetic populations. GBS can be used for the analysis of genomic selection (GS), genome-wide association study (GWAS) and constructing haplotype and genetic linkage maps in a variety of plant species. For efficiently dealing with plant GBS data, the TASSEL-GBS pipeline is one of the most popular choices for many researchers. TASSEL-GBS is JAVA based a software package to obtain genotyping data from raw GBS sequences. Here, we describe application of GBS and bioinformatics pipeline of TASSEL-GBS for analyzing plant genetics data.

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Complete Genome Sequence of Salmonella Typhimurium-Specific Phage vB_SalA_KFSST3 Possessing Antibiofilm Activity

  • Su-Hyeon Kim;Jaein Choe;Mi-Kyung Park
    • Microbiology and Biotechnology Letters
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    • v.52 no.3
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    • pp.339-341
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    • 2024
  • In a previous study, Salmonella Typhimurium-specific phage vB_SalA_KFSST3, which possess antibiofilm activity, was isolated and purified from wastewater used in slaughterhouses. This study aimed to perform bioinformatic analyses to investigate the genes associated with its antibiofilm activity. Phage genome consisted of a single chromosome of 156,555 bp with a GC content of 44.8%. Among its 202 open reading frames (ORFs), three tail spike proteins (TSPs; orf141, orf142, orf143) were identified with high confidence. All TSPs were predicted to encode putative depolymerase activities, including two endoglycosidases and one endorhamnosidase. The genome has been deposited in GenBank under the accession number PP_994976.1.

Genome-based identification of strain KCOM 1265 isolated from subgingival plaque at the species level

  • Park, Soon-Nang;Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.45 no.2
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    • pp.70-75
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    • 2020
  • The aim of this study was to identify strain KCOM 1265 isolated from subgingival plaque at the species level by comparing 16S ribosomal RNA gene (16S rDNA) and genome sequences. The whole genome of strain KCOM 1265 was extracted using the phenol-chloroform extraction method. 16S rDNA was amplified using polymerase chain reaction and sequenced using the dideoxy chain termination method. Pairwise genome comparison was performed using average nucleotide identity (ANI) and genome-to-genome distance (GGD) analyses. The data showed that the percent similarity of 16S rDNA sequence of strain KCOM 1265 was 99.6% as compared with those of Fusobacterium polymorphum ATCC 10953T and Fusobacterium hwasookii KCOM 1249T. The ANI values of strain KCOM 1265 with F. polymorphum ATCC 10953T and F. hwasookii KCOM 1249T were 95.8% and 93.0%, respectively. The GGD values of strain KCOM 1265 with F. polymorphum ATCC 10953T and F. hwasookii KCOM 1249T were 63.9% and 49.6%, respectively. These results indicate that strain KCOM 1265 belongs to F. polymorphum.

Genome-Based Insights into the Thermotolerant Adaptations of Neobacillus endophyticus BRMEA1T

  • Lingmin Jiang;Ho Le Han;Yuxin Peng;Doeun Jeon;Donghyun Cho;Cha Young Kim;Jiyoung Lee
    • Research in Plant Disease
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    • v.29 no.3
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    • pp.321-329
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    • 2023
  • The bacterium Neobacillus endophyticus BRMEA1T, isolated from the medicinal plant Selaginella involvens, known as its thermotolerant can grow at 50℃. To explore the genetic basis for its heat tolerance response and its potential for producing valuable natural compounds, the genomes of two thermotolerant and four mesophilic strains in the genus Neobacillus were analyzed using a bioinformatic software platform. The whole genome was annotated using RAST SEED and OrthVenn2, with a focus on identifying potential heattolerance-related genes. N. endophyticus BRMEA1T was found to possess more stress response genes compared to other mesophilic members of the genus, and it was the only strain that had genes for the synthesis of osmoregulated periplasmic glucans. This study sheds light on the potential value of N. endophyticus BRMEA1T, as it reveals the mechanism of heat resistance and the application of secondary metabolites produced by this bacterium through whole-genome sequencing and comparative analysis.

Assessment of Erythrobacter Species Diversity through Pan-Genome Analysis with Newly Isolated Erythrobacter sp. 3-20A1M

  • Cho, Sang-Hyeok;Jeong, Yujin;Lee, Eunju;Ko, So-Ra;Ahn, Chi-Yong;Oh, Hee-Mock;Cho, Byung-Kwan;Cho, Suhyung
    • Journal of Microbiology and Biotechnology
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    • v.31 no.4
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    • pp.601-609
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    • 2021
  • Erythrobacter species are extensively studied marine bacteria that produce various carotenoids. Due to their photoheterotrophic ability, it has been suggested that they play a crucial role in marine ecosystems. It is essential to identify the genome sequence and the genes of the species to predict their role in the marine ecosystem. In this study, we report the complete genome sequence of the marine bacterium Erythrobacter sp. 3-20A1M. The genome size was 3.1 Mbp and its GC content was 64.8%. In total, 2998 genetic features were annotated, of which 2882 were annotated as functional coding genes. Using the genetic information of Erythrobacter sp. 3-20A1M, we performed pan-genome analysis with other Erythrobacter species. This revealed highly conserved secondary metabolite biosynthesis-related COG functions across Erythrobacter species. Through subsequent secondary metabolite biosynthetic gene cluster prediction and KEGG analysis, the carotenoid biosynthetic pathway was proven conserved in all Erythrobacter species, except for the spheroidene and spirilloxanthin pathways, which are only found in photosynthetic Erythrobacter species. The presence of virulence genes, especially the plant-algae cell wall degrading genes, revealed that Erythrobacter sp. 3-20A1M is a potential marine plant-algae scavenger.

Evaluation and Genome Mining of Bacillus stercoris Isolate B.PNR1 as Potential Agent for Fusarium Wilt Control and Growth Promotion of Tomato

  • Rattana Pengproh;Thanwanit Thanyasiriwat;Kusavadee Sangdee;Juthaporn Saengprajak;Praphat Kawicha;Aphidech Sangdee
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.430-448
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    • 2023
  • Recently, strategies for controlling Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of Fusarium wilt of tomato, focus on using effective biocontrol agents. In this study, an analysis of the biocontrol and plant growth promoting (PGP) attributes of 11 isolates of loamy soil Bacillus spp. has been conducted. Among them, the isolates B.PNR1 and B.PNR2 inhibited the mycelial growth of Fol by inducing abnormal fungal cell wall structures and cell wall collapse. Moreover, broad-spectrum activity against four other plant pathogenic fungi, F. oxysporum f. sp. cubense race 1 (Foc), Sclerotium rolfsii, Colletotrichum musae, and C. gloeosporioides were noted for these isolates. These two Bacillus isolates produced indole acetic acid, phosphate solubilization enzymes, and amylolytic and cellulolytic enzymes. In the pot experiment, the culture filtrate from B.PNR1 showed greater inhibition of the fungal pathogens and significantly promoted the growth of tomato plants more than those of the other treatments. Isolate B.PNR1, the best biocontrol and PGP, was identified as Bacillus stercoris by its 16S rRNA gene sequence and whole genome sequencing analysis (WGS). The WGS, through genome mining, confirmed that the B.PNR1 genome contained genes/gene cluster of a nonribosomal peptide synthetase/polyketide synthase, such as fengycin, surfactin, bacillaene, subtilosin A, bacilysin, and bacillibactin, which are involved in antagonistic and PGP activities. Therefore, our finding demonstrates the effectiveness of B. stercoris strain B.PNR1 as an antagonist and for plant growth promotion, highlighting the use of this microorganism as a biocontrol agent against the Fusarium wilt pathogen and PGP abilities in tomatoes.

Genomic Tools and Their Implications for Vegetable Breeding

  • Phan, Ngan Thi;Sim, Sung-Chur
    • Horticultural Science & Technology
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    • v.35 no.2
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    • pp.149-164
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    • 2017
  • Next generation sequencing (NGS) technologies have led to the rapid accumulation of genome sequences through whole-genome sequencing and re-sequencing of crop species. Genomic resources provide the opportunity for a new revolution in plant breeding by facilitating the dissection of complex traits. Among vegetable crops, reference genomes have been sequenced and assembled for several species in the Solanaceae and Cucurbitaceae families, including tomato, pepper, cucumber, watermelon, and melon. These reference genomes have been leveraged for re-sequencing of diverse germplasm collections to explore genome-wide sequence variations, especially single nucleotide polymorphisms (SNPs). The use of genome-wide SNPs and high-throughput genotyping methods has led to the development of new strategies for dissecting complex quantitative traits, such as genome-wide association study (GWAS). In addition, the use of multi-parent populations, including nested association mapping (NAM) and multiparent advanced generation intercross (MAGIC) populations, has helped increase the accuracy of quantitative trait loci (QTL) detection. Consequently, a number of QTL have been discovered for agronomically important traits, such as disease resistance and fruit traits, with high mapping resolution. The molecular markers for these QTL represent a useful resource for enhancing selection efficiency via marker-assisted selection (MAS) in vegetable breeding programs. In this review, we discuss current genomic resources and marker-trait association analysis to facilitate genome-assisted breeding in vegetable species in the Solanaceae and Cucurbitaceae families.

Complete Mitochondrial Genome and Phylogenetic Analysis for the Korean Field Mouse Apodemus peninsulae Found on Baengnyeong Island in South Korea

  • Jung A Kim;Hye Sook Jeon;Seung Min Lee;Hong Seomun;Junghwa An
    • Proceedings of the National Institute of Ecology of the Republic of Korea
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    • v.4 no.2
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    • pp.69-71
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    • 2023
  • The Korean field mouse, Apodemus peninsulae mitochondrial genome has previously been reported for mice obtained from mainland Korea and China. In this investigation the complete mitochondrial genome sequence for a mouse obtained from Baengnyeong Island (BI) in South Korea was determined using high-throughput whole-genome sequencing for the first time. The circular genome was determined to be 16,268 bp in length. It was found to be composed of a typical complement gene that encodes 13 protein subunits of enzymes involved in oxidative phosphorylation, two ribosomal RNAs, 22 transfer RNAs, and one control region. Phylogenetic analysis involved 13 amino acid sequences and demonstrated that the A. peninsulae genome from BI was more closely grouped with two Korean samples (HQ660074 and JN546584) than the Chinese (KP671850) sample. This study verified the evolutionary status of A. peninsulae inhabiting the BI at the molecular level, and could be a significant supplement to the genetic background.

Draft Genome Sequence of Alternaria alternata JS-1623, a Fungal Endophyte of Abies koreana

  • Park, Sook-Young;Jeon, Jongbum;Kim, Jung A.;Jeon, Mi Jin;Jeong, Min-Hye;Kim, Youngmin;Lee, Yerim;Chung, Hyunjung;Lee, Yong-Hwan;Kim, Soonok
    • Mycobiology
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    • v.48 no.3
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    • pp.240-244
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    • 2020
  • Alternaria alternata JS-1623 is an endophytic fungus isolated from a stem tissue of Korean fir, Abies koreana. Ethyl acetate extracts of culture filtrates exhibited anti-inflammatory activity in LPS induced microglia BV-2 cell without cytotoxicity. Here we report a 33.67 Mb sized genome assembly of JS-1623 comprised of 13 scaffolds with N50 of 4.96 Mb, and 92.41% of BUSCO completeness. GC contents were 50.97%. Of the 11,197 genes annotated, gene families related to the biosynthesis of secondary metabolites or transcription factors were identified.