• Title/Summary/Keyword: RNA sequencing analysis

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Molecular Characterization of an Isolate of Bean Common Mosaic Virus First Identified in Gardenia Using Metatranscriptome and Small RNA Sequencing

  • Zhong-Tian Xu;Hai-Tao Weng;Jian-Ping Chen;Chuan-Xi Zhang;Jun-Min Li;Yi-Yuan Li
    • The Plant Pathology Journal
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    • v.40 no.1
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    • pp.73-82
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    • 2024
  • Gardenia (Gardenia jasminoides) is a popular and economically vital plant known for its ornamental and medicinal properties. Despite its widespread cultivation, there has been no documentation of plant viruses on gardenia yet. In the present study, gardenia leaves exhibiting symptoms of plant viral diseases were sampled and sequenced by both metatranscriptome and small RNA sequencing. As a consequence, bean common mosaic virus (BCMV) was identified in gardenia for the first time and named BCMV-gardenia. The full genome sequence of BCMV-gardenia is 10,054 nucleotides (nt) in length (excluding the poly (A) at the 3' termini), encoding a large polyprotein of 3,222 amino acids. Sequence analysis showed that the N-termini of the polyprotein encoded by BCMV-gardenia is less conserved when compared to other BCMV isolates, whereas the C-termini is the most conserved. Maximum likelihood phylogenetic analysis showed that BCMVgardenia was clustered closely with other BCMV isolates identified outside the leguminous plants. Our results indicated that the majority of BCMV-gardenia virus-derived small interfering RNAs (vsiRNAs) were 21 nt and 22 nt, with 21 nt being more abundant. The first nucleotide at the 5' termini of vsiRNAs derived from BCMV-gardenia preferred U and A. The ratio of vsiRNAs derived from sense (51.1%) and antisense (48.9%) strands is approaching, and the distribution of vsiRNAs along the viral genome is generally even, with some hot spots forming in local regions. Our findings could provide new insights into the diversity, evolution, and host expansion of BCMV and contribute to the prevention and treatment of this virus.

Comparison of ITS(Internal Transcribed Spacer) and 5.8S rDNA Sequences among varieties and Cultivars in Panax ginseng

  • Yang, Deok-Chun;Yang, Key-Jin;Yoon, Eui-Soo
    • Journal of Photoscience
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    • v.8 no.2
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    • pp.55-60
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    • 2001
  • Ginseng (Panax genus) is one of the most medicinally important genera and consists of highly regarded medicines. Among the species of Panax, the ginseng species is widely known to have most medicinal quality. P. ginseng has 3 varieties, Jakyung, Chunggyung and Hwangsook, discovered in nature with different colors of stem and fruit, Jakyung has two cultivars, Yunpoong and Chunpoong. Rigorous phylogenetic analysis of these varieties and cultivars has been conducted with sequencing of rDNA region. The sequences of ITS1, ITS2 of every varieties and cultivars within P. ginseng were identical. The sequence of 5.8S rDNAs of Hwangsook variety were different from the sequences of 5.8S rDNAs of others by only one base pair at nucleotide position 14. In phylogenetic analysis and predicted RNA secondary structure study, it is assumed that evolution has proceeded from Hwangsook to other varieties. recently.

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Examination of the xanthosine response on gene expression of mammary epithelial cells using RNA-seq technology

  • Choudhary, Shanti;Li, Wenli;Bickhart, Derek;Verma, Ramneek;Sethi, R.S.;Mukhopadhyay, C.S.;Choudhary, Ratan K.
    • Journal of Animal Science and Technology
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    • v.60 no.7
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    • pp.18.1-18.12
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    • 2018
  • Background: Xanthosine treatment has been previously reported to increase mammary stem cell population and milk production in cattle and goats. However, the underlying molecular mechanisms associated with the increase in stem cell population and milk production remain unclear. Methods: Primiparous Beetal goats were assigned to the study. Five days post-partum, one mammary gland of each goat was infused with xanthosine (TRT) twice daily ($2{\times}$) for 3 days consecutively, and the other gland served as a control (CON). Milk samples from the TRT and CON glands were collected on the 10th day after the last xanthosine infusion and the total RNA was isolated from milk fat globules (MEGs). Total RNA in MFGs was mainly derived from the milk epithelial cells (MECs) as evidenced by expression of milk synthesis genes. Significant differentially expressed genes (DEGs) were subjected to Gene Ontology (GO) terms using PANTHER and gene networks were generated using STRING db. Results: Preliminary analysis indicated that each individual goat responded to xanthosine treatment differently, with this trend being correlated with specific DEGs within the same animal's mammary gland. Several pathways are impacted by these DEGs, including cell communication, cell proliferation and anti-microbials. Conclusions: This study provides valuable insights into transcriptomic changes in milk producing epithelial cells in response to xanthosine treatment. Further characterization of DEGs identified in this study is likely to delineate the molecular mechanisms of increased milk production and stem or progenitor cell population by the xanthosine treatment.

RNA-Seq Transcriptome Analysis of the Cutlass Fish Reveals Photoreceptors Gene Expression in Peripheral Tissues (RNA-Seq transcriptome 분석을 통한 갈치 광수용체 유전자 탐색 및 mRNA 조직발현)

  • Hyeon, Ji-Yeon;Kim, Mun-Kwan;Lim, Bong-Soo;Byun, Jun-Hwan;Moon, Ji-Sung;Kang, Hyeong-Cheol;Hur, Sung-Pyo;Oh, Seong-Rip
    • Ocean and Polar Research
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    • v.39 no.2
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    • pp.149-158
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    • 2017
  • The opsin family of light sensitive proteins family makes up are the universal photoreceptor molecules of all visual systems in the vertebrates including teleosts. They can change their conformation from a resting state to a signaling state upon light absorption, which activates the G-protein coupled receptor, thereby resulting in a signaling cascade that produces physiological responses. However, this species is poorly characterized at molecular level due to little sequence information available in public databases. We have investigated the opsin family of nocturnal cutlass fish using the whole transcriptome sequencing method. The opsin genes were cloned and its expression in the tissues and organs were examined by qPCR. We cloned 6 opsin genes (RRH, Opn4, Rh1, Rh2, VA-opsin, and Opn3) in retina and brain tissue. It contained the seven presumed transmembrane domains that are characteristic of the G-protein-coupled receptor family. However, short wavelength sensitive pigment (SWS) and long wavelength sensitive pigment (LWS) were not detected in this study. The mRNA expression of the 6 photoreceptor genes were detected in retina and peripheral tissue. Our studies will lead to further investigation of the photic entrainment mechanism at molecular and cellular levels in cutlass fish and can be used in comparative studies of other fishes.

Profiling of glucose-induced transcription in Sulfolobus acidocaldarius DSM 639

  • Park, Jungwook;Lee, Areum;Lee, Hyun-Hee;Park, Inmyoung;Seo, Young-Su;Cha, Jaeho
    • Genes and Genomics
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    • v.40 no.11
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    • pp.1157-1167
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    • 2018
  • Sulfolobus species can grow on a variety of organic compounds as carbon and energy sources. These species degrade glucose to pyruvate by the modified branched Entner-Doudoroff pathway. We attempted to determine the differentially expressed genes (DEGs) under sugar-limited and sugar-rich conditions. RNA sequencing (RNA-seq) was used to quantify the expression of the genes and identify those DEGs between the S. acidocaldarius cells grown under sugar-rich (YT with glucose) and sugar-limited (YT only) conditions. The functions and pathways of the DEGs were examined using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Quantitative real-time PCR (qRT-PCR) was performed to validate the DEGs. Transcriptome analysis of the DSM 639 strain grown on sugar-limited and sugar-rich media revealed that 853 genes were differentially expressed, among which 481 were upregulated and 372 were downregulated under the glucose-supplemented condition. In particular, 70 genes showed significant changes in expression levels of ${\geq}$ twofold. GO and KEGG enrichment analyses revealed that the genes encoding components of central carbon metabolism, the respiratory chain, and protein and amino acid biosynthetic machinery were upregulated under the glucose condition. RNA-seq and qRT-PCR analyses indicated that the sulfur assimilation genes (Saci_2197-2204) including phosphoadenosine phosphosulfate reductase and sulfite reductase were significantly upregulated in the presence of glucose. The present study revealed metabolic networks in S. acidocaldarius that are induced in a glucose-dependent manner, improving our understanding of biomass production under sugar-rich conditions.

Therapeutic effects of paeoniflorin on irritable bowel syndrome in rats

  • Lei Wang;Jinyan Lei;Zeyu Zhao;Jianwei Jia;Li Wang
    • Journal of Veterinary Science
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    • v.24 no.3
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    • pp.23.1-23.16
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    • 2023
  • Background: Irritable bowel syndrome (IBS) is a functional bowel disorder (FBD). Objectives: To assess the therapeutic effects of paeoniflorin (PF) on IBS in rats. Method: Sixty male Sprague-Dawley rats were randomly divided into normal, model, positive drug, low-dose PF, medium-dose PF and high-dose PF groups (n = 10). After gavage for 2 consecutive weeks, the effect of PF on abdominal pain symptoms was assessed based on the abdominal withdrawal reflex (AWR) score, fecal water content and pathological changes in colon tissues. D-lactate, interleukin-1β (IL-1β), transforming growth factor-β (TGF-β) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay, and phosphorylated nuclear factor kappa B (p-NF-κB) p65 was detected by Western blotting. The abundance and diversity changes of intestinal flora were explored using 16S ribosomal RNA sequencing. Result: In PF groups, the mucosal morphology of colon tissues was intact, and the glands were arranged neatly and structured clearly, without obvious inflammatory cell infiltration. Compared with the model group, PF groups had significantly elevated pain threshold, and mRNA and protein levels of zonula occludens-1 (ZO-1) and occludin, decreased AWR score at 20 mmHg pressure, fecal water content, mRNA levels of IL-1β, TGF-β, and TNF-α, protein level of p-NF-κB p65 and level of serum D-lactate, and reduced levels of serum IL-1β, TGF-β, and TNF-α (p < 0.05, p < 0.01). PF groups had higher abundance of Lactobacillus, Akkermansia, Alistipes, and Bacteroides, but lower abundance of Desulfovibrio, Parasutterella, and Enterococcus than those of the model group. Conclusions: PF exerts therapeutic effects on IBS in rats probably by regulating the intestinal flora, and then up-regulating the expressions of ZO-1 and occludin in colon tissue while down-regulating the levels of IL-1β, TGF-β, TNF-α, D-lactate and p-NF-κB p65.

Generation of ints14 Knockout Zebrafish using CRISPR/Cas9 for the Study of Development and Disease Mechanisms

  • Ji Hye Jung;Sanghoon Jeon;Heabin Kim;Seung-Hyun Jung
    • Development and Reproduction
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    • v.27 no.4
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    • pp.205-211
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    • 2023
  • INTS14/VWA9, a component of the integrator complex subunits, plays a pivotal role in regulating the fate of numerous nascent RNAs transcribed by RNA polymerase II, particularly in the biogenesis of small nuclear RNAs and enhancer RNAs. Despite its significance, a comprehensive mutation model for developmental research has been lacking. To address this gap, we aimed to investigate the expression patterns of INTS14 during zebrafish embryonic development. We generated ints14 mutant strains using the CRISPR/Cas9 system. We validated the gRNA activity by co-injecting Cas9 protein and a single guide RNA into fertilized zebrafish eggs, subsequently confirming the presence of a 6- or 9-bp deletion in the ints14 gene. In addition, we examined the two mutant alleles through PCR analysis, T7E1 assay, TA-cloning, and sequencing. For the first time, we used the CRISPR/Cas9 system to create a model in which some sequences of the ints14 gene were removed. This breakthrough opens new avenues for in-depth exploration of the role of ints14 in animal diseases. The mutant strains generated in this study can provide a valuable resource for further investigations into the specific consequences of ints14 gene deletion during zebrafish development. This research establishes a foundation for future studies exploring the molecular mechanisms underlying the functions of ints14, its interactions with other genes or proteins, and its broader implications for biological processes.

Identification of Equine Heat Shock Proteins Gene and Their mRNA Expression Analysis after Exercise (말의 열충격 단백질(heat shock proteins)의 특성 구명과 운동 후 유전자의 발현 분석)

  • Cho, Hyun-Woo;Park, Jeong-Woong;Choi, Jae-Young;Sivakumar, S.;Kim, Nam-Young;Shin, Teak-Soon;Cho, Seong-Keun;Kim, Byeong-Woo;Cho, Byung-Wook
    • Journal of Life Science
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    • v.24 no.2
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    • pp.105-111
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    • 2014
  • The purpose of this study was to characterize equine heat-shock protein (Hsp) genes and analyze their expression pattern in various horse tissues and blood leukocytes after exercise. In a previous study, RNA sequencing of blood and skeletal muscles of thoroughbreds before and after exercise was performed using differently expressed gene (DEG) analysis. Three Hsp genes (HspH1, Hsp90${\alpha}$ and Hsp70) were selected by DEG analysis and were found to be differentially expressed in either blood or muscle. To validate and extend previous observations on these genes, we performed RT-PCR analyses of horse tissue as well as real-time qPCR analyses of blood leukocytes after exercise. mRNA expression of these Hsp genes was found to be ubiquitous in the analyzed tissues (including thyroid, colon, skeletal muscle, cecum, kidney, spinal cord, heart, and lung). In addition, Hsp mRNA expression of these genes in extracted whole blood increased after 120 minutes of exercise compared to the baseline condition. These results are in agreement with the results of human and other experimental animals, suggesting that regulatory mechanisms that are responsible for upregulation of Hsp gene transcription may be conserved among species. Further investigations to correlate Hsp gene expression patterns with athletic performance or recovery processes after exercise are warranted.

Metagenomic Analysis of Jang Using Next-generation Sequencing: A ComparativeMicrobial Study of Korean Traditional Fermented Soybean Foods (차세대 염기서열 분석을 활용한 장류의 메타지놈 분석 : 한국 전통 콩 발효식품에 대한 미생물 비교 연구)

  • Ranhee Lee;Gwangsu Ha;Ho Jin Jeong;Do-Youn Jeong;Hee-Jong Yang
    • Journal of Life Science
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    • v.34 no.4
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    • pp.254-263
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    • 2024
  • Korean jang is a food made using fermented soybeans, and the typical products include gochujang (GO), doenjang (DO), cheonggukjang (CH), and ganjang (GA). In this study, 16S rRNA metagenome analysis was performed on a total of 200 types of GO, DO, CH, and GA using next-generation sequencing to analyze the microbial community of fermented soybean foods and compare taxonomic (biomarker) differences. Alpha diversity analysis showed that in the CHAO index, the species richness index tended to be significantly higher compared to the DO and GA groups (p<0.001). The results of the microbial distribution analysis of the GO, DO, CH, and GA products showed that at the order level, Bacillales was the most abundant in the GO, DO, and CH groups, but Lactobacillales was most abundant in the GA group. Linear discriminant analysis effect (LEfSe) analysis was used to identify biomarkers at the family and species levels. Leuconostocaceae, Thermoactinomycetaceae, Bacillaceae, and Enterococcaceae appeared as biomarkers at the family level, and Bacillus subtilis, Kroppenstedtia sanguinis, Bacillus licheniformis, and Tetragenococcus halophilus appeared at the species level. Permutational multivariate analysis of variance (PERMANOVA) analysis showed that there was a significant difference in the microbial community structure of the GO, DO, CH, and GA groups (p=0.001), and the microbial community structure of the GA group showed the greatest difference. This study clarified the correlation between the characteristics of Korean fermented foods and microbial community distribution, enhancing knowledge of microorganisms participating in the fermentation process. These results could be leveraged to improve the quality of fermented soybean foods.

Bacterial Diversity of the Han River as Determined by 16S rRNA Gene Analysis (16S rRNA 유전자 계통분석에 의한 한강수계의 세균 다양성)

  • Han, Suk-Kyun;Lee, Il-Gyu;Ahn, Tae-Young
    • Korean Journal of Microbiology
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    • v.34 no.4
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    • pp.194-199
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    • 1998
  • Bacterial diversity was determined by amplification and sequencing of 16S rDNA at Tancheon and Jungrang in Han river. Twenty-seven clones constructed were divided 7 groups using RFLP. Fifteen clones were classified 4 groups in Tancheon and the group (HT-1 clone) including many clones was affiliated a high similarity with Aerobacter cryaerophilus (the class Proteobacteria including members of the delta subdivisions). The other two groups (HT-6 and HT-9 clone) including several clones were classified with the class Cytophagales in Tancheon. Twelve clones were classified 3 groups in Jungrang and the group (HJ-1 clone) including many clones was affiliated a high similarity with Sphingomonas sp. (the class Proteobacteria including members of the alpha subdivisions). As a whole results, the class Proteobacteria (alpha, beta and delta subdivision), the order Cytophagales, and the order Actinomycetales were detected.

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