• Title/Summary/Keyword: RNA-Seq. analysis

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Complete Mitochondrial Genome Sequences of Korean Phytophthora infestans Isolates and Comparative Analysis of Mitochondrial Haplotypes

  • Seo, Jin-Hee;Choi, Jang-Gyu;Park, Hyun-Jin;Cho, Ji-Hong;Park, Young-Eun;Im, Ju-Sung;Hong, Su-Young;Cho, Kwang-Soo
    • The Plant Pathology Journal
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    • v.38 no.5
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    • pp.541-549
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    • 2022
  • Potato late blight caused by Phytophthora infestans is a destructive disease in Korea. To elucidate the genomic variation of the mitochondrial (mt) genome, we assembled its complete mt genome and compared its sequence among different haplotypes. The mt genome sequences of four Korean P. infestans isolates were revealed by Illumina HiSeq. The size of the circular mt genome of the four major genotypes, KR_1_A1, KR_2_A2, SIB-1, and US-11, was 39,872, 39,836, 39,872, and 39,840 bp, respectively. All genotypes contained the same 61 genes in the same order, comprising two RNA-encoding genes, 16 ribosomal genes, 25 transfer RNA, 17 genes encoding electron transport and ATP synthesis, 11 open reading frames of unknown function, and one protein import-related gene, tatC. The coding region comprised 91% of the genome, and GC content was 22.3%. The haplotypes were further analyzed based on sequence polymorphism at two hypervariable regions (HVRi), carrying a 2 kb insertion/deletion sequence, and HVRii, carrying 36 bp variable number tandem repeats (VNTRs). All four genotypes carried the 2 kb insertion/deletion sequence in HVRi, whereas HVRii had two VNTRs in KR_1_A1 and SIB-1 but three VNTRs in US-11 and KR_2_A2. Minimal spanning network and phylogenetic analysis based on 5,814 bp of mtDNA sequences from five loci, KR_1_A1 and SIB-1 were classified as IIa-6 haplotype, and isolates KR_1_A2 and US-11 as haplotypes IIa-5 and IIb-2, respectively. mtDNA sequences of KR_1_A1 and SIB-1 shared 100% sequence identity, and both were 99.9% similar to those of KR_2_A2 and US-11.

Transcriptome analysis of the livers of ducklings hatched normally and with assistance

  • Liu, Yali;He, Shishan;Zeng, Tao;Du, Xue;Shen, Junda;Zhao, Ayong;Lu, Lizhi
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.6
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    • pp.773-780
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    • 2017
  • Objective: "Hatchability" is an important economic trait in domestic poultry. Studies on poultry hatchability focus mainly on the genetic background, egg quality, and incubation conditions, whereas the molecular mechanisms behind the phenomenon that some ducklings failed to break their eggshells are poorly understood. Methods: In this study, the transcriptional differences between the livers of normally hatched and assisted ducklings were systematically analyzed. Results: The results showed that the clean reads were de novo assembled into 161,804 and 159,083 unigenes (${\geq}200-bp$ long) by using Trinity, with an average length of 1,206 bp and 882 bp, respectively. The defined criteria of the absolute value of log2 fold-change ${\geq}1$ and false discovery rate${\leq}0.05$ were differentially expressed and were significant. As a result, 1,629 unigenes were identified, the assisted ducklings showed 510 significantly upregulated and 1,119 significantly down-regulated unigenes. In general, the metabolic rate in the livers of the assisted ducklings was lower than that in the normal ducklings; however, compared to normal ducklings, glucose-6-phosphatase and ATP synthase subunit alpha 1 associated with energy metabolism were significantly upregulated in the assisted group. The genes involved in immune defense such as major histocompatibility complex (MHC) class I antigen alpha chain and MHC class II beta chain 1 were downregulated in the assisted ducklings. Conclusion: These data provide abundant sequence resources for studying the functional genome of the livers in ducks and other poultry. In addition, our study provided insight into the molecular mechanism by which the phenomenon of weak embryos is regulated.

Identification of genes related to ER stress in bZIP28 gene transgenic potato plant

  • Kim, Dool Yi;Kim, Kyung Hwa;Choi, Man Soo;Ok, Hyun Choog;Kim, Jae Hyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.153-153
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    • 2017
  • Potato (Solanum tubersosum L.) is susceptible to various environmental stresses such as frost, high temperature, and drought. Enhancement of potato drought tolerance can reduce yield loss under drought that has negative effect on potato tuber growth. Genetic engineering can be utilized to achieve this goal, but such approaches using endogenous potato genes have rarely been applied. Since unpredictable global weather changes cause more severe and frequent water limiting conditions, improvement of potato drought tolerance can minimize such adverse effects under drought and can impact on sustainable potato production. Genetic engineering can be utilized to improve potato drought tolerance, but such approaches using endogenous potato genes have rarely been applied. We were obtained AtbZIP28 gene transgenic potato plants. It is identified transcript levels at various stress conditions, polyethylene glycol (PEG), NaCl, abscisic ${\underline{acid}}$ (ABA). Also, For identification to regulate ER stress response genes in AtbZIP28 gene transgenic potato plant, we screened seven potato genes from RNA-seq analysis under TM treatment. Five and two genes were up- and down-regulated by TM, respectively. Their expression patterns were re-examined at stress agents known to elicit TM, DTT, DMSO and salt stress.

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Regulation of hormone-related genes involved in adventitious root formation in sweetpotato

  • Nie, Hualin;Kim, Sujung;Lee, Yongjae;Park, Hyungjun;Lee, Jeongeun;Kim, Jiseong;Kim, Doyeon;Kim, Sunhyung
    • Journal of Plant Biotechnology
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    • v.47 no.3
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    • pp.194-202
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    • 2020
  • The sweetpotatoes (Ipomoea batatas) generate adventitious roots (ARs) from cut stems that develop into storage roots and make for an important means of propagation. However, few studies have investigated the hormones involved in AR development in sweetpotato. In this study, the expression patterns of hormone-related genes involved in AR formation were identified using the transcriptome data. RNA-seq data from stems grown for 0 and 3 days after cutting were analyzed. In addition, hormone-related genes were identified among differentially expressed genes (DEGs) and filtered genes, and cluster analysis was used to characterize expression patterns by function. Most hormone-related regulated genes expressed 3 days after growing the cut stems were abscisic acid (ABA)-related genes, followed by ethylene- and auxin-related genes. For ABA, the biosynthesis genes (including genes annotated to NINE-CIS-EPOXYCAROTENOID DIOXYGENASE 3 (NCED3)) and signal transduction and perception genes (including genes annotated to PROTEIN PHOSPHATASE 2Cs (PP2Cs)) tended to decrease. Expression patterns of auxin- and ethylene-related genes differed by function. These results suggest that ABA, auxin, and ethylene genes are involved in AR formation and that they may be regulated in a hormone function-dependent manner. These results contribute to the identification of hormone functions during AR formation and may contribute to understanding the mechanism of AR formation in the sweetpotato.

Functional analysis of a homologue of the FLORICAULA/LEAFY gene in litchi (Litchi chinensis Sonn.) revealing its significance in early flowering process

  • Ding, Feng;Zhang, Shuwei;Chen, Houbin;Peng, Hongxiang;Lu, Jiang;He, Xinhua;Pan, Jiechun
    • Genes and Genomics
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    • v.40 no.12
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    • pp.1259-1267
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    • 2018
  • Litchi (Litchi chinensis Sonn.) is an important subtropical fruit crop with high commercial value due to its high nutritional values and favorable tastes. However, irregular bearing attributed to unstable flowering is a major ongoing problem for litchi producers. Previous studies indicate that low-temperature is a key factor in litchi floral induction. In order to reveal the genetic and molecular mechanisms underlying the reproductive process in litchi, we had analyzed the transcriptome of buds before and after low-temperature induction using RNA-seq technology. A key flower bud differentiation associated gene, a homologue of FLORICAULA/LEAFY, was identified and named LcLFY (GenBank Accession No. KF008435). The cDNA sequence of LcLFY encodes a putative protein of 388 amino acids. To gain insight into the role of LcLFY, the temporal expression level of this gene was measured by real-time RT-PCR. LcLFY was highly expressed in flower buds and its expression correlated with the floral developmental stage. Heterologous expression of LcLFY in transgenic tobacco plants induced precocious flowering. Meantime, we investigated the sub-cellular localization of LcLFY. The LcLFY-Green fluorescent protein (GFP) fusion protein was found in the nucleus. The results suggest that LcLFY plays a pivotal role as a transcription factor in controlling the transition to flowering and in the development of floral organs in litchi.

De novo Assembly and Analysis of Amur Sturgeon (Acipenser schrenckii) Transcriptome in Response to Mycobacterium Marinum Infection to Identify Putative Genes Involved in Immunity

  • Zhang, Qianqian;Wang, Xiehao;Zhang, Defeng;Long, Meng;Wu, Zhenbing;Feng, Yuqing;Hao, Jingwen;Wang, Shuyi;Liao, Qian;Li, Aihua
    • Journal of Microbiology and Biotechnology
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    • v.29 no.8
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    • pp.1324-1334
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    • 2019
  • Fish mycobacteriosis is a common bacterial disease in many species of freshwater and marine fish and has caused severe loss of fish production. Mycobacterium marinum has been the most prevalent pathogen observed in several outbreaks of mycobacteriosis of farmed sturgeons in China. However, the immune responses and pathology of sturgeons in mycobacterial infection are rarely studied. Therefore, we used the Illumina RNA-seq method to analyze the transcriptome profile of Acipenser schrenckii challenged with Mycobacterium marinum. To begin, 168,220 non-redundant contigs were acquired from the infection and control groups, and among these, 33,225 contigs have acquired annotations. A total of 4,043 differently expressed (DE) contigs between the two groups were identified, and among these, 2479 were up-regulated and 1564 were down-regulated in the infected fish. A total of 1,340 DE contigs with acquired annotations in KEGG were enriched for 124 pathways including the TNF signaling pathway, and the Toll-like receptor signaling pathway. The roles of DE genes involved in significant pathways and other processes were discussed. The 2,209 DE contigs that have yet to acquire proper annotation may represent candidate genes associated with infection in sturgeons and are expected to serve as immunogenetic resources for further study. To our best knowledge, this is the first transcriptome study on sturgeons under bacterial infection.

Exploring effects of different male parent crossings on sheep muscles and related regulatory genes using mRNA-Seq

  • Shi, Jinping;Zhang, Quanwei;Song, Yali;Lei, Zhaomin;Fu, Lingjuan;Cheng, Shuru
    • Animal Bioscience
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    • v.35 no.8
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    • pp.1129-1140
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    • 2022
  • Objective: With improvements in living standards and increase in global population, the demand for meat products has been increasing; improved meat production from livestock could effectively meet this demand. In this study, we examined the differences in the muscle traits of different male crossbred sheep and attempted to identify key genes that regulate these traits. Methods: Dubo sheep×small-tailed Han sheep (DP×STH) and Suffolk×small-tailed Han sheep (SFK×STH) were selected to determine meat quality and production performance by Masson staining. Transcriptome sequencing and bioinformatic analysis were performed to identify differentially expressed genes (DEGs) related to meat quality. The presence of DEGs was confirmed by real-time polymerase chain reaction. Results: The production performance of SFK×STH sheep was better than that of DP×STH sheep, but the meat quality of DP×STH sheep was better than that of SFK×STH sheep. The muscle fiber diameter of DP×STH sheep was smaller than that of SFK×STH sheep. Twenty-two DEGs were identified. Among them, four gene ontology terms were related to muscle traits, and three DEGs were related to muscle or muscle fibers. There were no significant differences in the number of single nucleotide mutations and mutation sites in the different male parent cross combinations. Conclusion: This study provides genetic resources for future sheep muscle development and cross-breeding research.

Composition and functional diversity of bacterial communities during swine carcass decomposition

  • Michelle Miguel;Seon-Ho Kim;Sang-Suk Lee;Yong-Il Cho
    • Animal Bioscience
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    • v.36 no.9
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    • pp.1453-1464
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    • 2023
  • Objective: This study investigated the changes in bacterial communities within decomposing swine microcosms, comparing soil with or without intact microbial communities, and under aerobic and anaerobic conditions. Methods: The experimental microcosms consisted of four conditions: UA, unsterilized soil-aerobic condition; SA, sterilized soil-aerobic condition; UAn, unsterilized soil-anaerobic condition; and San, sterilized soil-anaerobic condition. The microcosms were prepared by mixing 112.5 g of soil and 37.5 g of ground carcass, which were then placed in sterile containers. The carcass-soil mixture was sampled at day 0, 5, 10, 30, and 60 of decomposition, and the bacterial communities that formed during carcass decomposition were assessed using Illumina MiSeq sequencing of the 16S rRNA gene. Results: A total of 1,687 amplicon sequence variants representing 22 phyla and 805 genera were identified in the microcosms. The Chao1 and Shannon diversity indices varied in between microcosms at each period (p<0.05). Metagenomic analysis showed variation in the taxa composition across the burial microcosms during decomposition, with Firmicutes being the dominant phylum, followed by Proteobacteria. At the genus level, Bacillus and Clostridium were the main genera within Firmicutes. Functional prediction revealed that the most abundant Kyoto encyclopedia of genes and genomes metabolic functions were carbohydrate and amino acid metabolisms. Conclusion: This study demonstrated a higher bacteria diversity in UA and UAn microcosms than in SA and SAn microcosms. In addition, the taxonomic composition of the microbial community also exhibited changes, highlighting the impact of soil sterilization and oxygen on carcass decomposition. Furthermore, this study provided insights into the microbial communities associated with decomposing swine carcasses in microcosm.

Effect of Elevated Temperature on Physiological and Molecular Responses and Photoassimilate Production of Rice Leaves During Early Seed Development

  • Jung-Il Cho;Yo-Han Yoo;Eun-Ji Kim;Hoejeong Jeong;Jae-Kyeong Baek;Wan-Gyu Sang;Sungyul Chang;Dongwon Kwon
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.107-107
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    • 2022
  • The increase in atmospheric temperature due to climate change prolongs the period of exposure to high-temperature environments during rice cultivation. In particular, high-temperature during early seed development greatly affects on the productivity and quality of rice. The high temperature at this time not only affects the transport and distribution of assimilates from leaves to seeds and the accumulation of starch in the seeds, but also affects the leaves, which are the production organs of assimilates, and increases the consumption of assimilation products due to an increase in respiration. Therefore, in this study, rice was grown in temperature gradient chambers(TGC) to analyze the effects of high temperature on physiological responses, assimilate production, and changes in gene expression in rice leaves. Analysis of chlorophyll and sugar contents and RNA-seq experiments were performed using flag leaves collected under normal and elevated temperature conditions, respectively, during the early seed development stage, and then these results were comprehensively discussed.

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Ginsenoside 20(S)-Rg3 reduces KIF20A expression and promotes CDC25A proteasomal degradation in epithelial ovarian cancer

  • Rong Zhang;Lei Li;Huihui Li;Hansong Bai;Yuping Suo;Ju Cui;Yingmei Wang
    • Journal of Ginseng Research
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    • v.48 no.1
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    • pp.40-51
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    • 2024
  • Background: Ginsenoside 20(S)-Rg3 shows promising tumor-suppressive effects in ovarian cancer via inhibiting NF-kB signaling. This study aimed to explore the downstream tumor suppressive mechanisms of ginsenoside Rg3 via this signaling pathway. Materials and methods: A systematical screening was applied to examine the expression profile of 41 kinesin family member genes in ovarian cancer. The regulatory effect of ginsenoside Rg3 on KIF20A expression was studied. In addition, we explored interacting proteins of KIF20A and their molecular regulations in ovarian cancer. RNA-seq data from The Cancer Genome Atlas (TCGA) was used for bioinformatic analysis. Epithelial ovarian cancer cell lines SKOV3 and A2780 were used as in vitro and in vivo cell models. Commercial human ovarian cancer tissue arrays were used for immunohistochemistry staining. Results: KIF20A is a biomarker of poor prognosis among the kinesin genes. It promotes ovarian cancer cell growth in vitro and in vivo. Ginsenoside Rg3 can suppress the transcription of KIF20A. GST pull-down and co-immunoprecipitation (IP) assays confirmed that KIF20A physically interacts with BTRC (β-TrCP1), a substrate recognition subunit for SCFβ-TrCP E3 ubiquitin ligase. In vitro ubiquitination and cycloheximide (CHX) chase assays showed that via interacting with BTRC, KIF20A reduces BTRC-mediated CDC25A poly-ubiquitination and enhances its stability. Ginsenoside Rg3 treatment partly abrogates KIF20A overexpression-induced CDC25A upregulation. Conclusion: This study revealed a novel anti-tumor mechanism of ginsenoside Rg3. It can inhibit KIF20A transcription and promote CDC25A proteasomal degradation in epithelial ovarian cancer.