Comprehensive Analysis of Transcriptomics and Metabolomics between the Heads and Tails of Angelica Sinensis: Genes Related to Phenylpropanoid Biosynthesis Pathway

2020 ◽  
Vol 23 ◽  
Author(s):  
Jie Yang ◽  
Chi Zhang ◽  
Wei-Hong Li ◽  
Tian-Er Zhang ◽  
Guang-Zhong Fan ◽  
...  
Keyword(s):  
Ferulic Acid ◽  
Metabolic Regulation ◽  
Comprehensive Analysis ◽  
Angelica Sinensis ◽  
Rna Seq ◽  
Targeted Metabolomics ◽  
Kegg Pathways ◽  
Qrt Pcr ◽  
Phenylpropanoid Biosynthesis ◽  
Combined Strategy

Background:: In Traditional Chinese Medicine (TCM), the heads and tails of Angelica sinensis (Oliv.) Diels (AS) is used in treating different diseases due to their different pharmaceutical efficacies. The underline mechanisms, however, have not been fully explored. Objective:: Novel mechanisms responsible for the discrepant activities between AS heads and tails were explored by a combined strategy of transcriptomes and metabolomics. Method:: Six pairs of the heads and tails of AS roots were collected in Min County, China. Total RNA and metabolites, which were used for RNA-seq and untargeted metabolomics analysis, were respectively isolated from each AS sample (0.1 g) by Trizol and methanol reagent. Subsequently, differentially expressed genes (DEGs) and discrepant pharmaceutical metabolites were identified for comparing AS heads and tails. Key DEGs and metabolites were quantified by qRT-PCR and targeted metabolomics experiment. Results:: Comprehensive analysis of transcriptomes and metabolomics results suggested that five KEGG pathways with significant differences included 57 DEGs. Especially, fourteen DEGs and six key metabolites were relation to the metabolic regulation of Phenylpropanoid biosynthesis (PB) pathway. Results of qRT-PCR and targeted metabolomics indicated that higher levels of expression of crucial genes in PB pathway, such as PAL, CAD, COMT and peroxidase in the tail of AS were positively correlated with levels of ferulic acid-related metabolites. The average content of ferulic acid in tails (569.58162.39 nmol/g) was higher than those in the heads (168.73  67.30 nmol/g) (P˂0.01); Caffeic acid in tails (3.82  0.88 nmol/g) vs heads (1.37  0.41 nmol/g) (P˂0.01), and Cinnamic acid in tails (0.24  0.09 nmol/g) vs heads (0.14  0.02 nmol/g) (P˂0.05). Conclusion:: Our work demonstrated that overexpressed genes and accumulated metabolites derived from PB pathway might be responsible for the discrepant pharmaceutical efficacies between AS heads and tails.

scholarly journals Comprehensive Analysis of Peripheral Exosomal circRNAs in Large Artery Atherosclerotic Stroke

2021 ◽  
Vol 9 ◽  
Author(s):  
Qi Xiao ◽  
Ruihua Yin ◽  
Yuan Wang ◽  
Shaonan Yang ◽  
Aijun Ma ◽  
...  
Keyword(s):  
Area Under The Curve ◽  
Comprehensive Analysis ◽  
Circular Rnas ◽  
Large Artery ◽  
Rna Seq ◽  
Roc Curve Analysis ◽  
Qrt Pcr ◽  
Model Combining ◽  
Novel Biomarkers ◽  
The Individual

Exosomes are crucial vehicles in intercellular communication. Circular RNAs (circRNAs), novel endogenous noncoding RNAs, play diverse roles in ischemic stroke. Recently, the abundance and stability of circRNAs in exosomes have been identified. However, a comprehensive analysis of exosomal circRNAs in large artery atherosclerotic (LAA) stroke has not yet been reported. We performed RNA sequencing (RNA-Seq) to comprehensively identify differentially expressed (DE) exosomal circRNAs in five paired LAA and normal controls. Further, quantitative real-time PCR (qRT-PCR) was used to verify the RNA-Seq results in a cohort of stroke patients (32 versus 32). RNA-Seq identified a total of 462 circRNAs in peripheral exosomes; there were 25 DE circRNAs among them. Additionally, circRNA competing endogenous RNA (ceRNA) network and translatable analysis revealed the potential functions of the exosomal circRNAs in LAA progression. Two ceRNA pathways involving 5 circRNAs, 2 miRNAs, and 3 mRNAs were confirmed by qRT-PCR. In the validation cohort, receiver operating characteristic (ROC) curve analysis identified two circRNAs as possible novel biomarkers, and a logistic model combining two and four circRNAs increased the area under the curve compared with the individual circRNAs. Here, we show for the first time the comprehensive expression of exosomal circRNAs, which displayed the potential diagnostic and biological function in LAA stroke.

scholarly journals Transcriptome Analysis and Identification of Insecticide Tolerance-Related Genes after Exposure to Insecticide in Sitobion avenae

Genes ◽  
2019 ◽  
Vol 10 (12) ◽  
pp. 951
Author(s):  
Ning Wei ◽  
Yongzhi Zhong ◽  
Lulu Lin ◽  
Minghui Xie ◽  
Guangling Zhang ◽  
...  
Keyword(s):  
Transcriptome Analysis ◽  
Expression Patterns ◽  
Sitobion Avenae ◽  
Rna Seq ◽  
Kegg Pathways ◽  
Qrt Pcr ◽  
Stress Response Genes ◽  
Grain Aphid ◽  
Insecticide Tolerance ◽  
Polymerase Chain

Aphids cause serious losses to the production of wheat. The grain aphid, Sitobion avenae, which is the dominant species of aphid in all wheat regions of China, is resistant to a variety of insecticides, including imidacloprid and chlorpyrifos. However, the resistance and mechanism of insecticide tolerance of S. avenae are still unclear. Therefore, this study employed transcriptome analysis to compare the expression patterns of stress response genes under imidacloprid and chlorpyrifos treatment for 15 min, 3 h, and 36 h of exposure. S. avenae adult transcriptome was assembled and characterized first, after which samples treated with insecticides for different lengths of time were compared with control samples, which revealed 60–2267 differentially expressed unigenes (DEUs). Among these DEUs, 31–790 unigenes were classified into 66–786 categories of gene ontology (GO) functional groups, and 24–760 DEUs could be mapped into 54–268 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Finally, 11 insecticide-tolerance-related unigenes were chosen to confirm the relative expression by quantitative real-time polymerase chain reaction (qRT-PCR) in each treatment. Most of the results between qRT-PCR and RNA sequencing (RNA-Seq) are well-established. The results presented herein will facilitate molecular research investigating insecticide resistance in S. avenae, as well as in other wheat aphids.

scholarly journals RcTGA1 and glucosinolate biosynthesis pathway involvement in the defence of rose against the necrotrophic fungus Botrytis cinerea

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Penghua Gao ◽  
Hao Zhang ◽  
Huijun Yan ◽  
Qigang Wang ◽  
Bo Yan ◽  
...  
Keyword(s):  
Disease Resistance ◽  
Gene Annotation ◽  
Infected Plant ◽  
Grey Mould ◽  
Cascade Reactions ◽  
Postharvest Quality ◽  
Rna Seq ◽  
Protein Activity ◽  
Phenylpropanoid Biosynthesis ◽  
And Cluster Analysis

Abstract Background Rose is an important economic crop in horticulture. However, its field growth and postharvest quality are negatively affected by grey mould disease caused by Botrytis c. However, it is unclear how rose plants defend themselves against this fungal pathogen. Here, we used transcriptomic, metabolomic and VIGS analyses to explore the mechanism of resistance to Botrytis c. Result In this study, a protein activity analysis revealed a significant increase in defence enzyme activities in infected plants. RNA-Seq of plants infected for 0 h, 36 h, 60 h and 72 h produced a total of 54 GB of clean reads. Among these reads, 3990, 5995 and 8683 differentially expressed genes (DEGs) were found in CK vs. T36, CK vs. T60 and CK vs. T72, respectively. Gene annotation and cluster analysis of the DEGs revealed a variety of defence responses to Botrytis c. infection, including resistance (R) proteins, MAPK cascade reactions, plant hormone signal transduction pathways, plant-pathogen interaction pathways, Ca2+ and disease resistance-related genes. qPCR verification showed the reliability of the transcriptome data. The PTRV2-RcTGA1-infected plant material showed improved susceptibility of rose to Botrytis c. A total of 635 metabolites were detected in all samples, which could be divided into 29 groups. Metabonomic data showed that a total of 59, 78 and 74 DEMs were obtained for T36, T60 and T72 (T36: Botrytis c. inoculated rose flowers at 36 h; T60: Botrytis c. inoculated rose flowers at 60 h; T72: Botrytis c. inoculated rose flowers at 72 h) compared to CK, respectively. A variety of secondary metabolites are related to biological disease resistance, including tannins, amino acids and derivatives, and alkaloids, among others; they were significantly increased and enriched in phenylpropanoid biosynthesis, glucosinolates and other disease resistance pathways. This study provides a theoretical basis for breeding new cultivars that are resistant to Botrytis c. Conclusion Fifty-four GB of clean reads were generated through RNA-Seq. R proteins, ROS signalling, Ca2+ signalling, MAPK signalling, and SA signalling were activated in the Old Blush response to Botrytis c. RcTGA1 positively regulates rose resistance to Botrytis c. A total of 635 metabolites were detected in all samples. DEMs were enriched in phenylpropanoid biosynthesis, glucosinolates and other disease resistance pathways.

scholarly journals Transcriptome Analysis of Responses to Dengue Virus 2 Infection in Aedes albopictus (Skuse) C6/36 Cells

Viruses ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 343
Author(s):  
Manjin Li ◽  
Dan Xing ◽  
Duo Su ◽  
Di Wang ◽  
Heting Gao ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Aedes Albopictus ◽  
Bioinformatics Analysis ◽  
Interaction Mechanism ◽  
Transcriptional Analysis ◽  
Functional Verification ◽  
Mosquito Vector ◽  
Rna Seq ◽  
Sequencing Data ◽  
Qrt Pcr

Dengue virus (DENV), a member of the Flavivirus genus of the Flaviviridae family, can cause dengue fever (DF) and more serious diseases and thus imposes a heavy burden worldwide. As the main vector of DENV, mosquitoes are a serious hazard. After infection, they induce a complex host–pathogen interaction mechanism. Our goal is to further study the interaction mechanism of viruses in homologous, sensitive, and repeatable C6/36 cell vectors. Transcriptome sequencing (RNA-Seq) technology was applied to the host transcript profiles of C6/36 cells infected with DENV2. Then, bioinformatics analysis was used to identify significant differentially expressed genes and the associated biological processes. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to verify the sequencing data. A total of 1239 DEGs were found by transcriptional analysis of Aedes albopictus C6/36 cells that were infected and uninfected with dengue virus, among which 1133 were upregulated and 106 were downregulated. Further bioinformatics analysis showed that the upregulated DEGs were significantly enriched in signaling pathways such as the MAPK, Hippo, FoxO, Wnt, mTOR, and Notch; metabolic pathways and cellular physiological processes such as autophagy, endocytosis, and apoptosis. Downregulated DEGs were mainly enriched in DNA replication, pyrimidine metabolism, and repair pathways, including BER, NER, and MMR. The qRT-PCR results showed that the concordance between the RNA-Seq and RT-qPCR data was very high (92.3%). The results of this study provide more information about DENV2 infection of C6/36 cells at the transcriptome level, laying a foundation for further research on mosquito vector–virus interactions. These data provide candidate antiviral genes that can be used for further functional verification in the future.

scholarly journals Integrated Transcriptomic and Un-Targeted Metabolomics Analysis Reveals Mulberry Fruit (Morus atropurpurea) in Response to Sclerotiniose Pathogen Ciboria shiraiana Infection

2020 ◽  
Vol 21 (5) ◽  
pp. 1789 ◽  
Author(s):  
Lijun Bao ◽  
Hongpeng Gao ◽  
Zelin Zheng ◽  
Xiaoxiao Zhao ◽  
Minjuan Zhang ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Early Stage ◽  
Metabolite Profile ◽  
Northwest China ◽  
Targeted Metabolomics ◽  
Mulberry Fruit ◽  
Phenylpropanoid Biosynthesis ◽  
Mapman Analysis ◽  
Mulberry Fruits ◽  
Prevention Methods

Mulberry sclerotiniose caused by Ciboria shiraiana is a devastating disease of mulberry (Morus alba L.) fruit in Northwest China. At present, no disease-resistant varieties are used in production, as the molecular mechanisms of this disease are not well understood. In this study, to explore new prevention methods and provide direction for molecular breeding, transcriptomic sequencing and un-targeted metabolomics were performed on healthy (CK), early-stage diseased (HB1), and middle-stage diseased (HB2) mulberry fruits. Functional annotation, gene ontology, a Kyoto encyclopedia of genes and genomes (KEGG) analysis, and a Mapman analysis of the differentially expressed genes revealed differential regulation of genes related to plant hormone signal transduction, transcription factors, and phenylpropanoid biosynthesis. A correspondence between the transcript pattern and metabolite profile was observed in the phenylpropanoid biosynthesis pathway. It should be noted that the log2 ratio of eugenol (isoeugenol) in HB1 and HB2 are 85 times and 23 times higher than CK, respectively. Our study shows that phenylpropanoid biosynthesis may play an essential role in response to sclerotiniose pathogen infection and eugenol(isoeugenol) enrichment in mulberry fruit, which may provide a novel method for mulberry sclerotiniose control.

Application of qRT-PCR and RNA-Seq analysis for the identification of housekeeping genes useful for normalization of gene expression values during Striga hermonthica development

2012 ◽  
Vol 40 (4) ◽  
pp. 3395-3407 ◽  
Author(s):  
M. Fernández-Aparicio ◽  
K. Huang ◽  
E. K. Wafula ◽  
L. A. Honaas ◽  
N. J. Wickett ◽  
...  
Keyword(s):  
Gene Expression ◽  
Housekeeping Genes ◽  
Striga Hermonthica ◽  
Rna Seq ◽  
Qrt Pcr

scholarly journals Extensive Metabolite Profiling in the Unexploited Organs of Black Tiger for Their Potential Valorization in the Pharmaceutical Industry

Life ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 544
Author(s):  
Jianfei Gao ◽  
Kangning Xiong ◽  
Wei Zhou ◽  
Weijie Li
Keyword(s):  
Acid Metabolism ◽  
Metabolite Profile ◽  
Industrial Applications ◽  
Targeted Metabolomics ◽  
Arginine Biosynthesis ◽  
Nutritional Values ◽  
Phenylpropanoid Biosynthesis ◽  
Tyrosine Metabolism ◽  
Organ Specific ◽  
Widely Targeted Metabolomics

Black tiger (Kadsura coccinea (Lem.)) has been reported to hold enormous pharmaceutical potential. The fruit and rhizome of black tiger are highly exploited in the pharmaceutical and other industries. However, the most important organs from the plant such as the leaf and stem are considered biowastes mainly because a comprehensive metabolite profile has not been reported in these organs. Knowledge of the metabolic landscape of the unexploited black tiger organs could help identify and isolate important compounds with pharmaceutical and nutritional values for a better valorization of the species. In this study, we used a widely targeted metabolomics approach to profile the metabolomes of the K. coccinea leaf (KL) and stem (KS) and compared them with the root (KR). We identified 642, 650 and 619 diverse metabolites in KL, KS and KR, respectively. A total of 555 metabolites were mutually detected among the three organs, indicating that the leaf and stem organs may also hold potential for medicinal, nutritional and industrial applications. Most of the differentially accumulated metabolites between organs were enriched in flavone and flavonol biosynthesis, phenylpropanoid biosynthesis, arginine and proline metabolism, arginine biosynthesis, tyrosine metabolism and 2-oxocarboxylic acid metabolism pathways. In addition, several important organ-specific metabolites were detected in K. coccinea. In conclusion, we provide extensive metabolic information to stimulate black tiger leaf and stem valorization in human healthcare and food.

scholarly journals RNA Sequencing (RNA-Seq) Based Transcriptome Analysis in Immune Response of Holstein Cattle to Killed Vaccine against Bovine Viral Diarrhea Virus Type I

Animals ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 344 ◽  
Author(s):  
Bryan Irvine Lopez ◽  
Kier Gumangan Santiago ◽  
Donghui Lee ◽  
Seungmin Ha ◽  
Kangseok Seo
Keyword(s):  
Immune Response ◽  
Enrichment Analysis ◽  
Receptor Interaction ◽  
Type I ◽  
Rna Seq ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Kegg Pathways ◽  
Viral Diarrhea Virus

Immune response of 107 vaccinated Holstein cattle was initially obtained prior to the ELISA test. Five cattle with high and low bovine viral diarrhea virus (BVDV) type I antibody were identified as the final experimental animals. Blood samples from these animals were then utilized to determine significant differentially expressed genes (DEGs) using the RNA-seq transcriptome analysis and enrichment analysis. Our analysis identified 261 DEGs in cattle identified as experimental animals. Functional enrichment analysis in gene ontology (GO) annotations and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways revealed the DEGs potentially induced by the inactivated BVDV type I vaccine, and might be responsible for the host immune responses. Our findings suggested that inactivated vaccine induced upregulation of genes involved in different GO annotations, including antigen processing and presentation of peptide antigen (via MHC class I), immune response, and positive regulation of interferon-gamma production. The observed downregulation of other genes involved in immune response might be due to inhibition of toll-like receptors (TLRs) by the upregulation of the Bcl-3 gene. Meanwhile, the result of KEGG pathways revealed that the majority of DEGs were upregulated and enriched to different pathways, including cytokine-cytokine receptor interaction, platelet activation, extracellular matrix (ECM) receptor interaction, hematopoietic cell lineage, and ATP-binding cassette (ABC) transporters. These significant pathways supported our initial findings and are known to play a vital role in shaping adaptive immunity against BVDV type 1. In addition, type 1 diabetes mellitus pathways tended to be significantly enriched. Thus, further studies are needed to investigate the prevalence of type 1 diabetes mellitus in cattle vaccinated with inactivated and live BVDV vaccine.

scholarly journals Ocena zmian profilu ekspresji genów kandydujących w podkładkach jabłoni o odmiennym stopniu tolerancji mrozowej

2020 ◽  
pp. 21-32
Author(s):  
Sylwia Keller-Przybyłkowicz ◽  
Mariusz Lewandowski
Keyword(s):  
De Novo ◽  
Rna Seq ◽  
Qrt Pcr

Celem przeprowadzonych badań była identyfikacja genów sprzężonych z cechą mrozoodporności podkładek jabłoni. Ocenę zmian w poziomie ekspresji wyizolowanych genów przeprowadzono metodami RNAseq i qRT-PCR, dla podkładek zróżnicowanych po względem stopnia tolerancji mrozowej: P 66 (tolerancyjna) i M.9 (wrażliwa).W wyniku przeprowadzonych odczytów sekwencji RNA (sekwencjonowanie de novo w systemie Illumina Solid) dla w/w podkładek zidentyfikowano około 167 milionów odczytów unikatowych sekwencji, z których do wstępnych badań weryfikacyjnych wytypowano 15 o zróżnicowanym profilu ekspresji. Sekwencje poddano adnotacji funkcjonalnej. Wytypowane geny kodują: białka strukturalne i integralne błon komórkowych i wakuoli komórkowych, czynników transkrypcyjnych, białek regulujących transport międzykomórkowy i wewnątrzkomórkowy, białek hydrolizujących wiązania C-O i C-N oraz białek wiążących makro- i mikroelementy. Celem weryfikacji typu regulacji sekwencji transkryptomu uzyskanych z sekwencjonowania nowej generacji (NGS), dla tych samych prób przeprowadzono ilościową analizę transkryptu genów (qRT-PCR). Spośród badanych genów, trzy reprezentowały identyczny typ regulacji w badanych układach eksperymentalnych RNA-seq i qRT-PCR. Wytypowane geny stanowią potencjalne sekwencje kandydujące do sporządzenia markerów funkcjonalnych, umożliwiających wczesną selekcję podkładek jabłoni tolerancyjnych na mróz.

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