scholarly journals Transcriptomic profiling uncovers novel players in innate immunity in Arabidopsis thaliana

2021 ◽  
Author(s):  
Mehdi Safaeizadeh ◽  
Thomas Boller ◽  
Claude Becker
Keyword(s):  
Innate Immunity ◽  
Differentially Expressed Genes ◽  
Differentially Expressed ◽  
In Planta ◽  
Susceptibility To Infection ◽  
Molecular Pattern ◽  
New Information ◽  
Analysis Technique ◽  
In The Wild ◽  
Molecular Patterns

AbstractIn this research a high-throughput RNA sequencing based transcriptome analysis technique (RNA-Seq) was used to evaluate differentially expressed genes (DEGs) in the wild type Arabidopsis seedling in response to flg22, a well-known microbe-associated molecular pattern (MAMP), and AtPep1, a well-known peptide representing an endogenous damage-associated molecular patterns (DAMP). The results of our study revealed that 1895 (1634 up-regulated and 261 down-regulated) and 2271 (1706 up-regulated and 565 down-regulated) significant differentially expressed genes in response to flg22 and AtPep1 treatment, respectively. Among significant DEGs, we observed that a number of hitherto overlooked genes have been found to be induced upon treatment with either flg22 or with AtPep1, indicating their possible involvement in innate immunity. Here, we characterized two of them, namely PP2-B13 and ACLP1. pp2-b13 and aclp1 mutants showed an increased susceptibility to infection by the virulent pathogen Pseudomomas syringae pv tomato mutant hrcC-, as evidenced by an increased growth of the pathogen in planta. Further we present evidence that the aclp1 mutant was deficient in ethylene production upon flg22 treatment, while the pp2-b13 mutant, was deficient in ROS production. The results from this research provide new information to a better understanding of the immune system in Arabidopsis.

scholarly journals Revealing Differentially Expressed Genes and Identifying Effector Proteins of Puccinia striiformis f. sp. tritici in Response to High-Temperature Seedling Plant Resistance of Wheat Based on Transcriptome Sequencing

mSphere ◽  
2020 ◽  
Vol 5 (3) ◽  
Author(s):  
Fei Tao ◽  
Yangshan Hu ◽  
Chang Su ◽  
Juan Li ◽  
Lili Guo ◽  
...  
Keyword(s):  
High Temperature ◽  
Differentially Expressed Genes ◽  
Puccinia Striiformis ◽  
Effector Proteins ◽  
Differentially Expressed ◽  
Content Type ◽  
Molecular Pattern ◽  
Seedling Plant ◽  
Effector Triggered Immunity ◽  
Pathogen Associated Molecular Pattern

ABSTRACT Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most important diseases of wheat (Triticum aestivum L.) globally. Recently, more aggressive Pst races have evolved to acquire new virulence profiles and are adapted better to high temperature than most of the previous races. Breeding cultivars with durable high-temperature seedling-plant (HTSP) resistance is an important strategy for controlling stripe rust. Understanding the mechanism of wheat HTSP resistance against Pst is important for more efficient breeding to improve host resistance. In the present study, transcriptomic analysis identified 25 Pst differentially expressed genes (DEGs) that were involved in the HTSP resistance in wheat cultivar Xiaoyan6 (XY6). Functional annotation indicated that these DEGs are related to membrane proteins, mRNA binding proteins, cell membrane transporters, and synthesis of cell nitrogen compounds. Among these DEGs, a candidate effector, PstCEP1 (PSTG_13342), was identified and cloned, and its function was verified. Barley stripe mosaic virus (BSMV)-mediated host-induced gene silencing (HIGS) of PstCEP1 reduced Pst virulence. Signal peptide verification and functional testing in Nicotiana benthamiana indicated that PstCEP1 is a secreted protein and has the function of suppressing programmed cell death (PCD). PstCEP1 as a candidate effector was further supported by type three secretion system (TTSS)-mediated overexpression responding to wheat HTSP resistance via affecting the pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI). IMPORTANCE In the present study, we performed transcriptomic analysis to identify differentially expressed genes and effector proteins of Puccinia striiformis f. sp. tritici (Pst) in response to the high-temperature seedling-plant (HTSP) resistance in wheat. Experimental validation confirmed the function of the highest upregulated effector protein, PstCEP1. This study provides a key resource for understanding the biology and molecular basis of Pst responses to wheat HTSP resistance, and PstCEP1 may be used in future studies to understand pathogen-associated molecular pattern-triggered immunity and effector-triggered immunity processes in the Pst-wheat interaction system.

scholarly journals Identification of Botrytis cinerea Genes Up-Regulated During Infection and Controlled by the Gα Subunit BCG1 Using Suppression Subtractive Hybridization (SSH)

2004 ◽  
Vol 17 (5) ◽  
pp. 537-546 ◽  
Author(s):  
Christian Schulze Gronover ◽  
Corinna Schorn ◽  
Bettina Tudzynski
Keyword(s):  
Botrytis Cinerea ◽  
Differentially Expressed Genes ◽  
Suppression Subtractive Hybridization ◽  
Target Genes ◽  
Subtractive Hybridization ◽  
Differentially Expressed ◽  
Wild Type ◽  
In Planta ◽  
Cell Wall Degrading Enzymes ◽  
Fungal Genes

The Gα subunit BCG1 plays an important role during the infection of host plants by Botrytis cinerea. Δbcg1 Mutants are able to conidiate, penetrate host leaves, and produce small primary lesions. However, in contrast to the wild type, the mutants completely stop invasion of plant tissue at this stage; secondary lesions have never been observed. Suppression subtractive hybridization (SSH) was used to identify fungal genes whose expression on the host plant is specifically affected in bcg1 mutants. Among the 22 differentially expressed genes, we found those which were predicted to encode proteases, enzymes involved in secondary metabolism, and others encoding cell wall-degrading enzymes. All these genes are highly expressed during infection in the wild type but not in the mutant. However, the genes are expressed in both the wild type and the mutant under certain conditions in vitro. Most of the BCG1-controlled genes are still expressed in adenylate cyclase (bac) mutants in planta, suggesting that BCG1 is involved in at least one additional signaling cascade in addition to the cAMP-depending pathway. In a second SSH approach, 1,500 clones were screened for those that are specifically induced by the wild type during the infection of bean leaves. Of the 22 BCG1-controlled genes, 11 also were found in the in planta SSH library. Therefore, SSH technology can be successfully applied to identify target genes of signaling pathways and differentially expressed genes in planta.

scholarly journals miR-146b-5p Plays a Critical Role in the Regulation of Autophagy in ∆per Brucella melitensis-Infected RAW264.7 Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-16 ◽  
Author(s):  
Jiao Hanwei ◽  
Xin Nie ◽  
Huapei Zhu ◽  
Baobao Li ◽  
Feng Pang ◽  
...  
Keyword(s):  
Differentially Expressed Genes ◽  
High Throughput ◽  
Brucella Melitensis ◽  
Critical Role ◽  
Differentially Expressed ◽  
Raw264.7 Cells ◽  
Luciferase Reporter ◽  
Qrt Pcr ◽  
Protective Antibodies ◽  
Molecular Patterns

Brucella-caused brucellosis is one of the most widespread worldwide zoonoses. Lipopolysaccharide (LPS) of Brucella, which functions as pathogen-associated molecular patterns (PAMPs), is an important virulence factor that elicits protective antibodies. Per of B. melitensis is involved in the biosynthesis of the O-side chain of LPS. Autophagy is a crucial element of the innate immune response against intracellular pathogens including Brucella. In this study, we observed that autophagy was inhibited in RAW264.7 cells infected with Brucella melitensis ∆per. And, a high-throughput array-based screen and qRT-PCR validation were performed to identify the differentially expressed miRNAs in RAW264.7 cells infected with B. melitensis M5-90 ∆per. The results suggested that mmu-miR-146a-5p, mmu-miR-155-5p, mmu-miR-146b-5p, and mmu-miR-3473a were upregulated and mmu-miR-30c-5p was downregulated. During B. melitensis M5-90 ∆per infection, the increased expression of miR-146b-5p inhibited the autophagy activation in RAW264.7 cells. Using a bioinformatics approach, Tbc1d14 was predicted to be a potential target of miR-146b-5p. The results of a luciferase reporter assay indicated that miR-146b-5p directly targeted the 3′-UTR of Tbc1d14, and the interaction between miR-146b-5p and the 3′-UTR of Tbc1d14 was sequence-specific. High-throughput RNA-Seq-based screening was performed to identify differentially expressed genes in Tbc1d14-expressing RAW264.7 cells, and these were validated by qRT-PCR. Among the differentially expressed genes, four autophagy associated genes, IFNγ-inducible p47 GTPase 1 (IIGP1), nuclear receptor binding protein 2 (Nrbp2), transformation related protein 53 inducible nuclear protein 1 (Trp53inp1), and immunity-related GTPase family M member 1 (Irgm1), were obtained. Our findings provide important insights into the functional mechanism of LPS of B. melitensis.

scholarly journals Overrepresentation Analyses of Differentially Expressed Genes in the Smut Fungus Ustilago bromivora during Saprophytic and in planta Growth

BIO-PROTOCOL ◽  
2017 ◽  
Vol 7 (15) ◽  
Author(s):  
Angelika Czedik-Eysenberg ◽  
Franziska Rabe ◽  
Heinz Ekker ◽  
Carmen Czepe ◽  
Armin Djamei
Keyword(s):  
Differentially Expressed Genes ◽  
Differentially Expressed ◽  
Smut Fungus ◽  
In Planta

scholarly journals PcGCE is a potent elicitor of defense responses in aspen

2021 ◽  
Author(s):  
Evgeniy N. Donev ◽  
Marta Derba-Maceluch ◽  
Xiaokun Liu ◽  
Henri Colyn Bwanika ◽  
Izabela Dobrowolska ◽  
...  
Keyword(s):  
Defense Responses ◽  
Immune Defense ◽  
Cellulose Content ◽  
In Planta ◽  
Molecular Pattern ◽  
Microbial Enzyme ◽  
Carbohydrate Complex ◽  
Premature Leaf Senescence ◽  
Molecular Patterns ◽  
Damage Associated Molecular Patterns

Using microbial enzymes in transgenesis is a powerful means to introduce new functionalities in plants. Glucuronoyl esterase (GCE) is a microbial enzyme hydrolyzing the ester bond between lignin and 4-O-methyl-α-D-glucuronic acid present as a side chain of glucuronoxylan. This bond mediates lignin-carbohydrate complex (LCC) formation, considered as crucial factor of lignocellulose recalcitrance to saccharification. Previous studies showed that hybrid aspen (Populus tremula L. x tremuloides Michx.) constitutively expressing Phanerochaete carnosa Burt GCE (PcGCE) had better efficiency of cellulose-to-glucose conversion but were stunned and had lower cellulose content indicating that more studies are needed to design strategy for deployment of this enzyme in planta. Here we report that the transgenic plants exhibit premature leaf senescence, increased accumulation of calcium oxalate crystals, tyloses and necrotic lesions and have strongly activated immune defense reactions as revealed by their altered profiles of transcriptomes, metabolomes and hormones in the leaves. To elucidate if these effects are triggered by damage-associated molecular patterns (DAMPs) or by PcGCE protein perceived as a pathogen-associated molecular pattern (PAMP), we ectopically expressed in aspen an enzymatically inactive PcGCES217A. The mutated PcGCE induced similar growth retardation, leaf necrosis and premature senescence as the active one, providing evidence that PcGCE protein is recognized as PAMP. Transcriptomics analysis of young expanding leaves of 35S:PcGCE plants identified several candidates for receptors of PcGCE, which were not expressed in developing wood tissues. Grafting experiments showed that PcGCE transcripts are not cell-to-cell mobile and that leaves augment systemic responses. In agreement, expressing PcGCE in developing wood by using the wood-specific promoter (WP), avoided all off-target effects. Moreover, WP:PcGCE lines grew better than control plants providing evidence that this strategy can be used in transgenic crops dedicated for biorefinery.

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