scholarly journals Expression analysis of defense-related genes in cucumber (Cucumis sativus L.) against Phytophthora melonis

2020 ◽  
Author(s):  
Lida Hashemi ◽  
Ahmad Reza Golparvar ◽  
Mehdi Nasr Esfahani ◽  
Maryam Golabadi
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Molecular Mechanisms ◽  
Crown Rot ◽  
Economic Losses ◽  
Post Inoculation ◽  
Expression Ratio ◽  
Phytophthora Melonis ◽  
Maximum Expression

AbstractPhytophthora melonis is the causal agent of damping-off or crown rot, one of the most destructive cucumber diseases that causes severe economic losses in Iran and some other parts of the world. Despite intense research efforts made in the past years, no permanent cure currently exists for this disease. With the aim to understand the molecular mechanisms of defense against P. melonis, root collars and leaves of four cucumber genotypes consisting of resistant Ramezz; moderately resistant Baby and very susceptible Mini 6-23 and Extrem, were monitored for quantitative gene expression analysis of five antifungal and/or anti-oomycete genes (CsWRKY20, CsLecRK6.1, PR3, PR1-1a and LOX1) at three points after inoculation with P. melonis. The gene expression analysis indicated that P. melonis strongly enhanced the expression of these genes after inoculation in both leaves and root collars. Further, not only the transcript levels of these genes were significantly higher in the resistant and moderately resistance genotypes, but also the time point of the highest relative expression ratio for the five genes was different in the four cucumber genotypes. CsWRKY20 and PR3 showed the maximum expression in Ramezz at 48 hours post inoculation (hpi) while CsLecRK6.1, and LOX1 showed the highest expression at 72 hpi. In addition, PR1-1a showed the maximum expression in the Baby at 72 hpi. Root collars responded faster than leaves and some responses were more strongly up-regulated in root collars than in leaves. The genes found to be involved in disease resistance in two different organs of cucumber after pathogen infection. The results suggest that increased expression of these genes led to activation of defense pathways and could be responsible for a reduced P. melonis colonization capacity in Ramezz and Baby. Overall, this work represents a valuable resource for future functional genomics studies to unravel the molecular mechanisms of C. sativus- P. melonis interaction.

scholarly journals Reference genes for gene expression analysis in the fungal pathogen Neonectria ditissima and their use demonstrating expression up-regulation of candidate virulence genes

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0238157
Author(s):  
Liz M. Florez ◽  
Reiny W. A. Scheper ◽  
Brent M. Fisher ◽  
Paul W. Sutherland ◽  
Matthew D. Templeton ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Reference Genes ◽  
Time Course ◽  
Gene Expression Analysis ◽  
Virulence Genes ◽  
Functional Characterization ◽  
Post Inoculation ◽  
In Planta ◽  
Regulation Of Expression

European canker, caused by the necrotrophic fungal phytopathogen Neonectria ditissima, is one of the most damaging apple diseases worldwide. An understanding of the molecular basis of N. ditissima virulence is currently lacking. Identification of genes with an up-regulation of expression during infection, which are therefore probably involved in virulence, is a first step towards this understanding. Reverse transcription quantitative real-time PCR (RT-qPCR) can be used to identify these candidate virulence genes, but relies on the use of reference genes for relative gene expression data normalisation. However, no report that addresses selecting appropriate fungal reference genes for use in the N. ditissima-apple pathosystem has been published to date. In this study, eight N. ditissima genes were selected as candidate RT-qPCR reference genes for gene expression analysis. A subset of the primers (six) designed to amplify regions from these genes were specific for N. ditissima, failing to amplify PCR products with template from other fungal pathogens present in the apple orchard. The efficiency of amplification of these six primer sets was satisfactory, ranging from 81.8 to 107.53%. Analysis of expression stability when a highly pathogenic N. ditissima isolate was cultured under 10 regimes, using the statistical algorithms geNorm, NormFinder and BestKeeper, indicated that actin and myo-inositol-1-phosphate synthase (mips), or their combination, could be utilised as the most suitable reference genes for normalisation of N. ditissima gene expression. As a test case, these reference genes were used to study expression of three candidate virulence genes during a time course of infection. All three, which shared traits with fungal effector genes, had up-regulated expression in planta compared to in vitro with expression peaking between five and six weeks post inoculation (wpi). Thus, these three genes may well be involved in N. ditissima pathogenicity and are priority candidates for further functional characterization.

scholarly journals Reference genes for gene expression analysis in the fungal pathogen Neonectria ditissima and their use demonstrating expression up-regulation of candidate virulence genes

2020 ◽  
Author(s):  
Liz M. Florez ◽  
Reiny W. A. Scheper ◽  
Brent M. Fisher ◽  
Paul W. Sutherland ◽  
Matthew D. Templeton ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Reference Genes ◽  
Time Course ◽  
Gene Expression Analysis ◽  
Virulence Genes ◽  
Functional Characterization ◽  
Post Inoculation ◽  
In Planta ◽  
Qrt Pcr

AbstractEuropean canker, caused by the necrotrophic fungal phytopathogen Neonectria ditissima, is one of the most damaging apple diseases worldwide. An understanding of the molecular basis of N. ditissima virulence is currently lacking. Identification of genes with an up-regulation of expression during infection, which are therefore probably involved in virulence, is a first step towards this understanding. Real-time quantitative reverse transcription PCR (qRT-PCR) can be used to identify these candidate virulence genes, but relies on the use of reference genes for relative gene expression data normalisation. However, no report that addresses selecting appropriate fungal reference genes for use in the N. ditissima-apple pathosystem has been published to date. In this study, eight N. ditissima genes were selected as candidate qRT-PCR reference genes for gene expression analysis. A subset of the primers (six) designed to amplify regions from these genes were specific for N. ditissima, failing to amplify PCR products with template from other fungal pathogens present in the apple orchard. The efficiency of amplification of these six primer sets was satisfactory, ranging from 81.8 to 107.53%. Analysis of expression stability when a highly pathogenic N. ditissima isolate was cultured under 10 regimes, using the statistical algorithms geNorm, NormFinder and BestKeeper, indicated that actin and myo-inositol-1-phosphate synthase (mips), or their combination, could be utilised as the most suitable reference genes for normalisation of N. ditissima gene expression. As a test case, these reference genes were used to study expression of three candidate virulence genes during a time course of infection. All three, which shared traits with fungal effector genes, had up-regulated expression in planta compared to in vitro with expression peaking between five and six weeks post inoculation (wpi). Thus, these three genes may well be involved in N. ditissima pathogenicity and are priority candidates for further functional characterization.

scholarly journals Identification of Genes Involved in the Response of Banana to Crown Rot Disease

2011 ◽  
Vol 24 (1) ◽  
pp. 143-153 ◽  
Author(s):  
Ludivine Lassois ◽  
Patrick Frettinger ◽  
Luc de Lapeyre de Bellaire ◽  
Philippe Lepoivre ◽  
Haissam Jijakli
Keyword(s):  
Gene Expression ◽  
Molecular Mechanisms ◽  
Cellulose Synthase ◽  
Crown Rot ◽  
Post Inoculation ◽  
Banana Fruit ◽  
High Susceptibility ◽  
Pathogenesis Related Protein ◽  
Host Pathogen ◽  
Rot Disease

Variations in banana susceptibility to crown rot disease have been observed but the molecular mechanisms underlying these quantitative host–pathogen relationships are still unknown. This study was designed to compare gene expression between crowns of banana fruit showing a high susceptibility (S+) and crowns showing a low susceptibility (S–) to the disease. Comparisons were performed at two situation times: i) between crowns (S+ and S–) collected 1 h before inoculation and ii) between crowns (S+ and S–) collected 13 days after inoculation. Gene expression comparisons were performed with cDNA-amplified fragment length polymorphism (AFLP) and results were confirmed by real-time reverse-transcription polymerase chain reaction. Among genes identified as differentially expressed between S+ and S– crowns, two were involved in signal transduction, three in proteolytic machinery, two had similarity to pathogenesis-related protein 14, one to a CCR4-associated factor protein, and one to a cellulose synthase. Paradoxically, the overexpression of the cellulose synthase gene was associated with banana showing a high susceptibility in both pre- and post-inoculation situations. Finally, the cDNA-AFLP identified a gene that seems to be associated with the quantitative banana responses to crown rot disease; this gene encodes a dopamine-β-monooxygenase, which is involved in the catecholamine pathway. To our knowledge, this work is the first to address both pre- and post-infection gene expression with the same host–pathogen combination and distinct susceptibility levels.

Gene expression analysis of molecular mechanisms of defense induced in Medicago truncatula parasitized by Orobanche crenata

2009 ◽  
Vol 47 (7) ◽  
pp. 635-641 ◽  
Author(s):  
José Vicente Die ◽  
Clara I. González Verdejo ◽  
Miguel Á. Dita ◽  
Salvador Nadal ◽  
Belén Román
Keyword(s):  
Gene Expression ◽  
Medicago Truncatula ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Molecular Mechanisms ◽  
Orobanche Crenata

scholarly journals 292 Differential gene expression analysis for piglets supplied dietary prebiotics and arachidonic acid for gastrointestinal disturbances

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 122-123
Author(s):  
Yuqing He ◽  
Sheila Jacobi ◽  
Christian Maltecca ◽  
Jack Odle
Keyword(s):  
Gene Expression ◽  
Arachidonic Acid ◽  
Expression Analysis ◽  
Fixed Effects ◽  
Gene Expression Analysis ◽  
Microbial Colonization ◽  
Economic Losses ◽  
Gi Tract ◽  
Swine Industry ◽  
Dss Colitis

Abstract Gastrointestinal (GI) disturbances cause significant economic losses in the swine industry. Dietary-based approaches have been well applied to control the inflammation and optimize microbial colonization of the GI tract. This study aimed to screen differentially expressed (DE) RNAs in piglets supplied with arachidonic acid (ARA) or prebiotics in an acute dextran sodium sulfate (DSS) colitis model. Suckling pigs (n = 48) from 6 litters were randomly assigned into 4 diet groups: 0.5% ARA, 0.5% ARA + PRE (4g/L galactooligosaccharide +4 g/L polydextrose), 2.5% ARA, and 2.5% ARA + PRE. On day 17–21, half of the pigs (n = 24) were treated with 0.625 g DSS/kg BW to induce colitis. RNA samples (n = 48) were isolated from the mucosal layer of GI tract on day 22 for the gene expression analysis via RNAseq. The GLIMMIX procedure of SAS (v.9.4) was used to fit the statistical models to gene counts, were the main effects and interactions of PRE, ARA, and DSS were fit in the model as fixed effects and litter as random. Genes with FDR < 0.05 were mapped to the KEGG pathway. A total of 133 DE genes (88 up-regulated, 45 down-regulated) were identified in pigs with colitis compared to healthy ones. PRE and ARA affected gene expression differently but with no interaction effect. PRE supplement decreased the total DE genes from 83 (59 up-regulated, 24 down-regulated) to 33 (16 up-regulated and 17 down-regulated) in pigs with colitis compared with healthy ones. A total of 37 DE genes (21 up-regulated, 16 down-regulated), and 49 DE genes (32 up-regulated, 17 down-regulated) were identified in pigs with colitis compared to healthy ones supplied with 0.5% and 2.5% ARA, respectively. DE genes identified in this study are involved in various signaling pathways with supplied prebiotics significantly changing the gene expression in pigs afflicted with colitis.

scholarly journals Selection of Reference Genes for Normalization of Real-Time PCR Data in Calliptamus italicus (Orthoptera: Acrididae) Under Different Temperature Conditions

2019 ◽  
Vol 19 (6) ◽  
Author(s):  
Hongxia Hu ◽  
Xiaofang Ye ◽  
Han Wang ◽  
Rong Ji
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Expression Analysis ◽  
Reference Genes ◽  
Gene Expression Analysis ◽  
Target Gene ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Quantitative Polymerase Chain Reaction ◽  
Target Gene Expression

Abstract Global warming has dominated worldwide climate change trends, and adaptability to high temperatures is the main factor underlying the spread of the pest Calliptamus italicus in Xinjiang Province, China. However, knowledge about the molecular mechanisms responsible for this adaptability and other related biological properties of C. italicus remain relatively unclear. Real-time quantitative polymerase chain reaction (RT-qPCR) is a key tool for gene expression analysis associated with various biological processes. Reference genes are necessary for normalizing gene expression levels across samples taken from specific experimental conditions. In this study, transcript level of five genes (GAPDH, 18S, TUB, ACT, and EF1α), commonly used as reference genes, were evaluated under nine different temperatures (27, 30, 33, 36, 39, 42, 45, 48, and 51°C) to assess their expression stability and further select the most suitable to be used on normalization of target gene expression data. Gene expression profiles were analyzed using geNorm, NormFinder, and BestKeeper software packages. The combined results demonstrated that the best-ranked reference genes for C. italicus are EF1α, GAPDH, and ACT under different thermal stress conditions. This is the first study that assesses gene expression analysis across a range of temperatures to select the most appropriate reference genes for RT-qPCR data normalization in C. italicus. These results should assist target gene expression analysis associated with heat stress in C. italicus.

Sign in / Sign up

Export Citation Format

Share Document