scholarly journals Validation of Reference Genes for Robust qRT-PCR Gene Expression Analysis in the Rice Blast Fungus Magnaporthe oryzae

PLoS ONE ◽  
2016 ◽  
Vol 11 (8) ◽  
pp. e0160637 ◽  
Author(s):  
Sarena Che Omar ◽  
Michael A. Bentley ◽  
Giulia Morieri ◽  
Gail M. Preston ◽  
Sarah J. Gurr
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Magnaporthe Oryzae ◽  
Reference Genes ◽  
Rice Blast ◽  
Gene Expression Analysis ◽  
Rice Blast Fungus ◽  
Qrt Pcr ◽  
Blast Fungus

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 Selection and validation of reference genes desirable for gene expression analysis by qRT-PCR in MeJA-treated ginseng hairy roots

PLoS ONE ◽  
2019 ◽  
Vol 14 (12) ◽  
pp. e0226168 ◽  
Author(s):  
Li Li ◽  
Kangyu Wang ◽  
Mingzhu Zhao ◽  
Shaokun Li ◽  
Yue Jiang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Hairy Roots ◽  
Reference Genes ◽  
Gene Expression Analysis ◽  
Qrt Pcr

scholarly journals Validation of reference genes for expression analysis in a murine trauma model combining traumatic brain injury and femoral fracture

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Ellen Otto ◽  
Paul Köhli ◽  
Jessika Appelt ◽  
Stefanie Menzel ◽  
Melanie Fuchs ◽  
...  
Keyword(s):  
Gene Expression ◽  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Reference Gene ◽  
Expression Analysis ◽  
Bone Fracture ◽  
Reference Genes ◽  
Gene Expression Analysis ◽  
Tissue Specific ◽  
Qrt Pcr

Abstract Systemic and local posttraumatic responses are often monitored on mRNA expression level using quantitative real-time PCR (qRT-PCR), which requires normalisation to adjust for confounding sources of variability. Normalisation requests reference (housekeeping) genes stable throughout time and divergent experimental conditions in the tissue of interest, which are crucial for a reliable and reproducible gene expression analysis. Although previous animal studies analysed reference genes following isolated trauma, this multiple-trauma gene expression analysis provides a notable study analysing reference genes in primarily affected (i.e. bone/fracture callus and hypothalamus) and secondarily affected organs (i.e. white adipose tissue, liver, muscle and spleen), following experimental long bone fracture and traumatic brain injury. We considered tissue-specific and commonly used top-ranked reference candidates from different functional groups that were evaluated applying the established expression stability analysis tools NormFinder, GeNorm, BestKeeper and RefFinder. In conclusion, reference gene expression in primary organs is highly time point as well as tissue-specific, and therefore requires careful evaluation for qRT-PCR analysis. Furthermore, the general application of Ppia, particularly in combination with a second reference gene, is strongly recommended for the analysis of systemic effects in the case of indirect trauma affecting secondary organs through local and systemic pathophysiological responses.

scholarly journals Investigating the cell and developmental biology of plant infection by the rice blast fungus Magnaporthe oryzae

2021 ◽  
pp. 103562
Author(s):  
Alice Bisola Eseola ◽  
Lauren S. Ryder ◽  
Míriam Osés-Ruiz ◽  
Kim Findlay ◽  
Xia Yan ◽  
...  
Keyword(s):  
Developmental Biology ◽  
Magnaporthe Oryzae ◽  
Rice Blast ◽  
Rice Blast Fungus ◽  
Plant Infection ◽  
Blast Fungus

scholarly journals Tanzawaic acids I–L: Four new polyketides from Penicillium sp. IBWF104-06

2014 ◽  
Vol 10 ◽  
pp. 251-258 ◽  
Author(s):  
Louis P Sandjo ◽  
Eckhard Thines ◽  
Till Opatz ◽  
Anja Schüffler
Keyword(s):  
Soil Sample ◽  
Magnaporthe Oryzae ◽  
Rice Blast ◽  
Rice Blast Fungus ◽  
Fungal Strain ◽  
Conidial Germination ◽  
Penicillium Sp ◽  
Culture Filtrates ◽  
Blast Fungus ◽  
New Compounds

Four new polyketides have been identified in culture filtrates of the fungal strain Penicillium sp. IBWF104-06 isolated from a soil sample. They are structurally based on the same trans-decalinpentanoic acid skeleton as tanzawaic acids A–H. One of the new compounds was found to inhibit the conidial germination in the rice blast fungus Magnaporthe oryzae at concentrations of 25 μg/mL.

scholarly journals Septin-Mediated Plant Cell Invasion by the Rice Blast Fungus, Magnaporthe oryzae

Science ◽  
2012 ◽  
Vol 336 (6088) ◽  
pp. 1590-1595 ◽  
Author(s):  
Y. F. Dagdas ◽  
K. Yoshino ◽  
G. Dagdas ◽  
L. S. Ryder ◽  
E. Bielska ◽  
...  
Keyword(s):  
Plant Cell ◽  
Cell Invasion ◽  
Magnaporthe Oryzae ◽  
Rice Blast ◽  
Rice Blast Fungus ◽  
Blast Fungus
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