70 research outputs found
CsPDR8 and CsPDR12, two of the 16 pleiotropic drug resistance genes in cucumber, are transcriptionally regulated by phytohormones and auxin herbicide in roots
Reference Gene Selection for Quantitative Real-time PCR Normalization in Quercus suber
The use of reverse transcription quantitative PCR technology to assess gene expression levels requires an accurate normalization of data in order to avoid misinterpretation of experimental results and erroneous analyses. Despite being the focus of several transcriptomics projects, oaks, and particularly cork oak (Quercus suber), have not been investigated regarding the identification of reference genes suitable for the normalization of real-time quantitative PCR data. In this study, ten candidate reference genes (Act, CACs, EF-1Îą, GAPDH, His3, PsaH, Sand, PP2A, Ă-Tub and Ubq) were evaluated to determine the most stable internal reference for quantitative PCR normalization in cork oak. The transcript abundance of these genes was analysed in several tissues of cork oak, including leaves, reproduction cork, and periderm from branches at different developmental stages (1-, 2-, and 3-year old) or collected in different dates (active growth period versus dormancy). The three statistical methods (geNorm, NormFinder, and CV method) used in the evaluation of the most suitable combination of reference genes identified Act and CACs as the most stable candidates when all the samples were analysed together, while Ă-Tub and PsaH showed the lowest expression stability. However, when different tissues, developmental stages, and collection dates were analysed separately, the reference genes exhibited some variation in their expression levels. In this study, and for the first time, we have identified and validated reference genes in cork oak that can be used for quantification of target gene expression in different tissues and experimental conditions and will be useful as a starting point for gene expression studies in other oaks
CsPDR8 and CsPDR12, two of the 16 pleiotropic drug resistance genes in cucumber, are transcriptionally regulated by phytohormones and auxin herbicide in roots
Quantitative RT-PCR analysis of differentially expressed genes in Quercus suber in response to Phytophthora cinnamomi infection
cDNA-AFLP methodology was used to gain insight into gene fragments differentially present in the mRNA profiles of Quercus suber roots infected with zoospores of Phytophthora cinnamomi at different post challenge time points. Fifty-three transcript-derived fragments (TDFs) were identified and sequenced. Six candidate genes were selected based on their expression patterns and homology to genes known to play a role in defence. They encode a cinnamyl alcohol dehydrogenase2 (QsCAD2), a protein disulphide isomerase (QsPDI), a CC-NBS-LRR resistance protein (QsRPc), a thaumatin-like protein (QsTLP), a chitinase (QsCHI) and a 1,3-β-glucanase (QsGlu). Evaluation of the expression of these genes by quantitative polymerase chain reaction (qPCR) revealed that transcript levels of QsRPc, QsCHI, QsCAD2 and QsPDI increased during the first 24 h post-inoculation, while those of thaumatin-like protein decreased. No differential expression was observed for 1,3-β-glucanase (QsGlu).Four candidate reference genes, polymerase II (QsRPII), eukaryotic translation initiation factor 5A (QsEIF-5A), β-tubulin (QsTUB) and a medium subunit family protein of clathrin adaptor complexes (QsCACs) were assessed to determine the most stable internal references for qRT-PCR normalization in the Phytophthora-Q. suber pathosystem in root tissues. Those found to be more stable, QsRPII and QsCACs, were used as internal reference in the present work.Knowledge on the Quercus defence mechanisms against biotic stress is scarce. This study provides an insight into the gene profiling of a few important genes of Q. suber in response to P. cinnamomi infection contributing to the knowledge of the molecular interactions involving Quercus and root pathogens that can be useful in the future to understand the mechanisms underlying oak resistance to soil-borne oomycetes.Peer Reviewe
Strategies in a metallophyte species to cope with manganese excess
The effect of exposure to high Mn concentration
was studied in a metallophyte species, Erica
andevalensis, using hydroponic cultures with a range
of Mn concentrations (0.06, 100, 300, 500, and
700 mg L-1). At harvest, biomass production, element
uptake, and biochemical indicators of metal
stress (leaf pigments, organic acids, amino acids,
phenols, and activities of catalase, peroxidase, superoxide
dismutase) were determined in leaves and roots.
Increasing Mn concentrations led to a decrease in
biomass accumulation, and tip leaves chlorosis was
the only toxicity symptom detected. In a similar way,
photosynthetic pigments (chlorophylls a and b, and
carotenoids) were affected by high Mn levels. Among
organic acids, malate and oxalate contents in roots
showed a significant increase at the highest Mn
concentration, while in leaves, Mn led to an increasing
trend in citrate and malate contents. An increase of Mn also induced an increase in superoxide dismutase
activity in roots and catalase activity in leaves. As
well, significant changes in free amino acids were
induced by Mn concentrations higher than
300 mg L-1, especially in roots. No significant
changes in phenolic compounds were observed in
the leaves, but root phenolics were significantly
increased by increasing Mn concentrations in treatments.
When Fe supply was increased 10 and 20 times
(7â14 mg Fe L-1 as Fe-EDDHA) in the nutrient
solutions at the highest Mn concentration
(700 mg Mn L-1), it led to significant increases in
photosynthetic pigments and biomass accumulation.
Manganese was mostly accumulated in the roots, and
the species was essentially a Mn excluder. However,
considering the high leaf Mn concentration recorded
without toxicity symptoms, E. andevalensis might be
rated as a Mn-tolerant speciesinfo:eu-repo/semantics/publishedVersio
Genome-wide identification, classification and transcriptional analysis of nitrate and ammonium transporters in Coffea
Selection and validation of reference genes for quantitative RT-PCR expression studies of the non-model crop Musa
Cucumber HMA5A and HMA5B are two tonoplast-localized homologs of Arabidopsis thaliana heavy metal ATPase HMA5 involved in cellular heavy metal homeostasis
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