Housekeeping gene selection for real-time RT-PCR normalization in potato during biotic and abiotic stress

Nicot, Nathalie; Hausman, Jean-François; Hoffmann, Lucien; Evers, Danièle

doi:10.1093/jxb/eri285

Cited by 1,172 publications

(916 citation statements)

References 45 publications

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“…qPCRs were performed in triplicate and included no-template controls using an ABI 7500 Real-Time PCR System (Applied Biosystems). Because several potential reference genes had previously been discovered for potato (27), we were able to select and validate two references genes that were deemed stable in our system using the geNorm function of the qbase PLUS software (Biogazelle, Zwijnaarde, Belgium): EF1-α and CyP. Target gene cycle thresholds (Cts) were normalized by subtracting the geometric mean of the Cts from both reference genes using the qbase PLUS software.…”

Section: Methodsmentioning

confidence: 99%

Long-Term Induction of Defense Gene Expression in Potato by Pseudomonas sp. LBUM223 and Streptomyces scabies

Arseneault¹,

Pieterse²,

Gérin-Ouellet³

et al. 2014

Phytopathology®

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Streptomyces scabies is a causal agent of common scab of potato, which generates necrotic tuber lesions. We have previously demonstrated that inoculation of potato plants with phenazine-1-carboxylic acid (PCA)-producing Pseudomonas sp. LBUM223 could significantly reduce common scab symptoms. In the present study, we investigated whether LBUM223 or an isogenic phzC-mutant not producing PCA could elicit an induced systemic resistance response in potato. The expression of eight defense-related genes (salicylic acid [SA]-related ChtA, PR-1b, PR-2, and PR-5; and jasmonic acid and ethylene-related LOX, PIN2, PAL-2, and ERF3) was quantified using newly developed TaqMan reversetranscription quantitative polymerase chain reaction assays in 5-and 10-week-old potted potato plants. Although only wild-type LBUM223 was capable of significantly reducing common scab symptoms, the presence of both LBUM223 and its PCA-deficient mutant were equally able to upregulate the expression of LOX and PR-5. The presence of S. scabies overexpressed all SA-related genes. This indicates that (i) upregulation of potato defense-related genes by LBUM223 is unlikely to contribute to common scab's control and (ii) LBUM223's capacity to produce PCA is not involved in this upregulation. These results suggest that a direct interaction occurring between S. scabies and PCA-producing LBUM223 is more likely involved in controlling common scab development.

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Section: Methodsmentioning

confidence: 99%

Long-Term Induction of Defense Gene Expression in Potato by Pseudomonas sp. LBUM223 and Streptomyces scabies

Arseneault¹,

Pieterse²,

Gérin-Ouellet³

et al. 2014

Phytopathology®

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“…The cDNA (1 µL) synthesized from the transgenic and untransformed control lines were taken as template for PCR using the GUS gene specific primers (5'-CCCTTACGCTGAAGAGATGC-3' and 5'-GAGCGTCGCAGAACATTACA-3') with same PCR condition. Actin was used as a loading control with PCR primers described by Nicot et al (2005). Reaction products were separated on 1% agarose gel stained with ethidium bromide and visualized under UV light.…”

Section: Reverse Transcriptase-pcr (Rt-pcr)mentioning

confidence: 99%

Effect of Explant, Genotype and Plant Growth Regulators on Regeneration and Agrobacterium-Mediated Transformation of Potato

Kamrani¹,

Ebadi²,

Shiri³

2015

J. of Agronomy

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A B S T R A C TThe objective of the present study is to develop an efficient protocol for plant regeneration and Agrobacterium-mediated transformation of potato cultivars Desiree, Agria and Marfona grown under Iranian agricultural conditions. The regeneration efficiency from internode, leaf and petiole explants was examined. The results from all Shoot Induction Medium (SIM) combinations and all cultivars together indicate highly efficient shoot regeneration from internode (4.56 shoots per explant). The response of petiole and leaf explants was lower, 3.83 and 2.55 shoots per explant, respectively. The highest efficiency of shoot regeneration was achieved with internode explants of cultivar Desiree (6.64 shoots per explant) on Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ) 1 mg LG 1 + 6-benzyladenine (BAP) 1 mg LG 1 . For plant transformation, internode explants were inoculated and co-cultivated with Agrobacterium tumefaciens strain LBA4404 harboring a binary vector pBI-121 containing β-glucuronidase (GUS) and nptII genes. Reverse transcriptase-polymerase chain reaction analysis and histochemical assay for β-glucuronidase indicated that the gene coding for this enzyme was integrated in the potato genome and could be expressed in potato tissues. The presence of nptII gene in the kanamycin resistant plants was verified by polymerase chain reaction analysis. The transformation frequency ranged from 22-42%.

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“…A 200-bp fragment of the ITS gene of V. dahliae was PCR-amplified using primers ITS1-F (Gardes and Bruns, 1993) and ST-VE1 (Lievens et al, 2006) on DNA samples using FirePol polymerase (Solis BioDyne). As an internal control, potato actin was amplified from the same templates using primers StActinF and StActinR (Nicot et al, 2005).…”

Section: Verticillium Dahliae Resistance Assaymentioning

confidence: 99%

The Effector SPRYSEC-19 ofGlobodera rostochiensisSuppresses CC-NB-LRR-Mediated Disease Resistance in Plants

et al. 2012

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The potato cyst nematode Globodera rostochiensis invades roots of host plants where it transforms cells near the vascular cylinder into a permanent feeding site. The host cell modifications are most likely induced by a complex mixture of proteins in the stylet secretions of the nematodes. Resistance to nematodes conferred by nucleotide-binding-leucine-rich repeat (NB-LRR) proteins usually results in a programmed cell death in and around the feeding site, and is most likely triggered by the recognition of effectors in stylet secretions. However, the actual role of these secretions in the activation and suppression of effector-triggered immunity is largely unknown. Here we demonstrate that the effector SPRYSEC-19 of G. rostochiensis physically associates in planta with the LRR domain of a member of the SW5 resistance gene cluster in tomato (Lycopersicon esculentum).

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Housekeeping gene selection for real-time RT-PCR normalization in potato during biotic and abiotic stress

Cited by 1,172 publications

References 45 publications

Long-Term Induction of Defense Gene Expression in Potato by Pseudomonas sp. LBUM223 and Streptomyces scabies

Long-Term Induction of Defense Gene Expression in Potato by Pseudomonas sp. LBUM223 and Streptomyces scabies

Effect of Explant, Genotype and Plant Growth Regulators on Regeneration and Agrobacterium-Mediated Transformation of Potato

The Effector SPRYSEC-19 ofGlobodera rostochiensisSuppresses CC-NB-LRR-Mediated Disease Resistance in Plants

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