Selection of internal control genes for quantitative real-time RT-PCR studies during tomato development process

Expósito-Rodríguez, Marino; Borges, Andrés A.; Borges-Pérez, Andrés; Pérez, Jose Antonio Pérez

doi:10.1186/1471-2229-8-131

Cited by 679 publications

(541 citation statements)

References 41 publications

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“…2008). We used the total RNA (1–10 mg) of C. korshinskii leaves from Huangling and Dalad Banner to construct each de novo library, and Cluster and Java Treeview to establish similar gene expression patterns via cluster analyses of gene expression patterns (Gibbons and Roth 2002; Ning et al.…”

Section: Methodsmentioning

confidence: 99%

Adaptional evolution of trichome in Caragana korshinskii to natural drought stress on the Loess Plateau, China

Ning

Wang

Zhou

et al. 2016

Ecology and Evolution

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Caragana korshinskii is commonly employed to improve drought ecosystems on the Loess Plateau, although the molecular mechanism at work is poorly understood, particularly in terms of the plant's ability to tolerate drought stress. Water is the most severe limiting factor for plant growth on the Loess Plateau. The trichome is known to play an efficient role in reducing water loss through decreasing the rate of transpiration, so in this study, we focused on the trichome‐related gene expression of ecological adaptation in C. korshinskii under low precipitation conditions. In order to explore the responses of trichomes to drought, we selected two experimental sites from wet to dry along the Loess Plateau latitude gradient for observation. Micro‐phenomena through which trichomes grew denser and larger under reduced precipitation were observed using a scanning electron microscope; de novo transcriptomes and quantitative PCR were then used to explore and verify gene expression patterns of C. korshinskii trichomes. Results showed that GIS2,TTG1, and GL2 were upregulated (as key positive‐regulated genes on trichome development), while CPC was downregulated (negative‐regulated gene). Taken together, our data indicate that downstream genes of gibberellin and cytokinin signaling pathways, alongside several cytoskeleton‐related genes, contribute to modulating trichome development to enhance transpiration resistance ability and increase the resistance to drought stress in C. korshinskii.

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Section: Methodsmentioning

confidence: 99%

Adaptional evolution of trichome in Caragana korshinskii to natural drought stress on the Loess Plateau, China

Ning

Wang

Zhou

et al. 2016

Ecology and Evolution

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“…Gene expression analyses of SlCLH1 to SlCLH4 during fruit ripening in wild-type tomato. SlTIP41 was used as a control (Expó sito- Rodríguez et al, 2008). Expression was analyzed with the number of PCR cycles as indicated.…”

Section: Discussionmentioning

confidence: 99%

Different Mechanisms Are Responsible for Chlorophyll Dephytylation during Fruit Ripening and Leaf Senescence in Tomato

et al. 2014

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Chlorophyll breakdown occurs in different green plant tissues (e.g. during leaf senescence and in ripening fruits). For different plant species, the PHEOPHORBIDE A OXYGENASE (PAO)/phyllobilin pathway has been described to be the major chlorophyll catabolic pathway. In this pathway, pheophorbide (i.e. magnesium-and phytol-free chlorophyll) occurs as a core intermediate. Most of the enzymes involved in the PAO/phyllobilin pathway are known; however, the mechanism of dephytylation remains uncertain. During Arabidopsis (Arabidopsis thaliana) leaf senescence, phytol hydrolysis is catalyzed by PHEOPHYTINASE (PPH), which is specific for pheophytin (i.e. magnesium-free chlorophyll). By contrast, in fruits of different Citrus spp., chlorophyllase, hydrolyzing phytol from chlorophyll, was shown to be active. Here, we enlighten the process of chlorophyll breakdown in tomato (Solanum lycopersicum), both in leaves and fruits. We demonstrate the activity of the PAO/phyllobilin pathway and identify tomato PPH (SlPPH), which, like its Arabidopsis ortholog, was specifically active on pheophytin. SlPPH localized to chloroplasts and was transcriptionally up-regulated during leaf senescence and fruit ripening. SlPPH-silencing tomato lines were impaired in chlorophyll breakdown and accumulated pheophytin during leaf senescence. However, although pheophytin transiently accumulated in ripening fruits of SlPPH-silencing lines, ultimately these fruits were able to degrade chlorophyll like the wild type. We conclude that PPH is the core phytol-hydrolytic enzyme during leaf senescence in different plant species; however, fruit ripening involves other hydrolases, which are active in parallel to PPH or are the core hydrolases in fruits. These hydrolases remain unidentified, and we discuss the question of whether chlorophyllases might be involved.

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“…The PCR reactions were performed in a 96-well thermocycler (Bio-Rad iCycler) by starting with 3 min at 95°C, followed by 45 cycles consisting of 15 s at 95°C and 60 s at 60°C and a melt curve from 65°C to 95°C in 0.5°C increments per 10 s to verify the presence of a specific product. TIP41, SAND, CAC, and Expressed (Expósito- Rodríguez et al, 2008) were used as reference genes. Reference gene stability was confirmed with geNorm software.…”

Section: Qpcr Analysismentioning

confidence: 99%

A Co-Opted Hormonal Cascade Activates Dormant Adventitious Root Primordia upon Flooding in Solanum dulcamara

et al. 2016

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Selection of internal control genes for quantitative real-time RT-PCR studies during tomato development process

Cited by 679 publications

References 41 publications

Adaptional evolution of trichome in Caragana korshinskii to natural drought stress on the Loess Plateau, China

Adaptional evolution of trichome in Caragana korshinskii to natural drought stress on the Loess Plateau, China

Different Mechanisms Are Responsible for Chlorophyll Dephytylation during Fruit Ripening and Leaf Senescence in Tomato

A Co-Opted Hormonal Cascade Activates Dormant Adventitious Root Primordia upon Flooding in Solanum dulcamara

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