Characterization of an allelic series in the <i>MONOPTEROS</i> gene of arabidopsis

Odat, Osama; Gardiner, Jason; Sawchuk, Megan G.; Verna, Carla; Donner, Tyler J.; Scarpella, Enrico

doi:10.1002/dvg.22729

Cited by 28 publications

(21 citation statements)

References 25 publications

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“…Total RNA was extracted as in (Chomczynski and Sacchi, 1987) from 4-day-old seedlings grown as in (Odat et al, 2014). RT-PCR was performed as in (Odat et al, 2014) with the “GN_qFb” and “GN_qRb” oligonucleotides (Table S19), and with the “ROC1 F” and “ROC1 R” oligonucleotides (Beeckman et al, 2002) (Table S19).…”

Section: Methodsmentioning

confidence: 99%

Coordination of Tissue Cell Polarity by Auxin Transport and Signaling

Verna

Ravichandran

Sawchuk

et al. 2019

Preprint

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Coordination of polarity between cells in tissues is key to multicellular organism development. In animals, coordination of this tissue cell polarity often requires direct cellcell interactions and cell movements, which are precluded in plants by a wall that separates cells and holds them in place; yet plants coordinate the polarity of hundreds of cells during the formation of the veins in their leaves. Overwhelming experimental evidence suggests that the plant signaling molecule auxin coordinates tissue cell polarity to induce vein formation, but how auxin does so is unclear. The prevailing hypothesis proposes that GNOM, a regulator of vesicle formation during protein trafficking, positions auxin transporters of the PIN-FORMED family to the correct side of the plasma membrane. The resulting cell-to-cell, polar transport of auxin would coordinate cell polarity and would induce vein formation. Here we tested this hypothesis by means of a combination of cellular imaging, molecular genetic analysis, and chemical induction and inhibition. Contrary to predictions of the hypothesis, we find that auxin-induced vein formation occurs in the absence of PIN-FORMED proteins or any known intercellular auxin transporter; that the auxin-transport-independent vein-patterning activity relies on auxin signaling; and that a GNOM-dependent signal that coordinates tissue cell polarity to induce vein formation acts upstream of both auxin transport and signaling. Our results reveal a synergism between auxin transport and signaling and their unsuspected control by GNOM in the coordination of cell polarity during vein patterning, one of the most spectacular and informative expressions of tissue cell polarization in plants.

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Section: Methodsmentioning

confidence: 99%

Coordination of Tissue Cell Polarity by Auxin Transport and Signaling

Verna

Ravichandran

Sawchuk

et al. 2019

Preprint

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“…7b). An extreme decrease in embryogenic capacity (over 80%) was observed in the culture of IZEs that had a strong, the so-called monopteros , mutant phenotype, which is characterised by a single cotyledon and the lack of the root meristem and hypocotyl (Odat et al 2014). …”

Section: Functional Test Of Se-transcribed Arfsmentioning

confidence: 99%

Expression profiling of AUXIN RESPONSE FACTOR genes during somatic embryogenesis induction in Arabidopsis

2017

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Key messageExtensive modulation of numerous ARF transcripts in the embryogenic culture of Arabidopsis indicates a substantial role of auxin signaling in the mechanism of somatic embryogenesis induction.AbstractSomatic embryogenesis (SE) is induced by auxin in plants and auxin signaling is considered to play a key role in the molecular mechanism that controls the embryogenic transition of plant somatic cells. Accordingly, the expression of AUXIN RESPONSE FACTOR (ARF) genes in embryogenic culture of Arabidopsis was analyzed. The study revealed that 14 of the 22 ARFs were transcribed during SE in Arabidopsis. RT-qPCR analysis indicated that the expression of six ARFs (ARF5, ARF6, ARF8, ARF10, ARF16, and ARF17) was significantly up-regulated, whereas five other genes (ARF1, ARF2, ARF3, ARF11, and ARF18) were substantially down-regulated in the SE-induced explants. The activity of ARFs during SE was also monitored with GFP reporter lines and the ARFs that were expressed in areas of the explants engaged in SE induction were detected. A functional test of ARFs transcribed during SE was performed and the embryogenic potential of the arf mutants and overexpressor lines was evaluated. ARFs with a significantly modulated expression during SE coupled with an impaired embryogenic response of the relevant mutant and/or overexpressor line, including ARF1, ARF2, ARF3, ARF5, ARF6, ARF8, and ARF11 were indicated as possibly being involved in SE induction. The study provides evidence that embryogenic induction strongly depends on ARFs, which are key regulators of the auxin signaling. Some clues on the possible functions of the candidate ARFs, especially ARF5, in the mechanism of embryogenic transition are discussed. The results provide guidelines for further research on the auxin-related functional genomics of SE and the developmental plasticity of somatic cells.Electronic supplementary materialThe online version of this article (doi:10.1007/s00299-017-2114-3) contains supplementary material, which is available to authorized users.

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“…Fluorophores were excited with the 488-nm line of a 30-mW Ar laser; GFP emission was collected with a BP 505-530 filter, and autofluorescence was collected between 550 and 754 nm. Mature leaves were fixed in 3 : 1 or 6 : 1 ethanol : acetic acid, rehydrated in 70% ethanol and in water, cleared briefly (few seconds to few minutes) -when necessary -in 0.4 M sodium hydroxide, washed in water, mounted in 80% glycerol or in 1 : 2 : 8 or 1 : 3 : 8 water : glycerol : chloral hydrate, and imaged as in (Odat et al, 2014). In the Fiji distribution (Schindelin et al, 2012) of ImageJ (Schindelin, Rueden, Hiner, & Eliceiri, 2015;Rueden et al, 2017;Schneider, Rasband, & Eliceiri, 2012), grayscaled RGB color images were turned into 8-bit images; when necessary, 8-bit images were combined into stacks, and maximum-intensity projection was applied to stacks; look-up-tables were applied to images or stacks, and brightness and contrast were adjusted by linear stretching of the histogram.…”

Section: Imagingmentioning

confidence: 99%

GAL4/GFP enhancer-trap lines for identification and manipulation of cells and tissues in developing Arabidopsis leaves

Amalraj

Govindaraju

Krishna

et al. 2019

Preprint

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AbstractUnderstanding developmental processes requires the unambiguous identification of cells and tissues, and the selective manipulation of the properties of those cells and tissues. Both requirements can most efficiently be satisfied through the use of GAL4/GFP enhancer-trap lines. No such lines are available, however, for the study of leaf development in the Columbia-0 reference genotype of Arabidopsis. Here we address this limitation by identifying and characterizing a set of GAL4/GFP enhancer-trap lines in the Columbia-0 background for the specific labeling of cells and tissues during leaf development, and for the targeted expression of genes of interest in those cells and tissues. By using one line in our set to resolve outstanding problems in leaf vein patterning, we show that these lines can be used to address key questions in plant developmental biology.

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Characterization of an allelic series in the MONOPTEROS gene of arabidopsis

Cited by 28 publications

References 25 publications

Coordination of Tissue Cell Polarity by Auxin Transport and Signaling

Coordination of Tissue Cell Polarity by Auxin Transport and Signaling

Expression profiling of AUXIN RESPONSE FACTOR genes during somatic embryogenesis induction in Arabidopsis

GAL4/GFP enhancer-trap lines for identification and manipulation of cells and tissues in developing Arabidopsis leaves

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