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Antagonistic peptide technology for functional dissection of CLE peptides revisited.

Czyzewicz N, Wildhagen M, Cattaneo P, Stahl Y, Pinto KG, Aalen RB, Butenko MA, Simon R, Hardtke CS, De Smet I - J. Exp. Bot. (2015)

Bottom Line: Based on the analyses, it was concluded that the antagonistic peptide approach is not the ultimate means to overcome redundancy or lack of loss-of-function lines.However, information collected using antagonistic peptide approaches (in the broad sense) can be very useful, but these approaches do not work in all cases and require a deep insight on the interaction between the ligand and its receptor to be successful.This, as well as peptide ligand structure considerations, should be taken into account before ordering a wide range of synthetic peptide variants and/or generating transgenic plants.

View Article: PubMed Central - PubMed

Affiliation: Division of Plant and Crop Sciences, School of Biosciences, University of Nottingham, Loughborough LE12 5RD, UK.

No MeSH data available.


CLE45 peptide treatment and pCLE45::CLE45 transgenic lines. (A) Sequence of synthetic CLE45 and mCLE45 peptides used. (B) Primary root length following treatment of wild-type seedlings with indicated concentrations of CLE45p or mCLE45p6Thr. The bar graph indicates the mean ± standard error. Statistical significance (Student’s t-test) compared with mock is indicated for each time point (DAG, days after germination): * P <0.01. (C) Primary root length of pCLE45::CLE45 lines. The bar graph indicates the mean ± standard error. Statistical significance (Student’s t-test) compared with Col-0 is indicated: * P <0.01. (D) Confocal images of primary root meristems of 7-d-old seedlings (propidium iodide-stained; composite images). The asterisks highlight the two protophloem strands that can be distinguished in wild-type (Col-0) grown on mock (left), but that do not develop when grown on 10nM CLE45p (middle). Protophloem strands also do not develop in wild-type seedlings that express a pCLE45::CLE45 transgene (right). Scale bar, 100 µm.
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Figure 2: CLE45 peptide treatment and pCLE45::CLE45 transgenic lines. (A) Sequence of synthetic CLE45 and mCLE45 peptides used. (B) Primary root length following treatment of wild-type seedlings with indicated concentrations of CLE45p or mCLE45p6Thr. The bar graph indicates the mean ± standard error. Statistical significance (Student’s t-test) compared with mock is indicated for each time point (DAG, days after germination): * P <0.01. (C) Primary root length of pCLE45::CLE45 lines. The bar graph indicates the mean ± standard error. Statistical significance (Student’s t-test) compared with Col-0 is indicated: * P <0.01. (D) Confocal images of primary root meristems of 7-d-old seedlings (propidium iodide-stained; composite images). The asterisks highlight the two protophloem strands that can be distinguished in wild-type (Col-0) grown on mock (left), but that do not develop when grown on 10nM CLE45p (middle). Protophloem strands also do not develop in wild-type seedlings that express a pCLE45::CLE45 transgene (right). Scale bar, 100 µm.

Mentions: Among many processes (Cock and McCormick, 2001; Fiume and Fletcher, 2012; Hirakawa et al., 2008; Okamoto et al., 2013), various CLE peptides affect primary and lateral root growth and development (Czyzewicz et al., 2015; Depuydt et al., 2013; Fiers et al., 2005; Hobe et al., 2003; Jun et al., 2010; Rodriguez-Villalon et al., 2014; Rodriguez-Villalon et al., 2015; Stahl et al., 2009). To build on previous work investigating CLE peptides in the context of lateral root development, primary root growth, root apical stem cell maintenance, and vascular development, putative antagonistic versions of CLV3, CLE1/4, CLE7, CLE26, CLE27, CLE40, and CLE45 peptides were designed—based on the findings by Song et al. (2013)—to further unravel CLE peptide function (Figs 2A, 3A, 4A). To assess the function of these mutated chemically synthesized CLE peptides with Gly/cysteine (Cys) to Ala or Gly/Cys to Thr substitutions (referred to as mCLEpAla6 or mCLEp6Thr, respectively), a number of biological assays were used.


Antagonistic peptide technology for functional dissection of CLE peptides revisited.

Czyzewicz N, Wildhagen M, Cattaneo P, Stahl Y, Pinto KG, Aalen RB, Butenko MA, Simon R, Hardtke CS, De Smet I - J. Exp. Bot. (2015)

CLE45 peptide treatment and pCLE45::CLE45 transgenic lines. (A) Sequence of synthetic CLE45 and mCLE45 peptides used. (B) Primary root length following treatment of wild-type seedlings with indicated concentrations of CLE45p or mCLE45p6Thr. The bar graph indicates the mean ± standard error. Statistical significance (Student’s t-test) compared with mock is indicated for each time point (DAG, days after germination): * P <0.01. (C) Primary root length of pCLE45::CLE45 lines. The bar graph indicates the mean ± standard error. Statistical significance (Student’s t-test) compared with Col-0 is indicated: * P <0.01. (D) Confocal images of primary root meristems of 7-d-old seedlings (propidium iodide-stained; composite images). The asterisks highlight the two protophloem strands that can be distinguished in wild-type (Col-0) grown on mock (left), but that do not develop when grown on 10nM CLE45p (middle). Protophloem strands also do not develop in wild-type seedlings that express a pCLE45::CLE45 transgene (right). Scale bar, 100 µm.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
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Figure 2: CLE45 peptide treatment and pCLE45::CLE45 transgenic lines. (A) Sequence of synthetic CLE45 and mCLE45 peptides used. (B) Primary root length following treatment of wild-type seedlings with indicated concentrations of CLE45p or mCLE45p6Thr. The bar graph indicates the mean ± standard error. Statistical significance (Student’s t-test) compared with mock is indicated for each time point (DAG, days after germination): * P <0.01. (C) Primary root length of pCLE45::CLE45 lines. The bar graph indicates the mean ± standard error. Statistical significance (Student’s t-test) compared with Col-0 is indicated: * P <0.01. (D) Confocal images of primary root meristems of 7-d-old seedlings (propidium iodide-stained; composite images). The asterisks highlight the two protophloem strands that can be distinguished in wild-type (Col-0) grown on mock (left), but that do not develop when grown on 10nM CLE45p (middle). Protophloem strands also do not develop in wild-type seedlings that express a pCLE45::CLE45 transgene (right). Scale bar, 100 µm.
Mentions: Among many processes (Cock and McCormick, 2001; Fiume and Fletcher, 2012; Hirakawa et al., 2008; Okamoto et al., 2013), various CLE peptides affect primary and lateral root growth and development (Czyzewicz et al., 2015; Depuydt et al., 2013; Fiers et al., 2005; Hobe et al., 2003; Jun et al., 2010; Rodriguez-Villalon et al., 2014; Rodriguez-Villalon et al., 2015; Stahl et al., 2009). To build on previous work investigating CLE peptides in the context of lateral root development, primary root growth, root apical stem cell maintenance, and vascular development, putative antagonistic versions of CLV3, CLE1/4, CLE7, CLE26, CLE27, CLE40, and CLE45 peptides were designed—based on the findings by Song et al. (2013)—to further unravel CLE peptide function (Figs 2A, 3A, 4A). To assess the function of these mutated chemically synthesized CLE peptides with Gly/cysteine (Cys) to Ala or Gly/Cys to Thr substitutions (referred to as mCLEpAla6 or mCLEp6Thr, respectively), a number of biological assays were used.

Bottom Line: Based on the analyses, it was concluded that the antagonistic peptide approach is not the ultimate means to overcome redundancy or lack of loss-of-function lines.However, information collected using antagonistic peptide approaches (in the broad sense) can be very useful, but these approaches do not work in all cases and require a deep insight on the interaction between the ligand and its receptor to be successful.This, as well as peptide ligand structure considerations, should be taken into account before ordering a wide range of synthetic peptide variants and/or generating transgenic plants.

View Article: PubMed Central - PubMed

Affiliation: Division of Plant and Crop Sciences, School of Biosciences, University of Nottingham, Loughborough LE12 5RD, UK.

No MeSH data available.