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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.


Distal root phenotypes after antagonistic peptide treatments. (A) Sequence of synthetic CLE and mCLE peptides used. (B–H) Distal root cell fates were analysed by mPSPI staining 5 d after germination in wild-type (Col-0) and cle40-2 mutant roots (C, D). Representative examples of Col-0 roots grown on media with 1 µM CLE40p (E), mCLE40p6Thr (F), CLV3p (G), and mCLV3p6Thr (H) are shown. Frequency of roots carrying starch granules in the designated domains is shown in (B). Arrowheads: blue, QC position; yellow, CSC position (D1); red, CC position (D2). Double yellow arrowheads indicate CSC fate in D2, whereas the lack of a yellow arrowhead indicates CC fate in D1 position. QC, quiescent centre position; D1, distal layer position one; D2, distal layer position two; CC, columella cell position. Scale bars represent 15 µm.
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Figure 3: Distal root phenotypes after antagonistic peptide treatments. (A) Sequence of synthetic CLE and mCLE peptides used. (B–H) Distal root cell fates were analysed by mPSPI staining 5 d after germination in wild-type (Col-0) and cle40-2 mutant roots (C, D). Representative examples of Col-0 roots grown on media with 1 µM CLE40p (E), mCLE40p6Thr (F), CLV3p (G), and mCLV3p6Thr (H) are shown. Frequency of roots carrying starch granules in the designated domains is shown in (B). Arrowheads: blue, QC position; yellow, CSC position (D1); red, CC position (D2). Double yellow arrowheads indicate CSC fate in D2, whereas the lack of a yellow arrowhead indicates CC fate in D1 position. QC, quiescent centre position; D1, distal layer position one; D2, distal layer position two; CC, columella cell position. Scale bars represent 15 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)

Distal root phenotypes after antagonistic peptide treatments. (A) Sequence of synthetic CLE and mCLE peptides used. (B–H) Distal root cell fates were analysed by mPSPI staining 5 d after germination in wild-type (Col-0) and cle40-2 mutant roots (C, D). Representative examples of Col-0 roots grown on media with 1 µM CLE40p (E), mCLE40p6Thr (F), CLV3p (G), and mCLV3p6Thr (H) are shown. Frequency of roots carrying starch granules in the designated domains is shown in (B). Arrowheads: blue, QC position; yellow, CSC position (D1); red, CC position (D2). Double yellow arrowheads indicate CSC fate in D2, whereas the lack of a yellow arrowhead indicates CC fate in D1 position. QC, quiescent centre position; D1, distal layer position one; D2, distal layer position two; CC, columella cell position. Scale bars represent 15 µm.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
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Figure 3: Distal root phenotypes after antagonistic peptide treatments. (A) Sequence of synthetic CLE and mCLE peptides used. (B–H) Distal root cell fates were analysed by mPSPI staining 5 d after germination in wild-type (Col-0) and cle40-2 mutant roots (C, D). Representative examples of Col-0 roots grown on media with 1 µM CLE40p (E), mCLE40p6Thr (F), CLV3p (G), and mCLV3p6Thr (H) are shown. Frequency of roots carrying starch granules in the designated domains is shown in (B). Arrowheads: blue, QC position; yellow, CSC position (D1); red, CC position (D2). Double yellow arrowheads indicate CSC fate in D2, whereas the lack of a yellow arrowhead indicates CC fate in D1 position. QC, quiescent centre position; D1, distal layer position one; D2, distal layer position two; CC, columella cell position. Scale bars represent 15 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.