A dual function of the IDA peptide in regulating cell separation and modulating plant immunity at the molecular level
- Section for Genetics and Evolutionary Biology, Department of Biosciences, University of Oslo, Norway
- Institute for Developmental Genetics and Cluster of Excellence on Plant Sciences, Heinrich Heine University, Germany
- Gregor Mendel Institute (GMI), Austrian Academy of Sciences, Vienna Biocenter (VBC), Austria
Figures
Figure 1 with 7 supplements
mIDA-induced [Ca2+]cyt release in Arabidopsis roots correlates with pHAE and pHSL2 activity.
(a) Normalized R-GECO1 fluorescence intensities (ΔF/F) were measured from regions of interest (ROI) (upper panel, outlined in red) in the meristematic and elongation zone of the root. Fluorescence …
Figure 1—figure supplement 1
IDA induces ROS production in Arabidopsis.
(a) ROS production detected by the luminol-based assay was monitored over time as RLU. ROS production from hae hsl2 leaf disks expressing 35 S:HAE-YFP in response to 1 μM mIDA (black) and in WT …
Figure 1—figure supplement 2
mIDA-induced [Ca2+]cyt release in Arabidopsis roots.
Shown are cytosolic calcium concentration ([Ca2+]cyt) dynamics in the ROI in response to 1 µM mIDA over time. Normalized R-GECO1 fluorescence intensities (ΔF/F) were measured from a region of …
Figure 1—figure supplement 3
flg22-induced [Ca2+]cyt release in Arabidopsis roots.
Shown are cytosolic calcium concentration ([Ca2+]cyt) dynamics in the ROI in response to 1 µM flg22 over time. Normalized R-GECO1 fluorescence intensities (ΔF/F) were measured from a region of …
Figure 1—figure supplement 4
The IDA triggered increase in [Ca2+]cyt is abolished in the presence of Ca2+ inhibitors.
(a) Increase in [Ca2+]cyt in 7 days old seedlings expressing the cytosolic localized Aequorin-based luminescence Ca2+ sensor (Aeq) measured in relative light units (RLU) treated with 1 μM mIDA …
Figure 1—video 1
[Ca2+]cyt dynamics in R-GECO1 expressing root tip in response to 1 µM mIDA.
Ten-days-old roots were treated with 1 µM mIDA and changes in [Ca2+]cyt was recorded over time. Response shown as normalized fluorescence intensities (∆F/F). Images were recorded with a frame rate …
Figure 1—video 2
[Ca2+]cyt dynamics in R-GECO1 expressing root tip in response to 1 µM flg22.
Ten days-old roots were treated with 1 µM flg22 and changes in [Ca2+]cyt was recorded over time. Response shown as normalized fluorescence intensities (∆F/F). Images were recorded with a frame rate …
Figure 1—video 3
[Ca2+]cyt dynamics in R-GECO1 expressing root tip in response to 1 mM eATP.
1 mM eATP induces a strong increase in [Ca2+]cyt and was added as a last treatment to all roots acting as a positive control. Response shown as normalized fluorescence intensities (∆F/F). Images …
Figure 2 with 6 supplements
mIDA-induced [Ca2+]cyt release in Arabidopsis abscission zones.
(a) Normalized R-GECO1 fluorescence intensities (ΔF/F) were measured from regions of interest (ROI) (outlined in red) in floral abscission zone (AZ)s. Shown are cytosolic calcium concentration ([Ca2+…
Figure 2—figure supplement 1
mIDA induces a Ca2+ response in flowers.
(a) Arabidopsis flowers and siliques at different developmental stages. Flowers along the main inflorescence are counted from the first flower with visible white petals at the top of the …
Figure 2—figure supplement 2
mIDA-induced [Ca2+]cyt release in Arabidopsis abscission zones.
Normalized R-GECO1 fluorescence intensities (ΔF/F) were measured from regions of interest (ROIs) in floral abscission zone (AZ)s. Shown are cytosolic calcium concentration ([Ca2+]cyt) dynamics in …
Figure 2—figure supplement 3
mIDAΔN69 does not induce a [Ca2+]cyt release in Arabidopsis abscission zones (AZ).
Normalized R-GECO1 fluorescence intensities (ΔF/F) were measured from regions of interest (ROI) (outlined in red) in the floral AZs. Shown are cytosolic calcium concentration ([Ca2+]cyt) dynamics in …
Figure 2—figure supplement 4
Investigation of CNGCs and RBOHs in the abscission process.
(a) Relative expression of CNGC1, CNGC2, CNGC4, CNGC5, CNGC6, CNGC9, CNGC12, RBOHD, and RBOHF in the floral organs during the onset of abscission (Cai and Lashbrook, 2008). X-axis represents the …
Figure 2—video 1
[Ca2+]cyt dynamics in R-GECO1 expressing AZ in response to 1 µM mIDA.
Flowers at position 6 were treated with 1 µM mIDA and changes in [Ca2+]cyt was recorded over time. Response shown as normalized fluorescence intensities (∆F/F). Images were recorded with a frame …
Figure 2—video 2
[Ca2+]cyt dynamics in R-GECO1 expressing AZ in response to to 1 µM of the inactive IDA peptide, IDAΔN69.
Flowers at position 6 were treated with 1 µM IDAΔN69 and changes in [Ca2+]cyt was recorded over time. Response shown as normalized fluorescence intensities (∆F/F). Images were recorded with a frame …
Figure 3
IDA is induced by biotic and abiotic stress.
Representative pictures of pIDA:GUS expression after 12 hr treatment with 1 µM flg22, chitin, NaCl, Mannitol and 1 µM IDAΔN69 in (a) cells surrounding emerging lateral roots, and (b) in the main …
Figure 4
mIDA induce callose deposition in Arabidopsis cotelydons expressing 35 S::HAE-YFP.
Callose deposition in Col-0 WT, 35 S:HAE in Col-0 and fls2 treated with water (mock treatment), 1 μM mIDA or 1 uM flg22. (a) Callose deposition could be observed in cotyledons of eight day old Col-0 …
Figure 5 with 2 supplements
mIDA induced transcription of defense-associated marker genes.
(a) Transcripts of FRK1, MYB51, and PEP3 in WT Col-0 seedlings exposed to 1 µM mIDA for 1 hr (red) and to 1 µM mIDA for 12 hr (blue) compared to untreated tissue (black). RNA levels were measured by …
Figure 5—figure supplement 1
mIDA induced enhanced transcription of defense-associated marker genes is only partially dependent on the HAE and HSL2 receptors.
RT-qPCR data showing transcription of FRK1, MYB51, and PEP3 in Col-0 WT and hae hsl2 seedlings exposed to 1 µM mIDA for 1 hr compared to untreated seedlings (No treatment). ACTIN was used to …
Figure 5—figure supplement 2
mIDA and flg22 induced expression of pMYB51:YFP.
Representative microscopic pictures of 7-days-old pMYB51:YFPN roots from seedlings transferred to liquid medium with 1 μM flg22 or 1 μM mIDA for 1, 3, 5, and 7 hr using a fluorescent microscope. …
Figure 6 with 1 supplement
mIDA and flg22 co-treatment enhances the transcription of defense-associated marker genes.
(a) RT-qPCR data showing transcription of FRK1, MYB51, and PEP3 in Col-0 WT seedlings exposed to 1 µM mIDA, 1 µM flg22 or a combination of 1 µM mIDA and 1 µM flg22 for 1 hr compared to untreated …
Figure 6—figure supplement 1
mIDA and PIP1 co-treatment do not enhances transcription of defense-associated marker genes.
RT-qPCR data showing transcription of FRK1, MYB51, and PEP3 in Col-0 WT seedlings exposed to 1 μM mIDA, 1 μM PIP1 or a combination of 1 μM mIDA and 1 μM PIP1 for 1 hr compared to untreated seedlings …
Figure 7 with 1 supplement
IDA regulates cell separation processes, but is also involved in major transcription of defense genes upon pathogen attack.
(a) IDA and the IDL peptides control cell separation processes during plant development and in response to abiotic and biotic stress (Butenko et al., 2003; Patharkar and Walker, 2016; Patharkar et …
Figure 7—figure supplement 1
HAE (yellow) and HSL2 (red) display very distinct repertoire of immune and growth-related interacting LRR-RKs.
HEA and HSL2 specific interacting partners determined using a sensitized high-throughput extracellular domain interaction assay (Smakowska-Luzan et al., 2018). HSL2 and HAE show distinct interacting …
Additional files
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Supplementary file 1
Peptide sequences.
- https://cdn.elifesciences.org/articles/87912/elife-87912-supp1-v1.docx
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Supplementary file 2
Primers sequences and function.
- https://cdn.elifesciences.org/articles/87912/elife-87912-supp2-v1.docx
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Supplementary file 3
Relative expression of genes of the CNGC and RBOH gene families during the onset of abscission (data from Cai and Lashbrook, 2008).
See Figure 2—figure supplement 1a for flower developmental stages (p2-p6). CNGC = CYCLIC NUCLEOTIDE GATED CHANNEL, RBOH = RESPIRATORY BURST OXIDASE PROTEIN, IDA = INFLORESCENCE DEFICIENT IN ABSCISSION, HSL2 = HAESA LIKE 2.
- https://cdn.elifesciences.org/articles/87912/elife-87912-supp3-v1.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/87912/elife-87912-mdarchecklist1-v1.docx
