Data from: Autoinhibitory calcium ATPases regulate the calcium gradient required for rapid polarized growth
Data files
Nov 03, 2025 version files 1.31 MB
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∆aca1_2_5-28.csv
23.69 KB
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∆aca1_5acaD464A-5.csv
39.09 KB
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actinMutant_6E.csv
132.76 KB
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actinWildtype_6E.csv
84.76 KB
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aip.csv
16.66 KB
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aipFigure7.csv
113.46 KB
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CalciumGradientQuantification.ijm
4.79 KB
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Data_Wrangle.Rmd
2.22 KB
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Figure2B.Rmd
5.64 KB
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Figure2D.Rmd
3.53 KB
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Figure4.Rmd
3.49 KB
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Figure5.Rmd
5.72 KB
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Figure6D.Rmd
2.56 KB
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Figure6E.Rmd
2.57 KB
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Figure7B.Rmd
3.54 KB
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Figure7D.Rmd
5.80 KB
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first10min.csv
33.78 KB
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LA)CSLD3_12001_Actin.csv
9.18 KB
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LA)CSLD3_12003_Actin.csv
9.64 KB
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LA)CSLD3_12005_Actin.csv
9.79 KB
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LA)CSLD3_12007_Actin.csv
9.06 KB
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LA)CSLD3_12009_Actin.csv
8.94 KB
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LA)CSLD3_12011_Actin.csv
10.34 KB
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LA)CSLD3_12013_Actin.csv
9.64 KB
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LA)CSLD3_12015_Actin.csv
10.54 KB
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LA)CSLD3_12017_Actin.csv
10.04 KB
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LA)CSLD3_12019_Actin.csv
10.64 KB
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LA)CSLD3_12021_Actin.csv
9.04 KB
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LA)CSLD3_12024_Actin.csv
9.27 KB
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LA)CSLD3_12025_Actin.csv
9.59 KB
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LA)CSLD3_12027_Actin.csv
10.36 KB
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LA)CSLD3_12029_Actin.csv
8.80 KB
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last10min.csv
33.78 KB
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MorphologyMacroV3.2.ijm
18.38 KB
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Mutant2_6D.csv
132.03 KB
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PMQuantificationMicrons_.ijm
2.37 KB
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pretreatment.csv
33.82 KB
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README.md
20.17 KB
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StatisticsMultipleComparisons_-_Sheet1.csv
25.23 KB
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strongQuadFigure7.csv
68.21 KB
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Wavelets_and_Timeseries_RevisedLW063021-pdf.R
10.54 KB
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weakQuadFigure7.csv
95.80 KB
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WildType.csv
16.66 KB
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Wildtype2_6D.csv
90.11 KB
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WTFigure7.csv
135.92 KB
Abstract
Polarized growth drives the morphogenesis of elongated cellular structures. In plants, polarized growth depends on actin and a tip-focused ionic calcium gradient. How the calcium gradient is maintained remains unclear. We discovered that autoinhibitory calcium ATPases (ACAs) redundantly contribute to the steepness of the calcium gradient. ACA1 and ACA2 localize to the subapical plasma membrane and ACA5 to the vacuole membrane, providing spatial regulation of calcium efflux. Tip-growing plant cells also exhibit apical calcium fluctuations. Even though Δaca1/2/5 cells have a diminished calcium gradient, they exhibit normal fluctuations and actin but have significantly reduced apical secretion. Furthermore, cells lacking apical actin retain a strong calcium gradient but have reduced apical secretion. Suppression of both the calcium gradient and apical actin dramatically impairs growth, supporting a model where two independent and parallel processes, the calcium gradient and apical actin, promote rapid polarized growth.
Dataset DOI: 10.5061/dryad.sn02v6xjp
Description of the data and file structure
Macros developed for Fiji:
- Calcium Gradient Quantification - takes the CFP and YFP images and calculates YFP/CFP ratio. Aligns the tips of the growing cell. Measures apical YFP/CFP levels. Measures the spatial gradient of YFP/CFP.
- Morphology Macro V3.2 - measures the chloroplast autofluorescence area for individual plants
- PMQuantificationMicrons - used to measure the levels of CSLD and actin at the cell tip.
R code:
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Data_Wrangle.Rmd: Aligning and normalizing data obtained from PMQuantificationMicrons.
Sample data files titled LA)CSLD3_1200x_Actin.csv included.
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Figure2B.Rmd: FDANOVA statistics
Data from Figure 2B included - first10min.csv, last10min.csv, pretreatment.csv
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Figure2D.Rmd: FDANOVA statistics
Data from Figure 2D included - aip.csv, WildType.csv
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Figure4.Rmd: Kruskal Wallis statistics with multiple comparisons
Data from Figure 4 included - StatisticsMultipleComparisons - Sheet1.csv
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Figure5.Rmd: FDANOVA statistics
Data from Figure 5B included - ∆aca125-28.csv, ∆aca1_5acaD464A-5.csv, wildtype.csv
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Figure6D.Rmd: FDANOVA statistics
Data from Figure 6D included - Mutant2_6D.csv, Wildtype2_6D.csv
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Figure6E.Rmd: FDANOVA statistics
Data from Figure 6E included - actinMutant_6E.csv, actinWildtype_6E.csv
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Figure7B.Rmd: Kruskal Wallis statistics with multiple comparisons
Data from Figure 7B included - StatisticsMultipleComparisons - Sheet1.csv
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Figure7D.Rmd: FDANOVA statistics
Data from Figure 7D included - aipFigure7.csv, strongQuadFigure7.csv, weakQuadFigure7.csv, WTFigure7.csv
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Wavelets and Timeseries RevisedLW063021-pdf.Rmd: Empirical mode decomposition code
Files and variables
File: Wavelets_and_Timeseries_RevisedLW063021-pdf.R
Description: Empirical mode decomposition code
File: Data_Wrangle.Rmd
Description: used to align data from PM quantification
File: LA)CSLD3_12001_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12005_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12007_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12003_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12013_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12011_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12017_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12024_Actin.csv
Description:
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12027_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12025_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12029_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12015_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12019_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12009_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: LA)CSLD3_12021_Actin.csv
Description: sample file for Data_Wrangle.Rmd
Variables
- First column is pixel position along the line
- Second column (value) is the measured intensity at that position.
File: Figure2B.Rmd
Description: FDANOVA statistics
File: first10min.csv
Description: data figure 2B
Variables
- index: pixel position
- microns: pixel position translated to microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- average: average intensity of cells 1-9
- standard error: standard error of the intensity of cells 1-9
File: last10min.csv
Description: data figure 2B
Variables
- index: pixel position
- microns: pixel position translated to microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- average: average intensity of cells 1-9
- standard error: standard error of the intensity of cells 1-9
File: pretreatment.csv
Description: data figure 2B
Variables
- index: pixel position
- microns: pixel position translated to microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- average: average intensity of cells 1-9
- standard error: standard error of the intensity of cells 1-9
File: aip.csv
Description: data figure 2D
Variables
- index: pixel position
- microns: pixel position translated to microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- average: average intensity of cells 1-6
- standard error: standard error of the intensity of cells 1-6
File: Figure2D.Rmd
Description: FDANOVA statistics
File: WildType.csv
Description: data figure 2D
Variables
- index: pixel position
- microns: pixel position translated to microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- average: average intensity of cells 1-6
- standard error: standard error of the intensity of cells 1-6
File: Figure4.Rmd
Description: Kruskall Wallis Statistics with multiple comparisons
File: StatisticsMultipleComparisons_-_Sheet1.csv
Description: Data for Figure 4B
Variables
- YCNLA: area of YCNLA plants
- aca1-8: area of ∆aca1-8
- aca1-13: area of ∆aca1-13
- aca2-40: area of ∆aca2-40
- aca2-d464a-14: area of ∆aca2-D464A-14
- aca5-3: area of ∆aca5-3
- aca5-7: area of ∆aca5-7
- aca1/2-2: area of ∆aca1/2-2
- aca1/2-19: area of ∆aca1/2-19
- aca1/5-7: area of ∆aca1/5-7
- aca1/5-9: area of ∆aca1/5-9
- aca2/5-109: area of ∆aca2/5-109
- aca1/2/5-28: area of ∆aca1/2/5-28
- aca1/2/5-36: area of ∆aca1/2/5-36
- trip-5: area of ∆aca1/5/aca2-D464-5
- trip-9: area of ∆aca1/5/aca2-D464-9
File: ∆aca1_2_5-28.csv
Description: Data for Figure 5B
Variables
- index: pixel position
- microns: pixel position translated to microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- cell10: intensity at that position for cell 10
- cell11: intensity at that position for cell 11
- average: average intensity of cells 1-11
- normalized: normalized average intensity of cells 1-11
- standard error: standard error of intensity of cells 1-11
File: ∆aca1_5acaD464A-5.csv
Description: Data for Figure 5B
Variables
- index: pixel position
- microns: pixel position translated to microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- cell10: intensity at that position for cell 10
- cell11: intensity at that position for cell 11
- cell12: intensity at that position for cell 12
- cell13: intensity at that position for cell 13
- cell14: intensity at that position for cell 14
- cell15: intensity at that position for cell 15
- cell16: intensity at that position for cell 16
- cell17: intensity at that position for cell 17
- cell18: intensity at that position for cell 18
- cell19: intensity at that position for cell 19
- cell20: intensity at that position for cell 20
- cell21: intensity at that position for cell 21
- cell22: intensity at that position for cell 22
- average: average intensity of cells 1-22
- normalized: normalized average intensity of cells 1-22
- standard error: standard error of intensity of cells 1-22
File: Figure5.Rmd
Description: FDANOVA statistics
File: Figure6D.Rmd
Description: FDANOVA statistics
File: Mutant2_6D.csv
Description: Data for Figure 6D
Variables
- index: pixel position
- microns: pixel position translated to microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- cell10: intensity at that position for cell 10
- cell11: intensity at that position for cell 11
- cell12: intensity at that position for cell 12
- cell13: intensity at that position for cell 13
- cell14: intensity at that position for cell 14
- cell15: intensity at that position for cell 15
- cell16: intensity at that position for cell 16
- cell17: intensity at that position for cell 17
- cell18: intensity at that position for cell 18
- cell19: intensity at that position for cell 19
- cell20: intensity at that position for cell 20
- cell21: intensity at that position for cell 21
- cell22: intensity at that position for cell 22
- average: average intensity of cells 1-22
- normalized: normalized average intensity of cells 1-22
- standard error: standard error of intensity of cells 1-22
File: Wildtype2_6D.csv
Description: Data for Figure 6D
Variables
- microns: position in microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- cell10: intensity at that position for cell 10
- cell11: intensity at that position for cell 11
- cell12: intensity at that position for cell 12
- cell13: intensity at that position for cell 13
- average: average intensity of cells 1-13
- standard error: standard error of intensity of cells 1-13
File: actinMutant_6E.csv
Description: Data for Figure 6E
Variables
- microns: position in microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- cell10: intensity at that position for cell 10
- cell11: intensity at that position for cell 11
- cell12: intensity at that position for cell 12
- cell13: intensity at that position for cell 13
- cell14: intensity at that position for cell 14
- cell15: intensity at that position for cell 15
- cell16: intensity at that position for cell 16
- cell17: intensity at that position for cell 17
- cell18: intensity at that position for cell 18
- cell19: intensity at that position for cell 19
- cell20: intensity at that position for cell 20
- cell21: intensity at that position for cell 21
- cell22: intensity at that position for cell 22
- average: average intensity of cells 1-22
- Standard error: standard error of intensity of cells 1-22
File: actinWildtype_6E.csv
Description: Data for Figure 6E
Variables
- microns: position in microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- cell10: intensity at that position for cell 10
- cell11: intensity at that position for cell 11
- cell12: intensity at that position for cell 12
- cell13: intensity at that position for cell 13
- average: average intensity of cells 1-13
- standard error: standard error of intensity of cells 1-13
File: Figure6E.Rmd
Description: FDANOVA statistics
File: Figure7B.Rmd
Description: Kruskal Wallis Statistics with multiple comparisons
File: aipFigure7.csv
Description: Data for Figure 7D
Variables
- index: pixel position
- microns: pixel position translated to microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- cell10: intensity at that position for cell 10
- cell11: intensity at that position for cell 11
- cell12: intensity at that position for cell 12
- Average: average intensity of cells 1-12
- standard error: standard error of intensity of cells 1-12
File: Figure7D.Rmd
Description: FDANOVA statistics
File: strongQuadFigure7.csv
Description: Data for Figure 7D
Variables
- micron: pixel position translated to microns
- Index: pixel position
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- average: average intensity of cells 1-6
- standard error: standard error of intensity of cells 1-6
File: weakQuadFigure7.csv
Description: Data for Figure 7D
Variables
- index: pixel position
- micron: pixel position translated to microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- cell10: intensity at that position for cell 10
- average: average intensity of cells 1-10
- standard error: standard error of intensity of cells 1-10
File: WTFigure7.csv
Description: Data for Figure 7D
Variables
- index: pixel position
- micron: pixel position translated to microns
- cell1: intensity at that position for cell 1
- cell2: intensity at that position for cell 2
- cell3: intensity at that position for cell 3
- cell4: intensity at that position for cell 4
- cell5: intensity at that position for cell 5
- cell6: intensity at that position for cell 6
- cell7: intensity at that position for cell 7
- cell8: intensity at that position for cell 8
- cell9: intensity at that position for cell 9
- cell10: intensity at that position for cell 10
- cell11: intensity at that position for cell 11
- cell12: intensity at that position for cell 12
- cell13: intensity at that position for cell 13
- cell14: intensity at that position for cell 14
- cell15: intensity at that position for cell 15
- Average: average intensity of cells 1-15
- Standard Error: standard error of intensity of cells 1-15
File: CalciumGradientQuantification.ijm
Description: Fiji Macro to generate calcium ratio image and to measure calcium gradient
File: MorphologyMacroV3.2.ijm
Description: Fiji Macro to quantify plant area based on chloroplast autofluorescence
File: PMQuantificationMicrons_.ijm
Description: Fiji Macro to quantify fluorescence at the cell tip
Code/software
Fiji Macros run in (Fiji is Just) ImageJ 2.14.0/1.54f:Java 1.8.0_202 [64bit]
R code runs in R Studio Version 2025.05.1+513 (2025.05.1+513)