This dataset corresponds to the final figures from ‘Next-generation ABACUS biosensors reveal cellular ABA dynamics driving root growth at low aerial humidity’ by James Rowe, Mathieu Grangé-Guermente, Marino Exposito-Rodriguez, Rinukshi Wimalasekera, Martin O. Lenz, Kartika N. Shetty, Sean R. Cutler, Alexander M. Jones.

Data was collected at Sainsbury Laboratory, Cambridge University between approximately Sept 2017 and March 2023.

Any queries on the data can be emailed to Alexander M Jones (alexander.jones@slcu.cam.ac.uk), the principal investigator on the project, or James Rowe (james.rowe@slcu.cam.ac.uk), the lead author.

Notes on the dataset:

All data is placed under a CC-BY-NC 4.0 International license.

Methods are detailed fully in the paper but include: Confocal and Lightsheet microscopy, Advanced image analysis, Arabidopsis root and stomatal phenotyping, fluorescent spectroscopy. Microscopy data was analysed with Fiji and FRETENATOR1.5 as detailed in the paper.

Due to size constraints we have been unable to include all of our analysis but we have tried to include original image files and csv files with the processed data. With the microscopy data, we have tried to include the Emission Ratio X1000 images and a Max z projection of emission ratio X1000 image where possible. Several experimental datasets have had to be split across several .zip files due to space constraints of the repository (Max 2gb per file).

We have provided microscopy data as .tif files, which should retain scale info etc. in the metadata tags, rather than the original proprietary .lif format in which they were acquired.

In some original data files ABACUS2-100n is referred to as ABACUS2-150n. The same change was based on further titrations which gave us a more accurate indication of the KD for this sensor.

Whilst we have included all experiments and replicates that are included in the paper, data storage constraints meant we could not place our independent repeats of the experiments on this drive. These other replicates can be made available on request.

Files are mostly collected into .zip/.7z files which can be opened with a (de)compression program such as 7-zip. These files are organised by Figure, where multiple figures use the same data, they are placed in the same .zip/.7z file.

Figure Data: Figure 1b d Ex Data Figure 1.csv (15.71 KB) Figure 1f Ex Data Figure 3.zip (1.14 GB) Figure 1g Supp Figure 7.xlsm (37.35 KB) Figure 1h LR phenotyping ABACUS2.csv (2.83 KB)

Figure 2a b.zip (96.61 MB) Figure 2c Supp Figure 9.zip (1.16 GB) Figure 2d Supp Figure 10.zip (416.26 MB) Figure 2e Supp Figure 11.zip (1.05 GB) Figure 2f g Supp Figure 13.zip (1.5 GB) Figure 2 h i Ex data Figure 6.zip.001 (1000 MB) (multipart archive) Figure 2 h i Ex data Figure 6.zip.002 (1000 MB) (multipart archive) Figure 2 h i Ex data Figure 6.zip.003 (929.19 MB) (multipart archive)

Figure 3a b Supp Figure 15.zip (49 MB) Figure 3c.zip (219.65 MB) Figure 3d e Ex Data Figure 8.zip (950.39 MB) Figure 3f Ex Data Figure 9.zip (1.85 GB) Figure 3g Ex Data Figure 10.7z (1.84 GB)

Figure 4 a d e.zip (110.56 KB) Figure 4b c.zip (291.47 MB)

Extended Figure Data: Ex data Figure 2.pse (4.5 MB) Ex Data Figure 4a 2023 02 06 ABACUS root growth assay 3DAT RoweNatP.csv (27.28 KB) Ex Data Figure 4b 2023 02 06 ABACUS root growth assay 6DAT RoweNatP.csv (25.63 KB) Ex Data Figure 5.zip (640 MB)

Supplemental Figure Data: Supp Figure 2.zip (8.56 MB) Supp Figure 3.zip (8.57 MB) Supp Figure 4.zip (1.09 KB) Supp Figure 8.csv (800 B) Supp Figure 12.zip (640 MB) Supp Figure 14.zip (87.11 MB) Supp Figure 16.zip (584.21 MB) Supp Figure 22.zip (275.08 KB)