Data from: Mismatch between pollen and pistil size causes asymmetric mechanical reproductive isolation across Phlox species
Data files
Sep 24, 2024 version files 212.90 KB
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Corollas.csv
6.25 KB
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IDs_batch1.csv
1.58 KB
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IDs_batch2.csv
2.15 KB
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IDs_batch3.csv
1.40 KB
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IDs_batch4.csv
2.14 KB
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ids.csv
1.42 KB
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Pollen_Size_Measurements.csv
120.90 KB
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README.md
6.54 KB
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Results_batch1.csv
7.73 KB
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Results_batch2.csv
16.41 KB
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Results_batch3.csv
8.26 KB
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Results_batch4.csv
13.98 KB
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Seed_Set_20231116.csv
9.53 KB
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Sides.csv
7.38 KB
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Styles.csv
7.25 KB
Abstract
Characterizing the mechanisms of reproductive isolation (RI) between lineages is key to determining how new species are formed and maintained. In flowering plants, interactions between the reproductive organs of the flower –the pollen and the pistil– serve as the last barrier to reproduction before fertilization. As such, these pollen-pistil interactions are both complex and important for determining a suitable mate. Here, we test whether differences in style length (a part of the pistil) generate a postmating prezygotic mechanical barrier between five species of perennial Phlox wildflowers with geographically overlapping distributions. We perform controlled pairwise reciprocal crosses between three species with long styles and two species with short styles to assess crossing success (seed set). We find that heterospecific seed set is broadly reduced compared to conspecific cross success and reveal a striking asymmetry in heterospecific crosses between species with different style lengths. To determine the mechanism underlying this asymmetric reproductive isolating barrier we assess pollen tube growth in vivo and in vitro. We demonstrate that pollen tubes of short-styled species do not grow long enough to reach the ovaries of long-styled species. We find that short-styled species also have smaller pollen and that both within and between species pollen diameter is highly correlated with pollen tube length. Our results support the hypothesis that the small pollen of short-styled species lacks resources to grow pollen tubes long enough to access the ovaries of the long-styled species, resulting in an asymmetrical, mechanical barrier to reproduction. Such reproductive isolating mechanisms, combined with additional pollen-pistil incompatibilities, may be particularly important for closely related species in geographic proximity that share pollinators.
README
Paper: Mismatch between pollen and pistil size causes asymmetric mechanical reproductive isolation across Phlox species
Authors: Anna F. Feller, Grace Burgin, Nia Lewis, Rohan Prabhu, Robin Hopkins
https://doi.org/10.5061/dryad.fttdz091s
This dataset contains the raw data files from experiments (described in detail in the paper) to assess seed set, floral morphology, pollen size, and pollen tube growth in and between six closely related species of Phlox flowers, as well as the R code files used to visualize and statistically analyze these data.
Description of the data and file structure
Data files:
- ids.csv - contains information on all individuals used in the experiments to assess seed set, floral morphology, pollen size, and pollen tube growth
- ID: individual ID, composed of 'species abbreviation-population number-individual number'
- species: scientific species name (without the preceding genus name 'Phlox')
- population ID: population ID, composed of 'species abbreviation-population number'
- species abbreviation - abbreviation of the species name (DIV: Phlox divaricata, GLA: Phlox glaberrima (ssp. interior), MAC: Phlox maculata, PAN: Phlox paniculata, PIL: Phlox pilosa (ssp. pilosa, North)
- lat: latitude
- lon: longitude
- Seed_Set_20231116.csv - contains the results of the crossing experiment to assess seed set
- female individual ID: the individual used as the pollen receiver in a cross (the ID is composed of 'species abbreviation-population number-individual number')
- female species abbr.: abbreviation of the species name of the individual used as the pollen receiver (DIV: Phlox divaricata, GLA: Phlox glaberrima (ssp. interior), MAC: Phlox maculata, PAN: Phlox paniculata, PIL: Phlox pilosa (ssp. pilosa, North)
- male individual ID: the individual used as the pollen donor in a cross (the ID is composed of 'species abbreviation-population number-individual number').
- male species abbr.: abbreviation of the species name of the individual used as the pollen donor (DIV: Phlox divaricata, GLA: Phlox glaberrima (ssp. interior), MAC: Phlox maculata, PAN: Phlox paniculata, PIL: Phlox pilosa (ssp. pilosa, North)
- cross name: 'ID of individual used as the pollen receiver x ID of individual used as the pollen donor'
- cross: 'abbreviation of the species name of the individual used as the pollen receiver x abbreviation of the species name of the individual used as the pollen donor'
- cross type: specifies if a cross was a cross within (heterospecific; 'het') or between (conspecific ; con') species
- crossed flowers: the number of crossed flowers per cross
- number of seeds: the number of seeds obtained in a cross
- Sides.csv, Styles.csv, Corollas.csv - contain the floral morphological measurements measured with ImageJ from 2D-scans of flowers (see Fig. 2 in paper)
- Individual: individual ID, composed of 'species abbreviation-population number-individual number'
- Calyx height (mm): height of the calyx in mm
- Tube length (mm): length of the corolla tube in mm
- Petal limb length (mm): length of a petal limb in mm
- Style length (mm): length of the style in mm
- Corolla Area (mm^2): area of the corolla in mm2 (square millimeters)
- Corolla radius (mm): radius if the corolla in mm
- Pollen_Size_Measurements.csv - contains the pollen diameter measurements measured in ImageJ from microscope images of pollen (the first column can be ignored)
- Area: pollen area in um2 (square micrometers)
- Perim.: pollen perimeter in um (micrometers)
- Filename: name of the input image file in tif format
- IDs_batch1.csv, IDs_batch2.csv, IDs_batch3.csv, IDs_batch4.csv - contain info on which sample is associated with which microscope images in the files containing in vitro pollen tube length measurements (the 'Results...' files below)
- Individual: individual ID, composed of 'species abbreviation-population number-individual number'
- SNAP#: number of the image file
- Results_batch1.csv, Results_batch2.csv, Results_batch3.csv, Results_batch4.csv - contain the pollen diameter and pollen tube length measurements of the in vitro experiment; each measurement comes on two lines (pollen diameter and then pollen tube length)
- Batch: number of batch (images were collected in four batches)
- Label: image file name
- Angle: angle of drawn line to measure pollen diameter (irrelevant, to be ignored)
- Length: the first of two lines of a measurement that always go together contains the pollen diameter measurement in um (micrometer), the second the pollen tube length measurement in um (micrometer)
Code/Software
Analysis scripts:
- Occurrence_Data.R - R script to plot occurrence data obtained from GBIF and locations of experimental samples; input files are 'ids.csv' (see Data files) and the GBIF occurrence data below
- Seed_Set.R - R script to visualize and analyze the data from the crossing experiments; input file is 'Seed_Set_20231116.csv'
- Floral_Measurements_and_Pollen_Size.R - R script to visualize and analyze the morphological data; input files are 'Sides.csv', 'Styles.csv', 'Corollas.csv', 'Pollen_Size_Measurements.csv'.
- Pollen_Tubes.R - R script to visualize and analyze the data from the in vitro pollen tube growth experiments; input files are 'IDs_batch1.csv', 'IDs_batch2.csv', 'IDs_batch3.csv', 'IDs_batch4.csv', 'Results_batch1.csv', 'Results_batch2.csv',' Results_batch3.csv', 'Results_batch4.csv'
These R scripts assume that all R scripts are in a folder called R/ANALYSIS, that all data files are in a folder called R/DATA (with the exception of the GBIF data, see 'Occurrence_Data.R' and Table S2 in the Supplementary Materials), and that there is a folder called R/OUTPUT to store plots and other output files.
Sharing/Access information
Occurrence data from GBIF (used for plotting Fig. 1 in paper with R-script 'Occurrence_Data.R'):