Summary of Experiment

This data repository contains data from three experiments. A summary of those experiments is below; additional details can be found in the associated manuscript.

1. Crossing surveys: To assess hybrid seed inviability within the widespread M. guttatus, we grew 3-5 individuals from 1 inbred line/maternal family for each of 23 populations of M. guttatus. Upon flowering, we crossed all accessions to each other in a nearly full diallel design, totaling 426 unique cross directions (out of a possible 506), with a mean of 4 replicate crosses per combination (range: 1-12, totaling 1,742 crosses). Based on the results of these crosses, we performed follow-up crossing surveys that incorporated other species within the complex: Mimulus decorus, M. corallinus, and M. glaucescens. Data from these crosses includes: Visual assessment of seed viability based on morphology (shriveled/flat corresponding to inviable, or round corresponding to viable); Germination rates on 0.6% agar media for 30 seeds per cross; Measurements of seed width.
2. Embryo Rescue: To determine if observed hybrid seed inviability was caused by defects in the endosperm rather than the embryo itself, we performed an embryo rescue experiment. Early developing seeds were grown on Murashige and Skoog media supplemented with 4 % sucrose to functionally compensate for the endosperm by providing external nutrition needed for growth. Seeds were scored for germination weekly for four weeks.
3. Complementation test: In order to assess whether the genetic basis of hybrid seed inviability with Northern Mimulus gutattus is shared between Southern M. decorus and M. corallinus, we performed a complementation test. We first crossed these two species reciprocally, then self-fertilized one reciprocal F1 from each cross direction to create reciprocal F2 populations. We planted 5 replicates of each parent, along with 15 F1s from each cross direction, and roughly 100 F2s from each cross direction. We performed an average of 3 replicate crosses per plant (range: 1-10), totaling 761 crosses and assessed the percentage of viable seed per fruit.

File Structure

1) Crossing survey data

File: heat_map_data.csv

This file contains the results of visual viability assessment from the Mimulus guttatus range-wide crossing survey.
The data table has 12 columns:

• female.pop: Line number of dam
• female.rep: Replicate number of dam
• female.type: Lineage identity of dam
• male.pop: Line number of sire
• male.rep: Replicate number of sire
• male.type: Lineage identity of sire
• total.seed: Number of seeds produced by cross
• number.inviable: Number of seeds assessed as inviable
• number.viable: Number of seeds assessed as inviable
• proportion.viable: number.viable divided by total.seed
• cross.type: lineage identity of each parent, with dam listed first
• counter: Initials of data collector
• notes: Notes on seed phenotypes

File: corallinusXdecorus_crosses.xlsx

This file contains the results of visual viability assessment from additional crosses completed using other members of the species complex.
The data table has 12 columns:

• female.pop: Line name of dam
• female.rep: Replicate number of dam
• female.type: Lineage identity of dam
• male.pop: Line name of sire
• male.rep: Replicate number of sire
• male.type: Lineage identity of sire
• total.seed: Number of seeds produced by cross
• number.inviable: Number of seeds assessed as inviable
• number.viable: Number of seeds assessed as inviable
• proportion.viable: number.viable divided by total.seed
• cross.type: lineage identity of each parent, with dam listed first
• counter: Initials of data collector
• notes: Notes on seed phenotypes

File: seed_sizes.txt

This file contains measurements of seed size.
The data table has 13 columns:

• Mat: Line name of dam
• Mat.rep: Replicate number of dam
• Pat: Line name of sire
• Pat.rep: Replicate number of sire
• cross.designation: Grouping of the crosses
• Mat.type: Lineage identity of dam
• Pat.type: Lineage identity of sire
• Mat.type.2: Lineage identity of dam, with M. glaucescens combined into the "Sierra" lineage
• Pat.type.2: Lineage identity of sire, with M. glaucescens combined into the "Sierra" lineage
• cross.specific: Lineage identity of each parent, with dam listed first
• cross: Lineage identity of each parent, with dam listed first
• image: Image number seed measurement was taken from
• seed.width: Seed width measured in mm

2) Embryo Rescue

File: embryo_rescue.xlsx

This file contains the results of the embryo rescue experiment.
The data table has four columns:

• line_ID: Names of the lines crossed, dam listed first
• cross: Lineage identity of lines crossed, dam listed first
• germ.percent: Percentage of seeds that germinated
• index: Whether the cross was part of the control ("Germination assay") or embryo rescue ("8 DAP")

3) Complementation Test

File: corallinusxSDec_Complementation_Test_seedcount.txt

This file contains results of visual viability assessment from complementation test crosses.
The data table has 12 columns:

• Plant: Line name of focal plant
• Rep: Replicate number of focal plant
• unique_ID: Line name and replicate number combined
• generation: Generation of focal plant (F1, F2, or parent line)
• ID: Cross ID
• inviable_seed_count: Number of inviable seeds
• viable_seed_count: Number of viable seeds
• total_seeds: Total number of seeds
• prop_viable: viable_seed_count divided by total_seeds

Population codes used across tables

• Mimulus corallinus = COR
• Northern Mimulus guttatus = NORTH = NGUT
• Southern Mimulus decorus = SDEC
• Northern Mimulus decorus = NDEC
• Sierra Mimulus guttatus = SIERRA
• Coastal Mimulus guttatus = COAST
• Mimulus glaucescens = GLAUC

Note: blank cells indicate 'not applicable'