Overview

Most of the analyses were conducted in R and are presented here as R Markdown (.Rmd) files. These files can be used to reproduce the figures and table found in the manuscript.

For Figure 1, the pollen diameters were calculated by averaging the diameter of microscope images of pollen grains using imageJ or equivalent image-analysis software with a microscope camera. Reference images of pollen types with scale bars can be found in the Pollen_Images.zip folder, with pollen types separated between subdirectories.

The .Rmd code is self-explanatory in most cases (i.e. pairwise.wilcox.test() is a pairwise wilcox test), but some comments are provided where necessary for clarity. Please note that text and formatting changes were applied to the generated figures after the data analysis step. Therefore, the .Rmd files contain out-of-date species names, for example H. scorpioides (meaning Helichrysum scorpioides) is Coronidium scorpioides in the final manuscript.

File Descriptions

Pollen_Images.zip

Reference images of pollen types with scale bars, with pollen types separated between subdirectories.

Incurviseta_floral_interactions.csv

This file includes all visitation data used in Visitation_data_analysis.rmd.

Columns:

• Site – subalpine or alpine
• Date – date of observation
• Time – time of day that observations occurred
• Plant Species – plant species I. cf. maculifrons was observed visiting
• Observed Visits – number of individuals observed visiting

Incurviseta_gut_and_body_pollen.csv

This file includes all body and gut pollen count data used in Analysing_gut_pollen.rmd and Visitation_data_analysis.rmd.

Columns:

• ID – unique identifier for each individual
• Location – where pollen was removed from (gut or body)
• Sex – usually unknown unless eggs/larvae were present in gut samples, in which case sex=female
• Site – subalpine or alpine
• Date – date the individual was collected
• Flower – flower individual was collected from
• PollenID – initial pollen identification
• PollenID2 – final pollen ID used for data analysis
• Count – number of pollen grains (gut pollen counts have been extrapolated by multiplying raw counts by four, as per the manuscript methodology)

Analysing_gut_pollen.Rmd

Reproduces the analyses for gut pollen, including figure creation and statistical tests presented in the manuscript.

Comparing_gut_pollen_to_body_pollen.Rmd

Reproduces statistical analyses comparing gut and body pollen data.
These analyses follow the approach outlined in:

Lysenkov, S. N. & Galinskaya, T. V. (2017). Comparison of the pollen content on the body and in the gut of hoverflies (Diptera: Syrphidae). Entomological Review, 97, 10–16.

Visitation_data_analysis.Rmd

Reproduces the visitation data analysis and generation of Table 1 in the manuscript, which describes the visitation of Incurviseta cf. maculifrons.