Code/software requirements

To view the analyses used in the manuscript, you will need RStudio and the appropriate packages described at the top of the script. To view raw data, Excel is recommended. Versions used in this manuscript are listed below.

Software: 
R - v. 4.4.3

Packages:

car - v. 3.1-3

emmeans - v. 1.11.2-8

glmmTMB - v. 1.1.10

logistf - v. 1.26.0

dplyr - v. 1.1.4

tidyr - v. 1.3.1

ggplot2 - v. 3.5.1

Description of the data and file structure

Playback data (playback_data.csv): To determine if the presence of another female influenced individual preferences in Enchenopa binotata, we exposed live female(s) to artificial playbacks of male advertisement signals. These signals, which varied in dominant signal frequency, were generated artificially and sent along the surface of a nannyberry plant(Virburnum lentago). Females were exposed to playbacks of artificial male advertisement signals in either pairs or alone. Each trial was exposed to 3 bouts of four advertisement signals at one of five frequencies (selected randomly). We then measured the number of responses and response length.

Mating duration data (live_behavior.csv): To determine if the likelihood of mating with a given male was influenced by the presence of another female, we recorded the live interactions of Enchenopa binotata females. One or two unmated females were placed on the plant followed by a male. Recordings were conducted both via laser vibrometry to capture substrate-borne vibrations (which Enchenopa use to communicate) and via video. All recordings were conducted for up to an hour, until an individual left the recording set, or mating occurred. We noted mating behaviors and successes, the time until stages of courtship occurred, and other types of substrate borne vibratisubstrate-bornewith this species.

Standard errors from mating duration data (live_se.csv): Following the experiment we calculated the standard error for the percentage of females that duetted, advanced, and mated, grouped according to treatment, in our study. These were calculated using the package emmeans and our models in R and have been kept as a separate file from the raw data.

Response length splines (Pfunc_spline_length.csv): These are cubic splines generated using Pfunc and used in our figure 2 for response.

Number of response splines (Pfunc_spline_resp.csv): These are cubic splines generated using Pfunc and used in our figure 2 for the number of Figures.

fem_comp_AMNAT.R: the annotated R script which contains the analyses used in our publication

Files and variables

File: playback_data.csv

Columns| units (if applicable)| description

ID| NA| number assigned to each trial

treat| number of females V number of males| Whether there were two females or one female in the trial

freq| Hz| frequency of the male advertisement signal

plant| NA| plant on which the individuals were reared

date| month/date/year| date of the recording 

temp| degrees Celsius| temperature at time of recording

bout_number| count| the order of the bout (1 through 3) played in the trial

responded| 1=Yes, 0=No| whether an individual responded to the bout

number_responses| count| number of responses

avg_bout_resp_length| milliseconds| average response length within bouts

response_length_1| milliseconds| response length to the first signal (if no response occurred, the cell was left empty)

response_length_2| milliseconds| response length to the second signal (if no response, cell was left empty)

response_length_3| milliseconds| response length to the third signal (if no response, cell was left empty)

response_length_4| milliseconds| response length to the fourth signal (if no response, cell was left empty)

File: live_behavior.csv

Columns| units (if applicable)| description

treatment| NA| Whether there were two females or one female in the trial

label| NA| name given to the recording file corresponding to the individual trial

male| NA| identifying name given to each male

femID_1| NA| identifying name given to each female

femID_2| NA| identifying name corresponding to the second female in two female trials. In 1-female trials where there is no second female, this cell is empty. 

site| NA| location the individual was collected from or whose parents came from

temp| degrees Celsius| temperature at time of recording

date| Month/day/year| date of the recording 

trial_length| h:m:s| how long the trial was recorded

unanswered_calls| count| number of male advertisement signals which were not met by a female response

double_response| count| instances where both females responded to the same advertisement signal

single_response| count| instances where only one female responded to an advertisement signal

total_response| count| total number of female responses within the trial (doubles were counted as one response)

spontaneous_response| count| female response signals produced in instances other than in response to a male advertisement signal

percent_answer| %| percent of male advertisement signals which received a responthatom a female

fem_wing_buzz| count| number of female wing buzzes

revs| count| number of male revs

knocks| count| number of male knocks

duetted| 1=Yes, 0=No| whether duetting occurred

Jumped| 1=Yes, 0=No| whether an individual left the recording set before the trial end

advancing_signal| 1=Yes, 0=No| whether a female produced an advancement signal

copulation| 1=Yes, 0=No| whether successful mating occurred

advancing_signal_time| seconds| time of receiving the advancement signal

percent_duet| %| percent of trials in which duetting occurredin  per treatment (as a result only two cells in the row are filled in)

percent_advance| %| percent trials which an advancing sign-in occurred per treatment (as a result only two cells in the row are filled in)

percent_mate| %| percent trials which mating occurred in peach treatment (as a result only two cells in the row are filled in)

File: live_se.csv

Columns| units (if applicable)| description

treatment| NA| Whether there were two females or one female in the trial

percent_mate| %| percent of individuals that mated during a trial in the live experiment

mated_se| NA| standard error of individuals that mated. Calculated from emmeans in R

percent_advance| %| percent of trials where an advancing signal was produced in the live experiment

advanced_se| NA| standard error of individuals that produced an advancing signal. Calculated from emmeans in R

percent_duet| %| percent of trials where a duet was established in the live experiment

duet_se| NA| standard error of individuals that duetted. Calculated from emmeans in R

File: Pfunc_spline_length.csv

Columns| units (if applicable)| description

freq| Hz| frequency of the male advertisement signal

X1V1 response| NA| cubic spline values generated by Pfunc as best fit of response length in the 1 female treatment

X2V1 response| NA| cubic spline values generated by Pfunc as best fit of response length in the 2 female treatment

File: Pfunc_spline_resp.csv

Columns| units (if applicable)| description

freq| Hz| frequency of the male advertisement signal

X1V1 response| NA| cubic spline values generated by Pfunc as best fit of response number in the 1 female treatment

X2V1 response| NA| cubic spline values generated by Pfunc as the best fit of the response number in the 2 female treatment