DOI: 10.5061/dryad.zs7h44jjj
title: Temperature influences the outcomes of an environmentally-acquired symbiosis

description: Pentatomomorphan bugs can form symbiotic associations with bacteria
 belonging to the supergenus Burkholderia sensu lato. This relationship has
 become a model for understanding environmental symbiont acquisition. Host
 insects can utilize various symbiont strains from across Burkholderia
 sensu lato, however, host colonization success and benefits conferred vary
 by bacterial clade. Therefore, we conducted a meta-analysis aimed at
 identifying candidate genes that underpin beneficial symbioses within this
 system. We scanned the entire Burkholderiaceae family for the presence of
 17 colonization-associated genes, as well as 88 candidate genes that are
 differentially expressed during symbiosis. There was no difference in the
 distribution of the 17 colonization-associated genes between symbiotic
 (Caballeronia and iPBE) and non-symbiotic lineages; however, there was a
 higher prevalence of the 88 candidate genes in the insect symbiont
 lineages. We subsequently analyzed the genomes of nine symbiotic
 Caballeronia strains that confer varying fitness benefits to their insect
 hosts. One symbiont strain was significantly worse, one was significantly
 better, and the remaining seven were intermediate in terms of conferred
 host fitness benefits. We found that strains possessing a higher number of
 the candidate genes conferred faster host development time. Furthermore,
 we identified two candidate genes that were missing in the least
 beneficial strain but present in the other eight, suggesting that these
 genes may be important in modulating symbiont quality. Our study suggests
 that the mechanisms required for host colonization are broadly distributed
 across Burkholderiaceae, but the genes that determine symbiont quality are
 more prevalent in insect-associated species. This work helps to identify
 genes that influence this highly specialized yet diverse symbiosis between
 Pentatomomorphan insects and Burkholderiaceae bacteria.
description: # Temperature influences the outcomes of an environmentally-acquired
 symbiosis
 [https://doi.org/10.5061/dryad.zs7h44jjj](https://doi.org/10.5061/dryad.zs7h44jjj) ## Description of the data and file structure This README file describes the data packages and code accompanying the above publication. ### Files and variables #### File: Stillson2023\_Adult\_weight\_data.csv **Description:** Fresh weights at adulthood of L. phyllopus subjected to six temperature treatments and colonized by one of six different Caballeronia isolates. Every row represents one insect. ##### Variables * BoxID: The cage in which an insect was reared. * BugID: Unique identification letter for each insect in a box. Letter was provided upon collection at adulthood. * Date: The date the insect was collected from its rearing cage. * Strains: Identity of the isolate a host was infected with. * Temp: The temperature in Celsius at which the insects were reared. * weight: Fresh weight in mg upon eclosion to adulthood. * sex: male (M) or female (F) * notes: Any notes about the individual. NA if there were no notes. #### File: Stillson2023\_Strain\_growth\_curves\_data.csv **Description:** Bacterial thermal growth curves subjected to six temperature treatments. Every row represents the maximum growth slope for one replicate per strain. ##### Variables * sid: Identity of the Caballeronia isolate. * rep: replicate number for the specified strain. * temp: The temperature in Celsius at which the strain was grown. * well: The plate well the culture was grown in. * slope: Maximum growth slope. * slope_lwr: Lower limit of the growth slope. * slope_upr: Upper limit of the growth slope. * intercept: Intercept of the growth curve region. * rsq: R-squared value of the growth curve region. * ln_od: Natural log of the optical density of the maximum growth curve region. #### File: Stillson2023\_Host\_titer\_data.csv **Description:** Symbiont titers of L. phyllopus subjected to six temperature treatments and colonized by one of six different Caballeronia isolates. Every row represents one insect. ##### Variables * Box ID: The cage in which an insect was reared. * Bug ID: Unique identification letter for each insect in a box. Letter was provided upon collection. * Date: The date the insect was collected from its rearing cage. * Age: The developmental staged of the host. 3 = third, 4 = fourth, 5 = fifth, Adult = Adult. * Strain: Identity of the isolate a host was infected with. * Temp: The temperature in Celsius at which the insects were reared. * titerCT1: Cycle threshold for first sample well. * titerCT2: Cycle threshold for second sample well. * meanCT: Mean cycle threshold for sample wells. * Intercept: Intercept generated from the standard curve. * Slope: Slope generated from the standard curve. * copy number: Calculated as 10^((meanCT – intercept)/slope) #### File: Stillson2023\_Strain\_phylogeny\_meta\_data.csv **Description:** Metadata for the tips of the Caballeronia 16S phylogeny in Figure S1. Isolates were Sanger-sequenced for this study. ##### Variables * Name: Name of the strain listed in the corresponding Newick phylogeny file. * new_name: The tip labels used on the phylogeny. These include the specific strain names and any named species used int eh phylogeny, followed by their strain names if applicable. * num: Used to apply colors to the tip labels. 1 = strains not used in generating growth curves, 2 = strains used to generate growth curves only, 3 = strains used to generate growth curves and used in the rearing experiment #### File: Stillson2023\_Strain\_phylogeny.newick **Description:** Phylogeny used to generate Figure S1. #### File: Stillson2023\_Mortality\_data.csv **Description:** Host survival of L. phyllopus subjected to six temperature treatments and colonized by one of six different Caballeronia isolates. Every row represents survival for each cage. ##### Variables * boxid: The cage in which an insect was reared. * strain: Identity of the isolate a host was infected with. * temp: The temperature in Celsius at which the insects were reared. * live3: The number of live third instars. * live4: The number of live fourth instars. * live5: The number of live fifth instars. * live6 : The number of live adults. Insects were collected and removed from the cages at this point. * dead3: The number of dead third instars. * dead4: The number of dead fourth instars. * dead5: The number of dead fifth instars. * totallive: Total number of insects alive in the cage. Includes adult insects that were removed during observations. * totaldead: Total number of insects that died in the cage. Includes all insects that died over the course of all observations. * totalbugs: Total number of insects in the cage. Includes adults and dead insects that were removed when found during daily observations. * sac3: The number of sacrificed third instars. * sac4: The number of sacrificed fourth instars. * sac5: The number of sacrificed fifth instars. * totalsac: Total number of sacrificed insects. #### File: Stillson2023\_Development\_time\_data.csv **Description:** Development times of L. phyllopus subjected to six temperature treatments and colonized by one of six different Caballeronia isolates. Every row represents one daily data collection for each cage. ##### Variables * boxid: The cage in which an insect was reared. * date: The date of data collection * strain: Identity of the isolate a host was infected with. * temp: The temperature in Celsius at which the insects were reared. * live3: The number of live third instars. * live4: The number of live fourth instars. * live5: The number of live fifth instars. * live6 : The number of live adults. Insects were collected and removed from the cages at this point. * dead3: The number of dead third instars. * dead4: The number of dead fourth instars. * dead5: The number of dead fifth instars. * totallive: Total number of insects alive in the cage. Includes adult insects that were removed during observations. * totaldead: Total number of insects that died in the cage. Includes all insects that died over the course of all observations. * totalbugs: Total number of insects in the cage. Includes adults and dead insects that were removed when found during daily observations. * Notes: Any notes about the cage. Start indicates initial observation for cage, retire indicates final observation, NA if there were no notes.