DOI: 10.5061/dryad.f4qrfj77b
title: Data from: Wolbachia feminises a spider host with assistance from co-infecting symbionts

description: Arthropods commonly harbor maternally-transmitted bacterial symbionts that
 manipulate host biology. Multiple heritable symbionts can co-infect the
 same individual, allowing these host-restricted bacteria to engage in
 cooperation or conflict, which can ultimately affect host phenotype. The
 spider Mermessus fradeorum is variably infected with up to five heritable
 symbionts: Rickettsiella (R), Tisiphia (T), and three strains of Wolbachia
 (W1-3). Quintuply infected spiders are feminized, causing genetic males to
 develop as phenotypic females and produce almost exclusively female
 offspring. We compared feminization rates (proportion of female offspring
 out of total) across nine infection combinations. We also compared
 feminization rates across a subset of three infection types (Uninfected,
 RTW1, and RTW123) in a second iteration of the experiment to validate
 coinfection effects on feminization strength observed in the first
 iteration of the feminization experiment. In both experiments we found
 that the Wolbachia strain W1 was required for feminization and this
 phenotype was strongest in quintuply infected spiders, implying a
 synergistic effect of coinfection on W1 feminization. Using digital PCR
 (dPCR), we estimated the amount of symbionts in different infection
 combinations to determine if a) symbiont titer changes across
 coinfections, and b) if W1 titer correlates with feminization strength. We
 extraction the DNA of adult females from the first feminization experiment
 and use species specific dPCR primers to analyze titer. We found limited
 changes in symbiont titer in different symbiont co-infections, suggestive
 of subtle effects of co-infection on the composition of the symbiont
 community. The synergistic effect of co-infection on Wolbachia
 feminization may help promote the spread of all five symbionts in spider
 populations. These results confirm the first instance of Wolbachia-induced
 feminization in spiders and demonstrate that co-infecting symbionts can
 improve the efficacy of symbiont-induced feminization.
description: # Data from: Wolbachia feminises a spider host with assistance from
 co-infecting symbionts Dataset DOI:
 [10.5061/dryad.f4qrfj77b](10.5061/dryad.f4qrfj77b) ## Description of the
 data and file structure This README file was generated on 2025-06-11 by
 Matthew Doremus. GENERAL INFORMATION 1\. Title of Dataset: Wolbachia
 feminises its spider host with assistance from co-infecting symbionts 2\.
 Author Information ``` A. Principal Contact Information Name: Matthew
 Doremus Institution: University of Illinois Urbana-Champaign Address:
 Urbana, IL USA Email: mdoremus@illinois.edu B. Co-investigator Contact
 Information Name: Jen White Institution: University of Kentucky Address:
 Lexington, KY USA Email: jenawhite@uky.edu C. Co-investigator Contact
 Information Name: Yuval Gottlieb Institution: Hebrew University of
 Jerusalem Address: Rehovot, Israel Email: gottlieb.yuval@mail.huji.ac.il
 ``` 3\. Date of data collection (single date, range, approximate date):
 2022-24 4\. Geographic location of data collection: Lexington, KY, USA
 (sex ratio experiments), Rehovot, Israel (digital PCR) 5\. Information
 about funding sources that supported the collection of the data: National
 Science Foundation (NSF) 1953223; Bi-national USA-Israel, Science
 Foundation (BSR) 201697; National Institute of Food and Agriculture, U.S.
 Department of Agriculture (USDA-NIFA) under Hatch 1020740 and 7007679;
 USDA-NIFA Fellowship 2023-67012-39352; Israel Science Foundation (ISF)
 2809/23 Foundation SHARING/ACCESS INFORMATION 1\. Licenses/restrictions
 placed on the data: 2\. Links to publications that cite or use the data:
 [https://doi-org.proxy2.library.illinois.edu/10.1111/1462-2920.70149](https://doi-org.proxy2.library.illinois.edu/10.1111/1462-2920.70149) 3\. Links to other publicly accessible locations of the data: None 4\. Links/relationships to ancillary data sets: None 5\. Was data derived from another source? No ``` A. If yes, list source(s): NA ``` 6\. Recommended citation for this dataset: Mackevicius-Dubickaja, V., Y. Gottlieb, J. A. White, and M. R. Doremus. 2025. “ *Wolbachia* Feminises a Spider Host With Assistance From Co-Infecting Symbionts.” *Environmental Microbiology* 27, no. 7: e70149. [https://doi-org.proxy2.library.illinois.edu/10.1111/1462-2920.70149](https://doi-org.proxy2.library.illinois.edu/10.1111/1462-2920.70149). ### Files and variables #### 1. File List: A) Sex_ratio1.csv B) Sex_ratio2.csv C) dPCR.csv 2\. Relationship between files, if important: Spider samples used in dPCR were the same females used in Sex_ratio1.csv. These samples share the same IDs in both experiments. Sex_ratio1.csv and Sex_ratio2.csv correspond with two independent experiments testing whether different symbiont combinations feminize their spider host. 3\. Additional related data collected that was not included in the current data package: None 4\. Are there multiple versions of the dataset? No ``` A. If yes, name of file(s) that was updated: NA i. Why was the file updated? NA ii. When was the file updated? NA ``` \######################################################################### DATA-SPECIFIC INFORMATION FOR: Sex_ratio1.csv 1\. Number of variables: 5 2\. Number of cases/rows: 116 3\. Variable List: * Female_ID: Code unique to each spider sample comrised of Infection (see below)+Replicate. * Female_Infection: Symbionts present in spider. U = uninfected (no symbiont), R = Rickettsiella (+), T = Tisiphia (+), W = Wolbachia (+). Spiders can have up to 3 strains of Wolbachia, strains are denoted by number (W1 = Wolbachia strain 1) * Female_Offspring: number of females that spider produced in their brood * Male_Offspring: number of males that spider produced in their brood * Total_Offspring: total number of offspring that spider produced in their brood * SR: sex ratio (Female_Offspring/Total_Offspring) 4\. Missing data codes: None 5\. Specialized formats or other abbreviations used: None \######################################################################### DATA-SPECIFIC INFORMATION FOR: Sex_ratio2.csv 1\. Number of variables: 5 2\. Number of cases/rows: 37 3\. Variable List: * Female_ID: Female_ID: Code unique to each spider sample comrised of Infection (see below)+Replicate. * Female_Infection: Symbionts present in spider. CC = uninfected (no symbiont), 3C = RTW1 (triple infection), 5C = RTW123 (quintuple infection) * Female_Offspring: number of females that spider produced in their brood * Male_Offspring: number of males that spider produced in their brood * Total_Offspring: total number of offspring that spider produced in their brood * SR: sex ratio (Female_Offspring/Total_Offspring) 4\. Missing data codes: NA (data not available) 5\. Specialized formats or other abbreviations used: None \######################################################################### DATA-SPECIFIC INFORMATION FOR: dPCR.csv Digital PCR (dPCR) was used to estimate the amount of symbionts cells in each female spider that laid egg masses. Spiders were collected from the same experiment as Sex_ratio1.csv and keep the same ID. 1\. Number of variables: 44 2\. Number of cases/rows: 108 3\. Variable List: * Female_ID: Code unique to each spider sample comrised of Infection (see below)+Replicate. Same individuals were used in Sex_ratio1.csv * Infection: Spiders had a combination of five heritable bacteria (symbionts): U = uninfected (no symbiont), R = Rickettsiella (+), T = Tisiphia (+), W = Wolbachia (+). Spiders can have up to 3 strains of Wolbachia, strains are denoted by number (W1 = Wolbachia strain 1) * Conc[cp/uL]: Concentration of DNA copies per microliter of sample - conc is presented for the host gene (18S) and each symbiont * valid: number of partitions ran in dPCR cycle - presented for the host gene (18S) and each symbiont * Positive: number of positive dPCR partitions - presented for the host gene (18S) and each symbiont * Negative: number of negative dPCR partitions (DNA not amplified) - presented for the host gene (18S) and each symbiont * λ : Value of the expectation for positive events (λ = -ln(Valid-Positive/Valid))) - presented for the host gene (18S) and each symbiont * CI (95%): Confidence interval for partions (samples with less than 100 unexpected positive partions and a high CI were considered negative) - presented for the host gene (18S) and each symbiont * M.fradeorum_18S_12ng: Original concentration of M. fradeorum (spider) 18S copies accounting for dilution factor. Only the host has this variable (replaced by symbiont titer for symbionts) * Symbiont titer: Final estimation of titer for normalized to M.fradeorum_18S_12ng value for that spider. Titer is presented for each symbiont. Symbionts used different single copy genes as targets for dPCR: Rickettsiella = recA, Tisiphia = rpoB, Wolbachia = wsp (with primers specific to each Wolbachia strain). 4\. Missing data codes: N/A (data not applicable) 5\. Specialized formats or other abbreviations used: Data sheet has sections for each symbiont with the above variable list: * *M fradeorum* (spider host) - columns C-I * *Rickettsiella* - J-P * *Tisiphia* - Q-W * *Wolbachia* 1 - X-AD * *Wolbachia* 2 - AE-AK * *Wolbachia* 3 - AL-AR