DOI: 10.5061/dryad.g4f4qrg1c
title: Fungus-farming termites can protect their crop by confining weeds with fungistatic soil boluses

description: The symbiotic agriculture of fungus-farming termites can collapse if they
 fail to prevent invading weeds. Previous studies suggest the role of
 symbiotic fungistatic microbes in bringing about weed control. However,
 how termites employ these microbes to suppress the fungal-weeds, without
 affecting the fungal-cultivar, remains unknown. Here we show that the
 fungus-farming termite, Odontotermes obesus, uses specific behaviors to
 remove, isolate and suppress the growth of the fungal weed Pseudoxylaria,
 by primarily encasing them with soil boluses containing fungistatic
 microbes. These behaviors efficiently suppress the weed without affecting
 the crop. This integration of specific behaviors with termite-derived
 microbes appears to be the proximate mechanism of how microbes are
 topically used by termites to confine the weed** while keeping the crop
 unaffected.
description: # Fungus-farming termites can protect their crop by confining weeds with
 fungistatic soil boluses ## [Access this dataset on Dryad
 \[**https://doi.org/10.5061/dryad.g4f4qrg1c**\]](https://doi.org/10.5061/dryad.g4f4qrg1c) Files in this dataset include: * Main figures (Figures 1- 4): As presented in the manuscript * Supplementary figures (Figs.S1-S18): As presented in the supplementary material * Supplementary tables (Table S1- Table S7): As presented in the supplementary material * Data files (Panchal_et_al_Data S1-S6.xlsx): Raw and processed data supporting analyses in the manuscript and is uploaded and available on Dryad. -  Data_S7.pdf: Raw data in the form of photographs of all behavioral assay plates uploaded and available on Zenodo. *Note: Main figures (Figures 1- 4), all the supplemental files including supplementary figures (Figs. S1-S18), supplementary tables (Table S1- Table S7) and Data_S7.pdf are available on Zenodo.  This README file describes the raw data files (Panchal_et_al_Data_S1-S6.xlsx) available on Dryad and (Data_S7.pdf) available on Zenodo. We have submitted our raw data: Area of comb scraped (%) for Figure 1C (**Panchal_et_al_Data_S1.xlsx**), Amount of soil bolus added for Figure 2C (**Panchal_et_al_Data_S2.xlsx**), Amount of soil bolus added for Figure 3C (**Panchal_et_al_Data_S3.xlsx**), Amount of soil bolus added for Figure S17 (**Panchal_et_al_Data_S4.xlsx**), microbiome of soil boluses shown in Figure 4B (**Panchal_et_al_Data_S5.xlsx**) and mycobiome of soil boluses shown in Figure 4A (**Panchal_et_al_Data_S6.xlsx**). ## Description of the data and file structure ### Panchal_et_al_Data_S1.xlsx *This file contains raw data for Figure 1C of the manuscript Data point: Individual assay plate FC: Fresh comb FC+P: Fresh comb with *Pseudoxylaria* 'FC' are the fresh fungus combs collected from the termite colonies and incubated in the lab to naturally grow the weedy fungus, *Pseudoxylaria* on them. 'FC+P' are the fungus combs with *Pseudoxylaria* tuft placed on them. A detailed methodology of fungus comb collection, incubation and growth of *Pseudoxylaria* is provided in the materials and methods to the main article under the section 'Collection of Combs, Termites and Setting up the Behavioral assays'. A pictorial representation of this methodology is given in Figure S2 of the manuscript. Area of comb scraped (%) on FC: Area of comb scraped by workers on fresh comb Area of comb scraped (%) on FC+P: Area of comb scraped by workers on fresh comb with *Pseudoxylaria* ### Panchal_et_al_Data_S2.xlsx *This file contains raw data for Figure 2C of the manuscript Data point: Individual assay plate FC: Fresh comb IC: Infected comb 'FC' are the fresh fungus combs collected from the termite colonies and incubated in the lab to naturally grow the weedy fungus, *Pseudoxylaria* on them. 'IC' are the severely infected combs with *Pseudoxylaria*. A detailed methodology of fungus comb collection, incubation and growth of *Pseudoxylaria* is provided in the materials and methods to the main article under the section 'Collection of Combs, Termites and Setting up the Behavioral assays'. A pictorial representation of this methodology is given in Figure S2 of the manuscript. Amount of soil boluses added (gram) on FC: Amount of soil boluses added by the workers on fresh comb Amount of soil boluses added (gram) on IC: Amount of soil boluses added by the workers on infected comb Outcome of behavioral response: Outcomes of worker behavior ·      Complete encasement: Infected comb entirely encased with soil boluses ·      Partial encasement: Some portions of the infected comb encased with soil boluses ·      No behavioral response: No discernible response from workers ### Panchal_et_al_Data_S3.xlsx *This file contains raw data for Figure 3C of the manuscript Data point: Individual assay plate FC: Fresh comb FC+IC: Fresh comb glued to *Pseudoxylaria* infected comb 'FC' are the fresh fungus combs collected from the termite colonies and incubated in the lab to naturally grow the weedy fungus, *Pseudoxylaria* on them. 'FC+IC' are the fresh fungus comb glued to *Pseudoxylaria* infected comb. A detailed methodology of fungus comb collection, incubation and growth of *Pseudoxylaria* is provided in the materials and methods to the main article under the section 'Collection of Combs, Termites and Setting up the Behavioral assays'. A pictorial representation of this methodology is given in Figure S2 of the manuscript. Amount of soil bolus added (gram) on FC: Amount of soil bolus added by the workers on fresh comb Amount of soil bolus added (gram) on FC+IC: Amount of soil bolus added by the workers on fresh comb glued to *Pseudoxylaria* infected comb Outcome of behavioral response: Outcomes of worker behavior ·      Complete separation with complete encasement: Complete separation of the glued comb pieces and complete encasement of the infected comb ·      Complete separation with partial encasement: Complete separation of the glued comb pieces but partial encasement of the infected comb ·      Partial separation with partial encasement: Partial separation of the glued comb pieces and partial encasement of the infected comb ·      Complete separation but no encasement: Complete separation of the glued comb pieces but no encasement of the infected comb ### Panchal_et_al_Data_S4.xlsx *This file contains raw data for Figure S17C of the manuscript Data point: Individual assay plate FC: Fresh comb FC+FC: Fresh comb glued to another fresh comb 'FC' are the fresh fungus combs collected from the termite colonies. 'FC+FC' is the fresh fungus comb glued to another fresh comb.* *A detailed methodology of fungus comb collection and incubation is provided in the materials and methods to the main article under the section 'Collection of Combs, Termites and Setting up the Behavioral assays'. A pictorial representation of this methodology is given in Figure S2 of the manuscript. Amount of soil bolus added (gram) on FC: Amount of soil bolus added by the workers on fresh comb Amount of soil boluses added (gram) on FC+FC: Amount of soil bolus added by the workers on the two fresh combs glued together Outcome of behavioral response: Outcomes of worker behavior ·      Complete separation: Complete separation of the two fresh combs glued together ·      Partial separation: Partial separation of the two fresh combs glued together ·      No separation: No discernible response from the workers ### Panchal_et_al_Data_S5.xlsx *This file contains raw data for Figure 4B of the manuscript *Data represents the taxonomic distribution and relative abundance of the bacterial OTUs (microbiome) present within the soil boluses for all the samples identified through Nanopore sequencing. A detailed methodology for sample preparation and sequencing has been provided in the supplementary text to the main article under the sections ‘Library preparation for Nanopore Sequencing’ and ‘Nanopore Sequence data generation and identification of Operational Taxonomic Units (OTUs)’ OTU: Operational Taxonomic Unit Domain: OTU at taxonomic rank ‘Domain’ Phylum: OTU at taxonomic rank ‘Phylum’ Class: OTU at taxonomic rank ‘Class’ Order: OTU at taxonomic rank ‘Order’ Family: OTU at taxonomic rank ‘Family’ Genus: OTU at taxonomic rank ‘Genus’ Mound SC3 Replicate 1: Sample 1 Mound SC3 Replicate 2: Sample 2 Mound SC4 Replicate 1: Sample 3 Mound SC4 Replicate 2: Sample 4 Mound SC4 Replicate 3: Sample 5 Mound HC Replicate 1: Sample 6 Mound HC Replicate 2: Sample 7 *Numeric values denote the relative abundance of the bacterial genera present across the samples ### Panchal_et_al_Data_S6.xlsx *This file contains raw data for Figure 4A of the manuscript *Data represents the taxonomic distribution and relative abundance of the fungal OTUs (mycobiome) present within the soil boluses for all the samples identified through Nanopore sequencing. A detailed methodology for sample preparation and sequencing has been provided in the supplementary text to the main article under the sections ‘Library preparation for Nanopore Sequencing’ and ‘Nanopore Sequence data generation and identification of Operational Taxonomic Units (OTUs)’ OTU: Operational Taxonomic Unit Phylum: OTU at taxonomic rank ‘Phylum’ Class: OTU at taxonomic rank ‘Class’ Order: OTU at taxonomic rank ‘Order’ Family: OTU at taxonomic rank ‘Family’ Genus: OTU at taxonomic rank ‘Genus’ Mound SC3: Sample 1 Mound SC4 Replicate 1: Sample 2 Mound SC4 Replicate 2: Sample 3 Mound HC Replicate 1: Sample 4 Mound HC Replicate 2: Sample 5 *Numeric values denote the relative abundance of the fungal genera present across the samples ### Data_S7.pdf *This file is available on Zenodo. However, the description of the data and file structure of Data_S7.pdf is provided here in this README file. *Data_S7.pdf file represents all data points in the form of photographs of all assay plates for the three behavioral assays depicted in Figures 1B, 2B, 3B and S17B of the manuscript. Data S7(i): Representation of all data points (photographs of all assay plates, N=57) of termite workers in response to introduced *Pseudoxylaria* tuft on the comb* Data S7(ii): Representation of all data points (photographs of all assay plates, N=101) of termite workers in response to extensive *Pseudoxylaria* infections. Data S7(iii): Representation of all data points (photographs of all assay plates, N=21) of major workers in saving healthy portions of the comb from *Pseudoxylaria* infections. Data S7(iv): Representation of all data points (photographs of all assay plates, N=10) of termite workers in response to two fresh combs attached together. ## Sharing/Access information Other publicly accessible locations of the data: **Panchal_et_al_Data_S5.xlsx**: Raw sequences generated from Nanopore are available at NCBI Sequence Read Archive (SRA) under BioProject PRJNA1119496 (BioSample accessions: SAMN41656945 - SAMN41656951) **Panchal_et_al_Data_S6.xlsx**: Raw sequences generated from Nanopore are available at NCBI Sequence Read Archive (SRA) under BioProject PRJNA1119511(BioSample accessions: SAMN41657358 - SAMN41657362) ## Data was derived from the following sources All data was derived from experiments and analyses conducted by the authors, as described in the manuscript.