From 19b4a3063cc16b961c2a9a46ca0414fa6d3ef008 Mon Sep 17 00:00:00 2001 From: Viktoria Petrova <vipet103@hhu.de> Date: Sat, 2 Nov 2024 14:19:50 +0100 Subject: [PATCH] add collection of root exudates assay and protocol --- assays/CollectionOfRootExudates/README.md | 0 .../CollectionOfRootExudates/dataset/.gitkeep | 0 .../CollectionOfRootExudates/isa.assay.xlsx | Bin 0 -> 6903 bytes .../protocols/.gitkeep | 0 .../CollectionOfRootExudatesProtocol.md | 3 +++ .../SynComInvasionExperiments/isa.assay.xlsx | Bin 6903 -> 6904 bytes 6 files changed, 3 insertions(+) create mode 100644 assays/CollectionOfRootExudates/README.md create mode 100644 assays/CollectionOfRootExudates/dataset/.gitkeep create mode 100644 assays/CollectionOfRootExudates/isa.assay.xlsx create mode 100644 assays/CollectionOfRootExudates/protocols/.gitkeep create mode 100644 assays/CollectionOfRootExudates/protocols/CollectionOfRootExudatesProtocol.md diff --git a/assays/CollectionOfRootExudates/README.md 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--- /dev/null +++ b/assays/CollectionOfRootExudates/protocols/CollectionOfRootExudatesProtocol.md @@ -0,0 +1,3 @@ +## Collection of root exudates + +*Arabidopsis* and *Lotus* plants were grown in a customized hydroponic system (original design by M. Peukert, University of Cologne, unpublished). This sterile growth setup consists of glass jars filled with glass beads and a stainless-steel mesh on top. Nutrient solution (modified 0.25× B&D medium, Fe-EDTA instead of Fe-citrate) was poured into the jars until the beads were covered in liquid and the liquid touched the metal mesh. We used the same medium for both plant species to allow for direct comparison of exudate composition, and to minimize differential effects on the bacterial community originating from different media types. We chose the *Lotus* B&D medium since *Arabidopsis* grew reasonably well in it. Sterilized and pregerminated seeds were placed onto the mesh, jars were put into sterile plastic boxes with filter lids (SacO2 microboxes) and plants were grown for 5 weeks. The medium containing root exudates was removed from the jars in the clean bench using a sterile metal needle and plastic syringe. After transfer to 50-ml Falcon tubes, exudates were frozen at −80 °C, freeze-dried until a volume of 2–3 ml was left, thawed and adjusted with sterile water to 5 ml. 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