Next to the inhibition experiments, bacterial volatiles emitted in monocultures and pairwise combinations were trapped and analyzed. For trapping of VOCs emitted by bacteria a volume of 100 μl of inoculation suspension was spread on 1/10th TSBA (20 mL) in glass Petri dishes designed for headspace volatile trapping (Garbeva et al., 2014b (link)). The Petri dishes were closed by a lid with an outlet connected to a steel trap containing 150 mg Tenax TA and 150 mg Carbopack B (Markes International, Ltd., Llantrisant, UK; Supplementary Figure S1 ). All treatments were inoculated in triplicate. The volatiles were collected after 48 and 72 h of incubation and the Tenax steel traps were stored at 4°C until GC-Q-TOF analysis.
Carbopack b
Manufactured by Markes International
Sourced in United Kingdom
Carbopack B is a porous polymer-based adsorbent material used in gas chromatography and thermal desorption applications. It is specifically designed for the collection and analysis of volatile organic compounds (VOCs) and other trace-level analytes in air samples.
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Lab products found in correlation
5 protocols using carbopack b
1
Bacterial Volatile Organic Compound Analysis
2
Volatile Organic Compound Analysis Protocol
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For analysis of volatile organic compounds, glass Petri dishes were used (Garbeva et al., 2014a ). The Petri dishes were closed by a lid with an outlet connected to a steel trap containing 150 mg Tenax TA and 150 mg Carbopack B (Markes International, Llantrisant, UK). The Tenax steel traps were collected after 72 h of incubation and stored at 4°C until GC‐Q‐TOF analysis. As controls glass Petri dishes containing 1/10th TSBA media without inoculated bacteria were used.
3
Volatile Organic Compounds Analysis Protocol
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For analysis and trapping of VOCs, two-compartment glass Petri-dishes80 (link) containing 1% LB agar were employed. A. niger was inoculated 24 h before addition of the bacterial strain on the left side of the two-compartment glass Petri dish and incubated overnight at 28 °C (Supplementary Fig. 2 ). After CoEvo 3 was incubated overnight at 28 °C in liquid LB medium, 2 µl of culture was spotted on the opposite compartment of the fungal culture. The glass Petri dishes were kept open for 25 min to allow droplets to dry. Plates were then closed by a lid with an outlet connected to a steel trap containing 150 mg Tenax TA and 150 mg Carbopack B (Markes International Ltd., Llantrisant, UK) and incubated at 28 °C. The Tenax steel traps were collected after 3 and 7 days of incubation and stored at 4 °C until GC-Q-TOF analysis. Glass Petri dishes containing LB agar medium but without inoculated bacteria or fungi served as controls.
4
Bacterial VOC Collection from Rhizosphere
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For the collection of bacterial VOCs, rhizospheric soil microcosms were established as indicated above in glass Petri-dishes with an exit at the top to which a steel trap filled with 150 mg Tenax TA and 150 mg Carbopack B (Markes International Ltd., Llantrisant, UK) could be fixed. In the rhizospheric soil microcosms, the bacterial strains Burkholderia sp. AD024, Dyella sp. AD056, Janthinobacterium sp. AD080, Pseudomonas sp. AD021, and Paenibacillus sp. AD087 were either incubated as monocultures or as mixtures of four (without Paenibacillus sp. AD087) or five strains. The growth of the strains was monitored by quantification of bacterial 16S rRNA genes from extracted DNA of soil samples (three per microcosm) taken at the beginning of the experiment and after 96 h incubation. Soil samples were stored at −80°C until nucleic acid extraction (see below). VOCs produced by the monocultures, bacterial mixtures and control (see above) were trapped for 24 h after 3 days of incubation. Traps were removed, capped and stored at 4°C until analysis.
5
Volatile Organic Compound Trapping in Plant Seedlings
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Four 20-days-old seedlings of each plant species were placed in individual 70 ml glass pots filled with sterilized soil (see choice experiment in soil). After 15 days, steel traps containing the volatile absorbants Tenax TA (150 mg) and Carbopack B (150 mg; Markes International Ltd., Llantrisant, United Kingdom) were attached at both sides of the glass pots (Supplementary Figure 1B ). After 24 h of incubation the traps were removed, capped and stored at 4°C until GC-QTOF analysis.
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