The hatching assays were conducted in a climatic chamber at 18°C, in dark. For this, 12‐well plates (Costar®) were used and a sieve with 20 µm pores was added in each flat‐bottomed well. Each root exudate was carbon dosed using a FLASH 2000 CHNS/O Analysers (Thermo Scientific™) and standardized to 30 mg of carbon per gram of dry matter with autoclaved permuted water. Three cysts of each population with 1.5 ml of root exudates were put per sieve, and due to the number of available cysts, four to five replicates were realized per population and root exudates except for two populations (P252 and P320) with two replicates. Overall, to test the 156 comparisons (13 G. pallida populations * 12 potato exudates), 672 hatching assays were performed (i.e., on average 4.3 replicates per treatment). The number of hatched J2s was counted at days 2, 4, 10, 15, and 30 after the beginning of assays, and at each count, root exudates were replaced with fresh root exudates. At the end of the hatching experiment, cysts were crushed and the number of unhatched viable eggs was counted, in order to calculate a hatching percentage.
Flash 2000 chns o analyser
Manufactured by Thermo Fisher Scientific
The FLASH 2000 CHNS/O Analyser is a laboratory instrument designed for the determination of carbon, hydrogen, nitrogen, sulfur, and oxygen content in a wide range of sample types. It utilizes a combustion-based analytical technique to perform these elemental analyses.
Automatically generated - may contain errors
Lab products found in correlation
12 well plate, by Corning (1 mentions)
400 mhz spectrometer, by Bruker (1 mentions)
Agilent 1260 infinity hplc, by Agilent Technologies (1 mentions)
Ir 435 spectrophotometer, by Shimadzu (1 mentions)
G1315d dad vl, by Agilent Technologies (1 mentions)
G1329, by Agilent Technologies (1 mentions)
G1311b quaternary pump, by Agilent Technologies (1 mentions)
4 protocols using flash 2000 chns o analyser
1
Potato Root Exudates Impact on Globodera pallida
2
Hatching Assays for Potato Cyst Nematodes
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The hatching assays were conducted in a climatic chamber, in the dark, at 18°C for Globodera pallida and Heterodera carotae and at 23°C for Heterodera schachtii. Root exudates from potato, tomato and broccoli were tested on G. pallida populations, sugar beet, brown mustard and broccoli on H. schachtii populations and the two carrots on H. carotae populations (see Ngala et al., 2021) . For this, 12-well plates (Costar®) with flat-bottomed wells, which contained a sieve with 170 µm pores, were used. Each sample of root exudates was carbon dosed using a FLASH 2000 CHNS/O Analysers (Thermo Scientific TM ) and standardised to 30 mg of carbon per g of dry matter with autoclaved permuted water. Three cysts of each population for each nematode species with 1.5 mL of root exudates were put per sieve and six replicates were realized per population and root exudates. The number of hatched J2s was counted at days 2, 4, 10, 15, 30, 45 and 60 for G. pallida and H. carotae and at days 1, 2, 4, 10, 15 and 30 for H. schachtii after the beginning of assays and at each count, root exudates were replaced with fresh root exudates. At the end of the hatching experiment, cysts were crushed and the number of unhatched viable J2s was counted, in order to calculate the hatching rate.
3
Spectroscopic Characterization of Organic Compounds
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Solvents, reagents and fine chemicals were purchased from Alfa Aesar and Sigma-Aldrich, and were used any without further purification. The NMR spectra were determined by Bruker 400 MHz spectrometer, and 13 C NMR spectra were run at 100 MHz in DMSO-d6. FLASH 2000 CHNS/O analyser, Thermo Scientific, was used for elemental analyses.
4
Characterization of Organic Compounds
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All chemicals and solvents were obtained from Sigma-Aldrich or Fisher Scientific. A Stuart melting point apparatus (SMP10) was used for determining uncorrected melting points. Infra-red spectra (IR) were recorded using KBr discs on a Shimadzu IR 435 spectrophotometer. Nuclear magnetic resonance (1H NMR and 13 C NMR) spectra were recorded on a Bruker spectrometer (400 MHz) using deuterated Pyridine (Pyridine-d5) as a solvent. Elemental analyses (C, H, N and S) were conducted on a FLASH 2000 CHNS/O Analyser (Thermo Scientific). In addition, compounds were found to be ≥95% pure by reversed phase HPLC analysis using Agilent 1260 infinity HPLC equipped with G1311B Quaternary pump, G1329 injector and G 1315 D DAD Vl detector. A G1316A C18 column (4.6 × 150 mm) was used. An injection volume of 0.5 ml (DMF and phosphate buffer pH 5 1:1), a flow rate of 1 ml/min and an isocratic elution of acetonitrile in water (1:1) were applied. The detection was done at a wavelength of 254 nm.
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