Four kinds of green deep eutectic solvent (DES) were synthesized, including choline chloride (ChCl)-urea, tetramethylammonium chloride (TMACl)-urea, tetrapropylammonium bromide (TPMBr)-urea and ChCl-methylurea. An aqueous two-phase system (ATPS) based ChCl-urea DES was studied for the first time for the extraction of bovine serum albumin (BSA). Single factor experiments proved that the extraction efficiency of BSA was influenced by the mass of the DES, concentration of K2HPO4 solution, separation time and extraction temperature. The optimum conditions were determined through an orthogonal experiment with the four factors described above. The results showed that under the optimum conditions, the average extraction efficiency could reach up to 99.94%, 99.72%, 100.05% and 100.05% (each measured three times). The relative standard deviations (RSD) of extraction efficiencies in precision, repeatability and stability experiments were 0.5533% (n = 5), 0.8306% (n = 5) and 0.9829% (n = 5), respectively. UV-vis and FT-IR spectra confirmed that there were no chemical interactions between BSA and the DES in the extraction process, and the CD spectra proved that the conformation of BSA did not change after extraction. The conductivity, DLS and TEM were combined to investigate the microstructure of the top phase and the possible mechanism for the extraction. The results showed that hydrophobic interactions, hydrogen bonding interactions and the salting-out effect played important roles in the transfer process, and the aggregation and surrounding phenomenon were the main driving forces for the separation. All of these results proved that ionic liquid (IL)-based ATPSs could potentially be substituted with DES-based ATPSs to offer new possibilities in the extraction of proteins.

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Sudden death syndrome (SDS) is an important soybean disease that potentially can be controlled by host plant resistance. In this study, over 800 soybean plant introductions (PIs), lines, and cultivars were screened for resistance to Fusarium solani. Of 728 PIs from China, PI 567.374 had mean foliar SDS severities significantly (P = 0.05) lower than PI 520.733 (resistant check) in both growth-chamber and greenhouse tests. In addition, PIs 567.315, 567.441C, 567.650B, and 567.664 had mean SDS severity ratings significantly (P = 0.05) lower than PI 520.733 in a growth-chamber test. Of 16 soybean cyst nematode-resistant entries tested, 5 had values lower than the resistant check, PI 520.733, with cv. Hartwig significantly lower in the first trial. In trial two, no entries were lower than the resistant check, although cvs. Bell and Hartwig were not significantly different from PI 520.733. In another experiment, few soybean cultivars or experimental lines had SDS severity ratings lower than PI 520.733 in any one of three trials. Some of the newly acquired PIs from China that exhibited low foliar SDS severity ratings may provide the sources of resistance needed to develop new SDS-resistant soybean breeding lines and cultivars.

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