A method was developed for the determination of matrine residues in quinoa (Chenopodium quinoa Willd.) plants and soil by liquid chromatography triple quadrupole tandem mass spectrometry (LC-MS/MS) with QuEChERS clean-up. Matrine from soil, quinoa roots, stems, leaves and seeds was extracted with 25% ammonia, 20 mL acetonitrile/methanol, salted with sodium chloride (NaCl) and purified with anhydrous magnesium sulfate (MgSO4), N-propyl ethylenediamine (PSA) and graphitized carbon black (GCB). Then a chromatographic separation was performed on a Shim-pack XR-ODS II (75 mm × 2.0 mm, i. d., 2.2 µm) column with a gradient elution of 5 mmol/L ammonium formate-methanol as the mobile phase and monitored in multiple reaction monitoring modes (MRM) in electrospray positive ionization mode. The results showed that in the range of 0.005~1 mg/L, the linear correlation coefficients of matrine in the five matrices were all above 0.999. The LOQs for soil, quinoa roots, stems, leaves and seeds were 0.005, 0.005, 0.01, 0.01 and 0.005 mg/kg, respectively. The mean recoveries ranged from 74.42% to 98.37%, with RSDs of 1.25–6.84% at the three concentration addition levels. The average intra-day and inter-day recoveries were 73.92–92.36% and 78.56–90.18%, respectively, with RSDs below 8.72% and 9.43%. The recoveries and reproducibility of the method were superior. The method was used to determine the actual samples, which indicated that the half-lives of matrine in quinoa seeds, leaves, stems and soil were 1.28–1.32, 1.03–1.21, 0.81–0.92 and 0.93–0.97 d. It has a half-life below 30 d, which is an easily dissipated pesticide. The method is simple, sensitive, accurate, reliable and applicable to a wide range of applications, and it can achieve the rapid multi-residue determination of matrine to a certain extent. Next Generation Sequencing was used to explore the effects of exposure to high and low doses of matrine on soil bacterial communities and the composition of the three soils in the Qinghai Province (Haixi, Haidong and Haibei). The results showed that the number of ASVs increased significantly after treatment with matrine at an effective dose of 0.1 mg/kg than after treatment with matrine at an effective dose of 5.0 mg/kg. Similarly, bacterial abundance was higher after 0.1 mg/kg of matrine treatment than after 5.0 mg/kg of matrine treatment. The inhibitory effect on some bacterial flora was enhanced with an increase in matrine application, while the inhibitory effect on bacterial flora was weakened with time. Applying a certain dose of matrine e changed the relative abundance of the dominant bacterial genera of the soil bacteria.
Halophilic microbial as prospective resources of biotechnology due to the advantages of flexible survivability. Qarhan Salt Lake is the second largest Salt Lake in the world which contains rich-unique extremophiles and deserved in-depth exploration. Present study first time isolated novel strain Halobacillus trueperi S61 from Qarhan Salt Lake and performed whole-genome sequencing through combined third-generation PacBio and second-generation Illumina technology. The whole genome of Halobacillus trueperi S61 identified 57549 total reads and consists a complete circular chromosome of 4047887 bp with 43.86% GC content without gaps. Total number of 139 non-coding RNA (included 86 tRNA, 30 rRNA and 23 sRNA), 16 gene islands with 260275 bp and two prophages (with 82682 length) were predicted. In addition, the whole genome of Halobacillus trueperi S61 summarized basic annotation for 3982 protein-coding genes, 3980, 3667, 2998 and 2303 unigenes were annotated with Nr, Swissport, KOG and KEGG database. Combined with advanced analysis, 561 carbohydrate enzymes and 4416 pathogen host interactions related genes were identified. The protein function of Halobacillus trueperi S61 was mainly focus on biological processes, and the protein function was mainly distributed in gene transcription and amino acids, and carbohydrates metabolism. The complete whole genome sequence assembly and annotation of novel strain Halobacillus trueperi S61 isolated from Qarhan Salt Lake mainly focus on protein biological processes and antibiotic resistance, provides a potential resource for biotechnology.
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