RESUMO
Pickle like odor (PLO) is one of the main defective flavors of Maotai flavor baijiu (MFB). Understanding and controlling the PLO compounds producing strains not only solves the problem of PLO from the source, but also ensures the high-quality production of MFB. However, the relevant research on PLO compounds producing strains has not been reported in MFB. In this study, we identified a Bacillus amyloliquefaciens ZZ7 with high yield of PLO compounds in the fermented grains of MFB, and measured its physiological characteristics. It produces 627 volatile compounds and 1,507 non-volatile compounds. There are 7 volatile sulfur compounds that cause the PLO, the content of dimethyl disulfide, dimethyl trisulfide, and dimethyl sulfur is relatively high, accounting for 89.43% of the total volatile sulfur compounds. The genome size of B. amyloliquefaciens ZZ7 is 3,902,720 bp with a GC content of 46.09%, and a total of 3,948 protein coding genes were predicted. Moreover, the functional annotation of coding genes and an assessment of the metabolic pathways were performed by genome annotation, showing it has strong ability to transport and metabolize amino acids and carbohydrates. Comprehensive genomic and metabolomic analysis, the metabolic pathway of PLO compounds of B. amyloliquefaciens ZZ7 was revealed, which mainly involves 12 enzymes including sulfate adenylyltransferase, cysteine synthase, cystathionine γ-synthase, etc. This work provides biological information support at both genetic and metabolic levels for the mechanism of B. amyloliquefaciens ZZ7 to synthesize PLO compounds, and provides a direction for the subsequent genetic modification of ZZ7 to solve PLO from the source in the MFB.
RESUMO
Ichthyophthirius multifiliis is a protozoan ciliate that causes white spot disease (also known as ichthyophthiriasis) in freshwater fish. Holland's spinibarbel (Spinibarbus hollandi) was less susceptible to white spot disease than grass carp (Ctenopharyngodon Idella). In this study, grass carp and Holland's spinibarbel are infected by I. multifiliis and the amount of infection is 10,000 theronts per fish. All grass carp died within 12 days after infection, and the survival rate of Holland's spinibarbel was more than 80%. In order to study the difference in sensitivity of these two fish species to I. multifiliis, transcriptome analysis was conducted using gill, skin, liver, spleen and head kidney of Holland's spinibarbel and grass carp at 48 h post-infection with I. multifiliis. A total of 489,296,696 clean reads were obtained by sequencing. A total of 105 significantly up-regulated immune-related genes were obtained by Gene Ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis in grass carp. Cluster of differentiation 40 (CD40), cluster of differentiation 80 (CD 80), tumor necrosis factor-alpha (TNF-α), toll-like receptor 4 (TLR-4), interleukin 1 beta (IL-1ß) and other inflammatory-related genes in grass carp were enriched in the cytokine-cytokine receptor interaction pathway and toll-like receptor pathway. In Holland's spinibarbel, a total of 46 significantly up-regulated immune-related genes were obtained by GO classification and KEGG pathway enrichment analysis. Immune-related genes, such as Immunoglobin heavy chain (IgH), cathepsin S (CTSS), complement C1q A chain (C1qA), complement component 3 (C3) and complement component (C9) were enriched in phagosome pathway, lysosome pathway and complement and coagulation concatenation pathway. C3 was significantly up-regulated in gill and head kidney. Fluorescence in situ hybridization (FISH) showed that the C3 gene was highly expressed in gill tissue of Holland's spinibarbel infected with I. multifiliis. A small amount of C3 gene was expressed in the gill arch of grass carp after infected with I. multifiliis. In conclusion, the severe inflammatory response in vivo after infecting grass carp with I. multifiliis might be the main cause of the death of grass carp. The extrahepatic expression of the gene of Holland's spinibarbel might play an important role in the immune defense against I. multifiliis.