As an indicator of the antioxidant capability of plants, catalase can detoxify reactive oxygen species (ROS) generated by environmental stresses. Sweet potato is one of the top six most important crops in the world. However, its catalases remain largely unknown. In this study, a catalase encoding gene, IbCAT2 (accession number: KY615708), was identified and cloned from sweet potato cv. Xushu 18. It contained a 1479 nucleotides' open reading frame (ORF). S-R-L, Q-K-L, and a putative calmodulin binding domain were located at the C-terminus of IbCAT2, which suggests that IbCAT2 could be a peroxisomal catalase. Next-generation sequencing (NGS) based quantitative analyses showed that IbCAT2 was mainly expressed in young leaves and expanding tuberous roots under normal conditions. When exposed to 10% PEG6000 or 200 mmol/L NaCl solutions, IbCAT2 was upregulated rapidly in the first 11 days and then downregulated, although different tissues showed different degree of change. Overexpression of IbCAT2 conferred salt and drought tolerance in Escherichia coli and Saccharomyces cerevisiae. The positive response of IbCAT2 to abiotic stresses suggested that IbCAT2 might play an important role in stress responses.
A bacterial strain, Streptomyces albogriseolus LBX-2, was isolated from a soil sample in Chengdu, China. S. albogriseolus LBX-2 is an aerobic and Gram-positive microorganism that is capable of using the polyethylene as the sole carbon source. Results of scanning electron microscopy and tensile tests indicated that S. albogriseolus LBX-2 could cause the damages to polyethylene (PE). Suspension culture of LBX-2 resulted in the weight loss in the PE powder over a 15-day period. The bacterial growth curve assay clearly demonstrated the utilization of n-hexadecane and n-octadecane for the strain LBX-2. Phylogenetic analysis showed that it was grouped in the same clade as S. albogriseolus belonging to Streptomyces. The complete genome of strain LBX-2 consists of a chromosome of 7,210,477 bp and a linear plasmid of 336,677 bp. Compared with other strains of Streptomyces, the genome size of S. albogriseolus LBX-2 was smaller than the average but its guanine and cytosine content (72.47%) was higher than the others. The Non-Redundant Protein Database (NR), Kyoto Encyclopedia of Genes and Genomes (KEGG), SwissProt, Gene Ontology (GO) and Clusters of Orthologous Groups (COG) annotations provided information on the specific functions of encoded proteins. A total of 21 monooxygenase and 22 dioxygenase genes were found in its genome. Synteny comparison with the genome of Streptomyces coelicolor A3(2) revealed a low overall genetic diversity between them. This study provides valuable information to reveal the underlying mechanisms on PE degradation by S. albogriseolus LBX-2.
Bacillus subtilis S1-4, isolated from chicken feather could efficiently degrade feathers by secreting several extracellular proteases. In order to get insight into the individual protease involved in keratin hydrolysis, a keratinase designed as BsKER71 was cloned and expressed in Bacillus subtilis WB600. In silico analysis revealed that BsKER71 protein contained a mature protein of 36.1 kDa. Further, purified BsKER71 could hydrolyze a variety of natural proteins, such as fibrous protein, collagen protein, casein, keratin and bovine serum albumin. In addition, this keratinase exhibited high enzyme activity in a wide range of pH and optimal pH of 10.0 and 9.0 in the hydrolysis of casein and keratin, respectively. Similarly, the optimal temperature was 55 °C and 50 °C for the hydrolysis of above two substrates, respectively. The hydrolytic activity was significantly inhibited by phenylmethanesulfonyl fluoride (PMSF), indicating the presence of serine residue in the active site. Moreover, ethylenediaminetetraacetic acid (EDTA) and phenanthroline moderately inhibited the hydrolytic activity. The catalytic activity was stimulated by Mg2+ and Ca2+, but greatly inhibited by Cu2+. Furthermore, several chemicals exhibited different effects on the hydrolysis of casein and keratin by BsKER71. These results provided a better understanding of BsKER71 from feather degrading bacterium B. subtilis S1-4.
Drought restricts the growth of alpine grassland vegetation. This study aimed to explore a new technical system to improve the drought resistance of forage grass. Qinghai cold-land Poa pratensis seedlings were used in the drought stress experiment. A combination of abscisic acid (ABA) and polyacrylamide (PAM) were used to affect the growth, leaf physiology, soil enzyme activity, and rhizosphere microbial diversity of P. pratensis. The fresh leaf weight and root surface area were significantly increased after ABA-PAM combined treatment, while root length was significantly reduced. Besides, the leaf catalase (CAT) and superoxide dismutase (SOD) enzyme activity, proline and chlorophyll content, increased after the treatment, while malondialdehyde (MDA) content decreased. The treatment also increased sucrase, urease, and alkaline protease activities in rhizosphere soil, while decreasing acid phosphatase and neutral phosphatase enzyme activities. ABA-PAM combined treatment enhanced the rhizosphere microbial community and forage drought resistance by altering the abundance of various dominant microorganisms in the rhizosphere soil. The relative abundances of Actinobacteria, Chloroflexi, and Acidobacteria decreased, while Proteobacteria, Firmicutes, and Ascomycota increased. Unlike the relative abundance of Gibberella that decreased significantly, Komagataeibacter, Lactobacillus, Pichia, and Dekkera were significantly increased. Single-factor collinearity network analysis revealed a close relationship between the different rhizosphere microbial communities of forage grass, after ABA-PAM treatment. This study implies that ABA-PAM combined treatment can improve the drought resistance of forages. Therefore, it provides a theoretical and practical basis for restoring drought-induced grassland degradation.
Main bearing that plays the role of supporting and making the cutter to rotate and tunnel is the core part of TBM. Because of the harsh working conditions and complex changeful construction, the axial and radiai load and environmental factors such as temperature of TBM main bearing are changing to make the fault of main bearing presenting randomness, and then the not easy identified fault may be produced. The traditional neural network model can not dynamic consider the cause of the reasons, parts and types, BP neural network fault diagnosis model based on fault reasons-signs matrix is presented in this papero Firstly, the faults are screened through the fault reasons-symptom matrix, and the neural network structure is designed according to the screening result, then, the fault type is identified by way of model training. According to TBM main bearing fault symptoms data provided by a heavy enterprises practical engineering and MATLAB simulation validation, the feasibility and superiority of this model method are proved.
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