Genome Resource of Sphingomonas carotinifaciens L9-754T, an Endophyte Isolated From Leaf Tissues of Jatropha curcas.

Sphingomonas carotinifaciens strain L9-754T (DSM 27347) is a gram-negative, chemoheterotrophic, and rod-shaped endophyte isolated from the stem tissues of Jatropha curcas L. This strain has putative in vitro antagonistic ability against the plant pathogenic fungus Magnaporthe grisea. A draft genome of L9-754T was obtained using the PacBio SMRT cell platform. By analyzing the genome of strain L9-754T, a gene cluster (GQR91_18700 - GQR91_18715) related to an antioxidant enzyme was identified in the obtained draft genome. The information obtained from the draft genome is expected to reveal the putative properties helpful in biocontrol applications.

Emerging of two new subgenotypes of porcine reproductive and respiratory syndrome viruses in Southeast China.

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the leading swine pathogens and causes major economic loss to the global swine industry. In this study, a total of 49 PRRSV isolates were collected from different swine herds in seven provinces in Southeast China from 2014 to 2015. All the ORF5 genes and some Nsp2 genes were sequenced. Phylogenetic analysis showed that all the isolates belonged to the North America genotype. Among them, five isolates formed a new subgenotype IV derived from highly pathogenic PRRSV (HP-PRRSV). Six isolates formed subgenotype III, which were closely related to the NADC30 strain in the US. These isolates formed 13 putative N-linked glycosylation site (NGS) patterns based on N30, 33, 34, 35, 44 and 51. There were fewer NGSs of isolates in subgenotype IV than in subgenotype III. This indicates that the two new subgenotypes of PRRSV strains with different NGS patterns were spreading in those regions of China. The genetic diversity should be considered for the control and prevention of this disease.

A Three-Dimensional Technique for The Visualization of Mitochondrial Ultrastructural Changes in Pancreatic Cancer Cells.

Understanding the dynamic features of the cell organelle ultrastructure, which is not only rich in unknown information but also sophisticated from a three-dimensional (3D) perspective, is critical for mechanistic studies. Electron microscopy (EM) offers good imaging depth and allows for the reconstruction of high-resolution image stacks to investigate the ultrastructural morphology of cellular organelles even at the nanometer scale; therefore, 3D reconstruction is gaining importance due to its incomparable advantages. Scanning electron microscopy (SEM) provides a high-throughput image acquisition technology that allows for reconstructing large structures in 3D from the same region of interest in consecutive slices. Therefore, the application of SEM in large-scale 3D reconstruction to restore the true 3D ultrastructure of organelles is becoming increasingly common. In this protocol, we suggest a combination of serial ultrathin section and 3D reconstruction techniques to study mitochondrial cristae in pancreatic cancer cells. The details of how these techniques are performed are described in this protocol in a step-by-step manner, including the osmium-thiocarbohydrazide-osmium (OTO) method, the serial ultrathin section imaging, and the visualization display.

Complete genome sequence of Achromobacter spanius type strain DSM 23806T, a pathogen isolated from human blood.

OBJECTIVES: Achromobacter spanius is a newly described, non-fermenting, Gram-negative, coccoid pathogen isolated from human blood. Whole-genome sequencing of the A. spanius type strain was performed to investigate the mechanism of pathogenesis of this strain at a genomic level. METHODS: The complete genome of A. spanius type strain DSM 23806T was sequenced using single-molecule real-time (SMRT) DNA sequencing. RESULTS: The complete genome of DSM 23806T consists of one circular DNA chromosome of 6425783bp with a G+C content of 64.26%. The entire genome contains 5804 predicted coding sequences (CDS) and 55 tRNAs. Genomic island (GI) analysis showed that this strain encodes several important pathogenesis- and resistance-related genes. CONCLUSIONS: These results strongly suggest that GIs provide some fitness advantages in A. spanius type strain DSM 23806T. This report provides an extensive understanding of A. spanius at a genomic level as well as an understanding of the evolution of A. spanius.

Complete genome sequence of Achromobacter insolitus type strain LMG 6003T, a pathogen isolated from leg wound.

Achromobacter insolitus is a newly described, Gram-negative, small (1-2 mm long) and coccoid pathogen that has been isolated from leg wound. Due to its potential threat to human beings, the type strain of this species LMG 6003T was completely sequenced in this study. The genomic analysis revealed that the genome of LMG 6003T consists of one circular DNA chromosome of 6 492 697 bp with a GC content of 65.10%. The entire genome contains 6208 predicted coding genes, 61 tRNAs and 13 rRNA genes. Comparative genome analysis between LMG 6003T and another selected 10 sequenced Achromobacter revealed that 725 genes only exist in LMG 6003T. In particular, these genes include several important pathogenic-related genes such as Type IV prepilin peptidase (TadV/CpaA), Type VI secretion lipoprotein (VasD) and type 1 fimbriae major subunit (FimA). Genomic island (GI) analysis also suggests the existence of GIs in LMG 6003T. All these results strongly suggest the unique genomic features in LMG 6003T against other Achromobacter species. This report will provide us with an extended understanding of A. insolitus at the genomic level and would be helpful for understanding the evolution of the Achromobacter genus.