Isolation, identification, and pathogenicity of Lelliottia amnigena causing soft rot of potato tuber in China.

Potato (Solanum tuberosum L.) is regarded as the fourth most important food crop because of its economic and nutritional benefits. This crop suffers significant annual losses due to a variety of phytopathogens. Bacterial soft rot disease is one of the most serious diseases that cause significant losses in potato yield all over the world. Therefore, identification of a soft rot pathogen is critical for easy control, as each pathogen has distinct ways of being controlled. Lelliottia amnigena is a subgroup of the genus Enterobacter with many species associated with crop plants, making its classification difficult and complex. Therefore, this study focused on the isolation and identification of a newly L. amnigena from rotten potato tuber obtained from the field after harvest, Lanzhou City, China. Four strains designated as PC2, PC3, PC4 and PC5 were isolated from the same rotting potato tuber. Pathogenicity test showed that strain PC3 induced soft rot symptoms on healthy potato tubers. Koch's postulates were confirmed by re-isolating the strain PC3 in the inoculated tubers. Strain PC3 showed a convex, oval and smooth colony, measuring 0.9-1.3 1.8-3.6 µm under the microscopic observation. Phylogenetic analysis based on 16S rRNA, rpoB and atpD genes showed that strain PC3 species was 99.44%, 97.24%, and 100%, closely related to L. amnigena with accession numbers 240-a-etp (MN208158.1), FDAARGOS (CP023529.1) and R-6 (MN658356.1), respectively. The bacterial strain (PC3) was deposited in the Genbank with the accession number SUB10508072 PC3 OK447935. To the best of our knowledge, this is the first report of L. amnigena causing soft rot on potato tubers in China.

The medusa of Aurelia coerulea is similar to its polyp in molecular composition and different from the medusa of Stomolophus meleagris in toxicity.

BACKGROUND: The incidents of Aurelia sp. stinging have recently increased because of a bloom in offshore area. However, their symptoms are much milder than those from another scyphozoan jellyfish, Stomolophus meleagris. METHODS: The molecular composition of the medusa and polyp of Aurelia coerulea was analyzed by sequencing the transcriptome and proteome. The toxicity of tentacle extract from A. coerulea medusa (A-TE) and S. meleagris medusa (S-TE) was measured by the survival rates of mice, their blood indexes, and integrity of red blood cells. RESULTS: The medusa and polyp of A. coerulea are similar in molecular composition, while their gene expressions are significantly different at both transcriptome and proteome levels. A-TE displayed no in vitro hemolysis and caused mild damage to the liver, heart and kidney instead of lethality. In contrast, S-TE showed strong hemolytic toxicity, and lethal effect with serious damage to the liver, heart and kidney. The toxin screening in the medusae showed that there were similar toxin categories though the number of toxin species in A. coerulea was larger than that in S. meleagris. Among them, lactotransferrin and venom prothrombin activator were the two predominant protein toxins in the medusae of A. coerulea and S. meleagris, respectively. CONCLUSIONS: A. coerulea medusa and polyp have similar molecular compositions, though there are observable morphological differences. The toxicity of A. coerulea medusa is significantly weaker than that of S. meleagris medusa of which the variation in toxin expressions is feasibly an important reason.

High-level expression and characterization of the Bacillus subtilis subsp. subtilis str. BSP1 YwaD aminopeptidase in Pichia pastoris.

Aminopeptidases are widely used for creating protein hydrolysates and peptide sequencing. The ywaD gene from a new Bacillus isolate, named Bacillus subtilis subsp. subtilis str. BSP1, was cloned into the yeast expression vector pHBM905A and expressed and secreted by Pichia pastoris strain GS115. The deduced amino acid sequence of the aminopeptidase encoded by the ywaD gene shared up to 98% identity with aminopeptidases from B. subtilis strains 168 and zj016. The yield (3.81 g/l) and specific activity (788 U/mg) of recombinant YwaD in high-density fermentation were extremely high. And 829.83 mg of the purified enzyme (4089.72 U/mg) were harvested. YwaD was glycosylated, and its activity decreased after deglycosylation, which was similar to that of the aminopeptidase from B. subtilis strain zj016. YwaD was most active toward l-arginine-4-nitroanilide. Moreover, it exhibited high resistance to carbamide, which was not true for aminopeptidases from B. subtilis strains 168 and zj016, which could simplify the purification of YwaD. Moreover, the expression and parts of characterization of the aminopeptidase from B. subtilis strain 168 in Pichia pastoris were added as supplementary material. The sequence and other characteristics of YwaD were compared with those of aminopeptidases from B. subtilis strains 168 and zj016, and they will provide a solid foundation for further research on the influence of amino acid mutations on the function of aminopeptidases.