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In the current study, we assessed bacterial diversity in the gut content of pond-reared grass carp (Ctenopharyngodon idellus), in the associated habitat environments (pond water and sediment) and in the ingested food (commercial feed and the reed Phragmites australis) by analysing 16S rDNA sequences from clone libraries. The highest bacterial diversity was observed in the gut content and was determined by the total number of operational taxonomic units, Shannon diversity index (H), Shannon equitability index (E H ), Coverage (C good ) and rarefaction curves calculated from the 16S rDNA gene libraries. Our data indicated that allochthonous gut microbes of grass carp were distinctively di¡erent from the corresponding environmental microbes. The pairwise similarity coe⁄cient (C s ) for microbe communities between gut content and ingested food was higher than for those between the gut content and habitats, indicating that the allochthonous microbiota identi¢ed in the intestines of grass carp were phylogenetically closer to those in the ingested food than to those in the habitat. Based on our study and previous research, we suggest that the digesta of grass carp harbours a microbiota phylogenetic core of Proteobacteria and Firmicutes and this observation deserves further investigations with respect to a potential pool of probiotics to grass carp.

Doenjang, a traditional Korean fermented soybean paste, is made by mixing and ripening meju with high salt brine (approximately 18%). Meju is a naturally fermented soybean block prepared by soaking, steaming, and molding soybean. To understand living bacterial community migration and the roles of bacteria in the manufacturing process of doenjang, the diversity of culturable bacteria in meju and doenjang was examined using media supplemented with NaCl, and some physiological activities of predominant isolates were determined. Bacilli were the major bacteria involved throughout the entire manufacturing process from meju to doenjang; some of these bacteria might be present as spores during the doenjang ripening process. Bacillus siamensis was the most populous species of the genus, and Bacillus licheniformis exhibited sufficient salt tolerance to maintain its growth during doenjang ripening. Enterococcus faecalis and Enterococcus faecium, the major lactic acid bacteria (LAB) identified in this study, did not continue to grow under high NaCl conditions in doenjang. Enterococci and certain species of coagulase-negative staphylococci (CNS) were the predominant acid-producing bacteria in meju fermentation, whereas Tetragenococcus halophilus and CNS were the major acid-producing bacteria in doenjang fermentation. We conclude that bacilli, LAB, and CNS may be the major bacterial groups involved in meju fermentation and that these bacterial communities undergo a shift toward salt-tolerant bacilli, CNS, and T. halophilus during the doenjang fermentation process.

Seed germination plays important roles in the establishment of seedlings and their subsequent growth; however, seed germination is inhibited by salinity, and the inhibitory mechanism remains elusive. Our results indicate that NaCl treatment inhibits rice seed germination by decreasing the contents of bioactive gibberellins (GAs), such as GA1 and GA4, and that this inhibition can be rescued by exogenous bioactive GA application. To explore the mechanism of bioactive GA deficiency, the effect of NaCl on GA metabolic gene expression was investigated, revealing that expression of both GA biosynthetic genes and GA-inactivated genes was up-regulated by NaCl treatment. These results suggest that NaCl-induced bioactive GA deficiency is caused by up-regulated expression of GA-inactivated genes, and the up-regulated expression of GA biosynthetic genes might be a consequence of negative feedback regulation of the bioactive GA deficiency. Moreover, we provide evidence that NaCl-induced bioactive GA deficiency inhibits rice seed germination by decreasing α-amylase activity via down-regulation of α-amylase gene expression. Additionally, exogenous bioactive GA rescues NaCl-inhibited seed germination by enhancing α-amylase activity. Thus, NaCl treatment reduces bioactive GA content through promotion of bioactive GA inactivation, which in turn inhibits rice seed germination by decreasing α-amylase activity via down-regulation of α-amylase gene expression.

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To increase the thermostability of Rhizomucor miehei lipase, the software Disulfide by Design was used to engineer a novel disulfide bond between residues 96 and 106, and the corresponding double cysteine mutants were constructed. The R. miehei lipase mutant could be expressed by Pichia pastoris in a free secreted form or could be displayed on the cell surface. The new disulfide bond spontaneously formed in the mutant R. miehei lipase. Thermostability was examined by measuring of hydrolysis activity using 4-nitrophenyl caprylate as a substrate. The engineered disulfide bond contributed to thermostability in the free form of the R. miehei lipase variant. The variant displayed on the yeast cell surface had significantly increased residual hydrolytic activity in aqueous solution after incubation at 60 degrees C for 5 h and increased synthetic activity in organic solvent at 60 degrees C. These results indicated that yeast surface display might improve the stability of R. miehei lipase, as well as amplifying the thermostability through the engineered disulfide bond.