Antimicrobial Activity and Identification of the Biosynthetic Gene Cluster of X-14952B From Streptomyces sp. 135.

The bacterial genus Streptomyces is an important source of antibiotics, and genome mining is a valuable tool to explore the potential of microbial biosynthesis in members of this genus. This study reports an actinomycete strain 135, which was isolated from Qinghai-Tibet Plateau in China and displayed broad antimicrobial activity. The fermentation broth of strain 135 displayed strong antifungal activity (>70%) against Sclerotinia sclerotiorum, Botrytis cinerea, Valsa mali, Phytophthora capsici, Glomerella cingulata, Magnaporthe grisea, Bipolaris maydis, Exserohilum turcicum in vitro, meanwhile possessed significant preventive and curative efficacy against S. sclerotiorum, Gaeumannomyces graminis, and P. capsici on rape leaves (54.04 and 74.18%), wheat (90.66 and 67.99%), and pepper plants (79.33 and 66.67%). X-14952B showed the greatest antifungal activity against S. sclerotiorum and Fusarium graminearum which the 50% inhibition concentration (EC50) were up to 0.049 and 0.04 µg/mL, respectively. Characterization of strain 135 using a polyphasic approach revealed that the strain displayed typical features of the genus Streptomyces. 16S rRNA gene sequencing and phylogenetic analysis demonstrated that the isolate was most closely related to and formed a clade with Streptomyces huasconensis HST28T (98.96% 16S rRNA gene sequence similarity). Average nucleotide identity (ANI) and DNA-DNA hybridization (DDH) values in strain 135 and related type strains were both below the threshold of species determination (91.39 and 56.5%, respectively). OrthoANI values between strain 135 and related type strains are under the cutoff of determining species (<95%). The biosynthetic gene cluster (BGC) designated to X-14952B biosynthesis was identified through genome mining and the possible biosynthesis process was deduced.

Optimization of a protective medium for enhancing the viability of freeze-dried Bacillus amyloliquefaciens B1408 based on response surface methodology.

Response surface methodology (RSM) is a commonly used system to optimize cryoprotectants of biocontrol strains when they are subjected to preparations. Various kinds of cryoprotectants and centrifugal conditions were tested to improve the survival of biocontrol agents after freeze-drying. To determine the optimum levels of incorporation of three cryoprotectants (glucose, trehalose and xylitol) in the freeze-drying process of strain Bacillus amyloliquefaciens B1408, a range of experiments based on Box-Behnken Design (BBD) were conducted. The results indicated that the suitable centrifugation conditions were 5000 r/min,10 min and the optimum concentrations of cryoprotectants were glucose 1.00%, trehalose 4.74% and xylitol 1.45%. The proven survival rate of cells after freeze-drying was 91.24%. These results convincingly demonstrated that freeze-drying could be used to preparation of biocontrol strain B1408. This study provides a theoretical basis for commercial possibilities and formulation development.