This present study deals with synthesis, characterization and antibacterial activity of cross-linked chitosan-glutaraldehyde. Results from this study indicated that cross-linked chitosan-glutaraldehyde markedly inhibited the growth of antibiotic-resistant Burkholderia cepacia complex regardless of bacterial species and incubation time while bacterial growth was unaffected by solid chitosan. Furthermore, high temperature treated cross-linked chitosan-glutaraldehyde showed strong antibacterial activity against the selected strain 0901 although the inhibitory effects varied with different temperatures. In addition, physical-chemical and structural characterization revealed that the cross-linking of chitosan with glutaraldehyde resulted in a rougher surface morphology, a characteristic Fourier transform infrared (FTIR) band at 1559 cm⁻¹, a specific X-ray diffraction peak centered at 2θ = 15°, a lower contents of carbon, hydrogen and nitrogen, and a higher stability of glucose units compared to chitosan based on scanning electron microscopic observation, FTIR spectra, X-ray diffraction pattern, as well as elemental and thermo gravimetric analysis. Overall, this study indicated that cross-linked chitosan-glutaraldehyde is promising to be developed as a new antibacterial drug.

The effects of temperature, agitation and aeration on glycoprotein GP-1 production by Streptomyces kanasenisi ZX01 in bench-scale fermentors were systematically investigated. The maximum final GP-1 production was achieved at an agitation speed of 200 rpm, aeration rate of 2.0 vvm and temperature of 30 °C. By using a dynamic gassing out method, the effects of agitation and aeration on volumetric oxygen transfer coefficient (k_La) were also studied. The values of volumetric oxygen transfer coefficient in the logarithmic phase increased with increase of agitation speed (from 14.53 to 32.82 h⁻¹) and aeration rate (from 13.21 to 22.43 h⁻¹). In addition, a successful scale-up from bench-scale to pilot-scale was performed based on volumetric oxygen transfer coefficient, resulting in final GP-1 production of 3.92, 4.03, 3.82 and 4.20 mg/L in 5 L, 15 L, 70 L and 500 L fermentors, respectively. These results indicated that constant volumetric oxygen transfer coefficient was appropriate for the scale-up of batch fermentation of glycoprotein GP-1 by Streptomyces kanasenisi ZX01, and this scale-up strategy successfully achieved 100-fold scale-up from bench-scale to pilot-scale fermentor.

Aims To extract and identify the metabolites of strain A217 as well as its antifungal spectrum and control effect on various plant pathogens. Methods and Results Strain A217 was identified as a Streptomyces spp which was most similar to Streptomyces lienomycini. An antimicrobial spectrum test indicated that strain A217 inhibited several plant pathogenic fungi and strong antibacterial effect such as Phytophthora capsici, Botrytis cinerea, Sclerotinia sclerotiorum, Fusarium oxysporum, Pseudomonas syringae and Xanthomonas campestris. An in vivo tissue test demonstrated that the fermentation broth of strain A217 exerted therapeutic and protective effects of 49.47% and 61.60%, respectively, on Sclerotinia sclerotiorum. Additionally, the fermentation broth of A217 exerted control effects on walnut black spot disease in walnut leaves and branches amounting to 79.33% and 81.52%, respectively. In a pot experiment, the fermentation broth exhibited a stronger protective and control effect (68.29%), as well as better bacteriostatic and disease control effects on Phytophthora blight of pepper, compared with Metalaxyl. Compounds possessing antifungal and antibacterial activities were obtained from the fermentation broth of strain A217, using column chromatography and HPLC. Chemical and structural analyses conducted using MS and NMR confirmed that these compounds were 1H‐pyrrole‐2‐carboxylic acid and 1H‐pyrrole‐2‐carboxamide. The EC₅₀ values of compound 1H‐pyrrole‐2‐carboxylic acid1 for S. sclerotiorum and P. capsici were 20.13 μg/mL and 50.36 μg/mL, respectively. Compound 1H‐pyrrole‐2‐carboxamide2 showed significant antibacterial activity against different plant pathogenic bacteria. The MIC values of Pseudomonas syringae, Xanthomonas campestris, and Xanthomonas campestris pv. jugiandis were 7.5 μg/mL, 30 μg/mL, and 15.0 μg/mL, respectively. Conclusions Actinomyces A217 fermentation products have a broad spectrum of bacteriostasis, and have good bacteriostasis activity to many plant pathogenic fungi and bacteria. Significance and Impact of Study The present study revealed a new antimicrobial producing strain of Streptomyces and its potential application as a biological control agent for plant diseases.

In order to develop a novel biofungicide, the antifungal activity and action mode of cuminic acid from the seed of Cuminum cyminum L. against Fusarium oxysporum f. sp. niveum (FON) on watermelon was determined systematically. In this study, the median effective concentration (EC₅₀) value for cuminic acid in inhibiting mycelial growth of FON was 22.53 μg/mL. After treatment with cuminic acid, the mycelial morphology was seriously influenced; cell membrane permeability and glycerol content were increased markedly, but pigment and mycotoxin (mainly fusaric acid) were significantly decreased. Synthesis genes of bikaverin (Bike1, Bike2 and Bike3) and fusaric acid (FUB1, FUB2, FUB3 and FUB4) both were downregulated compared with the control, as confirmed by quantitative RT-PCR. In greenhouse experiments, cuminic acid at all concentrations displayed significant bioactivities against FON. Importantly, significant enhancement of activities of SOD, POD, CAT and decrease of MDA content were observed after in vivo cuminic acid treatment on watermelon leaves. These indicated that cuminic acid not only showed high antifungal activity, but also could enhance the self-defense system of the host plant. Above all, cuminic acid showed the potential as a biofungicide to control FON.

Streptomyces kanasenisi ZX01 was found to produce a novel glycoprotein GP-1 previously, which was secreted into medium and had significant activity against tobacco mosaic virus. However, the low production of GP-1 by strain ZX01 limited its further studies. In order to improve the yield of GP-1, a series of statistical experimental design methods were applied to optimize medium of strain ZX01 in this work. Millet medium was chosen to be the optimal original medium for optimization. Soluble starch and yeast extract were identified as the optimal carbon and nitrogen source using one-factor-at-a-time method. Response surface methodology was used to optimize medium compositions (soluble starch, yeast extract and inorganic salts). A higher yield of GP-1 was 601.33 µg/L after optimization. The optimal compositions of medium were: soluble starch 13.61 g/L, yeast extract 4.19 g/L, NaCl 3.54 g/L, CaCO3 0.28 g/L, millet, 10 g/L. The yield of GP-1 in a 5 L fermentor using optimized medium was 2.54 mg/L, which is much higher than the result of shake flask. This work will be helpful for the improvement of GP-1 production on a large scale and lay a foundation for developing it to be a novel anti-plant virus agent.

Take-all, caused by Gaeumannomyces tritici, is one of the most important wheat root diseases worldwide, as it results in serious yield losses. In this study, G. tritici was transformed to express the hygromycin B phosphotransferase using a combined protoplast and polyethylene glycol (PEG)-mediated transformation technique. Based on a series of single-factor experimental results, three major factors—temperature, enzyme lysis time, and concentration of the lysing enzyme—were selected as the independent variables, which were optimized using the response surface methodology. A higher protoplast yield of 9.83 × 10⁷ protoplasts/mL was observed, and the protoplast vitality was also high, reaching 96.27% after optimization. Protoplasts were isolated under the optimal conditions, with the highest transformation frequency (46–54 transformants/μg DNA). Polymerase chain reaction and Southern blotting detection indicated that the genes of hygromycin phosphotransferase were successfully inserted into the genome of G. tritici. An optimised PEG-mediated protoplast transformation system for G. tritici was established. The techniques and procedures described will lay the foundation for establishing a good mutation library of G. tritici and could be used to transform other fungi.