Antibiotic Effects of Volatiles Produced by Bacillus tequilensis XK29 against the Black Spot Disease Caused by Ceratocystis fimbriata in Postharvest Sweet Potato.

Black spot disease caused by Ceratocystis fimbriata is destructive to the production, transportation, and storage of sweet potato. The antifungal effects of Bacillus tequilensis XK29 against C. fimbriata through volatile organic compounds (VOCs) were evaluated in this study. The activated carbon assay proved that XK29 could exert antibiotic effects through volatiles. By optimizing the wheat seed weight, inoculation method, concentration, volume, and time, the antifungal activity of XK29 was significantly improved. XK29 fumigation inhibited spore formation and germination and changed the cell morphology of C. fimbriata. During the storage of sweet potato tuber roots, XK29 effectively controlled black spot disease and reduced the weight loss and malondialdehyde content. Metabolomic analysis revealed that 21 volatile compounds were released from XK29. Isovaleric acid, isobutyric acid, and 2-methylbutanoic acid effectively inhibited the growth of C. fimbriata. These results indicate that B. tequilensis XK29 has a good potential to be developed as a microbial fumigation agent.

Antifungal effect of volatile organic compounds produced by Pseudomonas chlororaphis subsp. aureofaciens SPS-41 on oxidative stress and mitochondrial dysfunction of Ceratocystis fimbriata.

Ceratocystis fimbriata is the pathogen of black rot disease, which widely exists in sweet potato producing areas all over the world. The antifungal activity of volatile organic compounds (VOCs) released by Pseudomonas chlororaphis subsp. aureofaciens SPS-41 against C. fimbriata was reported in our previous study. In this study, we attempted to reveal the underlying antifungal mechanism of SPS-41 volatiles. Our results showed that the VOCs released by SPS-41 caused the morphological change of hyphae, destroyed the integrity of cell membrane, reduced the content of ergosterol, and induced massive accumulation of reactive oxygen species in C. fimbriata cells. Furthermore, SPS-41 fumigation decreased the mitochondrial membrane potential, acetyl-CoA and pyruvate content of C. fimbriata cells, as well as the mitochondrial dehydrogenases activity. In addition, the VOCs generated by SPS-41 reduced the intracellular ATP content and increased the extracellular ATP content of C. fimbriata. In summary, SPS-41 fumigation exerted its antifungal activity by inducing oxidative stress and mitochondrial dysfunction in C. fimbriata.

Coupling of capillary electrophoresis with electrospray ionization multiplexing ion mobility spectrometry.

In this work, ion mobility spectrometry (IMS) function as a detector and another dimension of separation was coupled with CE to achieve two-dimensional separation. To improve the performance of hyphenated CE-IMS instrument, electrospray ionization correlation ion mobility spectrometry is evaluated and compared with traditional signal averaging data acquisition method using tetraalkylammonium bromide compounds. The effect of various parameters on the separation including sample introduction, sheath fluid of CE and drift gas, data acquisition method of IMS were investigated. The experimental result shows that the optimal conditions are as follows: hydrodynamic sample injection method, the electrophoresis voltage is 10 kilo volts, 5 mmol/L ammonium acetate buffer solution containing 80% acetonitrile as both the background electrolyte and the electrospray ionization sheath fluid, the ESI liquid flow rate is 4.5 µL/min, the drift voltage is 10.5 kilo volts, the drift gas temperature is 383 K and the drift gas flow rate is 300 mL/min. Under the above conditions, the mixture standards of seven tetraalkylammoniums can be completely separated within 10 min both by CE and IMS. The linear range was 5-250 µg/mL, with LOD of 0.152, 0.204, 0.277, 0.382, 0.466, 0.623 and 0.892 µg/mL, respectively. Compared with traditional capillary electrophoresis detection methods, the developed CE-ESI-IMS method not only provide two sets of qualitative parameters including electrophoresis migration time and ion drift time, ion mobility spectrometer can also provide an additional dimension of separation and could apply to the detection ultra-violet transparent compounds or none fluorescent compounds.

Analysis of alkaloids from Peganum harmala L. sequential extracts by liquid chromatography coupled to ion mobility spectrometry.

An orthogonal two dimensional analysis method based on high performance liquid chromatography (HPLC) separation and electrospray ionization-ion mobility spectrometry (ESI-IMS) detection was developed for the analysis of alkaloid compounds from Peganum harmala L. seeds. Reverse phase (RP) and hydrophilic interaction chromatography (HILIC) were compared for the most optimal performance using three different chromatographic columns. The experimental results suggest that HILIC mode is a better option for combining with the ESI-IMS system for higher sensitivity and ease in hyphenating. Under optimized conditions, alkaloids from different extraction phases were determined by means of the established HPLC-IMS method. More compounds from Peganum harmala L. seed extracts were differentiated on the HPLC-ESI-IMS system by their retention time and drift time than by HPLC or ESI-IMS alone, and thirteen alkaloids were tentatively identified based on m/z and fragment ions using ultra-high-performance liquid chromatography tandem mass-spectrometry (UPLC-MS/MS). Hence, our results indicate that this method can be considered to be advantageous over traditional absorbance detection methods for resolving complex mixtures because of complementary separation steps, elevated peak capacity, and higher sensitivity.