Elucidation of the Initial Growth Process and the Infection Mechanism of Penicillium digitatum on Postharvest Citrus (Citrus reticulata Blanco).

Green mold disease, a common citrus post-harvest disease caused by Penicillium digitatum, has an unresolved initial infection mechanism. Understanding the infection mechanism leads to the development of potential controls and preventive measures against the disease. The present study aimed to delineate the infection mechanism by investigating spore germination, changes of organic molecules and enzyme activity, and differential expression of genes in the P. digitatum infection. P. digitatum spore germination was observed by a pathology section scanner and it was found that in vivo germination was 3 h behind the in vitro germination. In addition, cell wall degrading enzymes and soluble sugar and titratable acid content during the infection process measured dynamically. The level of pectinase reached its maximum of 6067 U/g before 48 hpi, while cellulase increased rapidly after 48 hpi. The soluble sugar and organic acid content increased considerably with the progression of the infection. The transcriptomic profile of P. digitatum before and after infection was analyzed by RNA-seq. The genes related to cell wall degrading enzymes were significantly up-regulated and annotated to participate in two major carbon source synthesis pathways. The study delineated the initial infection mechanism of P. digitatum which eventually opened the gate way for the development of new control strategies in the future.

Effect of ultrasonic treatment on total phenolic extraction from Lavandula pubescens and its application in palm olein oil industry.

The aims of the current study were to evaluate the best technique for total phenolic extraction from Lavandula pubescens (Lp) and its application in vegetable oil industries as alternatives of synthetic food additives (TBHQ and BHT). To achieve these aims, three techniques of extraction were used: ultrasonic-microwave (40 kHz, 50 W, microwave power 480 W, 5 min), ultrasonic-homogenizer (20 kHz, 150 W, 5 min) and conventional maceration as a control. By using the Folin-Ciocalteu method, the total phenolic contents (TPC) (mg gallic acid equivalent/g dry matter) were found to be 253.87, 216.96 and 203.41 for ultrasonic-microwave extract, ultrasonic-homogenizer extract and maceration extract, respectively. The ultrasonic-microwave extract achieved the higher scavenger effect of DPPH (90.53%) with EC50 (19.54 µg/mL), and higher inhibition of ß-carotene/linoleate emulsion deterioration (94.44%) with IC50 (30.62 µg/mL). The activity of the ultrasonic-microwave treatment could prolong the induction period (18.82 h) and oxidative stability index (1.67) of fresh refined, bleached and deodorized palm olein oil (RBDPOo) according to Rancimat assay. There was an important synergist effect between citric acid and Lp extracts in improving the oxidative stability of fresh RBDPOo. The results of this work also showed that the ultrasonic-microwave assisted extract was the most effective against Gram-positive and Gram-negative strains that were assessed in this study. The uses of ultrasonic-microwave could induce the acoustic cavitation and rupture of plant cells, and this facilitates the flow of solvent into the plant cells and enhances the desorption from the matrix of solid samples, and thus would enhance the efficiency of extraction based on cavitation phenomenon.

Effect of ultrasound and high hydrostatic pressure (US/HHP) on the degradation of dextran catalyzed by dextranase.

In our current research work, the effect of combination of ultrasonic irradiation and high hydrostatic pressure (US/HHP) on the enzymatic activity and enzymatic hydrolysis kinetic parameters of dextran catalytic by dextranase were investigated. Furthermore, the effects of US/HHP on the structure of dextranase were also discussed with the aid of fluorescence spectroscopy and circular dichroism (CD) spectroscopy. The maximum hydrolysis of dextran was observed under US (40 W at 25 kHz for 15 min) combined with HHP (400 MPa for 25 min), in which the hydrolysis of dextran increased by 163.79% compared with the routine thermal incubation at 50 °C. Results also showed that, Vmax and KM values, as well as, kcat of dextranase under US/HHP treatment were higher than that under US, HHP and thermal incubation at 50 °C, indicated that, the substrate is converted into the product at an increased rate when compared with the incubation at 50 °C. Compared to the enzymatic reaction under US, HHP, and routine thermal incubation, dextranase enzymatic reaction under US/HHP treatment showed decreases in Ea, ΔG and ΔH, however small increase in ΔS value was observed. In addition, fluorescence and CD spectra reflected that US/HHP treatment had increased the number of tryptophan on dextranase surface with increased α-helix by 19.80% and reduced random coil by 6.94% upon US/HHP-treated dextranase protein compared to the control, which were helpful for the improvement of its activity. These results indicated that, the combination of US and HHP treatments could be an effective method for improving the hydrolysis of dextran in many industrial applications including sugar manufacturing processes.