Metschnikowia citriensis FL01 antagonize Geotrichum citri-aurantii in citrus fruit through key action of iron depletion.

Metschnikowia citriensis FL01 has great potential for biocontrol applications for its excellent biocontrol efficacy on postharvest diseases of citrus fruit, and the iron depletion by pulcherriminic acid (PA) and then formation of insoluble pigment pulcherrimin had been speculated as an important action mechanism. To identify the genes involved in pulcherrimin synthesis and reutilization in M. citriensis FL01, we de novo assembled the genome of M. citriensis FL01 based on long-read PacBio sequencing. The final assembled genome consisted of 12 contigs with a genome size of 25.74 Mb, G + C content of 49.16% and 9310 protein-coding genes. The genome-wide BLAST of the PUL genes of M. pulcherrima APC 1.2 showed that the four PUL genes were clustered and located on Contig 4 of M. citriensis FL01. In order to further clarify the role of pulcherrimin pigment on biocontrol of M. citriensis FL01, CRISPR/cas9 technology was used to knock out PUL2 gene that was responsible for PA synthesis and the pigmentless mutants with stable phenotype were obtained. The mutant strains of M. citriensis FL01 lost the ability to produce pulcherrimin pigment, and simultaneously lost the ability to inhibit the growth of Geotrichum citri-aurantii in vitro. Moreover, the biocontrol efficacy of pigmentless mutant strains against sour rot was about 80% lower than that of wild-type M. citriensis FL01. These results directly proved that the iron depletion was an important mechanism of M. citriensis FL01.

Pichia galeiformis Induces Resistance in Postharvest Citrus by Activating the Phenylpropanoid Biosynthesis Pathway.

This study aimed to investigate the effect of Pichia galeiformis on disease resistance and elucidate the changes in phenylpropane biosynthesis treated by P. galeiformis in postharvest citrus. The results showed that P. galeiformis reduced the disease incidence and lesion diameters without direct contact with the pathogen Penicillium digitatum. Transcriptome analysis revealed that phenylpropanoid biosynthesis was triggered by P. galeiformis. Genes encoding phenylpropanoid biosynthesis were upregulated, including phenylalanine ammonia-lyase (PAL), 4-coumaroyl-CoA ligase (4CL), cinnamate-4-hydroxylase (C4H), peroxidase (POD), cinnamyl alcohol dehydrogenase (CAD), O-methyltransferase, and hydroxyl cinnamoyl transferase. Moreover, P. galeiformis increased the activity of PAL, 4CL, C4H, POD, polyphenol oxidase, and CAD in citrus pericarp. In addition, P. galeiformis treated citrus displayed higher levels of total phenolic compounds, flavonoid, and lignin and higher amounts of ferulic and sinapic acid. In conclusion, these results suggested that P. galeiformis could induce resistance through modulating the pathway of phenylpropanoid biosynthesis in postharvest citrus.

Redox-Active Separators for Lithium-Ion Batteries.

A bilayered cellulose-based separator design is presented that can enhance the electrochemical performance of lithium-ion batteries (LIBs) via the inclusion of a porous redox-active layer. The proposed flexible redox-active separator consists of a mesoporous, insulating nanocellulose fiber layer that provides the necessary insulation between the electrodes and a porous, conductive, and redox-active polypyrrole-nanocellulose layer. The latter layer provides mechanical support to the nanocellulose layer and adds extra capacity to the LIBs. The redox-active separator is mechanically flexible, and no internal short circuits are observed during the operation of the LIBs, even when the redox-active layer is in direct contact with both electrodes in a symmetric lithium-lithium cell. By replacing a conventional polyethylene separator with a redox-active separator, the capacity of the proof-of-concept LIB battery containing a LiFePO4 cathode and a Li metal anode can be increased from 0.16 to 0.276 mA h due to the capacity contribution from the redox-active separator. As the presented redox-active separator concept can be used to increase the capacities of electrochemical energy storage systems, this approach may pave the way for new types of functional separators.

Nonenzymatic biosensor based on Cu(x)O nanoparticles deposited on polypyrrole nanowires for improving detection range.

Cu(x)O (CuO and Cu2O composite) nanoparticles modified polypyrrole (PPy) nanowires were fabricated and used as a biosensor for detecting glucose (GLC). PPy nanowires were prepared through electrodeposition, while Cu(x)O nanoparticles were deposited on PPy nanowires by electrodeposition and electrochemical oxidation in situ. The scanning electron microscopy images showed the Cu(x)O nanoparticles aligned along the PPy nanowires uniformly and the average size of Cu(x)O nanoparticles is about 90 nm. The electrocatalytic activity of Cu(x)O/PPy/Au towards GLC was investigated under alkaline conditions using cyclic voltammetry and chronoamperometry. The sensor exhibited a linear range up to 8 mM of GLC, which is more than two times of most of the existing non-enzymatic GLC sensors based on CuO or Cu2O. The sensitivity of the sensor is 232.22 µAmM⁻¹ cm⁻² and detection limit is 6.2 µM (at signal/noise=3). Moreover, the sensor showed excellent selectivity, reproducibility and stability properties. These excellent performances make Cu(x)O/PPy/Au a good nonenzymatic GLC sensor.