A rich source of bioactive secondary metabolites from microorgannisms are widely used to control plant diseases in an eco-friendly way. To explore ideal candidates for prevention of pine wilt disease (PWD), a bacterial strain from rhizosphere of Pinus thunbergii, Lysinimonas M4, with nematicidal activity against pine wood nematode (PWN), Bursaphelenchus xylophilus, was isolated. Two nematicidal compounds were obtained from the culture of Lysinimonas M4 by silica gel chromatography based on bioactivity-guided fractionation and were subsequently identified as 2-coumaranone and cyclo-(Phe-Pro) by nuclear magnetic resonance (NMR) and mass spectrometry (MS). The 2-coumaranone and cyclo-(Phe-Pro) showed significant nematicidal activity against PWN, with LC₅₀ values at 24 h of 0.196 mM and 0.425 mM, respectively. Both compounds had significant inhibitory effects on egg hatching, feeding, and reproduction. The study on nematicidal mechanisms revealed that 2-coumaranone and cyclo-(Phe-Pro) caused the accumulation of reactive oxygen species (ROS) in nematodes, along with a notable decrease in CAT and POS activity and an increase in SOD activity in nematodes, which might contribute to the death of pine wood nematodes. Bioassay tests demonstrated that the two compounds could reduce the incidence of wilting in Japanese black pine seedlings. This research offers a new bacterial strain and two metabolites for biocontrol against PWN.

Pine wilt disease (PWD) caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus, is a globally distributed destructive disease of pine forest. To study the PWD pathogenic mechanism, the cinnamyl alcohol dehydrogenase gene (BxCAD-1) from B. xylophilus was selected. The BxCAD-1 gene was amplified by PCR from the cDNA library of B. xylophilus and cloned into the expression vector pET-15b to construct the recombinant vector pET-15b-BxCAD-1. The recombinant cinnamyl alcohol dehydrogenase protein was purified by Ni-NTA affinity chromatography from Escherichia coli BL21 (DE3) harboring pET-15b-BxCAD-1 induced by IPTG. The effects of pH, temperature, metal ions and substrates on the activity of BxCAD-1 were determined, showing the highest catalytic activity at pH 8.0 and 40 °C with cinnamyl alcohol as substrate and Zn²⁺ as an activator. To elucidate the functions of BxCAD-1 in B. xylophilus, the expression of the gene was down-regulated by RNA interference. Results showed that the movement, feeding, reproduction, spawning rate, hatching rate, lifespan, infectivity and sensitivity to ethanol decreased compared with negative controls. RNA interference also affected the development of B. xylophilus from the larval stage to the adult stage. In situ hybridization showed that the gene was expressed in the digestive tract of male and female adults. This study revealed a promising target for PWD control.

Alginate is the main structural polysaccharide in brown algal cell walls and is widely used in pharmaceuticals, textiles, cosmetics, feed, and other fields. Its tradition production methods feature low yield, high water consumption and environmental pollution, which need industrial upgrading. In this study, Aeromonas hydrophila W-02, a bacterial strain capable of simultaneously producing high levels of cellulase, pectinase, and glucanase, was isolated and identified from kelp humic liquid. Genome sequencing revealed that it has 5781 genes, 116 of which are related to glycoside hydrolysis. When A. hydrophila W-02 was applied to kelp for alginate extraction, the optimized bacterial method achieved a 38.21% yield, four times that of the traditional acid coagulation–acidification method (8.12%). Alginate was used to prepare alginate fiber and its performance indices were compared with those of commercial fibers. Scanning electron microscopy and mechanical properties analysis revealed no significant difference between alginate fibers from this novel process and commercial alginate. This study provides an efficient and green approach for alginate preparation and provides data to support the industrialization of high-efficiency bio-based alginate extraction.