Enhancing soil health and strawberry disease resistance: the impact of calcium cyanamide treatment on soil microbiota and physicochemical properties.

Introduction: Continuous strawberry cropping often causes soil-borne diseases, with 20 calcium cyanamide being an effective soil fumigant, pig manure can often be used as soil organic fertilizer. Its impact on soil microorganisms structure, however, remains unclear. Methods: This study investigated the effectiveness of calcium cyanamide and pig manure in treating strawberry soil, specifically against strawberry anthracnose. We examined the physical and chemical properties of the soil and the rhizosphere microbiome and performed a network analysis. Results: Results showed that calcium cyanamide treatment significantly reduces the mortality rate of strawberry in seedling stage by reducing pathogen abundance, while increasing actinomycetes and Alphaproteobacteria during the harvest period. This treatment also enhanced bacterial network connectivity, measured by the average connectivity of each Operational Taxonomic Unit (OTU), surpassing other treatments. Moreover, calcium cyanamide notably raised the levels of organic matter, available potassium, and phosphorus in the soil-key factors for strawberry disease resistance and yield. Discussion: Overall, applying calcium cyanamide to soil used for continuous strawberry cultivation can effectively decrease anthracnose incidence. It may be by changing soil physical and chemical properties and enhancing bacterial network stability, thereby reducing the copy of anthracnose. This study highlights the dual benefit of calcium cyanamide in both disease control and soil nutrient enhancement, suggesting its potential as a valuable tool in sustainable strawberry farming.

Anaerobic Degradation of Dicamba via a Novel Catabolic Pathway by a Consortium Enriched from Deep Paddy Soil.

Dicamba is widely used in the paddy field to control broadleaf weeds. Dicamba easily migrates to deep soil, which is anoxic; however, the anaerobic catabolism of dicamba in paddy soil is still unknown. In this study, an anaerobic dicamba-degrading consortium was enriched from deep paddy soil. The consortium completely degraded 0.83 mM dicamba within 7 days. Five metabolites were identified, one of which is a new metabolite, 2,5-dichlorophenol, and a novel anaerobic dicamba degradation pathway was proposed. 2.5 mM dicamba, 1.5-2.0% NaCl, and 20 mM electron acceptors Na2SO4, NaNO3, and FeCl3, and 0.5 mM or more of metabolites 3-CP and 2,5-DCP strongly inhibited the degradation efficiency. During enrichment, the microbial community of the consortium was significantly changed with OTU numbers, and diversity decreased. The study is valuable to elucidate the catabolism and ecotoxicology studies of dicamba in paddy soil and to facilitate the engineering application of anaerobic technology to treat dicamba-manufacturing wastewater.

Expression and Characterization of 3,6-Dihydroxy-picolinic Acid Decarboxylase PicC of Bordetella bronchiseptica RB50.

Picolinic acid (PA) is a typical mono-carboxylated pyridine derivative produced by human/animals or microorganisms which could be served as nutrients for bacteria. Most Bordetella strains are pathogens causing pertussis or respiratory disease in humans and/or various animals. Previous studies indicated that Bordetella strains harbor the PA degradation pic gene cluster. However, the degradation of PA by Bordetella strains remains unknown. In this study, a reference strain of genus Bordetella, B. bronchiseptica RB50, was investigated. The organization of pic gene cluster of strain RB50 was found to be similar with that of Alcaligenes faecalis, in which the sequence similarities of each Pic proteins are between 60% to 80% except for PicB2 (47% similarity). The 3,6-dihydroxypicolinic acid (3,6DHPA) decarboxylase gene (BB0271, designated as picCRB50) of strain RB50 was synthesized and over-expressed in E. coli BL21(DE3). The PicCRB50 showed 75% amino acid similarities against known PicC from Alcaligenes faecalis. The purified PicCRB50 can efficiently transform 3,6DHPA to 2,5-dihydroxypyridine. The PicCRB50 exhibits optimal activities at pH 7.0, 35 °C, and the Km and kcat values of PicCRB50 for 3,6DHPA were 20.41 ± 2.60 µM and 7.61 ± 0.53 S-1, respectively. The present study provided new insights into the biodegradation of PA by pathogens of Bordetella spp.

Paraflavisolibacter caeni gen. nov., sp. nov., a novel taxon within the family Chitinophagaceae isolated from sludge.

A Gram-stain-negative bacterium, designated strain LB-8T, was isolated from an activated sludge sample collected from a factory in Binzhou city, Shandong province, PR China. Cells of strain LB-8T were strictly aerobic, non-motile and rod-shaped. Growth occurred at 15-40 °C (optimum, 30 °C), at pH 6.0-9.0 (optimum, pH 7.0) and at 0-7.5 % (w/v) NaCl (optimum, 0.5 % NaCl). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain LB-8T formed a distinct phyletic branch within the family Chitinophagaceae and was most closely related to members of the genera Flavisolibacter, Cnuella and Paracnuella with 92.7-93.3 % 16S rRNA gene sequence similarities. The average amino acid identity values between strain LB-8T and its closed phylogenetic neighbours Flavisolibacter, Cnuella and Paracnuella were below 70 % supporting that strain LB-8T was a member of a novel genus. The predominant cellular fatty acids of LB-8T were iso-C15 : 0, anteiso-C15 : 0, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) and iso-C17 : 0 3-OH, and the only isoprenoid quinone was menaquinone-7 (MK-7). The major polar lipids of strain LB-8T were phosphatidylethanolamine, four unidentified aminolipids and two unidentified lipids. The genome size of strain LB-8T was 7.01 Mbp with 41.2 mol% G+C content. On the basis of the evidence presented in this study, strain LB-8T represents a novel species of a new genus in the family Chitinophagaceae, for which the name Paraflavisolibacter caeni gen. nov., sp. nov. (type strain LB-8T=GDMCC 1.3631T=KCTC 92688T) is proposed.

Quinolinic acid catabolism is initiated by a novel four-component hydroxylase QuiA in Alcaligenes faecalis JQ191.

Quinolinic acid (QA) is an essential nitrogen-containing aromatic heterocyclic compounds in organisms and it also acts as an important intermediate in chemical industry, which has strong neurotoxicity and cytotoxicity. The wide range of sources and applications caused the release and accumulation of QA in the environment which might poses a hazard to ecosystems and human health. However, few research on the degradation of QA by microorganisms and toxicity of QA and its metabolites were reported. Alcaligenes faecalis JQ191 could degrade QA but the genetic foundation of QA degradation has not been studied. In this study, the gene cluster quiA1A2A3A4 was identified from A. faecalis JQ191, which was responsible for the initial catabolism step of QA. The quiA1A2A3A4 gene cluster encodes a novel cytoplasmic four-component hydroxylase QuiA. The 1H nuclear magnetic resonance indicated that QuiA catalyzed QA to 6-hydroxyquinolinic acid (6HQA) and the H218O-labeling analysis confirmed that the hydroxyl group incorporating into 6HQA was derived from water. Toxicity tests showed that the QA could approximately inhibit 20%-80% growth of Chlorella ellipsoidea, and 6HQA could relieve at least 50% QA growth inhibition of Chlorella ellipsoidea, indicating that the 6-hydroxylation of QA by QuiA is a detoxification process. This research provides new insights into the metabolism of QA by microorganism and potential application in the bioremediation of toxic pyridine derivatives-contaminated environments.

Highly efficient degradation of cypermethrin by a co-culture of Rhodococcus sp. JQ-L and Comamonas sp. A-3.

Cypermethrin is an important synthetic pyrethroid pesticide that widely used to control pests in agriculture. However, extensive use has caused its residue and the metabolite 3-phenoxybenzoic acid (3-PBA) to seriously pollute the environments and agricultural products. In this study, a highly efficient cypermethrin-degrading bacterial consortium was acclimated from long-term pyrethroid-contaminated soil. Two strains, designated JQ-L and A-3, were screened from the consortium, and identified as Rhodococcus sp. and Comamonas sp., respectively. Strain JQ-L transformed 100 mg/L of cypermethrin to 3-PBA within 60 h of incubation; however, 3-PBA could not be further degraded by the strain. Strain A-3 utilized 3-PBA as sole carbon for growth, and completely degraded 100 mg/L of 3-PBA within 15 h of incubation. Co-culture of JQ-L and A-3 completely degraded 100 mg/L of cypermethrin within 24 h of incubation. Furthermore, a complete catabolic pathway of cypermethrin and the metabolite 3-PBA by the co-culture was proposed. This study provided a promising strategy for efficient elimination of cypermethrin residue-contaminated environments and agricultural products.

Cloning of a novel tetrahydrofolate-dependent dicamba demethylase gene from dicamba-degrading consortium and characterization of the gene product.

Dicamba, an important hormone-type systemic herbicide, is widely used to control more than 200 kinds of broadleaf weeds in agriculture. Due to its broad-spectrum, high efficiency and effectively killing glyphosate-resistant weeds, dicamba is considered as an excellent target herbicide for the engineering of herbicide-resistant crops. In this study, an efficient dicamba-degrading microbial consortium was enriched from soil collected from the outfall of a pesticide factory. The enriched consortium could almost completely degrade 500 mg/L of dicamba within 12 h of incubation. A novel tetrahydrofolate (THF)-dependent dicamba demethylase gene, named dmt06, was cloned from the total DNA of the enriched consortium. Dmt06 shared the highest identity (72.3%) with dicamba demethylase Dmt50 from Rhizorhabdus dicambivorans Ndbn-20. Dmt06 was expressed in Escherichia coli BL21 and purified to homogeneity using Co2+-charged nitrilotriacetic acid affinity chromatography. The purified Dmt06 catalyzed the transfer of methyl from dicamba to THF, generating the herbicidally inactive metabolite 3,6-dichlorosalicylate (3,6-DCSA) and 5-methyl-THF. The optimum pH and temperature for Dmt06 were detected to be 7.4 and 35°C, respectively. Under the optimal condition, the specific activity of Dmt06 reached 165 nmol/min/mg toward dicamba, which was much higher than that of Dmt and Dmt50. In conclusion, this study cloned a novel gene, dmt06, encoding an efficient THF-dependent dicamba demethylase, which was a good candidate for dicamba-resistant transgenic engineering.

Flow Effects on the Controlled Growth of Nanostructured Networks at Microcapillary Walls for Applications in Continuous Flow Reactions.

Low-cost microfluidic devices are desirable for many chemical processes; however, access to robust, inert, and appropriately structured materials for the inner channel wall is severely limited. Here, the shear force within confined microchannels was tuned through control of reactant solution fluid-flow and shown to dramatically impact nano- through microstructure growth. Combined use of experimental results and simulations allowed controlled growth of 3D networked Zn(OH)F nanostructures with uniform pore distributions and large fluid contact areas on inner microchannel walls. These attributes facilitated subsequent preparation of uniformly distributed Pd and PdPt networks with high structural and chemical stability using a facile, in situ conversion method. The advantageous properties of the microchannel based catalytic system were demonstrated using microwave-assisted continuous-flow coupling as a representative reaction. High conversion rates and good recyclability were obtained. Controlling materials nanostructure via fluid-flow-enhanced growth affords a general strategy to optimize the structure of an inner microchannel wall for desired attributes. The approach provides a promising pathway toward versatile, high-performance, and low-cost microfluidic devices for continuous-flow chemical processes.

Phenylobacterium kunshanense sp. nov., isolated from the sludge of a pesticide manufacturing factory.

A novel aerobic, Gram-stain-negative, motile bacterium, designated strain BUT-10(T), was isolated from the sludge of a pesticide manufacturing factory in Kunshan, China. Cells were rod-shaped (0.4-0.45×0.9-1.4 µm) and colonies were white, circular with entire edges and had a smooth surface. The strain grew at 25-37 °C, at pH 6.0-8.0 and with 0-0.5 % NaCl. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain BUT-10(T) was a member of the genus Phenylobacterium, and showed highest sequence similarities to Phenylobacterium muchangponense A8(T) (97.49 %), Phenylobacterium immobile DSM 1986(T) (97.14 %) and Phenylobacterium lituiforme FaiI3(T) (96.34 %). Major fatty acids (>5 %) were summed feature 8 (comprising C18 : 1ω7c and/or C18 : 1ω6c), C16 : 0 and summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c). The major isoprenoid quinone was ubiquinone-10. The DNA G+C content was 71.85 mol%. Strain BUT-10(T) showed low DNA-DNA relatedness with P. muchangponense A8(T) (15.7±2.9 %) and P. immobile DSM 1986(T) (12.8±1.1 %). On the basis of the phenotypic, phylogenetic and genotypic data, strain BUT-10(T) is considered to represent a novel species of the genus Phenylobacterium, for which the name Phenylobacterium kunshanense sp. nov. is proposed. The type strain is BUT-10(T) ( = CCTCC AB 2013085(T) = KCTC 42014(T)).