First report of 'Candidatus Phytoplasma asteris' associated with witches'-broom and plexus bud disease of Cerasus serrula in China.

Cherry blossoms (Cerasus serrula) are native to the temperate zone around the Himalayas in the northern hemisphere, mainly distributed in the west and southwest of China, including Yunnan, Sichuan and Tibet. Cherry has high ornamental, edible and medicinal value. In August 2022, we observed that Cherry trees exhibited witches' broom and plexus bud in Kunming City, Yunan Province, China. The symptoms consisted of many small branches with little leaves at the top of branches, stipule lobation, and clustered adventitious buds that are tumor-like on the branches that usually cannot sprout normally. As disease intensity increased, the branches dried up from the top to the bottom till the death of the whole plant. We named this disease C. serrula witches' broom disease (CsWB). We found CsWB in the areas of Panlong, Guandu, Xishan Districts in Kunming, where more than 17% of the plants we surveyed were infected. We collected 60 samples from across the three districts. These included 15 symptomatic and 5 asymptomatic plants per district. The lateral stem tissues were observed under a scanning electron microscope (Hitachi S-3000N). The nearly spherical bodies were found in the phloem cells of symptomatic plants. Total DNA extraction was conducted from 0.1 g tissue using the CTAB method (Porebski et al. 1997), ddH2O was used as the negative control, and Dodonaea viscose plants with witches' broom symptoms were used as the positive control. The nested PCR was used to amplify the 16S rRNA gene (Lee et al. 1993; Schneider et al. 1993) and PCR amplicon of 1.2 kb were amplified (GenBank accessions: OQ408098; OQ408099; OQ408100). The direct PCR specific to the ribosomal protein (rp) gene yielded amplicons of approximately 1.2 kb with primer pair rp(I)F1A and rp(I)R1A (Lee et a. 2003) (GenBank accessions: OQ410969; OQ410970; OQ410971). The fragment from 33 symptomatic samples was consistent with the positive control, and absent for asymptomatic samples, suggesting an association of phytoplasma with the disease. A BLAST analysis of the 16S rRNA sequences of CsWB phytoplasma showed that it has a 99.76% similarity with Trema laevigata witches' broom phytoplasma (GenBank accession MG755412). The rp sequence shared 99.75% identity with Cinnamomum camphora witches' broom phytoplasma (GenBank accession OP649594). An analysis with iPhyClassifier showed that the virtual RFLP pattern derived from the 16S rDNA sequence shares 99.3% similarity with that of the 'Ca. Phytoplasma asteris' reference strain (GenBank accession: M30790), and the virtual RFLP pattern derived from the fragment is identical (similarity coefficient 1.00) to the reference pattern of 16Sr group I, subgroup B (GenBank accession: AP006628). Thus, CsWB phytoplasma is identified as 'Ca. Phytoplasma asteris'-related strain belonging to sub-group 16SrI-B. The phylogenetic tree was constructed based on 16S rRNA gene and rp gene sequences by using MEGA version 6.0 (Tamura et al. 2013) with neighbor-joining (NJ) method and bootstrap support was estimated with 1000 replicates. The result indicated that the CsWB phytoplasma formed a subclade in 16SrI-B and rpI-B respectively. In addition, the clean 1-year-old C. serrula were tested positive for the phytoplasma using the nested PCR 30 days after being grafted with naturally infected twigs with CsWB symptoms. To the best of our knowledge, Cherry blossoms is a new host of 'Ca. Phytoplasma asteris'-related strains in China. The newly emerged disease is a threat to the ornamental value of cherry blossoms and the production of wood quality.

Effects of Different Salt Ion Concentrations in Immersion Vacuum Cooling on the Qualities of Spiced Chicken Drumsticks.

The traditional immersion vacuum cooling of meats can result in product defects. To optimize these processes, different salt ion concentrations in the immersion solution (0%, 3%, 5%, and 7%) were assessed, in relation to the cooling rate, cooling loss rate, color, texture, moisture status, and microstructure of chicken drumsticks. The cooling rate at 5% salt ion concentration was the fastest and most similar to the conventional vacuum cooling method, which can reduce the central temperature of drumsticks from 75 to 25 °C in 15 min. Immersion vacuum cooling did not cause weight loss and the 5% salt ion concentration was the best for weight maintenance, which can increase the weight of drumsticks by 2.3%. The L* and b* values first decreased and then increased with increasing salt ion concentrations, but not significantly. Hardness gradually decreased, whereas the low-field nuclear magnetic data showed that the transverse relaxation time of free water (T22) in the drumsticks increased from 200.01 ms to 237.79 ms with increasing salt ion concentrations. Scanning electron microscopy images revealed irregular and smaller pores between the muscle fibers with increasing salt ion concentrations. The 5% salt ion concentration in the immersion solution during vacuum cooling was optimal as it increased the cooling rate and improved the edible quality without cooling loss. Thus, adjusting the salt ion concentration of the immersion solution is a feasible way to improve economic benefits and quality characteristics of meat products.

Study on the roles of microorganisms and endogenous enzymes in the evolution of metabolic characteristics of lean portion during traditional Chinese bacon processing.

This study revealed the roles of microorganisms and endogenous enzymes in evolution of the metabolic characteristics of the lean portion of traditional Chinese bacon by exploring the changes in microbial succession, metabolic characteristics, lipid oxidation, lipid hydrolysis, lipoxygenase activity, lipohydrolase activity, protein degradation, and environmental factors during processing. The results showed that Proteobacteria and Firmicutes were the dominant phyla in the lean portion of traditional Chinese bacon, and 16 core genera (relative abundance > 1 %) were identified at the genus level. Significant differences were observed in the metabolic characteristics of different samples. A total of 23 differential metabolites were identified in positive and negative ion modes (P < 0.05, variable importance in the projection > 1.5). Peroxide value (POV), thiobarbituric acid reactive substances value (TBARS), proteolysis index (PI) and the degree of lipid hydrolysis increased during the processing. Phospholipids were the main hydrolysis substrates. Lipoxygenase activity increased first and then decreased, while lipohydrolase activity decreased gradually. The results of canonical correspondence analysis indicated that the salt content was the main environmental factor affecting the microbial composition. The results of Spearman correlation analysis and comprehensive discussion showed that the metabolic characteristics of the lean portion of traditional Chinese bacon during the curing period were mainly due to the catalysis of the endogenous enzymes in meat and the extracellular enzymes secreted by bacteria such as Brochothix, Acinetobacter, Psychrobacter, etc. The extracellular enzymes secreted by Salinivibrio, Vibrio, and Staphylococcus in light-fermented meat and final meat might have important contributions to the metabolic characteristics.

Comprehensive insights into the evolution of microbiological and metabolic characteristics of the fat portion during the processing of traditional Chinese bacon.

Both lean meat and fat together contribute to the consumer impression of traditional Chinese bacon. The quality of this meat product is mainly dependent on numerous biochemical reactions and microbes. For the first time, this study reveals the microbial community succession and metabolic characteristics of the fat portion during the processing of traditional Chinese bacon by high-throughput sequencing and untargeted metabolomics (ultra-high performance liquid chromatography, UHPLC). The results showed that Proteobacteria and Firmicutes were the predominant bacterial phyla in traditional Chinese bacon. At the genus level, 15 core genera (relative abundance >1%) were identified, including Salinivibrio, Staphylococcus, Brochothrix, Vibrio, Psychrobacter, Acinetobacter, Carnobacterium, Aeromonas, Pseudomona, Cobetia, Macrococcus, Enterobacter, Kocuria, unclassified-f-Vibrionaceae, and Empedobacter. Distinct separation between different groups were identified by principal component analysis, revealing significant differences in the metabolic characteristics of different samples. A total of 262 metabolites were identified, and most of the metabolites belonged to lipids and lipid-like molecules, of which 60 were identified as differential metabolites. Results of correlation analysis indicated that some differential metabolites were significantly positively correlated with dominant bacteria, such as Salinivibrio, Vibrio, Cobetia and Staphylococcus, which reinforced the fact that microorganisms have an indispensable role in the quality formation of traditional Chinese bacon.

Ternary flower-sphere-like MnO2-graphite/reduced graphene oxide nanocomposites for supercapacitor.

Chemical fabrication of a nanocomposite structure for electrode materials to regulate the ion diffusion channels and charge transfer resistances and Faradaic active sites is a versatile strategy towards building a high-performance supercapacitor. Here, a new ternary flower-sphere-like nanocomposite MnO2-graphite (MG)/reduced graphene oxide (RGO) was designed using the RGO as a coating for the MG. MnO2-graphite (MnO2-4) was obtained by KMnO4 oxidizing the pretreated graphite in an acidic medium (pH = 4). The GO coating was finally reduced by the NaBH4 to prepare the ternary nanocomposite MG. The microstructures and pore sizes were investigated by x-ray diffraction, scanning electron microscopy, thermogravimetric analysis, and nitrogen adsorption/desorption. The electrochemical properties of MG were systematically investigated by the cyclic voltammetry, galvanostatic charge-discharge, and electrochemical impedance spectroscopy in Na2SO4 solution. The MG as an electrode material for supercapacitor exhibits a specific capacitance of 478.2 and 454.6 F g-1 at a current density of 1.0 and 10.0 A g-1, respectively. In addition, the capacitance retention was 90% after 8,000 cycles. The ternary nanocomposite enhanced electrochemical performance originates from the specific flower-sphere-like morphology and coating architecture bringing higher specific surface area and lower charge transfer resistance (Rct).

Effects of Dazomet Fumigation on Soil Phosphorus and the Composition of phoD-Harboring Microbial Communities.

The soil phosphorus (P) cycle and P transformation are largely driven by the soil bacterial microbial community. However, little is known about the effects of dazomet (DZ) soil fumigation on soil P and soil microbial communities associated with P transformation. This research investigated P released from three farm soils as a result of DZ fumigation and changes in enzyme activity, gene abundance, and the encoding alkaline phosphatase PhoD microbial community. After DZ fumigation, we observed a briefly significant increase in the available P and the active P fractionation. The soil ALP activity, 16s rRNA abundance, and the phoD gene decreased significantly after DZ fumigation. The abundance and diversity of phoD-harboring microbes also decreased shortly after fumigation, increased significantly 14-28 days later, and then decreased again toward the end of the experimental period of 86 days. The shared OTUs between treatments became fewer with increasing time after fumigation. The changes in available P and the active P fractionation after DZ fumigation were significantly correlated with the abundance of the dominant phoD-harboring microbes. DZ fumigation promoted P mineralization in these farm soils and changed the composition of phoD-harboring microbial communities, which will benefit crops able to absorb and use P.

Comparison of headspace solid-phase microextraction and solvent extraction method for the simultaneous analysis of various soil fumigants in soil or water by gas chromatography-mass spectrometry.

The quantity of soil fumigants has increased globally that has focused attention on their environmental behavior. However, simultaneous analysis of traces of fumigant residues is often unreported because analysis methods are not readily available to measure them at low concentrations. In this study, typical solvent extraction methods were compared with headspace solid-phase microextraction methods. Both methods can be used for simultaneously measuring the concentrations of five commonly used soil fumigants in soil or water. The solvent extraction method showed acceptable recovery (76-103%) and intraday relative standard deviations (0.8-11%) for the five soil fumigants. The headspace solid-phase microextraction method also showed acceptable recovery (72-104%) and precision rates (1.3-17%) for the five soil fumigants. The solvent extraction method was more precise and more suitable for analyzing relatively high fumigant residue levels (0.05-5 µg/g) contained in multiple soil samples. The headspace solid-phase microextraction method, however, had a much lower limits of detection (0.09-2.52 µg/kg or µg/L) than the solvent extraction method (5.8-29.2 µg/kg), making headspace solid-phase microextraction most suitable for trace analysis of these fumigants. The results confirmed that the headspace solid-phase microextraction method was more convenient and sensitive for the determination of fumigants to real soil samples.

Comparative analysis of the effects of five soil fumigants on the abundance of denitrifying microbes and changes in bacterial community composition.

Soil fumigation is currently the most effective method for controlling soil-borne pests and diseases in high-value crops. To better understand the effect of chloropicrin (CP), dazomet (DZ), dimethyl disulfide (DMDS), allyl isothiocyanate (AITC) and 1,3-dichloropropene (1,3-D) fumigants on soil microorganisms, this study monitored changes in the diversity and community composition of soil bacteria involved in denitrification using real-time PCR and high-throughput gene sequencing techniques. These five fumigants significantly decreased the bacterial population size in some phyla including Proteobacteria, Chloroflexi and Acidobacteria, and increased the bacterial population size in other phyla such as Firmicutes, Gemmatimonadetes, Actinobacteria, Verrucomicrobia, Saccharibacteria and Parcubacteria. Although bacterial diversity declined after CP fumigation, it was briefly stimulated by the other four fumigants. Meanwhile, all five fumigants temporarily decreased populations of denitrifying bacteria containing the napA, narG, nirS or nirK enzyme-encoding genes. Denitrifiers bearing the cnorB, qnorB or nosZ genes were relatively stable following DZ and DMDS fumigation. However, cnorB and nosZ decreased initially following CP, AITC and 1,3-D fumigation. Simultaneously, the abundance of qnorB significantly increased in AITC and 1,3-D fumigated soils. These results showed that soil fumigation significantly shifted the abundance and community structure of denitrifying bacteria. This study will help to predict the response of different phyla of denitrifying bacteria to soil fumigation.

Chloropicrin fumigation alters the soil phosphorus and the composition of the encoding alkaline phosphatase PhoD gene microbial community.

The transformation of phosphorus (P) compounds in soil depends largely on soil microbial communities and is sensitive to agricultural practices. However, the effects of soil fumigation on soil P, and microbes involved in P transformation, are unknown. Our results showed that chloropicrin (CP) fumigation significantly increased the available-P, Leached-P and active-P fractionation (inorganic P extracted from H2O, NaHCO3 and NaOH) in Shangdong and Miyun soils in the early stages of culture, while soil alkaline phosphatase (ALP) activity and phoD gene abundance decreased significantly. Leached-P in fumigated soil was positively correlated with increased active-P fractionation, indicating that it was an important source of soil Leached-P after fumigation. The changes in P-fractionation, Leached-P and ALP after fumigation were also significantly correlated with the composition of the microbial communities. CP fumigation briefly stimulated an increase in the abundance and diversity of phoD-harboring microbial communities and promoted the mineralization process of soil P. PICRUSt metagenomic analysis showed an increase in the relative abundance of microorganisms with involved in carbohydrate/lipid transport and metabolism functions after fumigation. These results suggest CP fumigation altered soil P transformation and phoD-harboring microbes that might lead to an increased risk of P enrichment in waterways.

[Study on ultrasonic nebulizer sample feeding system for ICP-AES].

The sample feeding system of inductively coupled plasma atomic emission spectrometry (ICP-AES) is pneumatic nebulization system, but its efficiency is not good. The ultrasonic nebulization technology possesses advantages of high nebulization efficient and fine droplets, and it is free of blocking phenomenon. It has good application perspective in nebulization technology. In the present paper the authors study the working conditions of ultrasonic nebulizer such as carrier gas flow, injection time, injection rate and mode of washing that are likely to affect the detection results, and study the detecting conditions of several elements such as As and Se etc. that have poorly detection limits in normal ICP-AES methods. At the same time, the application of them in biochemical samples was studied. Testing results show that carrier gas flow, injection rate and injection time can greatly affect the intensity of spectral lines, and the ultrasonic nebulizer sample feeding system can increase the spectral line intensity and decrease the detection limit elements such as As, Pb, Se, Bi, Ge, Mo, Cd and Cu by about 10-25 times. Moreover, this ultrasonic nebulizer sample feeding system can reduce the time of memory effect by washing the sample cell.