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Abstract Seed coating with fungicides is a common practice in controlling seed-borne diseases, but conventional methods often result in high toxicity to plants and soil. In this study, a nanoparticle formulation was successfully developed using the metal–organic framework UiO-66 as a carrier of the fungicide ipconazole (IPC), with a tannic acid (TA)-ZnII coating serving as a protective layer. The IPC@UiO-66-TA-ZnII nanoparticles provided a controlled release, triggered and regulated by environmental factors such as pH and temperature. This formulation efficiently controlled the proliferation of Fusarium fujikuroi spores, with high penetration into both rice roots and fungal mycelia. The product exhibited high antifungal activity, achieving control efficacy rates of 84.09% to 93.10%, low biotoxicity, and promoted rice growth. Compared to the IPC flowable suspension formula, IPC@UiO-66-TA-ZnII improved the physicochemical properties and enzymatic activities in soil. Importantly, it showed potential for mitigating damage to beneficial soil bacteria. This study provides a promising approach for managing plant diseases using nanoscale fungicides in seed treatment. Graphical Abstract

Fungicides are an indispensable tool in plant disease control. Various modes of action (MOAs) have been identified in different fungicides to suppress plant pathogens. The combined use of fungicides with distinct MOAs has been recommended to prevent the development of pathogen resistance. In studying MOAs, metabolomics has been proven to be a robust and high-throughput method. Because metabolites are unique and distinct depending on the biological activities of an organism, MOAs can be identified and classified by establishing metabolic fingerprinting and metabolic profiles. Similarly, if fungicide resistance is developed in a pathogen, the metabolome will change, which can be identified. In this review, we have discussed the principles and advanced applications of metabolomics in the study of MOAs and resistance mechanisms of fungicides, and the potential of metabolic data in understanding the interaction between fungicides and pathogens. Challenges are also discussed in the application of metabolomics, improvement of the study on the mechanism of fungicides in their functions against pathogens and advancing the development of novel fungicides.

Late blight caused by Phytophthora infestans is the most devastating disease of potato. Phytophthora infestans produces many secondary metabolites and effector proteins, involved in the pathogenesis, which compromise host defense mechanisms. Pectinesterase (PE) is a cell wall degrading enzyme secreted by P. infestans to infect the host. To examine the role of PE in P. infestans, 15 strains of P. infestans were isolated from infected potato leaves in Yunnan, China. We analyzed the biological effects of exogenously added PE on P. infestans and its activity and gene expression after infection of potato using quantitative real-time polymerase chain reaction (RT-PCR). It was found that PE significantly promotes the growth of P. infestans, increases the weight of mycelium and the number of sporangia, and promotes the sporangial germination. PE accelerated the infection process of P. infestans on potato. The pathogenicity of P. infestans was positively correlated with PE activity and gene expression. PE is a key to the virulence difference of potato late blight.

IntroductionSoft rot Pectobacteriaceae (SRP) bacteria are globally dispersed pathogens that cause significant economic loss in potato and other crops. Our understanding of the SRP species diversity has expanded in recent years due to advances and adoption of whole-genome sequence technologies. There are currently 34 recognized SRP species that belong to the Dickeya and Pectobacterium genera.MethodsWe used whole-genome sequencing based analysis to describe the current distribution and epidemiology of SRP isolated from diseased potato samples obtained from commercial potato cropping systems in the United States. Our primary objectives in the present study were to: (1) identify the species of these SRP isolates recovered from potato samples across 14 states in the US, (2) describe the variation among SRP isolates from various US locations and track their temporal changes, and (3) evaluate the evolutionary relationships among these SRP isolates to deduce their source. We collected 118 SRP strains from diseased potato plants and tubers in 14 states between 2015 and 2022.ResultsWe identified three Dickeya and eight Pectobacterium species from diseased potato samples. Dickeya dianthicola, Pectobacterium parmentieri, P. carotovorum, and P. versatile appeared to be the predominant species, constituting 83% of the isolates. Furthermore, all D. dianthicola strains studied here as well as 90% of US D. dianthicola isolates sequenced to date exhibit significant clonality.DiscussionThe prevalence of this specific group of D. dianthicola, temporally and geographically, aligns with the occurrence of blackleg and soft rot outbreaks in the northeastern US after 2014. The genomic diversity observed in P. parmentieri implies multiple introductions to the US from at least four distinct sources, earlier than the arrival of the predominant group of D. dianthicola. In contrast, P. carotovorum and P. versatile appear to be widespread, long-term endemic strains in the US.

Soil microbiomes play crucial roles in pathogen suppression, nutrient mobilization, and maintenance of plant health. Their complexity and variability across spatial and temporal scales provide challenges for identifying common targets—microbial taxa or assemblages—for management in agricultural systems. To understand how microbiomes in potato production soils vary across growing regions and identify commonly distributed taxa among them, we compiled a continental-scale bacterial and eukaryotic amplicon dataset of over 1,300 communities with corresponding edaphic measurements from nine U.S. field sites. Field site explained most of the variance across bacterial and eukaryotic (predominantly fungal) communities, while pH and organic matter as well as nitrate, phosphate, and potassium concentrations also varied with community structure. Bacterial and eukaryotic potato soil microbiomes showed consistent phylum-level composition across locations at the continental scale, with regional-scale differences evident among genera and amplicon sequence variants (ASVs). Core community analysis identified 606 bacterial and 74 eukaryotic ASVs, which were present, but unequally distributed, across all nine field sites. Many of these core ASVs belonged to common soil genera, such as Bacillus and Mortierella, which may reveal the functional potential involved in maintaining soil health across regionally variable soil systems.

The orchard air-assisted sprayer exhibits strong droplet penetration, allowing uniform adhesion on both the front and back surfaces of leaves. The spray deposition and spray drift are influenced not only by the amount of pesticide but also by the airflow provided by the air-assisted system. To meet the requirements for regulating airflow and pesticide application in orchards, a method based on the iris structure for continuous adjustment of the air inlet area was proposed. An airflow control mechanism for orchard air-assisted sprayers, which is easy to install, was developed, and a circular recirculating pressure-stabilized spray system was designed. Additionally, an orchard sprayer supporting graded airflow control was developed. Experiments were conducted to assess airflow variation within the tree canopy and the spray deposition and spray drift under different airflow levels. The results showed that the average airflow attenuation rates in the canopy were 63.0%, 71.8%, and 82.5% for the leaf areas and canopy thicknesses of 826.5 cm2, 1409.8 cm2, and 1931.1 cm2, and 1.5 m, 2.0 m, and 2.5 m, respectively. Graded variable control of airflow based on canopy variation helps improve droplet uniformity within the canopy and reduce spray drift in non-target areas. When the airflow in the middle of the tree canopy exceeds 1.50 m/s, the spray deposition ratio on both sides of the leaves increases to 83.55%, and the coefficient of variation in the droplet deposition on both sides is less than 33.24%. These findings provide support for pesticide precision application and spray drift control in orchards.

Abstract Unmanned aerial vehicles (UAVs) have been increasingly employed for fungicide applications in plant disease control. However, due to weight limitations, the fungicides sprayed through UAVs must be in low volumes with high concentrations in many instances, which may result in potential phytotoxicity. Here we evaluate the safety of low-volume spray of chemicals on rice plants. The plants were sprayed with propiconazole emulsifiable concentrate (EC) at 250 g/L mixed with various adjuvants and applied at a low volume, which contained the fungicide at concentrations equivalent to or higher than that used in UAV application. The spray adjuvants included YS-20, Biaopu adjuvant, TriTek, Yipinsongzhi, AgriSolv-C100, and Hongyuyan. Potential phytotoxicity on rice plants was examined based on surface tension and crop growth. Additives suitable for a low-volume spray of propiconazole were also assessed on three rice varieties for phytotoxicity. The results showed that after 72 h of fungicide application at 2, 4, and 8 times the recommended dose of 7500 μg/mL for UAV spray, rice leaves exhibited abnormal growth, and the dry weight of rice significantly decreased 21 days after application. Phytotoxicity was evaluated on three rice varieties 5 days after spraying propiconazole EC at 2 × recommended dose with one of the spray adjuvants. The addition of 1% YS-20, Biaopu adjuvant, TriTek, and Yipinsongzhi significantly augmented the phytotoxicity. However, both AgriSolv-C100 and Hongyuyan significantly reduced the comprehensive index of phytotoxicity and, therefore, could be used for UAV applications.

Sunflower Verticillium Wilt (SVW) caused by Verticillium dahliae is a significant threat to sunflower production in China. This soilborne disease is difficult to control. It has been observed that delayed sowing reduces the severity of SVW on different varieties and across various locations. Soil was collected from multiple locations with different sowing dates to understand the underlying biological mechanisms driving this phenomenon. The soil bacterial community was characterized through 16S rRNA gene amplicon sequencing performed on the Illumina MiSeq platform, followed by comprehensive bioinformatics analysis. Microsclerotia numbers in soil were detected using both NP-10 selective medium and quantitative polymerase chain reaction (qPCR). By delaying the sowing date, the number of microsclerotia in soil and the biomass of V. dahliae colonized inside sunflower roots were reduced during the early developmental stages (V2–V6) of sunflowers. Amplicon sequencing revealed an increased abundance of bacterial genera, such as Pseudomonas, Azoarcus, and Bacillus in soil samples collected from delayed sowing plots. Five bacterial strains isolated from the delayed sowing plot exhibited strong antagonistic effects against V. dahliae. The result of the pot experiments indicated that supplying two different synthetic communities (SynComs) in the pot did increase the control efficiencies on SVW by 19.08% and 37.82% separately. Additionally, soil temperature and humidity across different sowing dates were also monitored, and a significant correlation between disease severity and environmental factors was observed. In conclusion, delayed sowing appears to decrease microsclerotia levels by recruiting beneficial rhizosphere bacteria, thereby reducing the severity of SVW.

To address the significant cutting resistance and fracture susceptibility of rotary blades, an innovative blade design was conceived to minimize resistance and enhance fracture resistance. By analyzing the interaction between the blade, soil, and root systems, an optimized design for the blade structure’s breakage resistance was developed. The theory of eccentric circular side cutting edges was applied to redesign the curve of the side cutting edge, and kinematic analysis was conducted to determine the optimal edge angle (26.57°). A flexible body model of corn residues was established, and cutting resistance measurements indicated a 15.1% reduction in cutting resistance. The breakage resistance of the rotary blade was validated using a discrete element method–finite element method (DEM–FEM) coupling approach. The results demonstrated the following: neck stress (−16.85%), specific strength efficiency (+9.72%), specific stiffness efficiency (+9.78%), fatigue life (+39.08%), and ultimate fracture stress (+20.16%), thereby meeting the design objectives. The comparison between field trial results and simulation data showed an error rate (

The widespread adoption of Internet of Things (IoT) applications has driven the demand for obtaining sensor data. Using unmanned aerial vehicles (UAVs) to collect sensor data is an effective means in scenarios with no ground communication facilities. In this paper, we innovatively consider an indeterministic data collection task in a UAV-assisted wide and sparse wireless sensor network, where the wireless sensor nodes (SNs) obtain effective data randomly, and the UAV has no pre-knowledge about which sensor has effective data. The UAV trajectories, SN serve scheduling and UAV-SN association are jointly optimized to maximize the amount of collected effective sensing data. We model the optimization problem and address the indeterministic effective indicator by introducing an effectiveness probability prediction model. The reformulated problem remains challenging to solve due to the number of constraints varying with the variable, i.e., the serve scheduling strategy. To tackle this issue, we propose a two-layer modified knapsack algorithm, within which a feasibility problem is resolved iteratively to find the optimal packing strategy. Numerical results demonstrate that the proposed scheme has remarkable advantages in the sum of effective data blocks, reducing the completion time for collecting the same ratio of effective data by nearly 30%.

Abstract In this paper, we concentrate on a non-orthogonal multiple access (NOMA)-enabled UAV data collection network for Internet of Things devices (IoTDs), where a unmanned aerial vehicle (UAV) is deployed as an aerial base station. During its flight period, the UAV can collect data from IoTDs and take advantage of the simultaneous wireless information and power transfer technology to charge the batteries of IoTDs. With the aid of NOMA, spectrum efficiency has been improved. We aim to prolong the lifetime of the IoT network, via jointly optimizing the UAV trajectory, the time allocation for information communication and wireless power transfer, the IoTDs’ transmit power, as well as the IoTDs’ group scheduling for NOMA. Then, we use the block coordinate decent and successive convex approximation techniques to tackle the non-convexity of the formulated problem. Numerical results show that the proposed solution increases the residual energy of the IoTDs, thus prolonging the lifetime of the network.

Abstract Aiming at the problems of low utilization rate of corn fertilizer, low precision of fertilization ratio, and time-consuming and laborious topdressing in the later stage, an U-shaped fertilization device with uniform fertilizer mechanism was designed. The device was mainly composed of uniform fertilizer mixing mechanism, fertilizer guide plate and fertilization plate. Compound fertilizer was applied on both sides and slow/controlled release fertilizer was applied at the bottom to form an U-shaped distribution of fertilizer around corn seeds. Through theoretical analysis and calculation, the structural parameters of the fertilization device were determined. Through the simulated soil tank test, the quadratic regression orthogonal rotation combination design was carried out on the main factors affecting the spatial stratification effect of fertilizer. The optimal parameters were obtained as follows: the stirring speed of the stirring structure was 300 r/min, the bending angle of the fertilization tube was 165°, and the operating speed of the fertilization device was 3 km/h. The results of bench verification test showed that under the optimized stirring speed and bending angle, the fertilizer particles were stirred evenly, and the average values of fertilizer flowing out of the fertilization tubes on both sides were 299.5 g and 297.4 g, respectively. The average fertilizer amounts of the three fertilizer outlets were 200.4 g, 203.2 g and 197.7 g, respectively, which met the agronomic requirements of 1:1:1 fertilization, and the variation coefficients of fertilizer amounts on both sides of the fertilizer pipe and each layer were less than 0.1% and 0.4%, respectively. The simulation results of the optimized U-shaped fertilization device can achieve the expected U-shaped fertilization effect around corn seeds. The results of field experiment showed that the U-shaped fertilization device could realize the U-shaped proportional application of fertilizer in soil. The distance between the upper end of fertilization on both sides and the distance between the base fertilizer and the surface were 87.3–95.2 mm and 197.8–206.0 mm, respectively. The transverse distance between the fertilizers on both sides was 84.3–99.4 mm, and the error with the designed theoretical fertilization was within 10 mm. Compared with the traditional side fertilization method, the number of corn roots increased by 5–6, the root length increased by 30–40 mm, and the yield increased by 9.9–14.8%.

Stem canker and black scurf of potatoes, caused by Rhizoctonia solani, are economically important diseases. Although the field application of sodium silicate has been shown to improve potato’s resistance against R. solani, the underlying mechanism remains unclear. In this study, we examined this resistance using transcriptomic analysis. Potato stems inoculated with R. solani were treated with sodium silicate, while a control group received no sodium silicate treatment. The plants were grown under natural environmental conditions at the farm of Inner Mongolia Agricultural University. Potato stems were sampled 4, 8, and 12 days after treatment. Total RNA was extracted using the TRIzol reagent and transformed into cDNA. The cDNA was sequenced, the reads were aligned, and the expression levels of genes were quantified and compared between the treated and control groups. A total of 1491 genes were identified as differentially expressed genes (DEGs). Furthermore, these DEGs were found to be involved in hydrolase activity, plant–pathogen interactions, hormone signal transduction, and the phenylpropanoid biosynthesis pathway. To confirm the up- and down-regulation of DEGs, quantitative real-time polymerase chain reaction (qRT-PCR) was performed on randomly selected genes. The results showed that the application of sodium silicate induces a complex defense network in potato plants involving physical barriers, innate immunity, phytohormone signaling, and various phenylpropanoid compounds to combat R. solani infection. This study provides valuable insights into the molecular mechanisms underlying sodium-silicate-induced resistance and its potential for reducing stem canker and black scurf in potato crops.

ABSTRACT Fungicides are an effective way to control gray mold of grapes, but the pathogen Botrytis cinerea can develop resistance, overcoming the effectiveness of a fungicide that is repeatedly applied. More importantly, the emergence of multidrug resistance (MDR) in the field, where multiple fungicides with different modes of action simultaneously lose their efficacies, is a significant concern. MDR is associated with ATP-binding cassette (ABC) transporters of the pathogen, and certain plant secondary metabolites (PSMs) stimulate the upregulation of ABC transporters, we hypothesized that the pathogen’s preadaptation to PSMs might contribute to MDR development. To test this in B. cinerea, ten PSMs, namely, resveratrol, reserpine, chalcone, flavanone, eugenol, farnesol, anethene, camptothecin, salicylic acid, and psoralen, were selected based on their association with ABC transporters involved in fungicide resistance. B. cinerea strain B05.10 was continuously transferred for 15 generations on potato dextrose agar amended with a PSM (PDAP), and sensitivities to PSMs and fungicides were examined on the 5th, 10th, and 15th generations. RNA was extracted from B. cinerea from the selected generations. After 15 generations of culture transfers, an up-regulation was observed in the expression of ABC transporter-encoding genes BcatrB, BcatrD, and BcatrK using quantitative polymerase chain reaction (qPCR). This upregulation was found to contribute to MDR of B. cinerea against two or more fungicides, among azoxystrobin, boscalid, fludioxonil, difenoconazole, prochloraz, and pyrimethanil. This finding was confirmed through genetic transformation. The decreased sensitivity of B. cinerea to fungicides was confirmed as a subsequent MDR phenotype after exposure to camptothecin, flavanone, and resveratrol. Besides, transcriptome analysis also revealed the upregulation of transcription factors related to ABC expression following resveratrol exposure. This suggests that PSMs contributed to inducing preadaptation of B. cinerea, leading to subsequent MDR.IMPORTANCEThe emergence of MDR in plant pathogens is a threat to plant disease management and leads to the use of excessive fungicides. Botrytis cinerea is of particular concern because its MDR has widely emerged in the field. Understanding its genesis is the first step for controlling MDR. In this study, the contribution of PSMs to MDR has been examined. Effective management of this pathogen in agroecosystems relies on a better understanding of how it copes with phytochemicals or fungicides.

This study examined the impact of sodium propionate concentration (0–40 g/L) on the methanogenic archaea in an inoculum which was cultured in basal nutrient medium, exploring its mechanisms and nonlinear stress intensity. The results indicated that at low concentrations, propionate-maintained homeostasis of the anaerobic digestion (AD) system and enriched Methanosaeta. However, when the concentration exceeded 16 g/L, the stability of the AD system was disrupted. The methanogenic pathway shifted towards a predominantly hydrogenotrophic pathway, resulting in a significant increase in methane yield. Below concentrations of 28 g/L, the AD system gradually enhanced its ability to utilize propionate in an orderly manner. At concentrations of 24–28 g/L, genera (e.g., Advenella and Methanosarcina) were enriched to adapt to the high-VFA environment. This was accompanied by a significant upregulation of genes related to the methylotrophic and hydrogenotrophic pathways, effectively mitigating propionate inhibition and enhancing methanogenesis. Conversely, excess concentrations (>30 g/L) suppressed methanogenesis-related genes and led to methane production arrest despite activating specialized propionate-metabolizing bacteria such as genus Pelotomaculum schinkii. As such, an increase in the stress intensity of propionate promotes a change in the metabolic pathways of methanogens and increases methane production; however, excessive sodium propionate was not conducive to maintaining the steady state of the system.

In order to improve the quality of transplanting devices and solve the problems of the poor effect on soil moisture conservation and more weeds easily growing due to the high mulching-film damage rate with an excessive number of hole openings, we developed a dibble-type transplanting device consisting of a dibble-type transplanting unit, a transplanting disc, and a dibble axis. The ADAMS software Adams2020 (64bit) was used to simulate and analyze the kinematic track of the transplanting device. The results of the analysis show that, when the hole opening of the envelope in the longitudinal dimension was the smallest, the transplanting characteristic coefficient was 1.034, the transplanting angle was 95°, and the transplanting frequency had no influence. With the help of the ANSYS WORKBENCH software Ansys19.2 (64bit), an analysis of the process of the formation of an opening in the mulching film and a mechanical simulation of this process were completed. The results indicate that, when the maximum shear stress of the mulching film was the smallest, the transplanting characteristic coefficient was 1.000, the transplanting frequency was 36 plants·min−1, and the transplanting angle was 95°. In addition, the device was tested in a film-breaking experiment on a soil-tank test bench to verify the hole opening in the mulching film. The bench test showed that, when the longitudinal dimension was the smallest, the transplanting characteristic coefficient was 1.034, the transplanting frequency was 36 plants·min−1, and the transplanting angle was 95°. When the lateral dimension was the smallest, the transplanting characteristic coefficient was 1.034, the transplanting frequency was 36 plants·min−1, and the transplanting angle was 90°. The theoretical analysis, kinematic simulation, and soil-tank test results were consistent, verifying the validity and ensuring the feasibility of the transplanting device. This study provides a reference for the development of transplanting devices.

The necrotrophic pathogen Botrytis cinerea infects a broad range of plant hosts and causes substantial economic losses to many crops. Although resistance to procymidone has been observed in the field, it remains uncertain why procymidone is usually involved in multidrug resistance (MDR) together with other fungicides. Nine mutants derived from the B. cinerea strain B05.10 through procymidone domestication exhibited high resistance factors (RFs) against both procymidone and fludioxonil. However, the fitness of the mutants was reduced compared to their parental strain, showing non-sporulation and moderate virulence. Furthermore, the RFs of these mutants to other fungicides, such as azoxystrobin, fluazinam, difenoconazole, and pyrimethanil, ranged from 10 to 151, indicating the occurrence of MDR. Transcriptive expression analysis using the quantitative polymerase chain reaction (qPCR) revealed that the mutants overexpressed ABC transporter genes, ranging from 2 to 93.7-fold. These mutants carried single-point mutations W647X, R96X, and Q751X within BcBos1 by DNA sequencing. These alterations in BcBos1 conferred resistance to procymidone and other fungicides in the mutants. Molecular docking analysis suggested distinct interactions between procymidone and Bos1 in the B. cinerea standard strain B05.10 or the resistant mutants, suggesting a higher affinity of the former towards binding with the fungicide. This study provides a comprehensive understanding of the biological characteristics of the resistant mutants and conducts an initial investigation into its fungicide resistance traits, providing a reference for understanding the causes of multidrug resistance of B. cinerea in the field.

This paper investigates a multiple unmanned aerial vehicle (multi-UAV) assisted backscatter communication network (BCN), where multiple UAVs are employed to transmit RF carriers to as well as collect data from multiple backscatter sensor nodes (BSNs) deployed on the ground. We formulate an optimization problem to maximize the max-min rate of the BCN by jointly optimizing three blocks of variables, i.e., the UAVs’ trajectories, the UAVs’ transmission power and the BSNs’ scheduling. The BSNs’ sequential energy constraints are innovatively considered in our work. However, the formulated optimization problem is difficult to be solved due to its non-convexity and combinatorial nature. To this end, we use the block coordinate descent (BCD) method and successive convex approximation (SCA) technique. Numerical results show the impact of the BSNs’ sequential energy constraint on the designed UAV trajectories and verify the gain of the proposed design in the max-min rate as compared to a benchmark scheme with UAV trajectory not optimized.

Fusarium root rot, caused by Fusarium spp. in alfalfa (Medicago sativa L.), adversely impacts alfalfa by diminishing plant quality and yield, resulting in substantial losses within the industry. The most effective strategy for controlling alfalfa Fusarium root rot is planting disease-resistant varieties. Therefore, gaining a comprehensive understanding of the mechanisms underlying alfalfa’s resistance to Fusarium root rot is imperative. In this study, we observed the infection process on alfalfa seedling roots infected by Fusarium acuminatum strain HM29-05, which is labeled with green fluorescent protein (GFP). Two alfalfa varieties, namely, the resistant ‘Kangsai’ and the susceptible ‘Zhongmu No. 1’, were examined to assess various physiological and biochemical activities at 0, 2, and 3 days post inoculation (dpi). Transcriptome sequencing of the inoculated resistant and susceptible alfalfa varieties were conducted, and the potential functions and signaling pathways of differentially expressed genes (DEGs) were analyzed through gene ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Meanwhile, a DEG co-expression network was constructed though the weighted gene correlation network analysis (WGCNA) algorithm. Our results revealed significant alterations in soluble sugar, soluble protein, and malondialdehyde (MDA) contents in both the ‘Kangsai’ and ‘Zhongmu No. 1’ varieties following the inoculation of F. acuminatum. WGCNA analysis showed the involvement of various enzyme and transcription factor families related to plant growth and disease resistance, including cytochrome P450, MYB, ERF, NAC, and bZIP. These findings not only provided valuable data for further verification of gene functions but also served as a reference for the deeper explorations between plants and pathogens.

Fusarium oxysporum f. sp. cucumerinum (Foc) is a prominent pathogen that adversely affects cucumber (Cucumis sativus) production. In the pathogen’s parasitic lifestyle, the pathogenesis and virulence evolution may be regulated by lysine acetylation, as demonstrated in many living organisms. However, its specific function in Foc remains poorly understood. In this study, the acetylome profiles of a mild virulence strain (foc-3b) and its derived virulence-enhanced strain (Ra-4) were analyzed before and post-inoculation on cucumber plants. In total, 10,664 acetylation sites were identified corresponding to 3874 proteins, and 45 conserved acetylation motifs were detected. Through comparison of the acetylomes, numerous differentially lysine-acetylated proteins were enriched in energy metabolism and protein processing processes, indicating the critical role of lysine acetylation during the transition from the saprotrophic lifestyle to the parasitic lifestyle. Comparative acetylome analyses on the two virulence-differentiated strains revealed that several differentially lysine-acetylated proteins were involved in pathways of defense response and energy metabolism. Ra-4 showed enhanced energy metabolism compared to foc-3b. This indicates that robust metabolic activity is required to achieve high virulence and facilitating adaptive evolution. Additionally, faster host responses are supported by an ample energy supply enhancing virulence. Thus, lysine acetylation plays a crucial role in the pathogenesis and virulence evolution of Foc.

Dickeya dianthicola (Samson) is an important pathogen causing blackleg disease of potato. Previous work suggested that insects might vector species of Pectobacteriaceae between plants but no conclusive work has confirmed this. Green peach aphids (Myzus persicae Sulzer) and Colorado potato beetles (Leptinotarsa decemlineata Say) are aggressive potato pests and related to known vectors of several species of bacteria other than D. dianthicola. This study sought to determine whether these insects vector D. dianthicola for potato infection. Neither insect species showed olfactory discrimination based on the presence of infection in laboratory tests but beetles were repelled by uninfected foliage treated with 2,3-butanediol, a primary Dickeya metabolite. Beetle recruitment to plants was not affected by their infection status; however, aphids preferred uninfected foliage when conspecifics were present. In the laboratory, neither insect acquired or transmitted D. dianthicola through feeding. In the field, neither insect’s abundance was significantly correlated with disease spread. Overall, this study did not find indications that D. dianthicola is vectored by the tested insect species. Therefore, efforts to limit Dickeya spread should focus on sanitation, water management, and seed screening—not on the control of these insect species.[Figure: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

In this paper, an online energy management strategy (EMS) for hybrid electric tracked vehicle (HETV) is developed based on deep deterministic policy gradient (DDPG) with time-varying weighting factor to further improve economic performance of HETV and reduce computational burden. The DDPG is applied to model the EMS problem for the target HETV. Especially, a time-varying weighting factor is introduced here to update old network parameters with experience learned from most recent cycle segment. Afterwards, simulation is conducted to verify the effectiveness and adaptability of the proposed method. Results show that DDPG-based EMS with online updating mechanism can achieve nearly 90% fuel economy performance as that of dynamic programming while computational time is greatly reduced. Finally, hardware-in-loop experiment is carried out to evaluate the real-world performance of the proposed method.

SYP-14288 is a fungicide as an uncoupler of oxidative phosphorylation, which is effective in controlling fungal pathogens like Rhizoctonia solani. To determine whether R. solani can develop SYP-14288 resistance and possibly multi-drug resistance (MDR), an SYP-14288-resistant mutant of R. solani X19-7 was generated from wild-type strain X19, and the mechanism of resistance was studied through metabolic and genetic assays. From metabolites of R. solani treated with SYP-14288, three compounds including M1, M2, and M3 were identified according to UPLC-MS/MS analysis, and M1 accumulated faster than M2 and M3 in X19-7. When X19-7 was treated by glutathione-S-transferase (GST) inhibitor diethyl maleate (DEM) and SYP-14288 together, or by DEM plus one of tested fungicides that have different modes of action, a synergistic activity of resistance occurred, implying that GSTs promoted metabolic resistance against SYP-14288 and therefore led to MDR. By comparing RNA sequences between X19-7 and X19, six cytochrome P450s (P450s) and two GST genes were selected as a target, which showed a higher expression in X19-7 than X19 both before and after the exposure to SYP-14288. Furthermore, heterologous expression of P450 and GST genes in yeast was conducted to confirm genes involved in metabolic resistance. In results, the P450 gene AG1IA_05136 and GST gene AG1IA_07383 were related to fungal resistance to multiple fungicides including SYP-14288, fluazinam, chlorothalonil, and difenoconazole. It was the first report that metabolic resistance of R. solani to uncouplers was associated with P450 and GST genes.

In response to pathogen infection, some plants increase production of secondary metabolites, which not only enhance plant defense but also induce fungicide resistance, especially multidrug resistance (MDR) in the pathogen through preadaptation. To investigate the cause of MDR in Botrytis cinerea, grapes ‘Victoria’ (susceptible to B. cinerea) and ‘Shine Muscat’ (resistant to B. cinerea) were inoculated into seedling leaves with B. cinerea, followed by extraction of metabolites from the leaves on days 3, 6, and 9 after inoculation. The extract was analyzed using gas chromatography/quadrupole time-of-flight mass (GC/QTOF) combined with solid-phase microextraction (SPME) for volatile and nonvolatile metabolomic components. Nonvolatile metabolites γ-aminobutyric acid (GABA), resveratrol, piceid, and some carbohydrates or amino acids, coupled with volatile metabolites β-ocimene, α-farnesene, caryophyllene, germacrene D, β-copaene, and alkanes, accumulated at a higher level in grape leaves infected with B. cinerea compared to in noninoculated leaves. Among the established metabolic pathways, seven had greater impacts, including aminoacyl-tRNA biosynthesis, galactose metabolism, valine, leucine, and isoleucine biosynthesis. Furthermore, isoquinoline alkaloid biosynthesis; phenylpropanoid biosynthesis; monobactam biosynthesis; tropane, piperidine, and pyridine alkaloid biosynthesis; phenylalanine metabolism; and glucosinolate biosynthesis were related to antifungal activities. Based on liquid chromatography/quadrupole time-of-flight mass (LC/QTOF) detection and bioassay, B. cinerea infection induced production of plant secondary metabolites (PSMs) including eugenol, flavanone, reserpine, resveratrol, and salicylic acid, which all have inhibitory activity against B. cinerea. These compounds also promoted overexpression of ATP-binding cassette (ABC) transporter genes, which are involved in induction of MDR in B. cinerea.

The high cost of acquiring training data in the field of emotion recognition based on electroencephalogram (EEG) is a problem, making it difficult to establish a high-precision model from EEG signals for emotion recognition tasks. Given the outstanding performance of generative adversarial networks (GANs) in data augmentation in recent years, this paper proposes a task-driven method based on CWGAN to generate high-quality artificial data. The generated data are represented as multi-channel EEG data differential entropy feature maps, and a task network (emotion classifier) is introduced to guide the generator during the adversarial training. The evaluation results show that the proposed method can generate artificial data with clearer classifications and distributions that are more similar to the real data, resulting in obvious improvements in EEG-based emotion recognition tasks.

Abstract Neonicotinoid insecticides are used for preventing insects from transmitting plant viruses. This group of chemicals are easily taken up by plants and translocated to different tissues and are applicable for soil treatment in sustained-release tablets, which greatly reduces environmental contamination compared to foliar spray. The goal of this study was to examine the efficacy of thiamethoxam, acetamiprid and nitenpyram in tablet formulation against whitefly, Bemisia tabaci, the vector of tomato chlorosis virus (ToCV) under laboratory and greenhouse conditions. Effective 50% lethal concentration (LC50) of thiamethoxam, acetamiprid, and nitenpyram on B. tabaci were 2.18, 0.46 and 0.18 μg/g plant tissue, respectively. In vitro test showed that tablets of 10 mg nitenpyram applied in seedbed with thiamethoxam transplanting treatment showed 80.9% control of B. tabaci on tomato and 75.5% of ToCV after 32 days of transplanting. Therefore, neonicotinoid tablets were an effective strategy of soil treatment in controlling viruliferous insects and ToCV with reduced environmental contamination.

Soil solarization using clear plastic is a promising weed management strategy for organic farms in the Northeast USA. Based on grower concerns that the practice might negatively affect beneficial soil microbiota, we conducted experiments to measure the effects of 2 and 4 weeks of solarization in a field and a closed greenhouse. Soil microbial communities were assayed by dilution plating on semi-selective agar media. Populations of general bacteria, general fungi, bacilli, and florescent pseudomonads were unaffected by field solarization, but fluorescent pseudomonads were reduced following greenhouse solarization. At plastic removal, soil biological activity was reduced non-significantly in the field and by 45% in the green- house. Soil biological activity fluctuated following field solarization, being significantly suppressed at 5 but not 14 days after plastic removal. In the greenhouse, biological activity remained suppressed up to 28 days after plastic removal. Solarization increased available nitrogen in the field and greenhouse. Four weeks of solarization reduced viability of buried weed seeds by 64% in the field and 98% in the greenhouse, indicating that the practice can cause substantial weed seed mortality. Maximum soil temperatures, measured at 10 cm depth under solarization, were 44◦ C in the field and 50◦ C in the greenhouse; temperatures were theoretically sufficient for the reduction of some soil borne pathogens. A subsequent experiment measured the effects of solarization and tarping (black plastic) on soil biological activity. During mulching, biological activity was unaffected by treatment, but 14 days after plastic removal, biological activity was reduced in the solarized treatment as compared with the control. Overall, these results suggest that solarization can deplete the weed seedbank. Although soil biological activity was reduced by solarization, it may bounce back after a period. Greenhouse solarization achieved higher temperatures and was more lethal to weed seeds and some microbiota than field solarization.

Soil treatment with disinfectants has been used for controlling soilborne phytopathogens. Besides suppressing specific pathogens, how these disinfectants impact soil health, especially soil microbial communities, is yet to be systemically determined. The objectives of this study were to examine the effects of three representative disinfectants, including the dazomet fumigant, fenaminosulf fungicide, and kasugamycin antibiotic on chemical properties, enzymatic activities, and microbial communities in soil for cucumber cultivation. Results showed that 14 days after soil treatment with these chemicals, residual content of dazomet and kasugamycin quickly declined in soil and were undetectable, while fenaminosulf residues were found at 0.48 ± 0.01 mg/kg. Total nitrogen and total carbon increased in soil after dazomet treatment. Urease and sucrase activities were significantly restrained after disinfectant application. The disinfectants did not significantly change the taxon of predominant bacteria and fungi but altered the relative abundance and diversity of soil microbiome, as well as microbial interspecific relationships. Moreover, cucumber cultivation enhanced the overall soil microbial diversity and enzymatic activities, which diminished the difference of soil microbiome among four treatments. The difference in soil microbial diversity among the four treatments became smaller after planting cucumber. Thus, soil microbial communities were affected by soil disinfectants and gradually recovered by cucumber application.

Northeast China is well known for cultivating ginseng (Panax ginseng and P. quinquefolius). Ginseng roots are threatened by the infection of the most notorious Cylindrocarpon-like fungi (CLF), which are a complex containing important soilborne pathogens. Although the disease is economically important, little is known about the genetic diversity and population structure of the pathogenic CLF complex. This knowledge will help in developing effective disease management strategies. To conduct this study, diseased ginseng roots were collected from 12 regions representing the main ginseng-growing areas in Northeast China, and CLF were isolated. A total of 169 isolates with CLF anamorph were identified in two Dactylonectria species and six Ilyonectria species using morphological and molecular methods. Cross pathogenicity tests showed that all species were pathogenic to both P. ginseng and P. quinquefolius, and most of them had slightly stronger aggressiveness in P. ginseng. The analysis of partial sequences of the Histone H3 gene generated a high level of genetic diversity and geographic differentiation. A total of 132 variable sites were detected in 169 sequences, which formed 20 haplotypes with a haplotype diversity of 0.824. Genetic differentiation was positively correlated with geographic distance. The geographic populations in the range of Changbai Mountain were distinctly discriminated from those in other non-Changbai Mountain populations. No significant genetic differentiation was found between ginseng hosts. Cylindrocarpon-like fungi causing ginseng root diseases are geographically correlated in the genetic distance in Northeast China and should be managed correspondingly.

The high-resolution terahertz spectra of the two structural isomers, mannose and galactose, have been measured by terahertz time-domain spectroscopy (THz-TDS) in the range of 0.5–4.0 THz at room temperature. Significant differences between these similar molecules have been found in their THz characteristic spectra, implying that THz-TDS is a powerful tool for identifying isomers. Structural analyses and normal mode calculations of the two systems were performed using solid-state density functional theory (DFT) with the PBE and PW91 density functionals as well as using gas-state DFT with B3LYP hybrid functional. Among these calculations, the solid-state simulated results obtained from the PBE method exhibit a good agreement with the experimentally measured spectra. According to the calculated results of PBE, the observed spectral features were assigned as primarily external lattice translations, deformations, and rotations with lesser contributions due to intramolecular motion of pyranose ring, CH2OH group, and hydroxyl groups.

The fungicide SYP-14288 has a high efficiency, low toxicity, and broad spectrum in inhibiting both fungi and oomycetes, but its mode of action (MoA) remains unclear on inhibiting fungi. In this study, the MoA was determined by analyzing the metabolism and respiratory activities of Rhizoctonia solani treated by SYP-14288. Wild-type strains and SYP-14288-resistant mutants of R. solani were incubated on potato dextrose agar amended with either SYP-14288 or one of select fungicides acting on fungal respiration, including complex I, II, and III inhibitors; uncouplers; and ATP synthase inhibitors. Mycelial growth was measured under fungicides treatments. ATP content was determined using an ATP assay kit, membrane potential of mitochondria was detected with the JC-1 kit, and respiratory rate was calculated based on the measurement of oxygen consumption of R. solani. A model of metabolic fingerprinting cluster was established to separate oxidation inhibitors and phosphorylation inhibitors. All the results together displayed a clear discrimination between oxidation inhibitors and phosphorylation inhibitors, and the latter inhibited ATP synthase production having or uncoupling activities. Based on the model, SYP-14288 was placed in phosphorylation inhibitor group, because it significantly reduced ATP content and membrane potential of mitochondria while increasing respiratory rate in R. solani. Therefore, the MoA of SYP-14288 on R. solani was confirmed to involve phosphorylation inhibition and possibly uncoupling activity.

Ceratocystis manginecans causes mango wilt with significant economic losses. In the infection court, cerato-platanin (CP) family proteins (CPPs) are believed to involve in pathogenesis but has not been determined in C. manginecans. To confirm this function, a CP protein (CmCP) of C. manginecans was characterized in this study. A protoplast of C. manginecans was prepared by treating its mycelia with driselase and lysing enzymes. The cmcp gene was edited using CRISPR/Cas-U6-1 expression vectors in 60% PEG and 50 μg/mL hygromycin B in the medium, resulting in mutants with cmcp deletion (Δcmcp). A complemented mutant (Δcmcp-C) was obtained by transforming cmcp to Δcmcp. Both Δcmcp and Δcmcp-C were characterized by comparing them with a wild-type strain on morphology, mycelial growth, conidial production and pathogenicity. Additionally, cmcp was transformed and expressed in Pichia pastoris, and the derived recombinant protein CmCP caused a severe necrosis on Nicotiana tabacum leaves. CmCP-treated plant leaves showed symptoms of hypersensitive response including electrolyte leakage, reactive oxygen species generation and overexpression of defense-related genes PR-1, PAD3, ERF1, HSR203J, and HIN1. All those results suggested that cmcp gene was required for the growth development of C. manginecans and functioned as a major pathogenicity factor in mango infection.

Soilborne diseases are a major constraining factor to soil health and plant health in potato production. In the toolbox of crop management, soil amendments have shown benefits to control these diseases and improve soil quality. Most amendments provide nutrients to plants and suppress multiple soilborne pathogens. Soil amendments are naturally derived materials and products and can be classified into fresh or living plants, organic or inorganic matters, and microbial supplements. Fresh plants have unique functions and continuously exude chemicals to interact with soil microbes. Organic and inorganic matter contain high levels of nutrients, including nitrogen and carbon that plants and soil microorganisms need. Soil microorganisms, whether being artificially added or indigenously existing, are a key factor in plant health. Microbial communities can be considered as a biological reactor in an ecosystem, which suppress soilborne pathogens in various mechanisms and turn soil organic matter into absorbable forms for plants, regardless of amendment types. Therefore, soil amendments serve as an energy input, nutrient source, and a driving force of microbial activities. Advanced technologies, such as microbiome analyses, make it possible to analyze soil microbial communities and soil health. As research advances on mechanisms and functions, amendment-based strategies will play an important role in enhancing soil health and disease suppression for better potato production.

An outbreak of bacterial soft rot and blackleg of potato has occurred since 2014 with the epicenter being in the northeastern region of the United States. Multiple species of Pectobacterium and Dickeya are causal agents, resulting in losses to commercial and seed potato production over the past decade in the Northeastern and North Central United States. To clarify the pathogen present at the outset of the epidemic in 2015 and 2016, a phylogenetic study was made of 121 pectolytic soft rot bacteria isolated from symptomatic potato; also included were 27 type strains of Dickeya and Pectobacterium species, and 47 historic reference strains. Phylogenetic trees constructed based on multilocus sequence alignments of concatenated dnaJ, dnaX and gyrB fragments revealed the epidemic isolates to cluster with type strains of D. chrysanthemi, D. dianthicola, D. dadantii, P. atrosepticum, P. brasiliense, P. carotovorum, P. parmentieri, P. polaris, P. punjabense, and P. versatile. Genetic diversity within D. dianthicola strains was low, with one sequence type (ST1) identified in 17 of 19 strains. Pectobacterium parmentieri was more diverse, with ten sequence types detected among 37 of the 2015–2016 strains. This study can aid in monitoring future shifts in potato soft rot pathogens within the U.S. and inform strategies for disease management.

The probenazole fungicide is used for controlling rice blast (Magnaporthe grisea) primarily by inducing disease resistance of the plant. To investigate the mechanism of induced plant defense, rice seedlings were treated with probenazole at 15 days post emergence, and non-treated plants were used for the control. The plants were infected with M. grisea 5 days after chemical treatment and incubated in a greenhouse. After 7 days, rice seedlings were sampled. The metabolome of rice seedlings was chemically extracted and analyzed using gas chromatography and mass spectrum (GC-MS). The GC-MS data were processed using analysis of variance (ANOVA), principal component analysis (PCA) and metabolic pathway elucidation. Results showed that probenazole application significantly affected the metabolic profile of rice seedlings, and the effect was proportionally leveraged with the increase of probenazole concentration. Probenazole resulted in a change of 54 metabolites. Salicylic acid, γ-aminobutyrate, shikimate and several other primary metabolites related to plant resistance were significantly up-regulated and some metabolites such as phenylalanine, valine and proline were down-regulated in probenazole-treated seedlings. These results revealed a metabolic pathway of rice seedlings induced by probenazole treatment regarding the resistance to M. grisea infection.

Dickeya and Pectobacterium spp. both cause blackleg and soft rot of potato, which can be a yield-reducing factor to potato production. The purpose of this study was to examine the interaction between these two bacterial genera causing potato infection, and subsequent disease development and yield responses under field conditions. Analysis of 883 potato samples collected in Northeastern USA using polymerase chain reaction determined that Dickeya dianthicola and P. parmentieri were found in 38.1% and 53.3% of all samples, respectively, and that 20.6% of samples contained both D. dianthicola and P. parmentieri. To further investigate the relationship between the two bacterial species and their interaction, field trials were established. Potato seed pieces of “Russet Burbank”, “Lamoka”, and “Atlantic” were inoculated with bacterial suspension of D. dianthicola at 107 colony-forming unite (CFU)/mL using a vacuum infiltration method, air dried, and then planted in the field. Two-year results showed that there was a high correlation (p < 0.01) between yield loss and percent of inoculated seed pieces. In a secondary field trial conducted in 2018 and 2019, seed pieces of potato “Shepody”, “Lamoka” and “Atlantic” were inoculated with D. dianthicola, P. parmentieri, or mixture of both species, and then planted. In 2019, disease severity index, as measured by the most sensitive variety “Lamoka”, was 16.2 with D. dianthicola inoculation, 10.4 with P. parmentieri, 25.4 with inoculation with both bacteria. Two-year data had a similar trend. Thus, D. dianthicola was more virulent than P. parmentieri, but the co-inoculation of the two species resulted in increased disease severity compared to single-species inoculation with either pathogen.

Pink rot (Phytophthora erythroseptica) of potato is a major concern in many potato production regions. The pathogen produces zoospores that serve as a primary inoculum for infection. To understand how the pink rot incidence is related to pathogen population, qualitative, and quantitative chemical analyses were conducted. It was demonstrated that P. erythroseptica zoospores required a minimal population of 103 zoospores/ml (threshold) for initiating germination and the subsequent infection; the percentage of zoosporic germination was positively correlated with the density of zoospores above the threshold. To elucidate the density-dependent behavior, zoospore exudate (ZE) was extracted from high-density (105/ml) zoospore suspension. Zoosporic inocula of P. erythroseptica at different concentrations were inoculated on potato tubers. Necrotic lesions were caused by inoculum with 100 zoospores per inoculation site; 5 zoospores per site did not cause lesions on the tuber. However, five zoospores did cause lesions when they were placed in ZE, suggesting ZE contained chemical compounds that regulate germination of zoospores. ZE was collected and analyzed using liquid chromatography mass spectroscopy (LC-MS). Results showed that the amino acid leucine was associated with zoosporic germination. Therefore, zoosporic germination and infection of P. erythroseptica were mediated by signaling molecules secreted from zoospores.

Potato common scab (PCS) is an economically important disease worldwide. In this study we demonstrated the possible role of Streptomyces violaceusniger AC12AB in controlling PCS. Isolates of Streptomyces scabies were obtained from CS infected tubers collected from Maine United States, which were confirmed by morphological and molecular analysis including 16S rRNA sequencing and RFLP analysis of amplified 16S-23S ITS. Pathogenicity assays related genes including txtAB, nec1, and tomA were also identified in all S. scabies strains through PCR reaction. An antagonistic bacterial strain was isolated from soil in Punjab and identified as S. violaceusniger AC12AB based on 16S rRNA sequencing analysis. Methanolic extract of S. violaceusniger AC12AB contained azalomycin RS-22A which was confirmed by 1H and 13C-NMR, 1H/1H-COSY, HMBC and HMQC techniques. S. violaceusniger AC12AB exhibited plant growth promotion attributes including Indole-3-acetic acid production with 17 μgmL-1 titers, siderophores production, nitrogen fixation and phosphates solubilization potential. When tubers were inoculated with S. violaceusniger AC12AB, significant (P < 0.05) PCS disease reduction up to 90% was observed in greenhouse and field trials, respectively. Likewise, S. violaceusniger AC12AB significantly (P < 0.05) increased potato crop up to 26.8% in field trial. Therefore, plant growth promoting S. violaceusniger AC12AB could provide a dual benefit by decreasing PCS disease severity and increasing potato yield as an effective and inexpensive alternative strategy to manage this disease.

Cucumber green mottle mosaic virus (CGMMV) is an economically important pathogen and causes significant reduction of both yield and quality of cucumber (Cucumis sativus). Currently, there were no satisfied strategies for controlling the disease. A better understanding of microRNA (miRNA) expression related to the regulation of plant-virus interactions and virus resistance would be of great assistance when developing control strategies for CGMMV. However, accurate expression analysis is highly dependent on robust and reliable reference gene used as an internal control for normalization of miRNA expression. Most commonly used reference genes involved in CGMMV-infected cucumber are not universally expressed depending on tissue types and stages of plant development. It is therefore crucial to identify suitable reference genes in investigating the role of miRNA expression. In this study, seven reference genes, including Actin, Tubulin, EF-1α, 18S rRNA, Ubiquitin, GAPDH and Cyclophilin, were evaluated for the most accurate results in analyses using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Gene expression was assayed on cucumber leaves, stems and roots that were collected at different days post inoculation with CGMMV. The expression data were analyzed using algorithms including delta-Ct, geNorm, NormFinder, and BestKeeper as well as the comparative tool RefFinder. The reference genes were subsequently validated using miR159. The results showed that EF-1α and GAPDH were the most reliable reference genes for normalizing miRNA expression in leaf, root and stem samples, while Ubiquitin and EF-1α were the most suitable combination overall.

Clavibacter michiganensis subsp. michiganensis (Cmm) is a seed-borne pathogen that causes bacterial canker disease of tomato. Cmm is typically detected in tomato seeds using quantitative real-time polymerase chain reaction (qPCR) combined with culture-based isolation. The viable but nonculturable (VBNC) state of Cmm may result in the underestimation or false negative detection of the pathogen. In the present study, propidium monoazide (PMA) and its improved structure PMAxx were used to pretreat Cmm prior to DNA extraction, followed by qPCR. Both PMA and PMAxx could bind to the chromosomal DNA of dead bacterial cells and therefore block DNA amplification by PCR. This effect, however, does not occur in living bacterial cells, as the chemicals cannot penetrate through the undamaged cell membrane. Both viable and dead Cmm cells were treated with PMA and PMAxx at various concentrations. With this treatment, the range of the cell population was determined for effective detection. PMAxx showed a better discrimination effect than PMA on the viable and dead cells of Cmm and was therefore used throughout the present study. VBNC cells of Cmm (108 CFU mL-1) was induced by 50 μM copper sulfate, which was detected at different sampling times up to a month by using both PMAxx-qPCR and flow cytometry assays. The optimal PMAxx concentration was 20 μM for detecting membrane-intact Cmm cells. High specificity and sensitivity were obtained at Cmm concentrations ranging from 103 to 107 CFU mL-1. The accurate and robust results of PMAxx-qPCR were confirmed by flow cytometry method to detect viable Cmm cells. Furthermore, the PMAxx-qPCR assay was successfully used in detecting VBNC Cmm cells in tomato seeds with as few as 10 seeds per set.

The genus Colletotrichum contains a wide variety of important plant pathogens, and Colletotrichum truncatum is one of the most prevalent species of Colletotrichum on chili in China. Demethylation-inhibitor fungicides (DMIs) are currently registered chemical agents for the management of the anthracnose disease caused by Colletotrichum spp. To assess the risk for DMI resistance development, 112 C. truncatum isolates were collected from infected pepper in 13 regions of China. The sensitivity of C. truncatum isolates to five DMI fungicides was determined based on mycelial growth inhibition assay. C. truncatum was sensitive to prochloraz, epoxiconazole, and difenoconazole, but not to tebuconazole or myclobutanil. Baseline sensitivity using the 112 C. truncatum isolates was established for the first three effective DMIs. Prochloraz, epoxiconazole, and difenoconazole EC50 values were 0.053 ± 0.023, 1.956 ± 0.815, and 1.027 ± 0.644 μg/ml, respectively. Eleven stable DMI-resistant mutants all exhibited lower fitness levels than their wild-type parents, suggesting a low risk of DMI resistance in C. truncatum. By inducing gene expression, CtCYP51 expression increased slightly in the resistant mutants as compared to wild-types when exposed to DMI fungicides and thus contributed at least partially to resistance. Molecular docking with CYP51 structure models was used to explain differential sensitivity of the DMI fungicides in C. truncatum. Our results suggest that the M376L/H373N mutations in CYP51 changed the conformation of DMIs in the binding pocket. These changes prevented the formation of the Fe – N coordinate bond between the heme iron active site and tebuconazole or myclobutanil, and apparently contributed to tebuconazole and myclobutanil insensitivity of C. truncatum.

Michael Albazzaz, Karthikeyan Narayanan, Jianjun Hao, Roma Andheri, Amsaveni Ramachandran, Sriram Ravindran, Anne GeorgeBrodie Tooth Development Genetics and Regenerative Medicine Research Laboratory, University of Illinois, Chicago, IL, USAAbstract: Dentin matrix protein 1 (DMP1) is a noncollagenous protein expressed in mineralized tissues such as bone, dentin, and cartilage. To investigate the role of DMP1 during bone formation, transgenic mice overexpressing DMP1 under the control of the CMV promoter were generated. These mice displayed an increased mineralization phenotype in bone. In addition, accelerated terminal differentiation of the epiphyseal growth plate chondrocytes were also observed. To investigate the potential role of DMP1 in osteoblast differentiation, bone marrow stem cells were stimulated with DMP1 and assayed for “early” and “late” markers for osteoblast differentiation. DMP1 treatment increased the expression of CBFA1, BMP2, COL1, and OCN within two days. An in vitro mineralized nodule formation assay demonstrated that the bone marrow stem cells could differentiate and form a mineralized matrix in the presence of DMP1. Together, these results support a model whereby DMP1 functions as a key regulatory molecule that is required for normal growth and development of bone and cartilage.Keywords: dentin matrix protein 1, mineralization, osteoblast, chondrocytes, transgenic mice

A variety of volatile organic compounds (VOCs) produced by Ceratocystis fimbriata have strong bioactivity against a wide range of fungi, bacteria and oomycetes. Mycelial growth, conidial production, and spore germination of fungi and oomycetes were significantly inhibited after exposure to cultures of C. fimbriata, and colony formation of bacteria was also inhibited. Two post-harvest diseases, peach brown rot caused by Monilinia fructicola and citrus green mold caused by Penicillium digitatum, were controlled during a 4-day storage by enclosing wound-inoculated fruits with 10 standard diameter Petri plate cultures of C. fimbriata in a 15 L box. The fruits were freshly inoculated at onset of storage and the cultures of C. fimbriata were 6 days old. Percentage of control was 92 and 97%, respectively. After exposure to C. fimbriata VOCs, severely misshapen hyphae and conidia of these two post-harvest pathogens were observed by scanning electron microscopy, and their pathogenicity was lost or greatly reduced.

Family with sequence similarity 20,-member C (FAM20C) is highly expressed in the mineralized tissues of mammals. Genetic studies showed that the loss-of-function mutations in FAM20C were associated with human lethal osteosclerotic bone dysplasia (Raine Syndrome), implying an inhibitory role of this molecule in bone formation. However, in vitro gain- and loss-of-function studies suggested that FAM20C promotes the differentiation and mineralization of mouse mesenchymal cells and odontoblasts. Recently, we generated Fam20c conditional knockout (cKO) mice in which Fam20c was globally inactivated (by crossbreeding with Sox2-Cre mice) or inactivated specifically in the mineralized tissues (by crossbreeding with 3.6 kb Col 1a1-Cre mice). Fam20c transgenic mice were also generated and crossbred with Fam20c cKO mice to introduce the transgene in the knockout background. In vitro gain- and loss-of-function were examined by adding recombinant FAM20C to MC3T3-E1 cells and by lentiviral shRNA-mediated knockdown of FAM20C in human and mouse osteogenic cell lines. Surprisingly, both the global and mineralized tissue-specific cKO mice developed hypophosphatemic rickets (but not osteosclerosis), along with a significant downregulation of osteoblast differentiation markers and a dramatic elevation of fibroblast growth factor 23 (FGF23) in the serum and bone. The mice expressing the Fam20c transgene in the wild-type background showed no abnormalities, while the expression of the Fam20c transgene fully rescued the skeletal defects in the cKO mice. Recombinant FAM20C promoted the differentiation and mineralization of MC3T3-E1 cells. Knockdown of FAM20C led to a remarkable downregulation of DMP1, along with a significant upregulation of FGF23 in both human and mouse osteogenic cell lines. These results indicate that FAM20C is a bone formation "promoter" but not an "inhibitor" in mouse osteogenesis. We conclude that FAM20C may regulate osteogenesis through its direct role in facilitating osteoblast differentiation and its systemic regulation of phosphate homeostasis via the mediation of FGF23.

Host plants and various fungicides inhibit plant pathogens by inducing the release of excessive reactive oxygen species (ROS) and causing DNA damage, either directly or indirectly leading to cell death. The mechanisms by which the oomycete Phytophthora sojae manages ROS stress resulting from plant immune responses and fungicides remains unclear. This study elucidates the role of histone acetylation in ROS-induced DNA damage responses (DDR) to adapt to stress. Mechanistically, the P. sojae DNA methyltransferase 1-associated protein (PsDMAP1) binds Tat-interactive protein 60 (PsTIP60) to comediate histone H4 acetylation on lysine 16 (H4K16ac). This regulation affects RNA polymerase II (pol II) recruitment, transcriptional induction of DDR-related genes, and the enrichment of histone H2Ax phosphorylated on serine 137 (γH2Ax) in response to both plant immunity and fungicide stress. The resulting H4K16ac serves as a crucial transgenerational epigenetic signal for virulence adaptation of P. sojae on plants, as a result of adaptation to ROS stress. [ABSTRACT FROM AUTHOR]