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Insects that evolved in mesic regions may have difficulty establishing in xeric regions. Rhagoletis pomonella (Walsh) (Diptera: Tephritidae) was introduced into drier western North America from mesic eastern North America while Rhagoletis indifferens Curran is native to western North America. Here, we predicted that R. indifferens survives water deprivation longer than R. pomonella, as R. indifferens is more abundant than R. pomonella in dry central Washington (WA) State, USA. Sweet and bitter cherry-origin R. indifferens and apple- and hawthorn-origin R. pomonella from xeric central or mesic western WA were provided water throughout or were water-deprived at 2-4 and 14-18 d old and held at 20°C or 30/31°C and daily survival recorded. At 20°C and 30°C, western WA apple-origin R. pomonella provided water survived longer than sweet cherry-origin R. indifferens. When water-deprived, however, 2-4 d old R. indifferens, although smaller, survived significantly longer than western WA apple-origin R. pomonella of the same age. This was also generally true for 14-18 d old flies, although differences were less often significant. Central WA large-thorn hawthorn-origin R. pomonella survived water deprivation significantly longer than western WA apple-origin R. pomonella, and as long as R. indifferens. Water-deprived flies of both species survived longer at 20°C than 30/31°C. Survival analyses suggest that low water availability rather than high temperature contributes to lower R. pomonella than R. indifferens abundances in central WA, with R. pomonella populations in that region differing from western WA R. pomonella with respect to tolerance of xeric climates., (Published by Oxford University Press on behalf of Entomological Society of America 2024.)

Pathogenesis-related protein 1 (PR-1) is an antimicrobial protein involved in systemic acquired resistance (SAR) in plants, but its regulatory role and interactions with other pathways remain unclear. In this study, we functionally characterize WsPR-1 gene of Withania somnifera in Nicotiana tabacum to elucidate its role in plant defense, growth, and development. Interestingly, transgenic tobacco plants with increased levels of cytokinin (CK) and decreased gibberellins (GAs) exhibited stunted shoot growth, an underdeveloped root system, modified leaf morphology, reduced seed pod production, and delayed leaf senescence. Transcriptional analysis revealed that WsPR-1 overexpression downregulated the GA 20-oxidase (GA20ox) gene involved in GA biosynthesis while upregulating GA 2-oxidase (GA2ox), a GA catabolic enzyme. Moreover, transcript levels of FRUITFULL (FUL) and LEAFY (NFL2) flowering genes exhibited a decrease in WsPR-1 plants, which could explain the delayed flowering and reduced seed pod development in transgenic plants. Confocal microscopy confirmed increased lignin deposition in stem cross-sections of WsPR-1 transgenic plants, supported by gene expression analysis and lignin content quantification. Additionally, our findings also suggest the involvement of Knotted1-like homeobox (KNOX) gene in enhancing cytokinin levels. This study highlights PR-1's regulatory role in plant growth and development, with potential to boost crop yields and enhance resilience., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Interspecific competition is an important factor in the population dynamics and geographical distribution of insect populations. Fall armyworm (FAW), Spodoptera frugiperda, an invasive species, and the Asian corn borer (ACB), Ostrinia furnacalis, a native pest species are major pests on maize in China, posing a threat to maize yield and grain quality. A series of laboratory, greenhouse, and field experiments were conducted to elucidate the competitive interactions between FAW and ACB. In the laboratory experiments, FAW exhibited aggressive behaviors more frequently when compared with ACB, while the latter species exhibited defense behaviors more frequently. Higher intraspecific competition was recorded in the FAW conspecific rather than in ACB, particularly in the 6th larval stage. FAW had a higher interspecific competitive advantage through intraguild predation over ACB, resulting in partial or complete displacement ACB when initially the ratio of the 2 species was 1 : 1. The interspecific competition also had significantly influenced on the population parameters, defensive enzymes, and nutrient of these 2 species. Competitive interaction proved that the response of superoxide, catalase, and soluble protein in FAW were significantly increased, whereas the total sugar content in both species was substantially decreased. Survival rate, and the plant damage that co-infested by both species varied significantly among the sequential combinations under greenhouse and field conditions. FAW consistently exhibited stronger intraspecific aggression than ACB under laboratory and field conditions when co-existing on the same feeding guild. These findings contribute to efforts toward the improvement of integrated pest management programs for FAW, in decision making for invasive and native pests' management strategies to reduce the high risks of FAW and ACB outbreaks., (© 2023 Institute of Zoology, Chinese Academy of Sciences.)

In the eastern coastal regions of Odisha, wilt caused by Fusarium oxysporum f. sp. capsici is an extremely damaging disease in chilli. This disease is very difficult to manage with chemical fungicides since it is soil-borne in nature. The natural rhizosphere soil of the chilli plant was used to isolate and test bacterial antagonists for their effectiveness and ability to promote plant growth. Out of the fifty-five isolates isolated from the rhizosphere of healthy chilli plants, five isolates, namely Iso 01, Iso 17, Iso 23, Iso 24, and Iso 32, showed their highly antagonistic activity against F. oxysporum f. sp. capsici under in vitro . In a dual culture, Iso 32 (73.3%) and Iso 24 (71.5%) caused the highest level of pathogen inhibition. In greenhouse trials, artificially inoculated chilli plants treated with Iso 32 (8.8%) and Iso 24 (10.2%) had decreased percent disease incidence (PDI), with percent disease reduction over control of 85.6% and 83.3%, respectively. Iso 32 and Iso 24 treated chilli seeds have shown higher seed vigor index of 973.7 and 948.8, respectively, as compared to untreated control 636.5. Furthermore, both the isolates significantly increased plant height as well as the fresh and dry weight of chilli plants under the rolled paper towel method. Morphological, biochemical, and molecular characterization identified Bacillus amyloliquefaciens (MH491049) as the key antagonist. This study demonstrates that rhizobacteria, specifically Iso 32 and Iso 24, can effectively protect chilli plants against Fusarium wilt while promoting overall plant development. These findings hold promise for sustainable and eco-friendly management of Fusarium wilt in chilli cultivation., Competing Interests: Ravinder Kumar is an Academic Editor for PeerJ., (© 2024 Shiva et al.)

Dormant microsclerotia play a vital role in the survival and spread of Verticillium longisporum , as they can stay viable in the soil and maintain their infectivity for many years. In our previous work, we revealed that soil bacterial volatiles are a key inhibitory factor causing microsclerotia dormancy in the soil. In this study, we further demonstrate that root exudates collected from both host and non-host plants can effectively rescue microsclerotia from bacterial suppression and initiate germination. To identify the specific compounds in root exudates responsible for microsclerotia germination, we fractionated the collected root exudates into polar and non-polar compounds. Subsequently, we conducted comprehensive bioassays with each fraction on germination-suppressed microsclerotia. The result revealed a pivotal role of primary metabolites in root exudates, particularly glutamic acid, in triggering microsclerotia germination and overcoming bacterial inhibition. Moreover, our studies revealed a decrease in inhibitory bacterial volatile fatty acids when bacteria were cultured in the presence of root exudates or glutamic acid. This suggests a potential mechanism, by which root exudates set-off bacterial suppression on microsclerotia. Here, we reveal for the first time that plant root exudates, instead of directly inducing the germination of microsclerotia, enact a set-off effect by counteracting the suppressive impact of soil bacteria on the microsclerotia germination process. This nuanced interaction advances our understanding of the multifaceted dynamics governing microsclerotia dormancy and germination in the soil environment., Importance: Our research provides first-time insights into the crucial interaction between plant root exudates and soil bacteria in regulating the germination of Verticillium longisporum microsclerotia, a significant structure in the survival and proliferation of this soil-borne pathogen. We describe so far unknown mechanisms, which are key to understand how root infections on oilseed rape can occur. By pinpointing primary metabolites in root exudates as key factors in overcoming bacteria-induced dormancy and promote microsclerotia germination, our study highlights the potential for exploiting plant - as well as soil microbe-derived - compounds to control V. longisporum . This work underscores the importance of elucidating the nuanced interactions within the soil ecosystem to devise innovative strategies for managing root infective plant diseases, thereby contributing to the resilience and health of cropping systems., Competing Interests: The authors declare no conflict of interest.

Fiorinia externa Ferris, elongate hemlock scale, was inadvertently introduced to North America from Japan. This insect is particularly problematic on hemlock, Tsuga spp., though it has been reported in association with several other conifers. The evidence that other conifers might be hosts, capable of supporting growing populations of the insect, has not been previously reviewed. Our review confirms that F. externa is an oligophagous pest of members of Pinaceae. Although species of Cupressaceae and Taxaceae have been reported as hosts of F. externa , they seem unable to support population growth of this pest. Evidence of the tree-killing potential of the insect, even on suitable hosts, is remarkably scant. The degree of pest risk posed by F. externa with respect to tree mortality in areas beyond the geographic range of hemlock seems modest, but uncertain., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Venette, Ambourn, Aukema, Jetton and Petrice.)

Extra virgin olive oil traceability and authenticity are important quality indicators, and are currently the subject of exhaustive research, for developing methods to secure olive oil origin-related issues. The aim of this study was the development of a classification model capable of olive cultivar identification based on olive oil chemical composition. To achieve our aim, 385 samples of two Greek and three Italian olive cultivars were collected during two successive crop years from different locations in the coastline part of western Greece and southern Italy and analyzed for their chemical characteristics. Principal Component Analysis showed trends of differentiation among olive cultivars within or between the crop years. Artificial intelligence model of the XGBoost machine learning algorithm showed high performance in classifying the five olive cultivars from the pooled samples., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Twelve Staphylococcus borealis strains, isolated in Canada and Poland from milk of cows with intramammary infections, were characterized phenotypically (biochemical reactions on ID 32 STAPH and Biolog Phenotype MicroArrays™ PM1 and PM2A, ability of biofilm production) and genotypically (random amplified polymorphic DNA). In addition, a genomic comparison was done with S. borealis strains of human and porcine origin using the multilocus sequence typing (MLST) technique. The bovine isolates showed a high degree of phenotypic and genotypic diversity, however, they could be differentiated from human strains by the negative test for urease (found in all but one bovine isolate examined with ID 32 STAPH) and positive reaction for D-galactose (on Biolog phenotype microarray PM1) and D-lactose (on both commercial systems). The MLST method, utilizing six concatenated genes of the total length of ∼2930 bp, revealed that bovine strains (irrespective of the country of origin) show a distinctly greater degree of mutual relationship than to the strains of human and porcine origin, suggesting that S. borealis has evolved independently in these hosts. In conclusion, bovine-specific S. borealis can be involved in intramammary infections in cattle., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The fall armyworm (FAW), Spodoptera frugiperda , an invasive pest originating from the Americas is a serious pest threatening cereal production and food security in Zambia. We studied the prevalence and abundance of natural enemies of FAW in three Agroecological regions (AERs I, II, and III) to identify those that could potentially serve as bio-control agents. Sampling of FAW parasitoids and predators was done along trunk roads at intervals of 10 km. Molecular sequence analysis and morphological characterization were used to identify natural enemies. Over 11 species of FAW natural enemies, including egg, egg-larval, and larval parasitoids, and predators, were identified in Zambia. The mean number of natural enemies and species richness was higher in AER I and IIa. Consequently, egg parasitism was highest in those two regions, at 24.5% and 12.2%, respectively. Larvae parasitism was highest in AER I (4.8%) and AER III (1.9), although no significant differences were observed. The most abundant and widely distributed parasitoid was Drino sp. (Diptera: Tachinidae), while Rhynocoris segmentarius (Germar) (Hemiptera: Reduviidae) and Belanogaster sp. (Hymenoptera: Vespidae) were the most prevalent predators. Our study reveals the presence of two natural enemies belonging to the genus Tiphia and Micromeriella , uncommon to FAW. Significant differences in the number of parasitoids were observed in polycropping, with the highest recovery of 12 ± 10% from maize + cowpeas + pumpkin and watermelon mixed cropping. The higher the rainfall, the lower the number of natural enemies recorded. Variations in rainfall patterns which affect FAW availability, cropping systems and the three AERs may explain natural enemies' species diversity in Zambia. The information provided in this study can aid the development of a national biological control programme for sustainable management of fall armyworm., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The authors SN, FK, PC, SS declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Chipabika, Sohati, Khamis, Chikoti, Copeland, Ombura, Kachapulula, Tonga, Niassy and Sevgan.)

Legumes are ubiquitous, low-cost meals that are abundant in protein, vitamins, minerals, and calories. Several biotic constraints are to blame for the global output of legumes not meeting expectations. Fungi, in particular, are substantial restrictions that not only hinder production but also pose a serious health risk to both human and livestock consumption. Athelia rolfsii (Syn. Sclerotium rolfsii) is a dangerous pathogenic fungus that attacks most crops, causing massive yield losses. Legumes are no longer immune to this dreadful fungus, which can potentially result in a 100% yield loss. The initial disease symptoms based on the formation of brown color lesions at the point of infection and further development of mycelia, followed by yellowing and wilting of the whole plant. To tackle such situation, various strategies, i.e., management in cultural practices, disease-free plant growth, genetic changes, crop hybridization and in vitro culture techniques have been undertaken. This present review encapsulates the entire situation, from sclerotial dissemination through infection development and control in legume crops, with the goal of developing a tangible understanding of sustainable legume production improvements. Further study in this area might be led in an integrated manner as a result of this information, which could contribute to a better understanding of the processes of disease incidence, resistance mechanism, and its control, and fostering greater inventiveness in the production of legumes., (© 2023. Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i.)

In the field of molecular genetics, DNA extraction protocols and kits are sample-specific and proprietary, preventing lateral distribution among similar facilities from different sectors to alleviate supply shortages during a crisis. Expanding upon previous fast extraction protocols such as alkaline- and detergent-based ones, the use of boiling-hot water to rupture cells, virions, and nuclei, as proposed during the COVID-19 pandemic, might alleviate shortages and costs. Different soft, relatively abundant (highly enriched), and uncomplicated (genomically homogenous and with few inhibitors) biosamples are collected in 1.5 mL tubes, mixed with boiling-hot water, and stirred vigorously, so as to have membranes lysed and proteins deactivated; mechanical disruption may be used as well if necessary. Incubation in boiling water bath for 20-30 min follows. Depending on sample type and quantity, which affects the total extraction volume, 2-5 μL are pipetted off for direct PCR and the same volume for two decimal serial dilutions. The latter are intended to optimize the crude extract to a workable DNA/inhibitor concentration balance for direct PCR. Uncomplicated, highly enriched samples such as mycelial growth in fruits and human swab samples can be processed, contrary to complicated samples such as blood and physically unyielding samples such as plant tissue. The extract can be used for immediate PCR in both benchtop and portable thermocyclers, thus allowing nucleic acid amplification tests (NAAT) being performed in resource-limited settings with low cost and waste footprint or during prolonged crises, where supply chain failures may occur. Key features DNA extraction from different sample types using only boiling water and occasional mechanical assistance. Crude extract serially diluted twice, 10- and 100-fold, to bypass purification and quantification steps. Direct PCR for 2-10 μL of crude lysate and dilutions (conditional to sample type and quantity) to enhance probability of workable DNA-inhibitors' concentrations. Lowers the cost and curtails the overall footprint of testing to increase sustainability in field operations and in standard lab environments under supply chain derailment., Competing Interests: Competing interestsThe authors declare no conflicts of interest or competing interests., (©Copyright : © 2023 The Authors; This is an open access article under the CC BY-NC license.)

Amino acids in variable positions of proteins may be correlated, with potential structural and functional implications. Here, we apply exact tests of independence in R × C contingency tables to examine noise-free associations between variable positions of the SARS-CoV-2 spike protein, using as a paradigm sequences from Greece deposited in GISAID ( N = 6,683/1,078 full length) for the period 29 February 2020 to 26 April 2021 that essentially covers the first three pandemic waves. We examine the fate and complexity of these associations by network analysis, using associated positions (exact P ≤ 0.001 and Average Product Correction ≥ 2) as links and the corresponding positions as nodes. We found a temporal linear increase of positional differences and a gradual expansion of the number of position associations over time, represented by a temporally evolving intricate web, resulting in a non-random complex network of 69 nodes and 252 links. Overconnected nodes corresponded to the most adapted variant positions in the population, suggesting a direct relation between network degree and position functional importance. Modular analysis revealed 25 k -cliques comprising 3 to 11 nodes. At different k -clique resolutions, one to four communities were formed, capturing epistatic associations of circulating variants (Alpha, Beta, B.1.1.318), but also Delta, which dominated the evolutionary landscape later in the pandemic. Cliques of aminoacidic positional associations tended to occur in single sequences, enabling the recognition of epistatic positions in real-world virus populations. Our findings provide a novel way of understanding epistatic relationships in viral proteins with potential applications in the design of virus control procedures. IMPORTANCE Paired positional associations of adapted amino acids in virus proteins may provide new insights for understanding virus evolution and variant formation. We investigated potential intramolecular relationships between variable SARS-CoV-2 spike positions by exact tests of independence in R × C contingency tables, having applied Average Product Correction (APC) to eliminate background noise. Associated positions (exact P ≤ 0.001 and APC ≥ 2) formed a non-random, epistatic network of 25 cliques and 1-4 communities at different clique resolutions, revealing evolutionary ties between variable positions of circulating variants and a predictive potential of previously unknown network positions. Cliques of different sizes represented theoretical combinations of changing residues in sequence space, allowing the identification of significant aminoacidic combinations in single sequences of real-world populations. Our analytic approach that links network structural aspects to mutational aminoacidic combinations in the spike sequence population offers a novel way to understand virus epidemiology and evolution., Competing Interests: The authors declare no conflict of interest.

The Japanese beetle, Popillia japonica (Newman, 1841) (Coleoptera: Scarabaeidae), was first detected in southern Washington State in 2020. Widespread trapping efforts ensued, and over 23,000 individuals were collected in both 2021 and 2022 in this region known for specialty crop production. The invasion of Japanese beetle is of major concern as it feeds on over 300 plant species and has shown an ability to spread across landscapes. Here, we created a habitat suitability model for Japanese beetle in Washington and used dispersal models to forecast invasion scenarios. Our models predict that the area of current establishment occurs in a region with highly suitable habitat. Moreover, vast areas of habitat that are likely highly suitable for Japanese beetle occur in coastal areas of western Washington, with medium to highly suitable habitat in central and eastern Washington. Dispersal models suggested that the beetle could spread throughout Washington within 20 years without management, which justifies quarantine and eradication measures. Timely map-based predictions can be useful tools to guide management of invasive species while also increasing citizen engagement to invaders., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Phaeoacremonium minimum is an important esca and Petri disease pathogen that causes dieback of grapevines in South Africa. Little is known regarding the reproductive strategy of the pathogen. Sexual reproduction could lead to a better adaptation of the pathogen to disease management strategies by combining alleles through recombination. The study aimed to investigate the genetic diversity and recombination potential of eight populations in the Western Cape, from six commercial vineyards and two nursery rootstock mother blocks. This was achieved by developing and applying nine polymorphic microsatellites and mating-type-specific markers. Thirty-seven genotypes were identified from 295 isolates. Populations were characterised by the same dominant genotype (MLG20 occurring 65.43%), low genotypic diversity (H) and high numbers of clones (81.36% of dataset). However, genotypes from the same sampling sites were not closely related based on a minimum spanning network and had high molecular variation within populations (94%), suggesting that multiple introductions of different genotypes occurred over time. Significant linkage disequilibrium among loci (r̅d) further indicated a dominant asexual cycle, even though perithecia have been observed in these four populations. The two rootstock mother blocks had unique genotypes and genotypes shared with the vineyard populations. Propagation material obtained from infected rootstock mother blocks could lead to the spread of more genotypes to newly established vineyards. Based on our results, it is important to determine the health status of rootstock mother blocks. Management strategies must focus on reducing aerial inoculum to prevent repeated infections and further spread of P. minimum genotypes., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The analysis of past epidemics and pandemics, either spontaneous or of human origin, may revise the physical history of microbiota and create a temporal context in our understanding regarding pathogen attributes like virulence, evolution, transmission and disease dynamics. The data of high-tech scientific methods seem reliable, but their interpretation may still be biased when tackling events of the distant past. Such endeavors should be adjusted to other cognitive resources including historical accounts reporting the events of interest and references in alien medical cultures and terminologies; the latter may contextualize them differently from current practices. Thus 'historical microbiology' emerges. Validating such resources requires utmost care, as these may be susceptible to different biases regarding the interpretation of facts and phenomena; biases partly due to methodological limitations.

The apple maggot fly, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae), was introduced from eastern North America into western North America via infested apples (Malus domestica Borkhausen) about 44 yr ago, where it subsequently adapted to 2 hawthorn species, Crataegus douglasii Lindley and C. monogyna Jacquin. Here, we test whether R. pomonella has also adapted to large-thorn hawthorn, Crataegus macracantha Loddiges ex Loudon, in Okanogan County, Washington State, USA. In 2020, 2021, and 2022, fruit of C. macracantha were shown to ripen in late September and were infested at rates from 0.7% to 3.0%. In laboratory rearing studies, large-thorn hawthorn flies from C. macracantha eclosed on average 9-19 days later than apple flies from earlier ripening apple (August-early September), consistent with large-thorn hawthorn flies having adapted to the later fruiting phenology of its host. In a laboratory no-choice test, significantly fewer (64.8%) large-thorn hawthorn than apple flies visited apples. In choice tests, greater percentages of large-thorn hawthorn than apple flies resided on and oviposited into C. macracantha versus apple fruit. Large-thorn hawthorn flies were also smaller in size than apple flies. Our results provide further support for the recursive adaptation hypothesis that R. pomonella has rapidly and independently specialized phenologically and behaviorally to different novel hawthorn hosts since its introduction into the Pacific Northwest of the USA, potentially leading to host race formation., (Published by Oxford University Press on behalf of Entomological Society of America 2023.)

The coronavirus disease-2019 (COVID-19) pandemic has raised the stakes for planetary health diagnostics. Because pandemics pose enormous burdens on biosurveillance and diagnostics, reduction of the logistical burdens of pandemics and ecological crises is essential. Moreover, the disruptive effects of catastrophic bioevents impact the supply chains in both highly populated urban centers and rural communities. One "upstream" focus of methodological innovation in biosurveillance is the footprint of Nucleic Acid Amplification Test (NAAT)-based assays. We report in this study a water-only DNA extraction, as an initial step in developing future protocols that may require few expendables, and with low environmental footprints, in terms of wet and solid laboratory waste. In the present work, boiling-hot distilled water was used as the main cell lysis agent for direct polymerase chain reactions (PCRs) on crude extracts. After evaluation (1) in blood and mouth swabs for human biomarker genotyping, and (2) in mouth swabs and plant tissue for generic bacterial or fungal detection, and using different combinations of extraction volume, mechanical assistance, and extract dilution, we found the method to be applicable in low-complexity samples, but not in high-complexity ones such as blood and plant tissue. In conclusion, this study examined the doability of a lean approach for template extraction in the case of NAAT-based diagnostics. Testing our approach with different biosamples, PCR settings, and instruments, including portable ones for COVID-19 or dispersed applications, warrant further research. Minimal resources analysis is a concept and practice, vital and timely for biosurveillance, integrative biology, and planetary health in the 21st century.

There is ample evidence that entomopathogenic fungi can be used as alternative biological control agents for the management of insect pests in storage facilities. As the market demands more environmentally friendly methods and chemical insecticides become increasingly obsolete, more studies are being conducted to evaluate new strains of entomopathogenic fungi for their efficacy in storage facilities. In this context, we tested ten species of fungi isolated from soil, belonging to the genera Cladosporium , Condenascus , Lecanicillium , and Penicillium , for their long-term effects on economically important beetle species. Whole wheat was directly sprayed with a conidial suspension of 10 8 spores/Ml of each of the tested fungi and then adults of Sitophilus granarius , S. oryzae , S. zeamais , Rhyzopertha dominica , and Trogoderma granarium were placed on the sprayed medium to study the mortality effects. Significantly higher mortality than the control was observed in all treatments. The lowest LT 50 (9.164 days) was observed in T. granarium infected with Penicillium goetzii . The isolate with the strongest results was L. dimorphum , which recorded remarkably low LT 50 values in S. oryzae (~11 days), R. dominica (~12 days), T. granarium (~10 days), and S. granarius (~13 days). However, for S. zeamais , it was more than 16 days. Our results confirm the existing literature on the efficacy of EPF on storage beetles, suggest the possible virulence of wild untested strains, and also highlight the importance of EPF specificity.

The fall armyworm (FAW) Spodoptera frugiperda is thought to have undergone a rapid 'west-to-east' spread since 2016 when it was first identified in western Africa. Between 2018 and 2020, it was recorded from South Asia (SA), Southeast Asia (SEA), East Asia (EA), and Pacific/Australia (PA). Population genomic analyses enabled the understanding of pathways, population sources, and gene flow in this notorious agricultural pest species. Using neutral single nucleotide polymorphic (SNP) DNA markers, we detected genome introgression that suggested most populations in this study were overwhelmingly C- and R-strain hybrids (n = 252/262). SNP and mitochondrial DNA markers identified multiple introductions that were most parsimoniously explained by anthropogenic-assisted spread, i.e., associated with international trade of live/fresh plants and plant products, and involved 'bridgehead populations' in countries to enable successful pest establishment in neighbouring countries. Distinct population genomic signatures between Myanmar and China do not support the 'African origin spread' nor the 'Myanmar source population to China' hypotheses. Significant genetic differentiation between populations from different Australian states supported multiple pathways involving distinct SEA populations. Our study identified Asia as a biosecurity hotspot and a FAW genetic melting pot, and demonstrated the use of genome analysis to disentangle preventable human-assisted pest introductions from unpreventable natural pest spread., (© 2023. Crown.)

The soil-borne pathogen Verticillium longisporum infects roots of its host plant, oilseed rape, and systemically colonizes stems where it finally forms microsclerotia at crop maturity. Once returned to the soil after harvest, microsclerotia undergo a stage of dormancy, in which they may survive for several years. Since there is neither efficient chemical control nor effective resistance in oilseed rape cultivars to control the disease, alternative control strategies may consist in regulating the germination and dormancy of microsclerotia in the soil. Therefore, a series of experiments were conducted to explore the effects of nutrients, soil moisture, and the soil microbiome on germination of dormant microsclerotia. Experiments with microsclerotia exposed in vitro to different nutrients indicated that under sterile conditions the stimulating effect of nutrients on microsclerotia germination was not enhanced as compared to water. Moreover, further assays revealed a strong inhibitory effect of unsterile soil on microsclerotia germination. Accordingly, oilseed rape plants inoculated with microsclerotia of V. longisporum showed severe infection with V. longisporum when grown in autoclaved soil, in contrast to plants grown in unsterile soil. These experiments indicate a crucial role of soil fungistasis and thus the soil microbiome on microsclerotia germination. Further bioassays demonstrated that viable soil bacteria obtained from the rhizosphere of oilseed rape plants and bulk field soil effectively inhibited microsclerotia germination, whereas dead bacteria and bacterial culture filtrates hardly suppressed germination. A putative inhibitory role of volatile organic compounds (VOCs) produced by soil bacteria was confirmed in two-compartment Petri dishes, where microsclerotia germination and colony growth were significantly inhibited. Bacterial VOCs were collected and analyzed by GC-MS. In total, 45 VOCs were identified, among which two acid and two alcohol compounds were emitted by all tested bacteria. A bioassay, conducted with corresponding pure chemicals in two-compartment Petri dishes, indicated that all acidic volatile compounds, including 3-methylbutanoic acid, 2-methylbutanoic acid, hexanoic acid, and 2-methylpropionic acid, induced strong inhibitory effects on microsclerotia. We conclude that bacterial acidic volatiles play a key role in the fungistatic effect on microsclerotia of V. longisporum in the soil and could thus be targeted for development of novel strategies to control this pathogen by artificially regulating dormancy of microsclerotia in soil., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Sarenqimuge, Rahman, Wang and von Tiedemann.)

Yellow (YR) and leaf (LR) rusts caused by Puccinia striiformis f. sp. tritici (Pst) and Puccinia triticina , respectively, are of utmost importance to wheat producers because of their qualitative and quantitative effect on yield. The search for new loci resistant to both rusts is an ongoing challenge faced by plant breeders and pathologists. Our investigation was conducted on a subset of 168 pre-breeding lines (PBLs) to identify the resistant germplasm against the prevalent local races of LR and YR under field conditions followed by its genetic mapping. Our analysis revealed a range of phenotypic responses towards both rusts. We identified 28 wheat lines with immune response and 85 resistant wheat genotypes against LR, whereas there were only eight immune and 52 resistant genotypes against YR. A GWAS (genome-wide association study) identified 190 marker-trait associations (MTAs), where 120 were specific to LR and 70 were specific to YR. These MTAs were confined to 86 quantitative trait loci (QTLs), where 50 QTLs carried MTAs associated with only LR, 29 QTLs carried MTAs associated with YR, and seven QTLs carried MTAs associated with both LR and YR. Possible candidate genes at the site of these QTLs are discussed. Overall, 70 PBLs carried all seven LR/YR QTLs. Furthermore, there were five PBLs with less than five scores for both LR and YR carrying positive alleles of all seven YR/LR QTLs, which are fit to be included in a breeding program for rust resistance induction.

Pseudophaeomoniella globosa has recently been identified as a pathogen contributing to olive trunk diseases in South Africa. Little is known regarding the biology and epidemiology of this pathogen. The aim of this study was to investigate whether olive pruning debris act as an inoculum source of P. globosa in established orchards. A nested species-specific PCR was developed for the detection of this pathogen on 138 samples of pruning debris collected from Paarl (40 wood pieces), Stellenbosch (42 wood pieces), and Worcester (56 pieces) in the Western Cape Province, South Africa. Spore washes were made from the samples (5 to 10 cm in length), after which the nested species-specific primers were used to determine the presence of P. globosa on the wood. P. globosa was detected on 37.5% of the pruning debris collected from Paarl, 61.9% from Stellenbosch, and 39.3% from Worcester. The pruning debris that tested positive for P. globosa were evaluated visually by microscopic observations for P. globosa pycnidia. Dark-brown to black pycnidia were found. Conidia from these pycnidia were measured, cultured, and confirmed as P. globosa by sequencing the internal transcribed spacer region. In this study, the pruning debris in established olive orchards were identified as inoculum sources of P. globosa. This study emphasizes the importance of additional means focused on reducing the inoculum sources of this pathogen in these orchards as an additional management strategy against olive trunk diseases.

Honeybush ( Cyclopia spp.) is an indigenous, leguminous member of the Cape fynbos biome growing in the coastal winter rainfall districts of the Western and Eastern Cape Provinces of South Africa (Joubert et al. 2011). Honeybush is used for the production of herbal teas and is harvested from wild-growing and cultivated plantations (du Toit et al. 1998). Very little is known regarding diseases caused by pathogens on this indigenous plant. Only one report of twig dieback on honeybush caused by several Diaporthe Nitschke species have been reported in South Africa (Smit et al. 2021). Several honeybush producers reported poor growth and dieback in their C. subternata plantations in the Western Cape Province, South Africa. Symptoms included twig dieback, branch dieback, death of branches as well as death of entire plants. In April 2008, branches from 8-year-old cultivated plants with dieback symptoms were collected in Stellenbosch. Fungal isolations were carried out from affected material as described by Van Niekerk et al. (2004) which consistently revealed the presence of a Botryosphaeriaceae species. Two isolates were grown on water agar with sterile pine needles and incubated at 25˚C using a 12-hour day/night cycle and near-ultraviolet light. Pycnidia formed after two weeks. Morphological characteristics similar to Neofusicoccum australe (Slippers, Crous & Wingfield) Crous, Slippers & Phillips were observed (Phillips et al. 2013). Conidia were hyaline, aseptate, fusiform with subtruncate bases (16.8-)18.8-22.1(-24.6) × (4.8-)5.3-6.1(-6.4) µm (n=50). Conidiogenous cells were holoblastic, hyaline and subcylindrical to flask-shaped tapering to the apex (11-15 × 2 µm) (n=10). Colonies on potato dextrose agar were light primrose turning olivaceous grey after 7 days with a light-yellow pigment diffusing into the medium. Mycelia was moderately dense with an appressed centre mat. The identity of the isolates was further confirmed by sequencing the ribosomal RNA Internal Transcribed Spacer (ITS) and the elongation factor 1-alpha (EF-1α) gene regions using primer pairs ITS4-ITS5 (White et al. 1990) and EF1-728F-EF1-986R (Alves et al. 2008), respectively. Sequences had a 100% similarity to N. australe ex-type CMW6837 isolate (accessions AY339262 and AY339270) (Slippers et al. 2004). Two isolates (STEU6554 and STEU6557) were deposited in the culture collection at the Department of Plant Pathology at Stellenbosch University and the sequences were submitted to GenBank with accession numbers ON745603, ON745604, ON746573 and ON746574. Pathogenicity tests using the two N. australe isolates were conducted by inoculating two shoots each of three field-grown C. subternata plants with a 4mm colonised potato dextrose agar (PDA) mycelium plug of each isolate on wounds made by a 4mm cork borer (Van Niekerk et al. 2004). A third shoot was inoculated with a uncolonized PDA plug as the negative control. After 12 weeks, brown-black lesions that were significantly longer (average 55.2 mm) than the uncolonized agar plug control (16.1 mm) were observed. Lesions were observed in all three plants. Neofusicoccum australe was re-isolated (van Niekerk et al. 2004) from all inoculated shoots confirming Koch's postulates. The economic impact and damages caused by N. australe as well as its incidence and severity on honeybush in South Africa is unknown. However, the pathogen caused dieback of entire branches and death of plants indicating that it could be an important pathogen of honeybush. Additionally, N. australe is one of the most important disease-causing Botryosphaeriaceae pathogens on a wide range of economical fruit and vine crops globally (Mojeremane et al. 2020). This is the first report of N. australe as a known pathogen causing decline and dieback of C. subternata in South Africa. References : Alves, A. et al. 2008. Fungal Divers. 28:1. du Toit, J. et al. 1998. J. Sustain. Agric. 12:67. Joubert, E. et al. 2011. S. Afr. J. Bot. 77:887. Mojeremane, K. et al. 2020. Phytopathol. Mediterr. 59:581. Phillips, A. J. et al. 2013. Stud. Mycol. 76:51. Slippers, B. et al. 2004. Mycologia 96:1030. Smit, L. et al. 2021. Eur. J. Plant Pathol. 161:565. van Niekerk, J. M. et al. 2004. Mycologia 96:781. White, T. J. et al. 1990. Pages 315 in: In PCR Protocols: A Guide to Methods and Applications. Academic Press Inc, USA. Declaration . The author(s) declare no conflict of interest Acknowledgments . This work benefitted from the financial support of the Agricultural Research Council, Infruitec-Nietvoorbij, South Africa.

The benchmark dose (BMD) approach is updated to create an international harmonizing process following rapid theoretical sophistication. We calculated the lower limit of BMD confidence interval (BMDL) for carcinogenicity based on 193 tumorigenicity bioassay data published in 50 pesticide risk assessment reports by the Food Safety Commission of Japan (FSCJ) to validate the appropriateness and necessity for the refinement of the FSCJ-established BMD guidance. Three well-known BMD software, PROAST, BMDS, and BBMD were used to compare their BMDLs with no-observed-adverse-effect levels (NOAELs) for carcinogenicity. Recently implemented methodologies such as model averaging or Bayesian inference were also used. Our results indicate that the BMD approach provides a point of departure similar to the NOAEL approach if the data used exhibit a clear dose-response relationship. In some cases, particularly in software with a frequentist approach, the calculation failed to provide BMDL or provided considerably lower BMDLs than NOAELs. However, most of the datasets that resulted in failed calculations or extremely low BMDLs exhibited unclear dose-response relationships, i.e., non-monotonous and sporadic responses. The expert review on the shape of the dose-response plot would help better apply the BMD approach. Furthermore, we observed that Bayesian approaches provided fewer failed or extreme BMD calculations than the frequentist approaches., (Copyright © 2022 Elsevier Inc. All rights reserved.)

This paper is the fourth contribution in the Genera of Phytopathogenic Fungi (GOPHY) series. The series provides morphological descriptions and information about the pathology, distribution, hosts and disease symptoms, as well as DNA barcodes for the taxa covered. Moreover, 12 whole-genome sequences for the type or new species in the treated genera are provided. The fourth paper in the GOPHY series covers 19 genera of phytopathogenic fungi and their relatives, including Ascochyta , Cadophora , Celoporthe , Cercospora , Coleophoma , Cytospora , Dendrostoma , Didymella , Endothia , Heterophaeomoniella, Leptosphaerulina , Melampsora , Nigrospora , Pezicula , Phaeomoniella , Pseudocercospora , Pteridopassalora, Zymoseptoria , and one genus of oomycetes, Phytophthora . This study includes two new genera, 30 new species, five new combinations, and 43 typifications of older names. Taxonomic novelties: New genera: Heterophaeomoniella L. Mostert, C.F.J. Spies, Halleen & Gramaje, Pteridopassalora C. Nakash. & Crous; New species: Ascochyta flava Qian Chen & L. Cai, Cadophora domestica L. Mostert, R. van der Merwe, Halleen & Gramaje, Cadophora rotunda L. Mostert, R. van der Merwe, Halleen & Gramaje, Cadophora vinacea J.R. Úrbez-Torres, D.T. O'Gorman & Gramaje, Cadophora vivarii L. Mostert, Havenga, Halleen & Gramaje, Celoporthe foliorum H. Suzuki, Marinc. & M.J. Wingf., Cercospora alyssopsidis M. Bakhshi, Zare & Crous, Dendrostoma elaeocarpi C.M. Tian & Q. Yang, Didymella chlamydospora Qian Chen & L. Cai, Didymella gei Qian Chen & L. Cai, Didymella ligulariae Qian Chen & L. Cai, Didymella qilianensis Qian Chen & L. Cai, Didymella uniseptata Qian Chen & L. Cai, Endothia cerciana W. Wang. & S.F. Chen, Leptosphaerulina miscanthi Qian Chen & L. Cai, Nigrospora covidalis M. Raza, Qian Chen & L. Cai, Nigrospora globospora M. Raza, Qian Chen & L. Cai, Nigrospora philosophiae-doctoris M. Raza, Qian Chen & L. Cai, Phytophthora transitoria I. Milenković, T. Májek & T. Jung, Phytophthora panamensis T. Jung, Y. Balci, K. Broders & I. Milenković, Phytophthora variabilis T. Jung, M. Horta Jung & I. Milenković, Pseudocercospora delonicicola C. Nakash., L. Suhaizan & I. Nurul Faziha, Pseudocercospora farfugii C. Nakash., I. Araki, & Ai Ito, Pseudocercospora hardenbergiae Crous & C. Nakash., Pseudocercospora kenyirana C. Nakash., L. Suhaizan & I. Nurul Faziha, Pseudocercospora perrottetiae Crous, C. Nakash. & C.Y. Chen, Pseudocercospora platyceriicola C. Nakash., Y. Hatt, L. Suhaizan & I. Nurul Faziha, Pseudocercospora stemonicola C. Nakash., Y. Hatt., L. Suhaizan & I. Nurul Faziha, Pseudocercospora terengganuensis C. Nakash., Y. Hatt., L. Suhaizan & I. Nurul Faziha, Pseudocercospora xenopunicae Crous & C. Nakash.; New combinations: Heterophaeomoniella pinifoliorum (Hyang B. Lee et al. ) L. Mostert, C.F.J. Spies, Halleen & Gramaje, Pseudocercospora pruni-grayanae (Sawada) C. Nakash. & Motohashi., Pseudocercospora togashiana (K. Ito & Tak. Kobay.) C. Nakash. & Tak. Kobay., Pteridopassalora nephrolepidicola (Crous & R.G. Shivas) C. Nakash. & Crous, Pteridopassalora lygodii (Goh & W.H. Hsieh) C. Nakash. & Crous; Typification: Epitypification: Botrytis infestans Mont., Cercospora abeliae Katsuki, Cercospora ceratoniae Pat. & Trab., Cercospora cladrastidis Jacz., Cercospora cryptomeriicola Sawada, Cercospora dalbergiae S.H. Sun, Cercospora ebulicola W. Yamam., Cercospora formosana W. Yamam., Cercospora fukuii W. Yamam., Cercospora glochidionis Sawada, Cercospora ixorana J.M. Yen & Lim, Cercospora liquidambaricola J.M. Yen, Cercospora pancratii Ellis & Everh., Cercospora pini-densiflorae Hori & Nambu, Cercospora profusa Syd. & P. Syd., Cercospora pyracanthae Katsuki, Cercospora horiana Togashi & Katsuki, Cercospora tabernaemontanae Syd. & P. Syd., Cercospora trinidadensis F. Stevens & Solheim, Melampsora laricis-urbanianae Tak. Matsumoto , Melampsora salicis-cupularis Wang, Phaeoisariopsis pruni-grayanae Sawada, Pseudocercospora angiopteridis Goh & W.H. Hsieh, Pseudocercospora basitruncata Crous, Pseudocercospora boehmeriigena U. Braun, Pseudocercospora coprosmae U. Braun & C.F. Hill, Pseudocercospora cratevicola C. Nakash. & U. Braun, Pseudocercospora cymbidiicola U. Braun & C.F. Hill, Pseudocercospora dodonaeae Boesew., Pseudocercospora euphorbiacearum U. Braun, Pseudocercospora lygodii Goh & W.H. Hsieh, Pseudocercospora metrosideri U. Braun, Pseudocercospora paraexosporioides C. Nakash. & U. Braun, Pseudocercospora symploci Katsuki & Tak. Kobay. ex U. Braun & Crous, Septogloeum punctatum Wakef.; Neotypification: Cercospora aleuritis I. Miyake; Lectotypification : Cercospora dalbergiae S.H. Sun, Cercospora formosana W. Yamam., Cercospora fukuii W. Yamam., Cercospora glochidionis Sawada, Cercospora profusa Syd. & P. Syd., Melampsora laricis-urbanianae Tak. Matsumoto , Phaeoisariopsis pruni-grayanae Sawada, Pseudocercospora symploci Katsuki & Tak. Kobay. ex U. Braun & Crous. Citation: Chen Q, Bakhshi M, Balci Y, Broders KD, Cheewangkoon R, Chen SF, Fan XL, Gramaje D, Halleen F, Horta Jung M, Jiang N, Jung T, Májek T, Marincowitz S, Milenković T, Mostert L, Nakashima C, Nurul Faziha I, Pan M, Raza M, Scanu B, Spies CFJ, Suhaizan L, Suzuki H, Tian CM, Tomšovský M, Úrbez-Torres JR, Wang W, Wingfield BD, Wingfield MJ, Yang Q, Yang X, Zare R, Zhao P, Groenewald JZ, Cai L, Crous PW (2022). Genera of phytopathogenic fungi: GOPHY 4. Studies in Mycology 101 : 417-564. doi: 10.3114/sim.2022.101.06., Competing Interests: The authors declare that there is no conflict of interest., (© 2022 Westerdijk Fungal Biodiversity Institute.)

Seasonal temperatures select for eclosion timing of temperate insects and their parasitoids. In western North America, the fruit fly Rhagoletis zephyria Snow (Diptera: Tephritidae) is parasitized by the hymenopterous wasps Utetes lectoides (Gahan), an egg parasite, and Opius downesi Gahan, a larval parasite (both Braconidae). Eclosion of wasps should be timed with the presence of susceptible fly stages, but reports indicate U. lectoides ecloses in the absence of flies under no-chill conditions. Based on this, we tested the hypotheses that chill durations and no-chill temperatures both differentially regulate eclosion times of R. zephyria and its parasitic wasps. When fly puparia were chilled at ~3°C for 130-180 d, U. lectoides and O. downesi always eclosed on average later than flies. However, after 180-d chill, flies eclosed on average earlier than after 130- and 150-d chill, whereas eclosion times of U. lectoides and O. downesi were less or not affected by chill duration. When fly puparia were exposed to 20-22°C (no chill), U. lectoides eclosed before flies, with 88.9% of U. lectoides versus only 0.61% of flies eclosing. Taken together, findings show that eclosion times of flies are more sensitive to changes in chill duration than those of wasps. Flies are less sensitive than wasps to no-chill in that most flies do not respond by eclosing after no-chill while most wasps do. Our results suggest that shorter winters and longer summers due to climate change could cause mismatches in eclosion times of flies and wasps, with potentially significant evolutionary consequences., (Published by Oxford University Press on behalf of Entomological Society of America 2022.)

The apple maggot, Rhagoletis pomonella (Walsh), was introduced into the apple-growing regions of the Pacific Northwest in the U.S.A. during the past 60-100 yr. Apple maggot (larvae, puparia, and adults) is difficult to distinguish from its morphologically similar sister species, Rhagoletis zephyria Snow, which is native and abundant in the Pacific Northwest. While morphological identifications are common practice, a simple, inexpensive assay based on genetic differences would be very useful when morphological traits are unclear. Here we report nucleotide substitution and insertion-deletion mutations in the nontranscribed spacer (NTS) of the ribosomal RNA gene cistron of R. pomonella and R. zephyria that appear to be diagnostic for these two fly species. Insertion-deletion variation is substantial and results in a 49 base-pair difference in PCR amplicon size between R. zephyria and R. pomonella that can be scored using agarose gel electrophoresis. PCR amplification and DNA sequencing of 766 bp of the NTS region from 38 R. pomonella individuals and 35 R. zephyria individuals from across their geographic ranges led to the expected PCR fragments of approx. 840 bp and 790 bp, respectively, as did amplification and sequencing of a smaller set of 26 R. pomonella and 16 R. zephyria flies from a sympatric site in Washington State. Conversely, 633 bp mitochondrial COI barcode sequences from this set of flies were polyphyletic with respect to R. pomonella and R. zephyria. Thus, differences in NTS PCR products on agarose gels potentially provide a simple way to distinguish between R. pomonella and R. zephyria., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America.)

The advances made by microbiome research call for new vocabulary and expansion of our thinking in microbiology. For example, the life-forms presenting in both unicellular and multicellular formats invite us to rethink microbial existence, organization, growth, pathogenicity, and therapeutics in the 21st century. A view of such populations as parts of single organisms with a loose, distributed multicellular organization, introduced here as a germ-ganism, rather than communities, might open up interesting prospects for diagnostics and therapeutics innovation. This study tested and further contextualized the concept of germ-ganism using solid cultures of bacteria and fungi. Based on our findings and the literature reviewed herein, we propose that germ-ganism has synergy with a systems medicine approach by broadening host-environment interactions from cells and microorganisms to a scale of biological ecosystems. Germ-ganism also brings about the possibility of studying the multilevel impacts of novel therapeutic agents within and across networks of microbial ecosystems. The germ-ganism would lend itself, in the long term, to a veritable biocybernetics system, while in the mid-term, we anticipate it will contribute to new diagnostics and therapeutics. Biosecurity applications would be immensely affected by germ-ganism. Industrial applications of germ-ganism are of interest as a more sustainable alternative to costly solutions such as tampered strains/microorganisms. In conclusion, germ-ganism is informed by lessons from microbiome research and invites rethinking microbial existence, organization, and growth as an organism. Germ-ganism has vast ramifications for understanding pathogenicity, and clinical, biosecurity, and biotechnology applications in the current historical moment of the COVID-19 pandemic and beyond.

In recent years, the number of apple trees affected by Botryosphaeria cankers and dieback has considerably increased in central Chile. This study aimed to identify the species of Botryosphaeriaceae associated with canker and dieback symptoms, estimate disease incidence and distributions, and study their pathogenicity and virulence on apple and other fruit crops. A field survey of 34 commercial orchards of apple (7 to 30 years old) was conducted in 16 localities, obtaining 270 symptomatic branch and trunk samples in 2017 and 2018 growing seasons. The incidence of Botryosphaeria canker and dieback ranged between 5 and 40%, and a total of 255 isolates of Botryosphaeriaceae spp. were obtained from 238 cankers. Morphological identification along with phylogenetic studies of the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) of the rDNA, part of the translation elongation factor 1-α ( tef1-α ), and part of the β-tubulin ( tub2 ) genes allowed us to identify Diplodia mutila ( n = 49 isolates), D. seriata ( n = 136 isolates), Lasiodiplodia theobromae ( n = 16 isolates), and Neofusicoccum arbuti ( n = 54 isolates). L. theobromae was isolated mainly from apple dieback from northern localities. All pathogens tested were pathogenic, causing canker and dieback symptoms on lignified twigs of apple, pear, walnut, and green grapevine shoots in the field. Isolates of N. arbuti were the most virulent, reproducing more severe cankers on the lignified tissues inoculated. This study reports, for the first time, D. mutila and L. theobromae associated with Botryosphaeria canker and dieback in Chile, and it is the first description of N. arbuti causing apple dieback worldwide.

The housefly, Musca domestica (Diptera: Muscidae), is capable of transmitting many pathogens that cause severe diseases in humans and animals. Mostly the management tactics rely on synthetic chemicals, but these chemicals creates lethal effects on biological ecosystem. For natural and safe options, bio-control strategy is one of the choice. The present study was a part of such effort to use this strategy and validate the biological performance of the potential pupal parasitoid Dirhinus giffardii (Silvestri) against house fly and provide alternative and safe control of filthy flies. This is the first report on parasitism potential and preference of D. giffardii against house fly. The D. giffardii, early reported as an effective pupal parasitoid of tephritid flies, here in the case presented, showed overall 70% reduction in the house fly population by parasitizing pupae. The parasitism efficiency and longevity of hymenopteran parasitoid was remarkably noted two-fold higher and one-fold more female production on house fly pupae as compared to primary hosts (Tephritids). Furthermore, sex ratio of the resultant progeny was also confirmed the dominancy of female by 74% as compared to males. Based on the novel findings we therefore conclude that D. giffardii is the best bio-control agent for controlling house flies., Competing Interests: The authors have declared that no competing interests exist.

The mitochondrial genome of the fungal pathogen Cercospora nicotianae was sequenced for the first time using a combination of Illumina and Nanopore sequencing technologies. The circular genome is 27,737 bp in length with G + C content of 27.43%, consisting of 15 protein-coding genes, 26 transfer RNA genes and 2 ribosomal RNA genes. Phylogenetic analysis shows that the C. nicotianae mtDNA is closely related to Pseudocercospora fijiensis ., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2021 Guizhou Academy of Tobacco Science. Published by Informa UK Limited, trading as Taylor & Francis Group.)

Photosynthesis in host plants is significantly reduced by many virus families. The early detection of viral infection before the onset of visual symptoms in both directly and systemically infected leaves is critical in crop protection. Viral pathogens cause a variety of symptoms through modifications of chloroplast structure and function and the response of the photochemistry process is immediate. Therefore, chlorophyll fluorescence monitoring has been extensively investigated the last two decades as a tool for timely assessment of pathogenic threats. Alternatively, the analysis of Chla fluorescence transients offers several interlinked parameters which describe the fate of excitation energy round and through the photosystems. Additionally, OJIP fluorescence transients and leaf reflectance spectra methodologies serve for rapid screening of large number of samples. The objective of the present study was to achieve early detection of viral infection, integrating the multiparametric information of the Chla fluorescence transients and of the leaf reflectance spectra into one photochemical performance index. Infection decreased the maximum quantum yield of PSII (F V /F M ), the effective quantum yield of PSII (Φ PSII ), the CO 2 assimilation rate (A) and the stomatal conductance (g s ) in the studied TMV-pepper plant pathosystem, while non-photochemical quenching (NPQ) increased. Some parameters from the OJIP transients and the leaf reflectance spectra were significantly affected 24 h after infection, while others modified three to five days later. Similar results were obtained from systemically infected leaves but with one to three days hysteresis compared to inoculated leaves. Differences between healthy and infected leaves were marginal during the first 24 h post infection. The Integrated Biomarker Response tool was used to create a photochemical infection index (PINFI) which integrates the partial effects of infection on each fluorescence and reflectance index. The PINFI, which to the best of our knowledge is the first photochemical infection index created by the IBR method, discriminated reliably between the infected and healthy leaves of pepper plants from the first 24 h after infection with the TMV., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

A recent olive trunk disease survey performed in the Western Cape Province, South Africa, identified several fungi associated with olive trunk disease symptoms, including species of Basidiomycota, Botryosphaeriaceae, Coniochaetaceae, Calosphaeriaceae, Diaporthaceae, Diatrypaceae, Phaeomoniellaceae, Phaeosphaeriaceae, Symbiotaphrinaceae, Togniniaceae, and Valsaceae. Many of the species recovered had not yet been reported from olive trees; therefore, the aim of this study was to determine their pathogenicity toward this host. Pathogenicity tests were first conducted on detached shoots to select virulent isolates, which were then used in field trials. During field trials, 2-year-old olive branches of 15-year-old trees were inoculated by inserting colonized agar plugs into artificially wounded tissue. Measurements were made of the internal lesions after 8 months. In total, 58 isolates were selected for the field trials. Species that formed lesions significantly larger than the control could be considered as olive trunk pathogens. These included Biscogniauxia rosacearum , Celerioriella umnquma , Coniochaeta velutina , Coniothyrium ferrarisianum , isolates of the Cytospora pruinosa complex, Didymocyrtis banksiae , Diaporthe foeniculina , Eutypa lata , Fomitiporella viticola , Neofusicoccum stellenboschiana , Neofusicoccum vitifusiforme , Neophaeomoniella niveniae , Phaeoacremonium africanum , Phaeoacremonium minimum , Phaeoacremonium oleae , Phaeoacremonium parasiticum , Phaeoacremonium prunicola , Phaeoacremonium scolyti , Phaeoacremonium spadicum , Pleurostoma richardsiae , Pseudophaeomoniella globosa , Punctularia atropurpurascens , Vredendaliella oleae , an undescribed Cytospora sp., Geosmithia sp., two undescribed Neofusicoccum spp., and four Xenocylindrosporium spp. Pseudophaeomoniella globosa can be regarded as one of the main olive trunk pathogens in South Africa because of its high incidence from olive trunk disease symptoms in established orchards and its high virulence in pathogenicity trials.

Digitalization provides access to an integrated network of unexploited big data with potential benefits for society and the environment. The development of smart systems connected to the internet of things can generate unique opportunities to strategically address challenges associated with the United Nations Sustainable Development Goals (SDGs) to ensure an equitable, environmentally sustainable, and healthy society. This perspective describes the opportunities that digitalization can provide towards building the sustainable society of the future. Smart technologies are envisioned as game-changing tools, whereby their integration will benefit the three essential elements of the food-water-energy nexus: (i) sustainable food production; (ii) access to clean and safe potable water; and (iii) green energy generation and usage. It then discusses the benefits of digitalization to catalyze the transition towards sustainable manufacturing practices and enhance citizens' health wellbeing by providing digital access to care, particularly for the underserved communities. Finally, the perspective englobes digitalization benefits by providing a holistic view on how it can contribute to address the serious challenges of endangered planet biodiversity and climate change., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Cactodera torreyanae Cid del Prado Vera & Subbotin, 2014 cysts were discovered during a Pale Potato Cyst Nematode (PCN) survey conducted by Minnesota Department of Agriculture as part of the Animal and Plant Health Inspection Service (APHIS) efforts to survey states for the presence of PCN. The soil samples were collected from a potato field, located in Karlstad, Kittson County, Minnesota, USA. Two out of 175 vials submitted for identification to the Mycology and Nematology Genetic Diversity and Biology Laboratory (MNGDBL) contained few cysts and juveniles of C. torreyanae . Cysts were dark brown in color, lemon-shaped to elongated with distinct vulval cone. Vulva with denticles present around fenestra, cyst length to width ratio between 1.6 and 2.3 and anus distinct. The juveniles had rounded stylet knobs, some sloping slightly posteriorly. The molecular analysis included sequence and phylogenetic analysis of ITS rRNA, D2-D3 expansion segments of 28S rRNA and COI of mtDNA genes. The nematode species was identified by both morphological and molecular means as Cactodera torreyanae . To the best of our knowledge this represents the first report of Cactodera torreyanae from the United States and first report of this cyst nematode species from potato fields. Definite host plant for this nematode remains unknown., (© 2021 Authors.)

Dieback and canker of young stone fruit trees can cause suboptimal growth and even death under severe conditions. One source of inoculum of canker pathogens could be through nursery trees harboring latent infections that would not be visible to inspections done according to the deciduous fruit scheme. The objectives of this study were to identify the canker and wood rot fungal pathogens present in nursery stone fruit trees as well as in propagation material and to evaluate their pathogenicity. Isolations were made from scion and rootstock propagation material and from certified nursery stone fruit trees. The plant material sampled did not have any external symptoms. The certified nursery trees when cross-sectioned displayed brown discoloration from the pruning wound, the bud union, and often the crown. Fungal species isolated were identified by sequencing of the relevant barcoding genes and phylogenetic analyses thereof. Canker- and wood rot-associated fungi were identified. Buds used for budding had low levels of infection, with 1.2% of dormant buds infected and 0.4% of green buds infected. The dormant rootstock shoots had a canker pathogen incidence of 6.2% before they were planted in the nursery fields and increased inasmuch as the ungrafted, rooted rootstock plants had 11.1% infection with canker and wood rot pathogens. Out of 1,080 nursery trees, the canker- and wood rot-associated fungi infected 21.8% of trees. The canker-causing pathogens that were isolated the most were Cadophora luteo-olivacea and Diplodia seriata . A low incidence of wood rot fungi was found, with only 1.5% of nursery trees infected. In total, 26 new reports of fungal species on stone fruit in South Africa were made. Of these, 22 have not been found on stone fruit worldwide. The pathogenicity trials' results confirmed the pathogenic status of these newly reported species. All of the isolates tested formed lesions significantly longer than the control, 4 months after wound inoculation of 2-year-old shoots of two plum orchards. Lasiodiplodia theobromae was the most virulent species on both plum cultivars. The results of this research showed that nursery stone fruit trees and propagation material can harbor latent infections. Different management practices need to be evaluated to prevent these infections to ensure healthier stone fruit nursery trees.

The South American tomato leaf miner, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), one of the most important invasive insect pests affecting tomato production worldwide, was for the first time detected in Myanmar. Preliminary surveys on pest occurrence on tomato crops in Myanmar suggested the presence of T. absoluta in Southern Shan State, but there was no official survey conducted until the end of 2019. Therefore, this study aimed to confirm the presence of T. absoluta in Myanmar. The presence of T. absoluta was specifically observed in the Southern Shan State, which is the largest tomato production area in Myanmar, where tomato is grown under two cultivation methods, floating and land cultivation. The highest T. absoluta infestation was recordedat Kalaw with (82%), followed by Inle Lake (i.e., floating cultivation) (20%) and Pin Ta Ya (10%). The amount of T. absoluta larvae was significantly higher in the lower third of the plants, followed by the middle section and upper section, respectively, in three fields surveyed. Potential and severe economic damage may be expected if management practices are not in place to reduce the presence of this invasive pest. It is of immediate importance that plant protection and quarantine offices of ASEAN member states coordinate their response to T. absoluta and build their capacity to monitor the pest and develop a strategy for when it arrives. In addition, a suitable management strategy is needed to reduce the occurrence of this invasive pest.

Nanotechnology is an avant-garde field of scientific research that revolutionizes technological advancements in the present world. It is a cutting-edge scientific approach that has undoubtedly a plethora of functions in controlling environmental pollutants for the welfare of the ecosystem. However, their unprecedented utilization and hysterical release led to a huge threat to the soil microbiome. Nanoparticles(NPs) hamper physicochemical properties of soil along with microbial metabolic activities within rhizospheric soils.Here in this review shed light on concentric aspects of NP-biosynthesis, types, toxicity mechanisms, accumulation within the ecosystem. However, the accrual of tiny NPs into the soil system has dramatically influenced rhizospheric activities in terms of soil properties and biogeochemical cycles. We have focussed on mechanistic pathways engrossed by microbes to deal with NPs.Also, we have elaborated the fate and behavior of NPs within soils. Besides, a piece of very scarce information on NPs-toxicity towards environment and rhizosphere communities is available. Therefore, the present review highlights ecological perspectives of nanotechnology and solutions to such implications. We have comprehend certain strategies such as avant-garde engineering methods, sustainable procedures for NP synthesis along with vatious regulatory actions to manage NP within environment. Moreover, we have devised risk management sustainable and novel strategies to utilize it in a rationalized and integrated manner. With this background, we can develop a comprehensive plan about NPs with novel insights to understand the resistance and toxicity mechanisms of NPs towards microbes. Henceforth, the orientation towards these issues would enhance the understanding of researchers for proper recommendation and promotion of nanotechnology in an optimized and sustainable manner., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)