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The phytopathogen Pantoea agglomerans belongs to the Bacteria, Proteobacteria, Gammaproteobacteria, Enterobacterales, Erwiniaceae in species classification. It causes disease symptoms in many plants such as corn, banana, and walnut. This study aimed to report the complete genome of P. agglomerans CHTF15, which represents the first whole-genome sequence of an isolate from diseased walnut leaves. The total length of the assembled genome was 4,820,607 bp, with an average GC content of 55.3%, including a circular chromosome and three circular plasmids, two of which were previously unreported sequences and one was announced previously. The CHTF15 genome helps understand the pathogenic mechanism of this important plant pathogen and provides an important theoretical basis for disease epidemic and field control. [Graphic: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 “No Rights Reserved” license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2023.

Chondrostereum purpureum, is a plant fungus causing silver leaf disease of plants, particularly of the rose family. Here we report a case of paratracheal abscess caused by C. purpureum. This is a first of its kind of a case wherein this plant fungus caused disease in a human. Conventional techniques (microscopy and culture) failed to identify the fungus. Only by sequencing, the identity of this unusual pathogen could be revealed. This case highlights the potential of environmental plant fungi to cause disease in humans and stresses the importance of molecular techniques to identify the causative fungal species.

ABSTRACTGolden rain trees (Koelreuteria paniculata) are largely cultivated because of their important ornamental, medicinal, and economic value. However, they are affected by canker and dieback disease to a large extent. To determine the fungi associated with canker and dieback disease of golden rain trees, isolations were obtained from diseased branches and twigs during 2019 and 2020 in greenbelts and nurseries in Beijing, China. Isolates were identified as six species (Allocryptovalsa castaneicola, Botryosphaeria dothidea, Cytospora koelreutericola sp. nov., Dothiorella acericola, Eutypella citricola, and Peroneutypa scoparia) based on morphological features and phylogenetic analyses of ITS, act, rpb2, tef1-α, and tub2. The results of pathogenicity tests indicated that all fungi produced discoloration and Botryosphaeria dothidea was highly aggressive to golden rain tree. In conclusion, this study explored the taxonomy, phylogeny, and pathogenicity of different fungal species associated with canker and dieback disease on golden rain tree and provided fundamental knowledge to improve disease management.

Abstract Over 110 million tons of nitrogen fertilizer every year is used for crop production. Scientists have dreamed of enabling rhizobial nitrogen fixation in non-leguminous crops to mitigate the increasing demand for nitrogen fertilizer. However, despite decades of research, rhizobial nitrogen fixation in non-host plants has not been demonstrated. Here, we reported that an N-fixing rhizobium and a clubroot pathogen Plasmodiophora brassicae exhibited a synergistic effect on fixing nitrogen in cruciferous plants. Rhizobia were found to invade P. brassicae-infected rapeseed (Brassica napus) roots in the field. The colonization of rhizobium on rapeseed roots was confirmed by co-inoculating Mesorhizobium huakuii with P. brassicae under controlled laboratory conditions. M. huakuii infection could alleviate clubroot symptoms and promote the growth of diseased rapeseeds. M. huakuii could fix nitrogen in P. brassicae-infected plants based on the results of 15N isotope dilution tests. The expression of homologs of legume genes required for symbiosis and early-nodulin genes was significantly upregulated in Arabidopsis during early infection by P. brassicae. More importantly, M. huakuii could even fix nitrogen in P. brassicae-resistant rapeseed cultivar and promote plant growth when co-inoculated with P. brassicae. Our findings provide a new avenue to understand the interaction of rhizobia with non-host plants, stimulate the exploration of fixing nitrogen in non-leguminous plants by nitrogen-fixing rhizobia, and develop a strategy for both disease control and nitrogen fixation on non-host crops.

ABSTRACT Genome evolution, and more specifically gene duplication, is a key process shaping host–microorganism interaction. The conserved paralogs usually provide an advantage to the bacterium to thrive. If not, these genes become pseudogenes and disappear. Here, we show that during the emergence of the genus Dickeya, the gene encoding the porin OmpF was duplicated. Our results show that the ompF2 expression is deleterious to the virulence of Dickeya dadantii, the agent causing soft rot disease. Interestingly, ompF2 is regulated while ompF is constitutive but activated by the EnvZ-OmpR two-component system. In vitro, acidic pH triggers the system. The pH measured in four eudicotyledons increased from an initial pH of 5.5 to 7 within 8 h post-infection. Then, the pH decreased to 5.5 at 10 h post-infection and until full maceration of the plant tissue. Yet, the production of phenolic acids by the plant’s defenses prevents the activation of the EnvZ-OmpR system to avoid the ompF2 expression even though environmental conditions should trigger this system. We highlight that gene duplication in a pathogen is not automatically an advantage for the infectious process and that, there was a need for our model organism to adapt its genetic regulatory networks to conserve these duplicated genes. IMPORTANCE Dickeya species cause various diseases in a wide range of crops and ornamental plants. Understanding the molecular program that allows the bacterium to colonize the plant is key to developing new pest control methods. Unlike other enterobacterial pathogens, Dickeya dadantii, the causal agent of soft rot disease, does not require the EnvZ-OmpR system for virulence. Here, we showed that during the emergence of the genus Dickeya, the gene encoding the porin OmpF was duplicated and that the expression of ompF2 was deleterious for virulence. We revealed that while the EnvZ-OmpR system was activated in vitro by acidic pH and even though the pH was acidic when the plant is colonized, this system was repressed by phenolic acid (generated by the plant’s defenses). These results provide a unique— biologically relevant—perspective on the consequence of gene duplication and the adaptive nature of regulatory networks to retain the duplicated gene.

The Gram-negative bacterium Xanthomonas campestris is one of the most problematic phytopathogens, and especially the pathovar campestris (Xcc) that causes a devastating plant disease known as black rot and it is of considerable interest to understand the molecular mechanisms that enable virulence and pathogenicity. Gram-negative bacteria depend on lipoproteins (LPs) that serve many important functions including control of cell shape and integrity, biogenesis of the outer membrane (OM) and establishment of transport pathways across the periplasm. The LPs are localized to the OM where they are attached via a lipid anchor by a process known as the localization of lipoprotein (Lol) pathway. Once a lipid anchor has been synthesized on the nascent LP, the Lol pathway is initiated by a membrane-bound ABC transporter that extracts the lipid anchor of the LP from the IM. The ABC extractor presents the extracted LP to the transport protein LolA, which binds the anchor and thereby shields it from the hydrophilic periplasmic milieu. It is assumed that LolA then carries the LP across the periplasm to the OM. At the periplasmic face of the OM, the LP cargo is delivered to LolB, which completes the Lol pathway by inserting the LP anchor in the inner leaflet of the outer membrane. Earlier studies have shown that loss of Xcc LolA or LolB leads to decreased virulence and pathogenicity during plant infection, which motivates studies to better understand the Lol system in Xcc. In this study, we report the first experimental structure of a complex between LolA and LolB. The crystal structure reveals a stable LolA-LolB complex in the absence of LP. The structural integrity of the LP-free complex is safeguarded by specific protein– protein interactions that do not coincide with interactions predicted to participate in lipid binding. The results allow us to identify structural determinants that enable Xcc LolA to dock with LolB and initiate LP transfer. [ABSTRACT FROM AUTHOR]

'Candidatus Liberibacter' is a group of bacterial species that are obligate intracellular plant pathogens and cause Huanglongbing disease of citrus trees and Zebra Chip in potatoes. Here, we examined the extent of intra- and interspecific genetic diversity across the genus using comparative genomics. Our approach examined a wide set of Liberibacter genome sequences including five pathogenic species and one species not known to cause disease. By performing comparative genomics analyses, we sought to understand the evolutionary history of this genus and to identify genes or genome regions that may affect pathogenicity. With a set of 52 genomes, we performed comparative genomics, measured genome rearrangement, and completed statistical tests of positive selection. We explored markers of genetic diversity across the genus, such as average nucleotide identity across the whole genome. These analyses revealed the highest intraspecific diversity amongst the 'Ca. Liberibacter solanacearum' species, which also has the largest plant host range. We identified sets of core and accessory genes across the genus and within each species and measured the ratio of nonsynonymous to synonymous mutations (dN/dS) across genes. We identified ten genes with evidence of a history of positive selection in the Liberibacter genus, including genes in the Tad complex, which have been previously implicated as being highly divergent in the 'Ca. L. capsica' species based on high values of dN. [ABSTRACT FROM AUTHOR]

The vascular plant pathogenic fungus Verticillium dahliae has to adapt to environmental changes outside and inside its host. V. dahliae harbors homologs of Neurospora crassa clock genes. The molecular functions and interactions of Frequency (Frq) and Frq-interacting RNA helicase (Frh) in controlling conidia or microsclerotia development were investigated in V. dahliae JR2. Fungal mutant strains carrying clock gene deletions, an FRH point mutation, or GFP gene fusions were analyzed on transcript, protein, and phenotypic levels as well as in pathogenicity assays on tomato plants. Our results support that the Frq–Frh complex is formed and that it promotes conidiation, but also that it suppresses and therefore delays V. dahliae microsclerotia formation in response to light. We investigated a possible link between the negative element Frq and positive regulator Suppressor of flocculation 1 (Sfl1) in microsclerotia formation to elucidate the regulatory molecular mechanism. Both Frq and Sfl1 are mainly present during the onset of microsclerotia formation with decreasing protein levels during further development. Induction of microsclerotia formation requires Sfl1 and can be delayed at early time points in the light through the Frq–Frh complex. Gaining further molecular knowledge on V. dahliae development will improve control of fungal growth and Verticillium wilt disease. [ABSTRACT FROM AUTHOR]

Non-ribosomal peptides (NRPs) are a diverse group of bioactive compounds synthesized by microorganisms, and their antimicrobial properties make them ideal candidates for use as biocontrol agents against pathogens. Non-ribosomal peptides produced by Plant-Growth-Promoting Bacteria (PGPB) have gained interest for the biocontrol of plants' bacterial and fungal pathogens. In this review, the structure and mode of action of NRPs, including their characterization and the characterization of NRP-producing microorganisms, are discussed. The use of NRPs in soilless agriculture and their potential as part of a sustainable plant disease control strategy are also highlighted. In addition, the review debates the commercial aspects of PGPB's formulations and their potential as a biocontrol agent. Overall, this review emphasizes the importance of NRPs derived from PGPB in the biocontrol of plant pathogens and their potential to be used as an environmentally friendly and sustainable plant disease control strategy. [ABSTRACT FROM AUTHOR]

Modern plant pathology relies on bioinformatics approaches to create novel plant disease diagnostic tools. In recent years, a significant amount of biological data has been generated due to rapid developments in genomics and molecular biology techniques. The progress in the sequencing of agriculturally important crops has made it possible to develop a better understanding of plant–pathogen interactions and plant resistance. The availability of host–pathogen genome data offers effective assistance in retrieving, annotating, analyzing, and identifying the functional aspects for characterization at the gene and genome levels. Physical mapping facilitates the identification and isolation of several candidate resistance (R) genes from diverse plant species. A large number of genetic variations, such as disease-causing mutations in the genome, have been identified and characterized using bioinformatics tools, and these desirable mutations were exploited to develop disease resistance. Moreover, crop genome editing tools, namely the CRISPR (clustered regulatory interspaced short palindromic repeats)/Cas9 (CRISPR-associated) system, offer novel and efficient strategies for developing durable resistance. This review paper describes some aspects concerning the databases, tools, and techniques used to characterize resistance (R) genes for plant disease management. [ABSTRACT FROM AUTHOR]

Calonectria leaf blight (CLB) is one of the most important diseases of Eucalyptus trees grown in plantations. This disease poses a serious threat to the sustainability of Eucalyptus plantations in southern China. To better understand the causal agents of CLB, we collected samples of diseased leaves and soil from Eucalyptus plantations from nine regions in Guangdong Province where the disease has become a serious problem. A total of 606 Calonectria isolates were purified from the samples, with 399 and 207 originating from diseased leaves and soils, respectively. Phylogenetic analyses utilizing six gene regions resolved 303 isolates in the C. kyotensis species complex and an equal number of isolates in the C. reteaudii species complex. These two complexes were represented by ten known Calonectria species, including C. aconidialis (12.0%), C. curvispora (0.3%), C. hongkongensis (24.8%), C. ilicicola (0.9%), and C. kyotensis (12.0%) in the C. kyotensis species complex, and C. crousiana (1.0%), C. Guangdongensis (0.3%), C. pseudoreteaudii (40.7%), C. queenslandica (7.3%), and C. reteaudii (0.7%) in the C. reteaudii species complex. Pathogenicity tests showed that all species were capable of causing disease on two tested Eucalyptus genotypes, albeit at varying degrees of aggressiveness. Most isolates (98.3%) in the C. reteaudii species complex were from the diseased leaves, indicating that species in this complex are the main causal agents of CLB outbreak. In addition, a significant number of the C. kyotensis species complex isolates (66.7%) from the soil samples could also cause the disease on Eucalyptus leaves. [ABSTRACT FROM AUTHOR]

The Xanthomonas oryzae pv. oryzae (Xoo) is a bacterial pathogen causing bacterial blight disease in rice, resulting in significant yield reductions of up to 50% in rice production. Despite its serious threat to food production globally, knowledge of its population structure and virulence evolution is relatively limited. In this study, we employed whole-genome sequencing to explore the diversity and evolution of Xoo in the main rice-growing areas of China over the past 30 years. Using phylogenomic analysis, we revealed six lineages. CX-1 and CX-2 primarily contained Xoo isolates from South China, while CX-3 represented Xoo isolates from North China. Xoo isolates belonging to CX-5 and CX-6 were the most prevalent across all studied areas, persisting as dominant lineages for several decades. Recent sporadic disease outbreaks were primarily caused by Xoo isolates derived from the two major lineages, CX-5 and CX-6, although Xoo isolates from other lineages also contributed to these outbreaks. The lineage and sub-lineage distributions of Xoo isolates were strongly correlated with their geographical origin, which was found to be mainly determined by the planting of the two major rice subspecies, indica and japonica. Moreover, large-scale virulence testing was conducted to evaluate the diversity of pathogenicity for Xoo. We found rapid virulence evolution against rice, and its determinant factors included the genetic background of Xoo, rice resistance genes, and planting environment of rice. This study provides an excellent model for understanding the evolution and dynamics of plant pathogens in the context of their interactions with their hosts, which are shaped by a combination of geographical conditions and farming practices. The findings of this study may have important implications for the development of effective strategies for disease management and crop protection in rice production systems. [ABSTRACT FROM AUTHOR]

Botrytis cinerea is a necrotrophic fungal pathogen with an extremely broad host range, causing significant economic losses in agricultural production. In this study, we discovered a culture filtrate of bacterial strain HK235, which was identified as Chitinophaga flava, exhibiting high levels of antifungal activity against B. cinerea. From the HK235 culture filtrate, we isolated a new antimicrobial peptide molecule designated as chitinocin based on activity-guided fractionation followed by characterization of the amino acid composition and spectroscopic analyses. The HK235 culture filtrate and chitinocin completely inhibited both conidial germination and mycelial growth of B. cinerea at a concentration of 20% and 200 mg/mL, respectively. In addition to antibiosis against B. cinerea, the active compound chitinocin had a broad antifungal and antibacterial activity in vitro. When tomato plants were treated with the culture filtrate and chitinocin, the treatment strongly reduced the development of gray mold disease in a concentration-dependent manner compared to the untreated control. Here, considering the potent antifungal property in vitro and in vivo, we present the biocontrol potential of C. flava HK235 for the first time. [ABSTRACT FROM AUTHOR]

Summary: Phytomyxea are intracellular biotrophic parasites infecting plants and stramenopiles, including the agriculturally impactful Plasmodiophora brassicae and the brown seaweed pathogen Maullinia ectocarpii. They belong to the clade Rhizaria, where phagotrophy is the main mode of nutrition. Phagocytosis is a complex trait of eukaryotes, well documented for free‐living unicellular eukaryotes and specific cellular types of animals. Data on phagocytosis in intracellular, biotrophic parasites are scant. Phagocytosis, where parts of the host cell are consumed at once, is seemingly at odds with intracellular biotrophy.Here we provide evidence that phagotrophy is part of the nutritional strategy of Phytomyxea, using morphological and genetic data (including a novel transcriptome of M. ectocarpii). We document intracellular phagocytosis in P. brassicae and M. ectocarpii by transmission electron microscopy and fluorescent in situ hybridization.Our investigations confirm molecular signatures of phagocytosis in Phytomyxea and hint at a small specialized subset of genes used for intracellular phagocytosis. Microscopic evidence confirms the existence of intracellular phagocytosis, which in Phytomyxea targets primarily host organelles.Phagocytosis seems to coexist with the manipulation of host physiology typical of biotrophic interactions. Our findings resolve long debated questions on the feeding behaviour of Phytomyxea, suggesting an unrecognized role for phagocytosis in biotrophic interactions. [ABSTRACT FROM AUTHOR]

The movement of pollen grains from anthers to stigmas, often by insect pollinator vectors, is essential for plant reproduction. However, pollen is also a unique vehicle for viral spread. Pollen‐associated plant viruses reside on the outside or inside of pollen grains, infect susceptible individuals through vertical or horizontal infection pathways, and can decrease plant fitness. These viruses are transferred with pollen between plants by pollinator vectors as they forage for floral resources; thus, pollen‐associated viral spread is mediated by floral and pollen grain phenotypes and pollinator traits, much like pollination. Most of what is currently known about pollen‐associated viruses was discovered through infection and transmission experiments in controlled settings, usually involving one virus and one plant species of agricultural or horticultural interest. In this review, we first provide an updated, comprehensive list of the recognized pollen‐associated viruses. Then, we summarize virus, plant, pollinator vector, and landscape traits that can affect pollen‐associated virus transmission, infection, and distribution. Next, we highlight the consequences of plant–pollinator–virus interactions that emerge in complex communities of co‐flowering plants and pollinator vectors, such as pollen‐associated virus spread between plant species and viral jumps from plant to pollinator hosts. We conclude by emphasizing the need for collaborative research that bridges pollen biology, virology, and pollination biology. [ABSTRACT FROM AUTHOR]

Pseudomonas syringae is a Gram-negative bacterium that infects a wide range of plants, causing significant economic losses in agricultural production. The pathogen exhibits a high degree of genetic and phenotypic diversity, which has led to the classification of P. syringae strains into different pathovars based on their host range and disease symptoms. Copper-based products have traditionally been used to manage infections in agriculture, but the emergence of copper-resistant strains has become a significant concern. Biological control is a promising strategy to manage P. syringae, as it offers an environmentally friendly and sustainable approach to disease management. The review includes an overview of the biology and epidemiology of P. syringae, and of the mechanisms of action of various biological control agents, mainly microorganisms (antagonistic bacteria, and fungi) and bacteriophages. Specifically, this review highlights the renewed interest in bacteriophages (bacteria-infecting viruses) due to their advantages over other eco-friendly management methods, thanks to their bactericidal properties and potential to target specific pathogenic bacteria. The potential benefits and limitations of biological control are also examined, along with research directions to optimize the use of this approach for the management of P. syringae. [ABSTRACT FROM AUTHOR]

Lethal bronzing (LB) is a fatal infection that affects over 20 species of palms (Arecaceae) and is caused by the phytoplasma 'Candidatus Phytoplasma aculeata'. This pathogen causes significant economic losses to landscape and nursery companies in Florida, USA. Recently, the vector was determined to be the planthopper Haplaxius crudus, which was more abundant on LB-infected palms. Herein, the volatile chemicals emitted from LB-infected palms were characterized using headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME/GC-MS). Infected Sabal palmetto were identified and confirmed as positive for LB via quantitative PCR. Healthy controls of each species were selected for comparison. All infected palms exhibited elevated levels of hexanal and E-2-hexenal. Threatened palms showed high releasing concentrations of 3-hexenal and Z-3-hexen-1-ol. The volatiles characterized herein are common green-leaf volatiles (GLVs) emitted by plants under stress. This study considers the first documented case of GLVs in palms attributed to phytoplasma infection. Due to the apparent attraction of LB-infected palms to the vector, one or several of the GLVs identified in this study could serve as a lure for the vector and supplement management programs. [ABSTRACT FROM AUTHOR]

Microbial pathogens can adapt rapidly to changing environments such as the application of pesticides or host resistance. Copy number variations (CNVs) are a major source of adaptive genetic variation for recent adaptation. Here, we analyse how a major fungal pathogen of barley, Rhynchosporium commune, has adapted to the host environment and fungicide applications. We screen the genomes of 125 isolates sampled across a worldwide set of populations and identify a total of 7,879 gene duplications and 116 gene deletions. Most gene duplications result from segmental chromosomal duplications. Although CNVs are generally under negative selection, we find that genes affected by CNVs are enriched in functions related to host exploitation (i.e., effectors and cell‐wall‐degrading enzymes). We perform genome‐wide association studies (GWAS) and identify a large segmental duplication of CYP51A that has contributed to the emergence of azole resistance and a duplication encompassing an effector gene affecting virulence. We show that the adaptive CNVs were probably created by recently active transposable element families. Moreover, we find that specific transposable element families are important drivers of recent gene CNV. Finally, we use a genome‐wide single nucleotide polymorphism data set to replicate the GWAS and contrast it with the CNV‐focused analysis. Together, our findings show how extensive segmental duplications create the raw material for recent adaptation in global populations of a fungal pathogen. [ABSTRACT FROM AUTHOR]

Summary: Testing effector knockout strains of the Pseudomonas syringae pv. actinidiae biovar 3 (Psa3) for reduced in planta growth in their native kiwifruit host revealed a number of nonredundant effectors that contribute to Psa3 virulence. Conversely, complementation in the weak kiwifruit pathogen P. syringae pv. actinidifoliorum (Pfm) for increased growth identified redundant Psa3 effectors.Psa3 effectors hopAZ1a and HopS2b and the entire exchangeable effector locus (ΔEEL; 10 effectors) were significant contributors to bacterial colonisation of the host and were additive in their effects on virulence. Four of the EEL effectors (HopD1a, AvrB2b, HopAW1a and HopD2a) redundantly contribute to virulence through suppression of pattern‐triggered immunity (PTI).Important Psa3 effectors include several redundantly required effectors early in the infection process (HopZ5a, HopH1a, AvrPto1b, AvrRpm1a and HopF1e). These largely target the plant immunity hub, RIN4.This comprehensive effector profiling revealed that Psa3 carries robust effector redundancy for a large portion of its effectors, covering a few functions critical to disease. [ABSTRACT FROM AUTHOR]

Fusarium graminearum (F. graminearum) is a filamentous fungus that infects cereals such as corn, wheat, and barley, with serious impact on yield as well as quality when the grain is contaminated with mycotoxins. Despite the huge impact of F. graminearum on food security and mammalian health, the mechanisms used by F. graminearum to export virulence factors during infection are not fully understood and may involve non-classical secretory pathways. Extracellular vesicles (EVs) are lipid-bound compartments produced by cells of all kingdoms that transport several classes of macromolecules and are implicated in cell–cell communication. EVs produced by human fungal pathogens carry cargo that facilitate infection, leading us to ask whether plant fungal pathogens also deliver molecules that increase virulence via EVs. We examined the metabolome of the EVs produced by F. graminearum to determine whether they carry small molecules that could modulate plant–pathogen interactions. We discovered that EVs from F. graminearum were produced in liquid medium-containing inducers of trichothecene production, but in lower quantities compared to other media. Nanoparticle tracking analysis and cryo-electron microscopy revealed that the EVs were morphologically similar to EVs from other organisms; hence, the EVs were metabolically profiled using LC-ESI-MS/MS. This analysis revealed that EVs carry 2,4-dihydroxybenzophenone (BP-1) and metabolites that have been suggested by others to have a role in host–pathogen interactions. BP-1 reduced the growth of F. graminearum in an in vitro assay, suggesting that F. graminearum might use EVs to limit metabolite self-toxicity. [ABSTRACT FROM AUTHOR]

IntroductionMany Gram-negative plant- and animal-pathogenic bacteria employ type IV secretion (T4S) systems to transport proteins or DNA/protein complexes into eukaryotic or bacterial target cells. T4S systems have been divided into minimized and expanded T4S systems and resemble the VirB/VirD4 T4S system from the plant pathogen Agrobacterium tumefaciens and the Icm/Dot T4S system from the human pathogen Legionella pneumophila, respectively. The only known plant pathogen with both types of T4S systems is Xanthomonas euvesicatoria which is the causal agent of bacterial spot disease on pepper and tomato plants.Results and discussionIn the present study, we show that virB/virD4 and icm/dot T4S genes are expressed and encode components of oligomeric complexes corresponding to known assemblies of VirB/VirD4 and Icm/Dot proteins. Both T4S systems are dispensable for the interaction of X. euvesicatoria with its host plants and do not seem to confer contact-dependent lysis of other bacteria, which was previously shown for the chromosomally encoded VirB/VirD4 T4S system from Xanthomonas axonopodis pv. citri. The corresponding chromosomal T4S gene cluster from X. euvesicatoria is incomplete, however, the second plasmid-localized vir gene cluster encodes a functional VirB/VirD4 T4S system which contributes to plasmid transfer. In agreement with this finding, we identified the predicted relaxase TraI as substrate of the T4S systems from X. euvesicatoria. TraI and additional candidate T4S substrates with homology to T4S effectors from X. axonopodis pv. citri interact with the T4S coupling protein VirD4. Interestingly, however, the predicted C-terminal VirD4-binding sites are not sufficient for T4S, suggesting the contribution of additional yet unknown mechanisms to the targeting of T4S substrates from X. euvesicatoria to both VirB/VirD4 and Icm/Dot T4S systems.

Having bacteria grown in pure culture has been the foundation of bacteriology, by allowing a wide range of microbiological studies towards understanding the functionality of a specific bacterium. However, most bacteria have not been axenically cultured to date, thus hindering the understanding of their role in the context of their host or environment. One of these unculturable bacteria are the recently emergent plant pathogens ‘Candidatus Liberibacter spp.’. This group is comprised by dynamic psyllid-vectored, phloem-limited plant pathogens and endophytes that harm a wide range of economically important crops worldwide. ‘Candidatus Liberibacter asiaticus’ (CLas) is associated with Huanglongbing (HLB) in most of the main citrus-producing areas globally, a disease causing severe economic damages. Although the establishment of axenic cultures of CLas remains a major scientific challenge, many research groups have devoted efforts to culture this bacterium to aid in elucidating its virulence mechanism and contribute towards effective HLB management. This led to the development of innovative systems to culture and grow CLas, however different authors have approached the concepts of bacterial culture and axenic culture in different manners, leading to confusion in the terminology used. In this review, we provide the scientific definitions of important terms in bacteriology, while critically reviewing the contribution of each of these important CLas culturing studies.

The Gram-negative bacterium Xanthomonas campestris is one of the most problematic phytopathogens, and especially the pathovar campestris (Xcc) that causes a devastating plant disease known as black rot and it is of considerable interest to understand the molecular mechanisms that enable virulence and pathogenicity. Gram-negative bacteria depend on lipoproteins (LPs) that serve many important functions including control of cell shape and integrity, biogenesis of the outer membrane (OM) and establishment of transport pathways across the periplasm. The LPs are localized to the OM where they are attached via a lipid anchor by a process known as the localization of lipoprotein (Lol) pathway. Once a lipid anchor has been synthesized on the nascent LP, the Lol pathway is initiated by a membrane-bound ABC transporter that extracts the lipid anchor of the LP from the IM. The ABC extractor presents the extracted LP to the transport protein LolA, which binds the anchor and thereby shields it from the hydrophilic periplasmic milieu. It is assumed that LolA then carries the LP across the periplasm to the OM. At the periplasmic face of the OM, the LP cargo is delivered to LolB, which completes the Lol pathway by inserting the LP anchor in the inner leaflet of the outer membrane. Earlier studies have shown that loss of Xcc LolA or LolB leads to decreased virulence and pathogenicity during plant infection, which motivates studies to better understand the Lol system in Xcc. In this study, we report the first experimental structure of a complex between LolA and LolB. The crystal structure reveals a stable LolA-LolB complex in the absence of LP. The structural integrity of the LP-free complex is safeguarded by specific protein–protein interactions that do not coincide with interactions predicted to participate in lipid binding. The results allow us to identify structural determinants that enable Xcc LolA to dock with LolB and initiate LP transfer.

‘Candidatus Liberibacter’ is a group of bacterial species that are obligate intracellular plant pathogens and cause Huanglongbing disease of citrus trees and Zebra Chip in potatoes. Here, we examined the extent of intra- and interspecific genetic diversity across the genus using comparative genomics. Our approach examined a wide set of Liberibacter genome sequences including five pathogenic species and one species not known to cause disease. By performing comparative genomics analyses, we sought to understand the evolutionary history of this genus and to identify genes or genome regions that may affect pathogenicity. With a set of 52 genomes, we performed comparative genomics, measured genome rearrangement, and completed statistical tests of positive selection. We explored markers of genetic diversity across the genus, such as average nucleotide identity across the whole genome. These analyses revealed the highest intraspecific diversity amongst the ‘Ca. Liberibacter solanacearum’ species, which also has the largest plant host range. We identified sets of core and accessory genes across the genus and within each species and measured the ratio of nonsynonymous to synonymous mutations (dN/dS) across genes. We identified ten genes with evidence of a history of positive selection in the Liberibacter genus, including genes in the Tad complex, which have been previously implicated as being highly divergent in the ‘Ca. L. capsica’ species based on high values of dN.

Abstract The main concern of today's agricultural production around the world is a cleaner and greener approach to crop production. Streptomyces species with distinct characteristics can be found in soil. The current research focuses on Streptomyces species isolated from marine sediment samples. To evaluate the morphology and biochemical properties of the isolated Streptomyces strain ukm1, it was grown in Starch Casein Nitrate (SCN) agar medium. Morphological, biochemical, and 16s rRNA analysis were performed on the strain. Streptomyces sp. was identified based on these findings. The production of bioactive metabolites by strain nkm1 was carried out in five different fermentative media. After 10 days of incubation, M6 medium was found to be efficient and was extracted with ethyl acetate. The MIC values revealed that the ethyl acetate extract inhibited the growth of plant pathogenic fungi such as Phytophthora palmivora (31.25 g/ml), Aspergillus flavus (15.62 g/ml), Rhizoctonia solani (31.25 g/ml), and Cladosporium herbarum (31.25 g/ml). The GC-MS analysis of the ethyl acetate extract revealed 26 components, the major volatile bioactive compound being nonadecane.

In this case report, we present a renal transplant recipient who developed an infection due to a plant pathogen – Pantoea agglomerans. This pathogen is a rare organism with no case reports in a renal transplant recipient from India so far. A 50-year-old renal transplant recipient presented with fever for 20 days associated with productive cough and upper back pain. She had received her graft kidney 8 months earlier in a deceased donor program in her local state. In the immediate postoperative period, she had one episode of a mixed rejection, which required a course of antithymocyte globulin and plasmapheresis. On examination, tenderness was noted in her upper back over the cervical and thoracic vertebrae. Her computed tomography scan showed a collection over the cervical vertebrae and consolidation in the lung. Blood culture grew P. agglomerans. P. agglomerans is a rare plant pathogen usually occurring after trauma involving organic debris being deposited within the body. Spontaneously occurring infection is seen only in special situations such as immunocompromised states, malignancy, or excessive antacid use. It has been known to cause lung infections, bone and joint infections, and fever with systemic signs. Spontaneously occurring infections like in this patient are very rare with a handful of case reports worldwide. In an immunocompromised host, a multitude of infections can occur. This case report highlights the importance of removing unnecessary medication from our transplant recipients and optimizing immunosuppression while pursuing the underlying microbiologic cause of infection aggressively.

The genus Cladosporium (Cladosporiaceae, Capnodiales) is a large genus of Ascomycota. Although the genus is mostly reported as saprobes from a wide range of substrates with a worldwide distribution, members of this genus comprise infectious agents in animals and plants. Of those, Cladosporium anthropophilum is a common saprophytic fungus and has been found to be a human opportunistic pathogen and plant pathogen. The complete mitochondrial genome of C. anthropophilum is characterized through the de novo assembly of Illumina sequencing data. The mitochondrial genome is a circular molecule of 35,937 bp with 30.23% GC content and has a total of 47 genes including 16 protein-coding genes, 29 transfer RNA genes, and two ribosomal RNA genes. Based on protein-coding sequences of the mitochondrial genome sequence, a phylogenetic tree was constructed to demonstrate the phylogenetic relationship of C. anthropophilum and its related genera.

Salix babylonica L. is a popular ornamental tree species in China and widely cultivated in Asia, Europe, and North America. Anthracnose in S. babylonica poses a serious threat to its growth and reduces its medicinal properties. In 2021, a total of 55 Colletotrichum isolates were isolated from symptomatic leaves in three provinces in China. Phylogenetic analyses using six loci (ITS, ACT, CHS-1, TUB2, CAL, and GAPDH) and a morphological characterization of the 55 isolates showed that they belonged to four species of Colletotrichum, including C. aenigma, C. fructicola, C. gloeosporioides s.s., and C. siamense. Among them, C. siamense was the dominant species, and C. gloeosporioides s.s. was occasionally discovered from the host tissues. Pathogenicity tests revealed that all the isolates of the aforementioned species were pathogenic to the host, and there were significant differences in pathogenicity or virulence among these isolates. The information on the diversity of Colletotrichum spp. that causes S. babylonica anthracnose in China is new. [ABSTRACT FROM AUTHOR]

Himalayan balsam (Impatiens glandulifera) is one of the most invasive weeds across Europe. The rust fungus, Puccinia komarovii var. glanduliferae has been introduced as a biological control agent, but success has been patchy. Here, we investigated whether mycorrhizal and endophytic fungi can affect rust efficacy and plant growth. Over three experiments, we found that AM fungi and the rust alone or together consistently reduced plant growth, but this depended on the identity of species in the AM inoculum. Meanwhile, AM fungi increased infection frequency of the endophyte Colletotrichum acutatum. Rust inoculation had no detrimental effects on mycorrhizal colonisation or C. acutatum infection, but the latter two fungi reduced rust sporulation. However, plant size was reduced when all three fungal types were present, suggesting that a combined fungal inoculum offers a promising approach for the control of this weed. [ABSTRACT FROM AUTHOR]

Evidence that climate change will impact the ecology and evolution of individual plant species is growing. However, little, as yet, is known about how climate change will affect interactions between plants and their pathogens. Climate drivers could affect the physiology, and thus demography, and ultimately evolutionary processes affecting both plant hosts and their pathogens. Because the impacts of climate drivers may operate in different directions at different scales of infection, and, furthermore, may be nonlinear, abstracting across these processes may mis-specify outcomes. Here, we use mechanistic models of plant-pathogen interactions to illustrate how counterintuitive outcomes are possible, and we introduce how such framing may contribute to understanding climate effects on plant-pathogen systems. We discuss the evidence-base derived from wild and agricultural plant-pathogen systems that could inform such models, specifically in the direction of estimates of physiological, demographic and evolutionary responses to climate change. We conclude by providing an overview of knowledge gaps and directions for future research in this important area. [ABSTRACT FROM AUTHOR]

To successfully colonize the host, phytopathogens have developed a large repertoire of components to both combat the host plant defense mechanisms and to survive in adverse environmental conditions. Microbial proteases are predicted to be crucial components of these systems. In the present work, we aimed to identify active secreted proteases from the oomycete Aphanomyces euteiches, which causes root rot diseases on legumes. Genome mining and expression analysis highlighted an overrepresentation of microbial tandemly repeated proteases, which are upregulated during host infection. Activity Based Protein Profiling and mass spectrometry (ABPP-MS) on apoplastic fluids isolated from pea roots infected by the pathogen led to the identification of 35 active extracellular microbial proteases, which represents around 30% of the genes expressed encoding serine and cysteine proteases during infection. Notably, eight of the detected active secreted proteases carry an additional Cterminal domain. This study reveals novel active modular extracellular eukaryotic proteases as potential pathogenicity factors in Aphanomyces genus. [ABSTRACT FROM AUTHOR]

Scolecobasidium is cosmopolitan and includes species that inhabit a wide range of ecosystems including soil, water, air, plant and cold-blooded vertebrates. During a fungal survey from mangrove, strains of Scolecobasidium occurring on leaf spots of true mangrove plants, Aegiceras corniculatum and Acanthus ebracteatus, were isolated from Futian Mangrove in Shenzhen and the Qi'ao-Dangan Island Mangrove in Zhuhai, China. Unlike most species in Scolecobasidium that produce dark conidia, our strains are characterized by hyaline to pale brown conidia and inconspicuous thread-like sterigmata. Further detailed morphological comparison and multi-locus (LSU, ITS, tub2, tef1-a) phylogenetic analyses revealed these collections as two new taxa, namely S. acanthi sp. nov. and S. aegiceratis sp. nov. We further emend the generic description of Scolecobasidium, propose one new combination, S. terrestre comb. nov., and clarify the taxonomic status of S. constrictum. [ABSTRACT FROM AUTHOR]

Golden rain trees (Koelreuteria paniculata) are largely cultivated because of their important ornamental, medicinal, and economic value. However, they are affected by canker and dieback disease to a large extent. To determine the fungi associated with canker and dieback disease of golden rain trees, isolations were obtained from diseased branches and twigs during 2019 and 2020 in greenbelts and nurseries in Beijing, China. Isolates were identified as six species (Allocryptovalsa castaneicola, Botryosphaeria dothidea, Cytospora koelreutericola sp. nov., Dothiorella acericola, Eutypella citricola, and Peroneutypa scoparia) based on morphological features and phylogenetic analyses of ITS, act, rpb2, tef1-α, and tub2. The results of pathogenicity tests indicated that all fungi produced discoloration and Botryosphaeria dothidea was highly aggressive to golden rain tree. In conclusion, this study explored the taxonomy, phylogeny, and pathogenicity of different fungal species associated with canker and dieback disease on golden rain tree and provided fundamental knowledge to improve disease management. [ABSTRACT FROM AUTHOR]

Phosphite has been sprayed on threatened natural plant communities in southwestern Australia for the past 25 years to mitigate the impact of the globally widespread plant pathogen, Phytophthora cinnamomi. Of concern is that this may result in an increase in phosphorus concentrations in naturally nutrient deficient soils, causing the loss of plant species that are sensitive to increased phosphorus levels. The impact of aerial spraying phosphite on soil and Proteaceae species richness and abundance was measured at three sites within the Southeast Coastal Province of Western Australia. Healthy and diseased vegetation which had been sprayed with phosphite for 14–26 years was compared with nonsprayed healthy and diseased vegetation. Contrary to expectations, there was no evidence that spraying is doing harm to Proteaceae species at the three sites studied. Healthy‐nonsprayed and Healthy‐sprayed areas had generally similar levels of soil total P, and this was associated with similar values for Proteaceae species richness and abundance in these areas. Phosphite spraying of diseased vegetation benefited Proteaceae plant diversity and abundance by preventing the disappearance of some species. Given these results, it was recommended that phosphite spraying be continued as a management action for controlling Phytophthora cinnamomi while research into alternative solutions continues. [ABSTRACT FROM AUTHOR]

A contemporary agricultural production system relying on heavy usage of agrochemicals represents a questionable outlook for sustainable food supply in the future. The visible negative environmental impacts and unforeseen consequences to human and animal health have been requiring a shift towards the novel eco-friendly alternatives for chemical pesticides for a while now. Microbial-based biocontrol agents have shown a promising potential for plant disease management. The bacteria of the genus Bacillus have been among the most exploited microbial active components due to several highly efficient mechanisms of action against plant pathogens, as well as a palette of additional plant-beneficial mechanisms, together with their suitable properties for microbial biopesticide formulations. Among other bioactive metabolites, volatile organic compounds (VOCs) have been investigated for their biocontrol applications, exhibiting the main advantage of long-distance effect without the necessity for direct contact with plants or pathogens. The aim of this study is to give an overview of the state-of-the-art in the field of Bacillus-based VOCs, especially in terms of their antibacterial, antifungal, and nematicidal action as the main segments determining their potential for biocontrol applications in sustainable agriculture. [ABSTRACT FROM AUTHOR]

Soil borne pathogens are significant contributor of plant yield loss globally. The constraints in early diagnosis, wide host range, longer persistence in soil makes their management cumbersome and difficult. Therefore, it is crucial to devise innovative and effective management strategy to combat the losses caused by soil borne diseases. The use of chemical pesticides is the mainstay of current plant disease management practices that potentially cause ecological imbalance. Nanotechnology presents a suitable alternative to overcome the challenges associated with diagnosis and management of soil-borne plant pathogens. This review explores the use of nanotechnology for the management of soil-borne diseases using a variety of strategies, such as nanoparticles acting as a protectant, as carriers of actives like pesticides, fertilizers, antimicrobials, and microbes or by promoting plant growth and development. Nanotechnology can also be used for precise and accurate detection of soil-borne pathogens for devising efficient management strategy. The unique physico-chemical properties of nanoparticles allow greater penetration and interaction with biological membrane thereby increasing its efficacy and releasability. However, the nanoscience specifically agricultural nanotechnology is still in its toddler stage and to realize its full potential, extensive field trials, utilization of pest crop host system and toxicological studies are essential to tackle the fundamental queries associated with development of commercial nano-formulations. [ABSTRACT FROM AUTHOR]

Pineapple is one of the most economically important fruits in tropical countries, particularly in Thailand. Canned pineapple is currently Thailand's main exported commodity to many countries, including the United States, Russia, Germany, Poland, and Japan. Fungal diseases are considered a permanent threat to fruits in the pre- and post-harvest stages, leading to considerable economic losses. Fungal disease is one of the primary causes of massive yield losses in pineapples around the world. Colletotrichum species are the most common fungal pathogens affecting different tropical fruits. Although there are many reports regarding Colletotrichum species associated with pineapple, they do not have molecular data to confirm species identification. However, the occurrence of Colletotrichum species on pineapple has not been reported in Thailand so far. In this study, we isolated and identified Colletotrichum fructicola on pineapple in northern Thailand and have proven its pathogenicity to the host. This is the first report of the occurrence of Colletotrichum in pineapple, based on morpho-molecular approaches. [ABSTRACT FROM AUTHOR]

Melatonin is a multifunctional and ubiquitous molecule. In animals, melatonin is a hormone that is involved in a wide range of physiological activities and is also an excellent antioxidant. In plants, it has been considered a master regulator of multiple physiological processes as well as of hormonal homeostasis. Likewise, it is known for its role as a protective biomolecule and activator of tolerance and resistance against biotic and abiotic stress in plants. Since infections by pathogens such as bacteria, fungi and viruses in crops result in large economic losses, interest has been aroused in determining whether melatonin plays a relevant role in plant defense systems against pathogens in general, and against viruses in particular. Currently, several strategies have been applied to combat infection by pathogens, one of them is the use of eco-friendly chemical compounds that induce systemic resistance. Few studies have addressed the use of melatonin as a biocontrol agent for plant diseases caused by viruses. Exogenous melatonin treatments have been used to reduce the incidence of several virus diseases, reducing symptoms, virus titer, and even eradicating the proliferation of viruses such as Tobacco Mosaic Virus, Apple Stem Grooving Virus, Rice Stripe Virus and Alfalfa Mosaic Virus in tomato, apple, rice and eggplant, respectively. The possibilities of using melatonin as a possible natural virus biocontrol agent are discussed. [ABSTRACT FROM AUTHOR]

Phytophthora are plant pathogens that damage agricultural products. Lycosides (1a -d), found in vegetable juice, have the potential to curb the rapid outbreak and crop damage caused by the asexual reproduction of Phytophthora. Here, aglycones 2a, b with slightly higher activity than lycosides were synthesized as a diastereomeric mixture (mix - 2) possessing activity (IC50 = 4.1 µm) comparable with that of lycosides. The importance of the cyclohexanone structure and side-chain length was demonstrated via structure -activity relationship analysis using synthetic intermediates. In addition, the action mechanism of lycosides was investigated using transcriptome analysis, which revealed a contribution to proline biosynthesis inhibition, a process crucial for the asexual reproduction of Phytophthora. These findings indicate that lycosides (and aglycone) are environmentally benign agents that can be used for protecting agricultural products from Phytophthora pathogens. [ABSTRACT FROM AUTHOR]

Simple Summary: The bacterium Streptomyces scabiei is the main causative agent of common scab disease on economically important root and tuber crops. Our work investigated if and how this pathogen uses multiple sugar transport systems to sense the presence of a living plant host through perception and import of cello-oligosaccharides, the elicitors that trigger the onset of the pathogenic lifestyle of S. scabiei. Plant colonization by Streptomyces scabiei, the main cause of common scab disease on root and tuber crops, is triggered by cello-oligosaccharides, cellotriose being the most efficient elicitor. The import of cello-oligosaccharides via the ATP-binding cassette (ABC) transporter CebEFG-MsiK induces the production of thaxtomin phytotoxins, the central virulence determinants of this species, as well as many other metabolites that compose the 'virulome' of S. scabiei. Homology searches revealed paralogues of the CebEFG proteins, encoded by the cebEFG2 cluster, while another ABC-type transporter, PitEFG, is encoded on the pathogenicity island (PAI). We investigated the gene expression of these candidate alternative elicitor importers in S. scabiei 87-22 upon cello-oligosaccharide supply by transcriptomic analysis, which revealed that cebEFG2 expression is highly activated by both cellobiose and cellotriose, while pitEFG expression was barely induced. Accordingly, deletion of pitE had no impact on virulence and thaxtomin production under the conditions tested, while the deletion of cebEFG2 reduced virulence and thaxtomin production, though not as strong as the mutants of the main cello-oligosaccharide transporter cebEFG1. Our results thus suggest that both ceb clusters participate, at different levels, in importing the virulence elicitors, while PitEFG plays no role in this process under the conditions tested. Interestingly, under more complex culture conditions, the addition of cellobiose restored thaxtomin production when both ceb clusters were disabled, suggesting the existence of an additional mechanism that is involved in sensing or importing the elicitor of the onset of the pathogenic lifestyle of S. scabiei. [ABSTRACT FROM AUTHOR]

The Dickeya and Pectobacterium bacterial species cause blackleg and soft-rot diseases on potato plants and tubers. Prophylactic actions are essential to conserve a high quality of seed potato tubers. Biocontrol approaches are emerging, but we need to know how efficient biocontrol agents are when facing the natural diversity of pathogens. In this work, we sampled 16 production fields, which were excluded from the seed tuber certification scheme, as well as seven experimental parcels, which were planted with seed tubers from those production fields. We collected and characterized 669 Dickeya and Pectobacterium isolates, all characterized using nucleotide sequence of the gapA gene. This deep sampling effort highlighted eleven Dickeya and Pectobacterium species, including four dominant species namely D. solani, D. dianthicola, P. atrosepticum and P. parmentieri. Variations in the relative abundance of pathogens revealed different diversity patterns at a field or parcel level. The Dickeya-enriched patterns were maintained in parcels planted with rejected seed tubers, suggesting a vertical transmission of the pathogen consortium. Then, we retained 41 isolates representing the observed species diversity of pathogens and we tested each of them against six biocontrol agents. From this work, we confirmed the importance of prophylactic actions to discard contaminated seed tubers. We also identified a couple of biocontrol agents of the Pseudomonas genus that were efficient against a wide range of pathogen species. [ABSTRACT FROM AUTHOR]

Fossils and other preserved specimens are integral for informing timing and evolutionary history in every biological system. By isolating a plant pathogen genome from herbarium-preserved diseased grapevine material from 1906 (Herb_1906), we were able to answer questions about an enigmatic system. The emergence of Pierce's disease (PD) of grapevine has shaped viticultural production in North America; yet, there are uncertainties about the geographic origin of the pathogen (Xylella fastidiosa subsp. fastidiosa , Xff) and the timing and route of its introduction. We produced a high-quality, de novo genome assembly of this historical plant pathogen and confirmed degradation patterns unique to ancient DNA. Due to the inclusion of the Herb_1906 sample, we were able to generate a significant temporal signal in the genomic data. This allowed us to build a time-calibrated phylogeny, where we estimate the introduction of Xff into the US between 1734 and 1741 CE, an earlier time frame than previously inferred. In a large collection of >300 Xff genomes, the Herb_1906 sample was genetically most similar to a small population from Northern California but not basal to the entire Xff California clade. Based on phylogenetic placement and a phylogeographic reconstruction, our data support a single introduction of Xff into the Southeastern US from Central America, with multiple subsequent introductions into California. [Display omitted] • The pathogen X. fastidiosa was recovered from a 1906 grapevine herbarium sample • Deep sequencing enabled the de novo assembly of a high-quality aDNA pathogen genome • The PD-causing pathogen was likely introduced multiple times into California • Time calibration pushed back the date of introduction to the US to 1734–1741 CE Donegan et al. leverage the genome of a century-old specimen of the pathogen Xylella fastidiosa to unravel the emergence of Pierce's disease of grapevine across North America. The results recalibrate the timeline of pathogen introduction and challenge the assumption about a single introduction of X. fastidiosa from Central America through California. [ABSTRACT FROM AUTHOR]