Colonization Of Host Plants Research Articles

Phosphonates of Pectobacterium atrosepticum: Discovery and Role in Plant-Pathogen Interactions.

Many phytopathogens' gene products that contribute to plant-pathogen interactions remain unexplored. In one of the most harmful phytopathogenic bacterium Pectobacterium atrosepticum (Pba), phosphonate-related genes have been previously shown to be among the most upregulated following host plant colonization. However, phosphonates, compounds characterized by a carbon-phosphorus bond in their composition, have not been described in Pectobacterium species and other phytopathogenic bacteria, with the exception of Pseudomonas syringae and Pantoea ananatis. Our study aimed to determine whether Pba synthesizes extracellular phosphonates and, if so, to analyze their physiological functions. We demonstrated that Pba produces two types of extracellular phosphonates: 2-diethoxyphosphorylethanamine and phenylphosphonic acid. Notably, such structures have not been previously described among natural phosphonates. The production of Pba phosphonates was shown to be positively regulated by quorum sensing and in the presence of pectic compounds. Pba phosphonates were found to have a positive effect on Pba stress resistance and a negative effect on Pba virulence. The discovered Pba phosphonates are discussed as metabolites that enable Pba to control its "harmful properties", thereby maintaining its ecological niche (the host plant) in a relatively functional state for an extended period.

Philaenus tesselatus (Hemiptera: Aphrophoridae), the main potential vector of Xylella fastidiosa in Morocco: seasonal abundance, phenology and host-plant colonization by nymphs

Summary The bacterium Xylella fastidiosa presents a potential risk to the Moroccan flora, particularly for the citrus, olive, almond and grapevine industries. Philaenus tesselatus, the meadow spittlebug, is the most important Moroccan spittlebug, though recorded with uneven occurrence and considered to be the main potential insect vector of X. fastidiosa. A three-year field survey (2019 to 2021) was conducted at three sites in the provinces of Larache and Kenitra in northwestern Morocco, to study seasonal abundance, phenology and host-plant colonization by nymphs. Philaenus tesselatus is a univoltine species. Nymphs occurred in groundcover from end of mid-March to mid-May, with abundance peaking between end of March and mid-May. Emergence of teneral adults occurred between April and May, depending on the year. Adults were present on ground vegetation from March until early August, with the highest abundances surveyed by sweep net in April, and occurred in canopy in low abundance between the end of May and mid-September, depending on site. Both nymphal and adult abundances were significantly higher in Larache than at the other two sites. Philaenus tesselatus, the meadow spittlebug, is a polyphagous species, recorded on nine botanical families of host plant species including Asteraceae, Poaceae, Fabaceae, Polygonaceae, Apiaceae, Primulaceae, Urticaceae, Papaveraceae and Malvaceae, and 37 host plant species, of which 29 were identified. The Asteraceae were best represented. Carduus pycnocephalus, Vulpia geniculata, Anthoxanthum odoratum and Cistus salviifolius were the most suitable for this spittlebug. Avena barbata, L. foemina, Scolymus hispanicus and S. oleraceus were dominant in the herbaceous layer. Philaenus tesselatus preferred to feed on the mid-section of the host plants. A single foam can host zero to five individuals, but foams with one or two nymphs were most frequent. Implications of our findings for the setting up of a survey program to control X. fastidiosa and its vectors in case the bacterium is introduced to Morocco are discussed.

Unveiling genomic features linked to traits of plant growth-promoting bacterial communities from sugarcane

Microorganisms are ubiquitous, and those inhabiting plants have been the subject of several studies. Plant-associated bacteria exhibit various biological mechanisms that enable them to colonize host plants and, in some cases, enhance their fitness. In this study, we describe the genomic features predicted to be associated with plant growth-promoting traits in six bacterial communities isolated from sugarcane. The use of highly accurate single-molecule real-time sequencing technology for metagenomic samples from these bacterial communities allowed us to recover 17 genomes. The taxonomic assignments for the binned genomes were performed, revealing taxa distributed across three main phyla: Bacillota, Bacteroidota, and Pseudomonadota, with the latter being the most representative. Subsequently, we functionally annotated the metagenome-assembled genomes (MAGs) to characterize their metabolic pathways related to plant growth-promoting traits. Our study successfully identified the enrichment of important functions related to phosphate and potassium acquisition, modulation of phytohormones, and mechanisms for coping with abiotic stress. These findings could be linked to the robust colonization of these sugarcane endophytes.

Root endophytic bacterial communities are shaped by the specific microbiota associated to mycorrhizal symbionts

Abstract Background and aims Arbuscular mycorrhizal fungi (AMF) are beneficial soil microorganisms establishing mutualistic symbioses with most crop plants and promoting plant growth and health. AMF beneficial activities are complemented by their associated microbiota, leading to synergistic interactions positively affecting plant performance. In this work we assessed whether AMF may act as drivers of root bacterial endophytes, facilitating root colonization of host plants by their associated bacteria. Methods Two AMF isolates were used, Funneliformis mosseae from Indiana (USA) and Septoglomus sp. from Tuscany (Italy) in an original experimental microcosm system, utilizing micropropagated plants of Prunus persica x Prunus amygdalus inoculated with either intact or mechanically crushed AMF spores, the former able and the latter unable to establish the symbiosis. Spore and root endophytic bacterial communities diversity were analysed by Illumina Miseq sequencing. Results This study revealed that AMF with their associated bacteria can shape the root endophytic bacterial communities, inducing differential recruitment depending on the composition of spore-associated microbiota. Such data were consistent between two AMF isolates, associated with diverse bacterial communities, as shown by PERMANOVA, Bray Curtis dissimilarity, hierarchical clustering and indicator species analyses. Moreover, specific bacterial taxa were found exclusively in mycorrhizal roots. Our findings suggested also a differential recruitment depending on the ability of AMF to establish mycorrhizal symbioses. Conclusion This work revealed that AMF represent drivers of the endophytic bacterial communities diversity and composition, facilitating root colonization of host plants by their associated bacteria, that become an integral part of the root microbiome as endophytes.

A comprehensive comparative genomic analysis revealed that plant growth promoting traits are ubiquitous in strains of Stenotrophomonas.

Stenotrophomonas strains, which are often described as plant growth promoting (PGP) bacteria, are ubiquitous in many environments. A total of 213 genomes of strains of Stenotrophomonas were analyzed using comparative genomics to better understand the ecological roles of these bacteria in the environment. The pan-genome of the 213 strains of Stenotrophomonas consists of 27,186 gene families, including 710 core gene families, 11,039 unique genes and 15,437 accessory genes. Nearly all strains of Stenotrophomonas harbor the genes for GH3-family cellulose degradation and GH2- and GH31-family hemicellulose hydrolase, as well as intact glycolysis and tricarboxylic acid cycle pathways. These abilities suggest that the strains of this genus can easily obtain carbon and energy from the environment. The Stenotrophomonas strains can respond to oxidative stress by synthesizing catalase, superoxide dismutase, methionine sulfoxide reductase, and disulfide isomerase, as well as managing their osmotic balance by accumulating potassium and synthesizing compatible solutes, such as betaine, trehalose, glutamate, and proline. Each Stenotrophomonas strain also contains many genes for resistance to antibiotics and heavy metals. These genes that mediate stress tolerance increase the ability of Stenotrophomonas strains to survive in extreme environments. In addition, many functional genes related to attachment and plant colonization, growth promotion and biocontrol were identified. In detail, the genes associated with flagellar assembly, motility, chemotaxis and biofilm formation enable the strains of Stenotrophomonas to effectively colonize host plants. The presence of genes for phosphate-solubilization and siderophore production and the polyamine, indole-3-acetic acid, and cytokinin biosynthetic pathways confer the ability to promote plant growth. These strains can produce antimicrobial compounds, chitinases, lipases and proteases. Each Stenotrophomonas genome contained 1-9 prophages and 17-60 genomic islands, and the genes related to antibiotic and heavy metal resistance and the biosynthesis of polyamines, indole-3-acetic acid, and cytokinin may be acquired by horizontal gene transfer. This study demonstrates that strains of Stenotrophomonas are highly adaptable for different environments and have strong potential for use as plant growth-promoting bacteria.

Surviving the Potato Stems: Differences in Genes Required for Fitness by Dickeya dadantii and Dickeya dianthicola.

Bacteria belonging to the genus Dickeya cause blackleg and soft rot symptoms on many plant hosts, including potato. Although there is considerable knowledge about the genetic determinants that allow Dickeya to colonize host plants, as well as the genes that contribute to virulence, much is still unknown. To identify the genes important for fitness in potato stems, we constructed and evaluated randomly barcoded transposon mutant (RB-TnSeq) libraries of Dickeya dadantii and Dickeya dianthicola. We identified 169 and 157 genes important for growth in D. dadantii and D. dianthicola in stems, respectively. This included genes related to metabolic pathways, chemotaxis and motility, transcriptional regulation, transport across membranes, membrane biogenesis, detoxification mechanisms, and virulence-related genes, including a potential virulence cluster srfABC, c-di-GMP modulating genes, and pectin degradation genes. When we compared the results of the stem assay with other datasets, we identified genes important for growth in stems versus tubers and in vitro conditions. Additionally, our data showed differences in fitness determinants for D. dadantii and D. dianthicola. These data provide important insights into the mechanisms used by Dickeya when interacting with and colonizing plants and thus might provide targets for management.

MoMkk1 and MoAtg1 dichotomously regulating autophagy and pathogenicity through MoAtg9 phosphorylation in Magnaporthe oryzae.

Autophagy is a central biodegradation pathway critical in eliminating intracellular cargo to maintain cellular homeostasis and improve stress resistance. At the same time, the key component of the mitogen-activated protein kinase cascade regulating cell wall integrity signaling MoMkk1 has an essential role in the autophagy of the rice blast fungus Magnaporthe oryzae. Still, the mechanism of how MoMkk1 regulates autophagy is unclear. Interestingly, we found that MoMkk1 regulates the autophagy protein MoAtg9 through phosphorylation. MoAtg9 is a transmembrane protein subjected to phosphorylation by autophagy-related protein kinase MoAtg1. Here, we provide evidence demonstrating that MoMkk1-dependent MoAtg9 phosphorylation is required for phospholipid translocation during isolation membrane stages of autophagosome formation, an autophagic process essential for the development and pathogenicity of the fungus. In contrast, MoAtg1-dependent phosphorylation of MoAtg9 negatively regulates this process, also impacting growth and pathogenicity. Our studies are the first to demonstrate that MoAtg9 is subject to MoMkk1 regulation through protein phosphorylation and that MoMkk1 and MoAtg1 dichotomously regulate autophagy to underlie the growth and pathogenicity of M. oryzae.IMPORTANCEMagnaporthe oryzae utilizes multiple signaling pathways to promote colonization of host plants. MoMkk1, a cell wall integrity signaling kinase, plays an essential role in autophagy governed by a highly conserved autophagy kinase MoAtg1-mediated pathway. How MoMkk1 regulates autophagy in coordination with MoAtg1 remains elusive. Here, we provide evidence that MoMkk1 phosphorylates MoAtg9 to positively regulate phospholipid translocation during the isolation membrane or smaller membrane structures stage of autophagosome formation. This is in contrast to the negative regulation of MoAtg9 by MoAtg1 for the same process. Intriguingly, MoMkk1-mediated MoAtg9 phosphorylation enhances the fungal infection of rice, whereas MoAtg1-dependant MoAtg9 phosphorylation significantly attenuates it. Taken together, we revealed a novel mechanism of autophagy and virulence regulation by demonstrating the dichotomous functions of MoMkk1 and MoAtg1 in the regulation of fungal autophagy and pathogenicity.

Subtilisin-like proteases from Fusarium graminearum induce plant cell death and contribute to virulence.

Fusarium head blight (FHB), caused by Fusarium graminearum, causes huge annual economic losses in cereal production. To successfully colonize host plants, pathogens secrete hundreds of effectors that interfere with plant immunity and facilitate infection. However, the roles of most secreted effectors of F. graminearum in pathogenesis remain unclear. We analyzed the secreted proteins of F. graminearum and identified 255 candidate effector proteins by liquid chromatography-mass spectrometry (LC-MS). Five subtilisin-like family proteases (FgSLPs) were identified that can induce cell death in Nicotiana benthamiana leaves. Further experiments showed that these FgSLPs induced cell death in cotton (Gossypium barbadense) and Arabidopsis (Arabidopsis thaliana). A signal peptide and light were not essential for the cell death-inducing activity of FgSLPs. The I9 inhibitor domain and the entire C-terminus of FgSLPs were indispensable for their self-processing and cell death-inducing activity. FgSLP-induced cell death occurred independent of the plant signal transduction components BRI-ASSOCIATED KINASE 1 (BAK1), SUPPRESSOR OF BIR1 1 (SOBIR1), ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1), and PHYTOALEXIN DEFICIENT 4 (PAD4). Reduced virulence was observed when FgSLP1 and FgSLP2 were simultaneously knocked out. This study reveals a class of secreted toxic proteins essential for F. graminearum virulence.

Unraveling the functional characteristics of endophytic bacterial diversity for plant growth promotion and enhanced secondary metabolite production in Pelargonium graveolens

The rich diversity of microbial endophytic communities associated with plants, often referred to as the second genome, serves as a compelling illustration of efficient co-evolution. This noteworthy partnership plays a pivotal role in sustaining plant well-being and enhancing plant adaptability across diverse habitats. Therefore, examining the diversity of endophytic microbes associated with their particular host plant is valuable for gaining insights into the vast spectrum of plant-microbe interactions. The present experiments aimed at investigating the bacterial endophytic diversity in both root and shoot tissues of Pelargonium graveolens, employing culture dependent and culture independent high-throughput metagenomics approach. A total of 614 and 620 operational taxonomic units (OTUs), encompassing 291 and 229 genera, were identified in the shoot and root tissues of P. graveolens, respectively. Furthermore, the subsequent classification of OTUs revealed 15 highly abundant phyla, with Proteobacteria dominating both root and shoot tissues. Notably, an exceptionally high abundance of Firmicutes phyla was observed in the shoot compared to the root. Additionally, 30 bacterial endophytes from the root, stem, petiole, and leaves were isolated and molecularly characterized, unveiling a consistent pattern of diversity distribution between the root and shoot of P. graveolens. Upon screening all isolates for plant growth promoting traits, Pseudomonas oryzihabitans was found to be positive for major biochemical test like nitrogen fixation, phosphate solubilization etc. and on inoculation resulted in about two-fold increase in content of essential oil accompanied by a significant rise in the geraniol and citronellol content. Diving deep into the genetic constitution of P. oryzihabitans unveiled a substantial number of genes directly and indirectly contributing to the endophyte’s capability in colonizing host plants effectively. In summary, data obtained from metagenomics and culture dependent approaches including glass house trials suggest potential bacterial endophytes suitable for field applications for yield enhancement and in planta secondary metabolite enhancement investigations.

Unraveling pathogen deceptive disguise: from modules to mimicry

Pathogens rely on their effector proteins to colonize host plants. These effectors have diverse functions. A recent study by Li et al. highlights the significance of protein modularity in generating functional diversity among Phytophthora effectors. It underscores the sophisticated tactics that phytopathogens adopt to alter host cellular processes.

Resilience of root and soil bacteria to drought stress depends on host plant's colonization affinity towards arbuscular mycorrhiza fungi

Water deficit is one of the most important climate events that has strong effect on agricultural ecosystem functionality comprising soil microbial communities and their functions. Arbuscular mycorrhizal fungi (AMF) are widely known for their roles in combating drought, including facilitation of drought-tolerant bacteria. However, differences in cultivar/variety affinity for mycorrhization have never been considered as influencing factors. In the present study, we evaluated the influence of mycorrhizal affinity of two durum wheat (T. turgidum subsp. Durum (Desf.)) varieties, Iride and Ramirez (high and low, respectively) on root and soil bacteriomes under well-watered and drought conditions. We used the 16S metagenomics approach (amplicon sequencing) to assess the bacterial communities of root and soil samples. The suppression effect of drought was evident across a wide range of bacterial taxa, including drought-tolerant taxa, especially in the non-inoculated plants. Nevertheless, the protective effect of AMF was also shown, especially in the Iride variety (high AMF colonization affinity) in both compartments (root and soil), as the relative abundance of drought-depleted taxa, such as Planctomycetes, Bacteroidetes and Verrucomicrobia, was either similar under well-watered and water deficit conditions or increased under water deficit conditions. Moreover, drought reduced the network complexity of root and soil bacteria, especially in Ramirez variety which has a lower AMF colonization affinity. Together, our results suggest that not only AMF colonization, but also host plant colonization affinity is one of the regulating factors in alleviating drought-induced changes in wheat plants by altering plant-fungal-bacterial interactions.

VmPacC-mediated pH regulation of Valsa mali confers to host acidification identified by comparative proteomics analysis

Apple valsa canker caused by the Ascomycete fungus Valsa mali is one of the most serious diseases of apple, resulting in huge economic losses in the apple-growing area of China. Previous study found that the pathogen could acidify the infected tissues to make lower ambient pH (from 6.0 to 3.5) for their successfully colonization. The pH signaling transcription factor VmPacC is required for acidification of its environment and for full virulence in V. mali. It is known that the functional cooperation of proteins secreted by V. mali plays pivotal role in its successful colonization of host plants. In this study, we used tandem mass tag (TMT) labeling coupled with LC-MS/MS-based quantitative proteomics to analyze the VmPacC-mediated pH regulation in V. mali, focusing on differentially expressed proteins (DEPs). We identified 222 DEPs specific to VmPacC deletion, and 921 DEPs specific to different pH conditions (pH 6.0 and 3.4). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses indicated that these DEPs were mainly involved in pathways associated with carbon metabolism, biosynthesis of antibiotics, citrate cycle (TCA cycle), glycolysis/gluconeogenesis, glutathione metabolism, ribosomes, and pentose phosphate pathways. Additionally, we identified 119 DEPs that were shared among the VmPacC deletion mutant and different pH conditions, which were mainly related to energy metabolism pathways, providing the energy required for the hyphal growth and responses to environmental stresses. A protein-protein interaction (PPI) network analysis indicated that most of the shared proteins were mapped to an interaction network with a medium confidence score of 0.4. Notably, one uncharacterized protein (KUI69106.1), and two known proteins (heat shock protein 60 (KUI73579.1), aspartate aminotransferase (KUI73864.1)) located in the core of the network were highly connected (with ≥ 38 directed edges) with the other shared DEPs. Our results suggest that VmPacC participates in the pathogen’s regulation to ambient pH through the regulation of energy metabolism pathways such as the glycolysis/gluconeogenesis pathway and TCA cycle. Finally, we proposed a sophisticated molecular regulatory network to explain pH decrease in V. mali. Our study, by providing insights into V. mali regulating pH, helps to elucidate the mechanisms of host acidification during pathogen infection.

The Novel Effector Ue943 Is Essential for Host Plant Colonization by Ustilago esculenta.

The smut fungus Ustilago esculenta obligately parasitizes Zizania latifolia and induces smut galls at the stem tips of host plants. Previous research identified a putative secreted protein, Ue943, which is required for the biotrophic phase of U. esculenta but not for the saprophytic phase. Here, we studied the role of Ue943 during the infection process. Conserved homologs of Ue943 were found in smut fungi. Ue943 can be secreted by U. esculenta and localized to the biotrophic interface between fungi and plants. It is required at the early stage of colonization. The Ue943 deletion mutant caused reactive oxygen species (ROS) production and callose deposition in the host plant at 1 and 5 days post inoculation, which led to failed colonization. The virulence deficiency was restored by overexpressing gene Ue943 or Ue943:GFP. Transcriptome analysis further showed a series of changes in plant hormones following ROS production when the host plant was exposed to ΔUe943. We hypothesize that Ue943 might be responsible for ROS suppression or avoidance of recognition by the plant immune system. The mechanism underlying Ue943 requires further study to provide more insights into the virulence of smut fungi.

Quorum sensing activities and genomic insights of plant growth-promoting rhizobacteria isolated from Assam tea.

Secretion of quorum sensing (QS) molecules is important for the effective colonization of host plants by plant growth-promoting rhizobacteria. The current study aims at the isolation and characterization of tea rhizo bacteria, which produce the QS molecules, acyl homoserine lactone (AHLs), along with multiple plant growth-promoting (PGP) activities. Thirty-one isolates were isolated from the tea rhizosphere, and screening for PGP activities resulted in the selection of isolates RTE1 and RTE4 with multiple PGP traits, inhibiting the growth of tea fungal pathogens. Both isolates also showed production of AHL molecules when screened using two biosensor strains, Chromobacterium violaceum CV026 and Escherichia coli MT 102(jb132). The isolates identified as Burkholderia cepacia RTE1 and Pseudomonas aeruginosa RTE4 based on genome-based analysis like phylogeny, dDDH, and fastANI calculation. Detailed characterization of AHLs produced by the isolates using reverse-phase TLC, fluorometry, and LC-MS indicated that the isolate RTE1 produced a short chain, C8, and a long chain C12 AHL, while RTE4 produced short-chain AHLs C4 and C6. Confocal microscopy revealed the formation of thick biofilm by RTE1 and RTE4 (18 and 23μm, respectively). Additionally, we found several genes involved in QS, and PGP, inducing systemic resistance (ISR) activities such as lasI/R, qscR, pqq, pvd, aldH, acdS, phz, Sod, rml, and Pch, and biosynthetic gene clusters like N-acyl homoserine lactone synthase, terpenes, pyochelin, and pyocyanin. Based on the functional traits like PGP, biofilm formation and production of AHL molecules, and genetic potential of the isolates B. cepacia RTE1 and P. aeruginosa RTE4 appear promising candidates to improve the health and growth of tea plantations.

Profiling the Pyrenophora tritici-repentis secretome: The Pf2 transcription factor regulates the secretion of the effector proteins ToxA and ToxB.

The global wheat disease tan spot is caused by the necrotrophic fungal pathogen Pyrenophora tritici-repentis (Ptr) which secretes necrotrophic effectors to facilitate host plant colonization. We previously reported a role of the Zn2 Cys6 binuclear cluster transcription factor Pf2 in the regulation of the Ptr effector ToxA. Here, we show that Pf2 is also a positive regulator of ToxB, via targeted deletion of PtrPf2 which resulted in reduced ToxB expression and defects in conidiation and pathogenicity. To further investigate the function of Ptr Pf2 in regulating protein secretion, the secretome profiles of two Δptrpf2 mutants of two Ptr races (races 1 and 5) were evaluated using a SWATH-mass spectrometry (MS) quantitative approach. Analysis of the secretomes of the Δptrpf2 mutants from in vitro culture filtrate identified more than 500 secreted proteins, with 25% unique to each race. Of the identified proteins, less than 6% were significantly differentially regulated by Ptr Pf2. Among the downregulated proteins were ToxA and ToxB, specific to race 1 and race 5 respectively, demonstrating the role of Ptr Pf2 as a positive regulator of both effectors. Significant motif sequences identified in both ToxA and ToxB putative promoter regions were further explored via GFP reporter assays.

Essential Oil Trunk Injection Into Orchard Trees: Consequences on the Performance and Preference of Hemipteran Pests.

Apples and pears are among the most widely cultivated fruit species in the world. Pesticides are commonly applied using ground sprayers in conventional orchards; however, most of it will not reach the target plant, increasing the contamination of nontarget organisms such as natural predators, pollinators, and decomposers. Trunk injection is an alternative method of pesticide application that could reduce risks to beneficials and workers. Essential oils represent a 'green' alternative to pesticides due to their reported insecticidal, antimicrobial, antiviral, nematicidal, and antifungal properties. The aim of this study was, therefore, to evaluate the impact that the injection of a cinnamon essential oil solution into the trunk of apple and pear trees could have on their respective pests, Dysaphis plantaginea and Cacopsylla pyri, respectively. The feeding behavior (preference), the life history traits (performance), and the timing of this effect were measured. The injection of an essential oil emulsion in trees impacted hemipteran host-plant colonization, as for both species a modification of their preference and of their performance was observed. The feeding behavior of D. plantaginea was altered as a significantly lower proportion of aphids ingested phloem sap on injected trees, suggesting that the aphids starved to death. On the contrary, the feeding behavior of the psyllids was little changed compared to the control condition, implying that the observed mortality was due to intoxication. The results presented here could theoretically be used to control these two orchard hemipteran pests, although the effectiveness in real conditions still has to be demonstrated.

Identification, molecular characterization, and plant growth promoting activities of endophytic fungi of Jasminum sambac, Camellia sinensis, and Ocimum basilicum

ObjectivesEndophytic fungi are widely known to reside in plant tissues causing no harm to host plants or even no noticeable change. They may colonize host plants for a part of their life cycle or may complete the entire life cycle since host provide a variety of novel metabolites. Despite being in a close relationship with medicinal host plants, the diversity and metabolic spectrum of endophytic fungi have not been researched well. Their diversity and ecological niche as plant symbionts make them attractive targets in the search for novel biochemicals. MethodsWe aimed to isolate, purify, and characterize the endophytic fungal population of medicinal host plants cultivated in Riyadh, Saudi Arabia. The goal was achieved through morphological identification and internal transcribed spacer (ITS) regions in rRNA analyses to determine phylogeny and diversity. Three aromatic and medicinal plants Jasminum sambac, Camellia sinensis, and Ocimum basilicum collected from the Riyadh region, Saudi Arabia were investigated. In total, 84 isolates were purified and they were grouped into 20 operational taxonomic units (OTUs) as per their sequences of ITS regions in rRNA. ResultsTwenty species of endophytic fungi were grouped in 12 genera i.e. Neopestalotiopsis, Trichoderma, Fusarium, Colletotrichum, Myrothecium, Chaetomium, Alternaria, Phoma, Curvularia, Cladosporium, Neodidymelliopsis, and Aspergillus and all isolates belonged to Ascomycota phylum. J. sambac was found dominant among other and had a relative frequency of 27%. C. sinensis was next with 18.7% relative frequency. The diversity was prominently recorded in leaf organs over stem and roots while roots exhibited the lowest diversity. Isolates also produced indole-3-acetic acid (IAA), 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase enzyme, and siderophores with variable magnitudes that could be assigned to their wide range of metabolic variations from species to species. ConclusionConclusively, J. sambac, C. sinensis, and O. basilicum are a good source of endophytic fungi with certain plant growth-promoting traits. Overall, Alternaria was found as the most predominant genus in terms of colonization rate. Further deternminations are required to screen the beneficial compounds released by these endophytic fungi.

Differences in Oxidative Stress Markers and Antioxidant Enzyme Activities in Black Bean Aphid Morphs (Aphis fabae Scop.) Fed on the Primary Host Viburnum opulus L.

Changes in the level of oxidative stress markers-superoxide anion radical (O2-), hydrogen peroxide (H2O2) and malondialdehyde (MDA) and the activity of antioxidant enzymes-superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) in the black bean aphid occurring on the primary host (viburnum plants) were studied. Among the aphid morphs, the lowest contents of O2-, H2O2 and MDA were noted for winged adults (alatae), which were also characterized by the highest activity of antioxidant enzymes. These metabolic features indicate the adaptation of winged morphs to the colonization of new host plants. During spring migration, an increase in the content of oxidative stress markers and antioxidant enzyme activities in wingless females (fundatrigeniae) was observed. The significance of the biochemical adaptation of the black bean aphid to its winter host is discussed.

Comprehensive genome-wide identification and expression profiling of ADF gene family in Citrus sinensis, induced by endophytic colonization of Beauveria bassiana

Endophytic entomopathogenic species are known to systematically colonize host plants and form symbiotic associations that benefit the plants they live with. The actin-depolymerizing factors (ADFs) are a group of gene family that regulate growth, development, and defense-related functions in plants. Systematic studies of ADF family at the genome-wide level and their expression in response to endophytic colonization are essential to understand its functions but are currently lacking in this field. 14ADF genes were identified and characterized in the Citrus sinensis genome. The ADF genes of C. sinensis were classified into five groups according to the phylogenetic analysis of plant ADFs. Additionally, the cis-acting analysis revealed that these genes play essential role in plant growth/development, phytohormone, and biotic and abiotic responses; and the expression analysis showed that the symbiotic interactions generate a significant expression regulation level of ADF genes in leaves, stems and roots, compared to controls; thus enhancing seedlings' growth. Additionally, the 3D structures of the ADF domain were highly conserved during evolution. These results will be helpful for further functional validation of ADFs candidate genes and provide important insights into the vegetative growth, development and stress tolerance of C. sinensis in responses to endophytic colonization by B. bassiana.

Host plant phenology shapes aphid abundance and interactions with ants

Phenological mismatch can occur when plants and herbivores differentially respond to changing phenological cues, such as temperature or snow melt date. This often shifts herbivore feeding to plant stages of lower quality. How herbivores respond to plant quality may be also mediated by temperature, which could lead to temperature‐by‐phenology interactions. We examined how aphid abundance and mutualism with ants were impacted by temperature and host plant phenology. In this study system, aphids Aphis asclepiadis colonize flowering stalks of the host plant, Ligusticum porteri. Like other aphids, abundance of this species is dependent on ant protection. To understand how host plant phenology and temperature affect aphid abundance, we used a multiyear observational study and a field experiment. We observed 20 host plant populations over five years (2017–2021), tracking temperature and snow melt date as well as host plant phenology and insect abundance. We found host plant and aphid phenology to differentially respond to temperature and snow melt timing. Early snow melt accelerated host plant phenology to a greater extent than aphid phenology, which was more responsive to temperature. Both the likelihood of aphid colony establishment and ant recruitment were reduced when aphids colonized host plants at post‐flowering stages. In 2019, we experimentally accelerated host plant phenology by advancing snow melt date by two weeks. We factorially combined this treatment with open top warming chambers surrounding aphid colonies. Greatest growth occurred for colonies under ambient temperatures when they occurred on host plants at the flowering stage. Altogether, our results suggest that phenological mismatch with host plants can decrease aphid abundance, and this effect is exacerbated by temperature increases and changes to the ant–aphid mutualism.