Colonize Plant Tissues Research Articles

Trichoderma posible micoparásito de Sporisorium reilianum y su influencia en el rendimiento de maíz

<div class="page" title="Page 1"><div class="layoutArea"><div class="column"><p><strong>Objetivo - </strong><span>evaluar el efecto de cepas de </span><em>Trichoderma </em><span>en el control de </span><em>Sporisorium reilianum </em><span>y aumentar el rendimiento en un cultivo de maíz en el Valle del Mezquital, Hidalgo.</span></p><p><strong>Método - </strong><span>se desarrollaron experimentos </span><span><em>in vitro</em> e <em>in vivo</em></span><span>, las variables evaluadas fueron colonización por </span><em>Trichoderma </em><span>en cultivos duales, Incidencia, colonización de tejidos vegetales, altura y rendimiento. Los resultados serán relevantes para las regiones templadas donde se establezca maíz. </span></p><p><span><strong>Resultados - </strong></span><span>se demostró </span><em>in vitro </em><span>que el patógeno fue controlado principalmente por las cepas C5THBr y C3VM14. En incidencia en el cultivo, el tratamiento con el cual se obtuvo el mejor resultado fue con la cepa C4TVBr-no nativa (8.31%). Los tejidos fueron colonizados por </span><em>Trichoderma </em><span>desde un 19.99% (hojas) hasta un 53.3% (raíces). A excepción de C3VM14 todos los tratamientos tuvieron efecto positivo en altura de plantas. Con la cepa C5THBr-no nativa hubo un aumento de rendimiento de un 38.8%. </span></p><p><strong>Limitaciones - </strong><span>se requieren estudios detallados sobre los mecanismos de acción de las cepas en estudio. </span></p><p><strong>Principales hallazgos - </strong><span>las cepas de </span><span><em>Trichoderma</em> </span><span>tanto nativas como no nativas pueden tener un efecto en el control de </span><em>Sporisorium reilianum </em><span>así como en el aumento del rendimiento del cultivo del maíz. </span></p></div></div></div>

MRNA extraction of Xanthomonas fragariae in strawberry and validation of reference genes for the RT-qPCR for study of bacterial gene expression

This is the first study which describes a unique procedure of isolating of high-quality, intact RNA from strawberry leaves of Xanthomonas fragariae, three most suitable reference genes, crucial for the normalization of RT-qPCR data for this pathogen and accurate expression analysis of target genes. In our study, various mathematic algorithms: NormFinder geNorm, BestKeeper, the delta CT method, RefFinder were adopted for validation of most stable reference genes from nine candidate genes (ffh, glyA, gyrA, gyrB, proC, pykA, recA, rpoB, rpoD). The analyses allowing to select three most suitable pioneer reference genes, gyrB, ffh, and pykA, that we recommend for the normalization of RT-qPCR data and for the study of the expression of target genes in Xf. Moreover, their combination as references allowed for an accurate expression analysis and computation of the fold change of the flhF and iroN2 genes in Xf. These two genes are important for the success of the colonization of plant tissue and pathogenicity and sequences of primers designed to study these genes, are presented.

The critical role of biofilms in bacterial vascular plant pathogenesis

Bacterial life is a combination of two lifestyles, mobile and social. In the social lifestyle, cells are usually embedded in a self‐produced matrix and attached to biotic or abiotic surfaces. These communities can be organized as either single or multilayered structures termed biofilms. Biofilms evolved to cope with the harsh environmental conditions that bacteria encounter within the host, mostly from the host’s defence response. In plant pathogenic bacteria, biofilms participate in the whole process of pathogenicity, from the first step of invasion to the full colonization of plant tissues. The specific role that biofilms play in the pathogenicity process of plant bacterial pathogens is poorly understood. In this review, the role of biofilms in the pathogenic process of major vascular plant pathogens is examined. In addition, quorum sensing signals and components that are essential for biofilm formation and therefore, for pathogenesis, are addressed. Although, in certain systems, further research is required, experimental evidence in the literature indicates that biofilms are, in most cases, essential for pathogenesis.

English

Research institutions have emphasized alternative disease control agents because they represent a great option with beneficial effects on human and animal health and ecosystem balance. Despite the studies already done on botanical fungicides, their effective use requires knowledge about the applicability of natural products to different production systems. The main objective of this study is to evaluate the efficacy of essential oils of Ocimum pupuraceus L., Cymbopogon nardus (L.) Rendle, Cymbopogon citratus (DC.) Stapf, and Lippia sidoides Cham. on the inhibition of mycelial growth and germination of conidia of B. maydis. The other objectives of this study were to perform gas chromatographic and phytotoxicity analyses, and test the control of bipolaris leaf spot using essential oils of L. sidoides applied as a preventive and curative agent. Among the treatments studied, the essential oil of L. sidoides is effective in inhibiting mycelial growth and conidial germination at the concentrations of 5 and 1%, respectively. The main constituent of the oil is thymol (92.68%). The concentration range 0.75-3% of L. sidoides essential oil is phytotoxic to maize plants. Lower values of the area under the progress curve of bipolaris leaf spot are observed at concentrations 0.1-0.5%, when the essential oil was applied as a preventive agent prior to the colonization of plant tissues by the pathogen. The application of the oil as a curative to plants with the disease also shows efficacy at the concentration 0.1%, reducing the severity by more than 54%. These results demonstrate the potential effects of L. sidoides essential oil on preventive and curative control of bipolaris leaf spot in maize and mycelial growth. Key words: Medicinal plants, alternative control, phytopathogens, Zea mays, Bipolaris maydis, Lippia sidoides.

Improved Methods for Treatment of Phytopathogenic Biofilms: Metallic Compounds as Anti-Bacterial Coatings and Fungicide Tank-Mix Partners.

Fungi and bacteria cause disease issues in cultivated plants world-wide. In most cases, the fungi and bacteria colonize plant tissues as biofilms, which can be very challenging to destroy or eradicate. In this experiment, we employed a novel (biofilm) approach to crop disease management by evaluating the efficacies of six fungicides, and four silver-based compounds, versus biofilms formed by fungi and bacteria, respectively. The aim was to identify combinations of fungicides and metallic cations that showed potential to improve the control of white mold (WM), caused by the ascomycete fungus Sclerotinia sclerotiorum, and to evaluate novel high valency silver compounds as seed coatings to prevent biofilm formation of four bacterial blight pathogens on dry bean seeds. Our results confirmed that mature fungal biofilms were recalcitrant to inactivation by fungicides. When metallic cations were added to the fungicides, their efficacies were improved. Some improvements were statistically significant, with one combination (fluazinam + Cu2+) showing a synergistic effect. Additionally, coatings with silver compounds could reduce bacterial blight biofilms on dry bean seeds and oxysilver nitrate was the most potent inhibitor of bacterial blight.

Non rhizobial endophytic bacteria from chickpea (Cicer arietinum L.) tissues and their antagonistic traits

Bacteria that colonize plant tissues other than rhizobia and are beneficial for plant growth referred to non rhizobial plant growth-promoting endophytic bacteria (PGPEB). This study was designed to assay the biocontrol activity of plant growth promoting endophytic bacterial isolates those found positive for P. solubilization, ACC deaminase, Indole acetic acid and Gibberelic acid production. These bacterial isolates were obtained from chickpea (Cicer arietinum L.) tissues (roots and nodules). In a previous study a total of 263 non rhizobial endophytic bacterial isolates were isolated. Out of 263 isolates, 64.5% and 34.5% were Gram positive and negative, respectively. Further for biochemical characterization, catalase, oxidase, citrate utilization, nitrate reduction, methyl red and Voges Proskauer’s tests, were performed. On the basis of P solubilization, ACC deaminase, Indole acetic acid and Gibberelic acid production 75 potential isolates were selected and screened for their biocontrol activity viz. (production of cell wall degrading enzymes, production of HCN and fluorescent pigment). Out of 75 isolates, only 29 isolates produced cellulase, 64 isolates were able to produce protease and 28 were positive for both cellulose and protease. Of 75 endophytic isolates 12 isolates (7 from root tissue and 5 from nodules tissue, respectively) were positive for HCN production and 16 isolates were found to be fluorescent pigment producer under µv ligh. As chemical fertilizers and pesticides have detrimental effects on the environment. So these bacterial endophytic isolates will be used not only as a biofertilizer because of their plant growth promotional activities but also used as an alternative of synthetic chemicals for control of several plant diseases.

Identification of Endophytic Fungi from Fruits and Seeds of Jambolana (Syzygium cumini L.) Skeels

The endophytes are microorganisms that colonize plant tissue without causing infections. One of endophytic microorganisms, fungi, comprises a reliable source of genetic diversity and has been applied in pharmacology. In this study, the endophytic fungi was isolated from fruit and seed of Syzygium cumini and identified based on ITS-rDNA. Five fungal isolates were obtained from fruit and seed of Jambolana, two isolates from fruit and three isolates from seed. Based on ITS-rDNA analysis showed that that endophytic fungus isolates from jambolana (S. cumini L.) Skeels was identified as Neofusicoccum parvum (F1 isolates), Pestalotiopsis vismiae (F2 isolates), Phomopsis sp. (S1 and S3 isolates), Colletotrichum fructicola (S2 isolates). All the isolates are belonged to Ascomycota.

Broad-specificity GH131 β-glucanases are a hallmark of fungi and oomycetes that colonize plants.

Plant-tissue-colonizing fungi fine-tune the deconstruction of plant-cell walls (PCW) using different sets of enzymes according to their lifestyle. However, some of these enzymes are conserved among fungi with dissimilar lifestyles. We identified genes from Glycoside Hydrolase family GH131 as commonly expressed during plant-tissue colonization by saprobic, pathogenic and symbiotic fungi. By searching all the publicly available genomes, we found that GH131-coding genes were widely distributed in the Dikarya subkingdom, except in Taphrinomycotina and Saccharomycotina, and in phytopathogenic Oomycetes, but neither other eukaryotes nor prokaryotes. The presence of GH131 in a species was correlated with its association with plants as symbiont, pathogen or saprobe. We propose that GH131-family expansions and horizontal-gene transfers contributed to this adaptation. We analysed the biochemical activities of GH131 enzymes whose genes were upregulated during plant-tissue colonization in a saprobe (Pycnoporus sanguineus), a plant symbiont (Laccaria bicolor) and three hemibiotrophic-plant pathogens (Colletotrichum higginsianum, C. graminicola, Zymoseptoria tritici). These enzymes were all active on substrates with β-1,4, β-1,3 and mixed β-1,4/1,3 glucosidic linkages. Combined with a cellobiohydrolase, GH131 enzymes enhanced cellulose degradation. We propose that secreted GH131 enzymes unlock the PCW barrier and allow further deconstruction by other enzymes during plant tissue colonization by symbionts, pathogens and saprobes.

Arabidopsis Histone Deacetylase 5 is Required for Pseudomonas‐Induced Deacetylation of Histone H3 at Lysine 9

The gram‐negative plant pathogen Pseudomonas syringae pv. tomato DC3000 (Pto DC3000) depends on a type III secretion system (T3SS) to colonize plant tissue, and cause disease. The T3SS facilitates injection of virulence proteins called effector proteins (T3Es) directly into host cells. The T3Es have been shown to target components of plant immunity and are essential for Pto to cause disease. We have been interested in determining the extent that Pto T3Es are involved in modulating host chromatin to favor pathogenesis. Through Immunoblot and Chromatin Immunoprecipitation analyses we found a rapid deacetylation of host histone H3 lysine 9 (H3K9) along innate immunity related genes in response to Pto. This deacetylation was dependent on both T3Es and the host histone deacetylase HDA5. Moreover, Arabidopsis plants lacking HDA5 had an enhanced resistance to Pto infections and produced more callose in response to Pathogen Associated Molecular Pattern exposure. Moreover, through analyses of specific T3E Pto mutants and transient and transgenic expression of specific T3Es, we found that only a small subset of the T3Es are needed for the HDA5‐triggered deacetylation. Collectively our data show a Pto T3E‐induced deacetylation by HDA5 along immunity related genes in Arabidopsis.Support or Funding InformationThis work was supported by NSF award #1557417, NSF award # 0940177 and NIGMS award # 8P20GM103427This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Fungal adaptation to plant defences through convergent assembly of metabolic modules.

The ongoing diversification of plant defence compounds exerts dynamic selection pressures on the microorganisms that colonize plant tissues. Evolutionary processes that generate resistance towards these compounds increase microbial fitness by giving access to plant resources and increasing pathogen virulence. These processes entail sequence-based mechanisms that result in adaptive gene functions, and combinatorial mechanisms that result in novel syntheses of existing gene functions. However, the priority and interactions among these processes in adaptive resistance remain poorly understood. Using a combination of molecular genetic and computational approaches, we investigated the contributions of sequence-based and combinatorial processes to the evolution of fungal metabolic gene clusters encoding stilbene cleavage oxygenases (SCOs), which catalyse the degradation of biphenolic plant defence compounds known as stilbenes into monophenolic molecules. We present phylogenetic evidence of convergent assembly among three distinct types of SCO gene clusters containing alternate combinations of phenolic catabolism. Multiple evolutionary transitions between different cluster types suggest recurrent selection for distinct gene assemblages. By comparison, we found that the substrate specificities of heterologously expressed SCO enzymes encoded in different clusters types were all limited to stilbenes and related molecules with a 4'-OH group, and differed modestly in substrate range and activity under the experimental conditions. Together, this work suggests a primary role for genome structural rearrangement, and the importance of enzyme modularity, in promoting fungal metabolic adaptation to plant defence chemistry.

EXPERIMENTAL EVIDENCES ON A CROP PLANT ROLE IN EPIDEMIOLOGY OF SAPRONOTIC (SOIL-BORNE) BACTERIAL INFECTIONS

Specific epidemiology of sapronotic (soil-borne) bacteria is characterized from the ecological point of view. The characteristic feature of soil-borne pathogens is an ability to exist autonomously in the environment. This analytical review is focused on crops as alternative hosts for a number of soil-borne pathogenic bacteria (Yersinia, Salmonella, Listeria, Escherichia etc). Published experimental results evidence capabilities of human and animal pathogens to colonize plant tissues. Novel approaches are discussed to minimize risks of infection spreading with crops. These approaches include an analysis of wild plant natural resistance to pathogenic bacteria and a construction of transgenic plant crops expressing antimicrobial peptides. Multiple studies are cited that established wild plants used in traditional medicine as a source for obtaining molecules effective against resistant pathogens. The review includes recent author results on activity of wild plant extracts against Listeria and toxin-producing Escherichia.

Mucor sp.-An endophyte of Brassicaceae capable of surviving in toxic metal-rich sites.

The aim of this study was to assess the biodiversity of endophytic fungi from Arabidopsis arenosa growing on a post mining waste dump and to evaluate their role in plant adaptation to metal toxicity. Severeal of the fungi were beneficial for the plant. Among them, a fungus belonging to the Mucor genus, was found to interact with a broad range of plants, including Brassicaceae metallophytes. Mucor sp. was shown to be highly tolerant to elevated levels of Zn, Cd, and Pb and to accelerate plant-host growth under either toxic-metal stress or control conditions. When inoculated with Mucor sp., A. arenosa under toxic-metal stress acquired more N and showed significantly down-regulated catalase activity, which suggests suppression of toxic-metal-induced oxidative stress. We used the model plant-A. thaliana to evaluate the dynamics of plant-tissue colonization by the fungus as monitored with qPCR and to analyze the host's transcriptome response during early stages of the interaction. The results revealed the induction of a plant-defense and stress-related response on the 5th day of co-culture, which was in accord with the decrease of fungal abundance in shoots on the 6th day of interaction. Presented results demonstrate the importance of endophytic fungi in plant toxic-metal tolerance.

Fungicide tolerant Bradyrhizobium japonicum mitigate toxicity and enhance greengram production under hexaconazole stress

Bacterial strain RV9 recovered from greengram nodules tolerated 2400μg/mL of hexaconazole and was identified by 16S rDNA sequence analysis as Bradyrhizobium japonicum (KY940048). Strain RV9 produced IAA (61.6μg/mL), ACC deaminase (51.7mg/(protein·hr)), solubilized TCP (105μg/mL), secreted 337.6μg/mL EPS, and produced SA (52.2μg/mL) and 2,3-DHBA (28.3μg/mL). Exopolysaccharides produced by strain RV9 was quantified and characterized by SEM, AFM, EDX and FTIR. Beyond tolerance limit, hexaconazole caused cellular impairment and reduced the viability of strain RV9 revealed by SEM and CLSM. Hexaconazole distorted the root tips and altered nodule structure leading thereby to reduction in the performance of greengram. Also, the level of antioxidant enzymes, proline, TBARS, ROS and cell death was increased in hexaconazole treated plants. CLSM images revealed a concentration dependent increase in the characteristic green and blue fluorescence of hexaconazole treated roots. The application of B. japonicum strain RV9 alleviated the fungicide toxicity and improved the measured plant characteristics. Also, rhizobial cells were localized inside tissues as revealed by CLSM. Colonization of B. japonicum strain RV9 decreased the levels of CAT, POD, APX, GPX and TBARS by 80%, 5%, 13%, 13% and 19%, respectively over plants grown at 80μg/(hexaconazole·kg) soil. The ability to detoxify hexaconazole, colonize plant tissues, secrete PGP bioactive molecules even under fungicide pressure and its unique ability to diminish oxidative stress make B. japonicum an attractive choice for remediation of fungicide polluted soils and to concurrently enhance greengram production under stressed environment.

Antagonism and antibiotic resistance drive a species-specific plant microbiota differentiation in Echinacea spp

A key factor in the study of plant-microbes interactions is the composition of plant microbiota, but little is known about the factors determining its functional and taxonomic organization. Here we investigated the possible forces driving the assemblage of bacterial endophytic and rhizospheric communities, isolated from two congeneric medicinal plants, Echinacea purpurea (L.) Moench and Echinacea angustifolia (DC) Heller, grown in the same soil, by analysing bacterial strains (isolated from three different compartments, i.e. rhizospheric soil, roots and stem/leaves) for phenotypic features such as antibiotic resistance, extracellular enzymatic activity, siderophore and indole 3-acetic acid production, as well as cross-antagonistic activities. Data obtained highlighted that bacteria from different plant compartments were characterized by specific antibiotic resistance phenotypes and antibiotic production, suggesting that the bacterial communities themselves could be responsible for structuring their own communities by the production of antimicrobial molecules selecting bacterial-adaptive phenotypes for plant tissue colonization.

Phytophthora palmivora establishes tissue-specific intracellular infection structures in the earliest divergent land plant lineage

The expansion of plants onto land was a formative event that brought forth profound changes to the earth's geochemistry and biota. Filamentous eukaryotic microbes developed the ability to colonize plant tissues early during the evolution of land plants, as demonstrated by intimate, symbiosis-like associations in >400 million-year-old fossils. However, the degree to which filamentous microbes establish pathogenic interactions with early divergent land plants is unclear. Here, we demonstrate that the broad host-range oomycete pathogen Phytophthora palmivora colonizes liverworts, the earliest divergent land plant lineage. We show that P. palmivora establishes a complex tissue-specific interaction with Marchantia polymorpha, where it completes a full infection cycle within air chambers of the dorsal photosynthetic layer. Remarkably, P. palmivora invaginates M. polymorpha cells with haustoria-like structures that accumulate host cellular trafficking machinery and the membrane syntaxin MpSYP13B, but not the related MpSYP13A. Our results indicate that the intracellular accommodation of filamentous microbes is an ancient plant trait that is successfully exploited by pathogens like P. palmivora.

2-Tridecanone impacts surface-associated bacterial behaviours and hinders plant-bacteria interactions.

Surface motility and biofilm formation are behaviours which enable bacteria to infect their hosts and are controlled by different chemical signals. In the plant symbiotic alpha-proteobacterium Sinorhizobium meliloti, the lack of long-chain fatty acyl-coenzyme A synthetase activity (FadD) leads to increased surface motility, defects in biofilm development and impaired root colonization. In this study, analyses of lipid extracts and volatiles revealed that a fadD mutant accumulates 2-tridecanone (2-TDC), a methylketone (MK) known as a natural insecticide. Application of pure 2-TDC to the wild-type strain phenocopies the free-living and symbiotic behaviours of the fadD mutant. Structural features of the MK determine its ability to promote S. meliloti surface translocation, which is mainly mediated by a flagella-independent motility. Transcriptomic analyses showed that 2-TDC induces differential expression of iron uptake, redox and stress-related genes. Interestingly, this MK also influences surface motility and impairs biofilm formation in plant and animal pathogenic bacteria. Moreover, 2-TDC not only hampers alfalfa nodulation but also the development of tomato bacterial speck disease. This work assigns a new role to 2-TDC as an infochemical that affects important bacterial traits and hampers plant-bacteria interactions by interfering with microbial colonization of plant tissues.

Diversity and symbiotic divergence of endophytic and non-endophytic rhizobia of Medicago sativa

Knowledge of rhizobium diversity is helping to enable the utilization of rhizobial resources. To analyze the phenotypic and genetic diversity and the symbiotic divergence of rhizobia of Medicago sativa, 30 endophytic and non-endophytic isolates were collected from different parts of five alfalfa varieties in three geographic locations in Gansu, China. Numerical analyses based on 72 phenotypic properties and restriction fragment length polymorphism (RFLP) fingerprinting indicated the abundant phenotypic and genetic diversity of the tested strains. According to the phylogenetic analysis of 16S RNA, atpD, glnII, and recA gene sequences, Rhizobium and Ensifer were further classified into four different genotypes: Rhizobium radiobacter, Rhizobium sp., Rhizobium rosettiformans, and Ensifer meliloti. The differences in architecture and functioning of the rhizobial genomes and, to a lesser extent, environment diversification helped explain the diversity of tested strains. The tested strains exhibited similar symbiotic feature when inoculated onto M. sativa cvs. Gannong Nos. 3 and 9 and Qingshui plants for the clustering feature of their parameter values. An obvious symbiotic divergence of rhizobial strains was observed in M. sativa cvs. Longzhong and WL168HQ plants because of the scattered parameter values. Their symbiotic divergence differed according to alfalfa varieties, which indicated that the sensitivity of different alfalfa varieties to rhizobial strains may differ. Most of the tested strains exhibited plant growth-promoting traits including phosphate solubilization and production of indole-3-acetic acid (IAA) when colonizing plant tissues and soil.

BigR is a sulfide sensor that regulates a sulfur transferase/dioxygenase required for aerobic respiration of plant bacteria under sulfide stress

To cope with toxic levels of H2S, the plant pathogens Xylella fastidiosa and Agrobacterium tumefaciens employ the bigR operon to oxidize H2S into sulfite. The bigR operon is regulated by the transcriptional repressor BigR and it encodes a bifunctional sulfur transferase (ST) and sulfur dioxygenase (SDO) enzyme, Blh, required for H2S oxidation and bacterial growth under hypoxia. However, how Blh operates to enhance bacterial survival under hypoxia and how BigR is deactivated to derepress operon transcription is unknown. Here, we show that the ST and SDO activities of Blh are in vitro coupled and necessary to oxidize sulfide into sulfite, and that Blh is critical to maintain the oxygen flux during A. tumefaciens respiration when oxygen becomes limited to cells. We also show that H2S and polysulfides inactivate BigR leading to operon transcription. Moreover, we show that sulfite, which is produced by Blh in the ST and SDO reactions, is toxic to Citrus sinensis and that X. fastidiosa-infected plants accumulate sulfite and higher transcript levels of sulfite detoxification enzymes, suggesting that they are under sulfite stress. These results indicate that BigR acts as a sulfide sensor in the H2S oxidation mechanism that allows pathogens to colonize plant tissues where oxygen is a limiting factor.

Escherichia coli O157:H7 Converts Plant-Derived Choline to Glycine Betaine for Osmoprotection during Pre- and Post-harvest Colonization of Injured Lettuce Leaves.

Plant injury is inherent to the production and processing of fruit and vegetables. The opportunistic colonization of damaged plant tissue by human enteric pathogens may contribute to the occurrence of outbreaks of foodborne illness linked to produce. Escherichia coli O157:H7 (EcO157) responds to physicochemical stresses in cut lettuce and lettuce lysates by upregulation of several stress response pathways. We investigated the tolerance of EcO157 to osmotic stress imposed by the leakage of osmolytes from injured lettuce leaf tissue. LC-MS analysis of bacterial osmoprotectants in lettuce leaf lysates and wound washes indicated an abundant natural pool of choline, but sparse quantities of glycine betaine and proline. Glycine betaine was a more effective osmoprotectant than choline in EcO157 under osmotic stress conditions in vitro. An EcO157 mutant with a deletion of the betTIBA genes, which are required for biosynthesis of glycine betaine from imported choline, achieved population sizes twofold lower than those of the parental strain (P < 0.05) over the first hour of colonization of cut lettuce in modified atmosphere packaging (MAP). The cell concentrations of the betTIBA mutant also were 12-fold lower than those of the parental strain (P < 0.01) when grown in hypertonic lettuce lysate, indicating that lettuce leaf cellular contents provide choline for osmoprotection of EcO157. To demonstrate the utilization of available choline by EcO157 for osmoadaptation in injured leaf tissue, deuterated (D-9) choline was introduced to wound sites in MAP lettuce; LC-MS analysis revealed the conversion of D9-choline to D-9 glycine betaine in the parental strain, but no significant amounts were observed in the betTIBA mutant. The EcO157 ΔbetTIBA-ΔotsBA double mutant, which is additionally deficient in de novo synthesis of the compatible solute trehalose, was significantly less fit than the parental strain after their co-inoculation onto injured lettuce leaves and MAP cut lettuce. However, its competitive fitness followed a different time-dependent trend in MAP lettuce, likely due to differences in O2 content, which modulates betTIBA expression. Our study demonstrates that damaged lettuce leaf tissue does not merely supply EcO157 with substrates for proliferation, but also provides the pathogen with choline for its survival to osmotic stress experienced at the site of injury.

Manipulation of Bryophyte Hosts by Pathogenic and Symbiotic Microbes.

The colonization of plant tissues by pathogenic and symbiotic microbes is associated with a strong and directed effort to reprogram host cells in order to permit, promote and sustain microbial growth. In response to colonization, hosts accommodate or sequester invading microbes by activating a set of complex regulatory programs that initiate symbioses or bolster defenses. Extensive research has elucidated a suite of molecular and physiological responses occurring in plant hosts and their microbial partners; however, this information is mostly limited to model systems representing evolutionarily young plant lineages such as angiosperms. The extent to which these processes are conserved across land plants is therefore poorly understood. In this review, we outline key aspects of host reprogramming that occur during plant–microbe interactions in early diverging land plants belonging to the bryophytes (liverworts, hornworts and mosses). We discuss how further knowledge of bryophyte–microbe interactions will advance our understanding of how plants and microbes co-operated and clashed during the conquest of land.