The need for plants to defend themselves, communicate, and somehow contribute to the social life in their ecosystems has triggered the evolution of an astonishing number of diverse chemicals, some of ...which involved in plant–plant interactions. In the present study, specific aspects of allelopathy are investigated. A combination of bioassays and metabolomics was used in order to study the chemical interactions occurring between three donor species of Mediterranean area (
Arbutus unedo
,
Medicago minima
,
Myrtus communis
) and a receiving species (
Aegilops geniculata
). The biochemical changes occurring in the receiving plant upon the treatments with the donor extracts were studied. Oxidative stress and altered water balance were found to be the major changes in the receiving plant. Putative allelochemicals synthesized by the donor plants were also identified and it was shown that their activity was enhanced by co-occurring metabolites. This study provides evidence that metabolite mixtures are to be taken into consideration for allelopathic activity. Furthermore, not only it reports the chemicals responsible for the activity in the specific system, but it also shows that the response of the receiving plant to the treatment with extracts from donor plants is comparable to the response to other stresses.
'Candidatus Phytoplasma ulmi' is the agent associated with elm yellows and has been categorised in the European Union as a quarantine pathogen. For central and northern European countries, ...information on the occurrence and distribution of the pathogen and its impact on elms is scarce, so a survey of native elm trees has been conducted in Germany.
About 6500 samples from Ulmus minor, Ulmus laevis and Ulmus glabra, were collected nationwide. Phytoplasma detection was performed by applying a universal 16Sr DNA-based quantitative PCR (qPCR) assay and a novel 'Ca. P. ulmi' specific qPCR assay targeting the 16S-23S spacer region. Both assays revealed that 28% of the samples were infected by 'Ca. P. ulmi', but infection rates of the elm species and regional incidences differed. The phytoplasma presence in the trees was not correlated to disease-specific symptoms. The survey identified a regional disparity of infection which was high in east, south and central Germany, whereas only a few infected sites were found in the western and northern parts of the country. Monitoring the seasonal titre of 'Ca. P. ulmi' in an infected tree by qPCR revealed a high colonisation in all parts of the tree throughout the year.
'Ca. P. ulmi' is widely present in elms in Germany. The rare occurrence of symptoms indicates either a high degree of tolerance in elm populations or a low virulence of pathogen strains enabling high infection rates in a long-living host.
Selection pressure exerted by insects and microorganisms shapes the diversity of plant secondary metabolites. We identified a metabolic pathway for glucosinolates, known insect deterrents, that ...differs from the pathway activated by chewing insects. This pathway is active in living plant cells, may contribute to glucosinolate turnover, and has been recruited for broad-spectrum antifungal defense responses. The Arabidopsis CYP81F2 gene encodes a P450 monooxygenase that is essential for the pathogen-induced accumulation of 4-methoxyindol-3-ylmethylglucosinolate, which in turn is activated by the atypical PEN2 myrosinase (a type of β-thioglucoside glucohydrolase) for antifungal defense. We propose that reiterated enzymatic cycles, controlling the generation of toxic molecules and their detoxification, enable the recruitment of glucosinolates in defense responses.
Phytoplasmas are insect-transmitted, uncultivable bacterial plant pathogens that cause diseases in hundreds of economically important plants. They represent a monophyletic group within the class ...Mollicutes (trivial name mycoplasmas) and are characterized by a small genome with a low GC content, and the lack of a firm cell wall. All mycoplasmas, including strains of 'Candidatus (Ca.) Phytoplasma asteris' and 'Ca. P. australiense', examined so far have circular chromosomes, as is the case for almost all walled bacteria.
Our work has shown that 'Ca. Phytoplasma mali', the causative agent of apple proliferation disease, has a linear chromosome. Linear chromosomes were also identified in the closely related provisional species 'Ca. P. pyri' and 'Ca. P. prunorum'. The chromosome of 'Ca. P. mali' strain AT is 601,943 bp in size and has a GC content of 21.4%. The chromosome is further characterized by large terminal inverted repeats and covalently closed hairpin ends. Analysis of the protein-coding genes revealed that glycolysis, the major energy-yielding pathway supposed for 'Ca. P. asteris', is incomplete in 'Ca. P. mali'. Due to the apparent lack of other metabolic pathways present in mycoplasmas, it is proposed that maltose and malate are utilized as carbon and energy sources. However, complete ATP-yielding pathways were not identified. 'Ca. P. mali' also differs from 'Ca. P. asteris' by a smaller genome, a lower GC content, a lower number of paralogous genes, fewer insertions of potential mobile DNA elements, and a strongly reduced number of ABC transporters for amino acids. In contrast, 'Ca. P. mali' has an extended set of genes for homologous recombination, excision repair and SOS response than 'Ca. P. asteris'.
The small linear chromosome with large terminal inverted repeats and covalently closed hairpin ends, the extremely low GC content and the limited metabolic capabilities reflect unique features of 'Ca. P. mali', not only within phytoplasmas, but all mycoplasmas. It is expected that the genome information obtained here will contribute to a better understanding of the reduced metabolism of phytoplasmas, their fastidious nutrition requirements that prevented axenic cultivation, and the mechanisms involved in pathogenicity.
Beewolf digger wasps cultivate specific symbiotic bacteria (Streptomyces spp.) that are incorporated into the larval cocoon for protection against pathogens. We identified the molecular basis of this ...protective symbiosis in the natural context and demonstrate that the bacteria produce a 'cocktail' of nine antibiotic substances. The complementary action of all symbiont-produced antibiotics confers a potent antimicrobial defense for the wasp larvae that parallels the 'combination prophylaxis' known from human medicine.
Norway spruce (Picea abies) forests suffer periodic fatal attacks by the bark beetle Ips typographies and its fungal associate, Ceratocystis polonica. Norway spruce protects itself against fungal and ...bark beetle invasion by the production of terpenoid resins, but it is unclear whether resins or other defenses are effective against the fungus. We investigated stilbenes, a group of phenolic compounds found in Norway spruce bark with a diaryl-ethene skeleton with known antifungal properties. During C. polonica infection, stilbene biosynthesis was up-regulated, as evidenced by elevated transcript levels of stilbene synthase genes. However, stilbene concentrations actually declined during infection, and this was due to fungal metabolism. C. polonica converted stilbenes to ring-opened, deglycosylated, and dimeric products. Chromatographie separation of C. polonica protein extracts confirmed that these metabolites arose from specific fungal enzyme activities. Comparison of C. polonica strains showed that rapid conversion of host phenolics is associated with higher virulence. C. polonica is so well adapted to its host's chemical defenses that it is even able to use host phenolic compounds as its sole carbon source.
To assess the influence of dissolved organic matter (DOM) on the acid–base system of the Baltic Sea, 19 stations along the salinity gradient from Mecklenburg Bight to the Bothnian Bay were sampled in ...November 2011 for total alkalinity (AT), total inorganic carbon concentration (CT), partial pressure of CO2 (pCO2), and pH. Based on these data, an organic alkalinity contribution (Aorg) was determined, defined as the difference between measured AT and the inorganic alkalinity calculated from CT and pH and/or CT and pCO2. Aorg was in the range of 22–58μmolkg−1, corresponding to 1.5–3.5% of AT. The method to determine Aorg was validated in an experiment performed on DOM-enriched river water samples collected from the mouths of the Vistula and Oder Rivers in May 2012. The Aorg increase determined in that experiment correlated directly with the increased DOC concentration caused by enrichment of the >1kDa DOM fraction. To examine the effect of Aorg on calculations of the marine CO2 system, the pCO2 and pH values measured in Baltic Sea water were compared with calculated values that were based on the measured alkalinity and another variable of the CO2 system, but ignored the existence of Aorg. Large differences between measured and calculated pCO2 and pH were obtained when the computations were based on AT and CT. The calculated pCO2 was 27–56% lower than the measured value whereas the calculated pH was overestimated by more than 0.4 pH units. Since biogeochemical models are based on the transport and transformations of AT and CT, the acid–base properties of DOM should be included in calculations of the CO2 system in DOM-rich basins like the Baltic Sea. In view of our limited knowledge about the composition and acid/base properties of DOM, this is best achieved using a bulk dissociation constant, KDOM, that represents all weakly acidic functional groups present in DOM. Our preliminary results indicated that the bulk KDOM in the Baltic Sea is 2.94·10−8molkg−1. Although this KDOM has no thermodynamic meaning, it can be a useful tool in numerical studies as it allows an approximation of Aorg in seawater.
•Organic alkalinity (Aorg) constitutes 1.5–3.5% of AT in the Baltic Sea.•Ignoring Aorg causes significant uncertainty in the calculations of pH and pCO2.•The highest uncertainty occurs when pH and pCO2 are calculated from CT and AT.•Bulk dissociation constant of DOM (KDOM) in the Baltic Sea amounts to 2.94·10−8molkg−1.
Biologische Bundesanstalt für Land- und Forstwirtschaft, Institut für Pflanzenschutz im Obstbau, D-69221 Dossenheim, Germany
Correspondence Erich Seemüller E.Seemueller{at}bba.de
Apple proliferation ...(AP), pear decline (PD) and European stone fruit yellows (ESFY) are among the most economically important plant diseases that are caused by phytoplasmas. Phylogenetic analyses revealed that the 16S rDNA sequences of strains of each of these pathogens were identical or nearly identical. Differences between the three phytoplasmas ranged from 1·0 to 1·5 % of nucleotide positions and were thus below the recommended threshold of 2·5 % for assigning species rank to phytoplasmas under the provisional status Candidatus . However, supporting data for distinguishing the AP, PD and ESFY agents at the species level were obtained by examining other molecular markers, including the 16S23S rDNA spacer region, protein-encoding genes and randomly cloned DNA fragments. The three phytoplasmas also differed in serological comparisons and showed clear differences in vector transmission and host-range specificity. From these results, it can be concluded that the AP, PD and ESFY phytoplasmas are coherent but discrete taxa that can be distinguished at the putative species level, for which the names Candidatus Phytoplasma mali, Candidatus Phytoplasma pyri and Candidatus Phytoplasma prunorum, respectively, are proposed. Strains AP15 R , PD1 R and ESFY-G1 R were selected as reference strains. Examination of available data on the peach yellow leaf roll (PYLR) phytoplasma, which clusters with the AP, PD and ESFY agents, confirmed previous results showing that it is related most closely to the PD pathogen. The two phytoplasmas share 99·6 % 16S rDNA sequence similarity. Significant differences were only observed in the sequence of a gene that encodes an immunodominant membrane protein. Until more information on this phytoplasma is available, it is proposed that the PYLR phytoplasma should be regarded as a subtype of Candidatus Phytoplasma pyri.
Abbreviations: AP, apple proliferation; BWB, buckthorn witches-broom; ESFY, European stone fruit yellows; IMP, immunodominant membrane protein; PD, pear decline; PYLR, peach yellow leaf roll; SpaWB, spartium witches-broom
The GenBank/EMBL/DDBJ accession numbers for the P1/P7 amplimers (16S rRNA gene and 16S23S rDNA spacer region) of phytoplasma strains AP15 R , AP1/93, PD1 R , ESFY-G1 R and ESFY-G2 are AJ542541 , AJ542542 , AJ542543 , AJ542544 and AJ542545 , respectively.
Restriction maps of rDNA obtained from P1/P7 PCR products from the apple proliferation, pear decline and European stone fruit yellows phytoplasmas are available as supplementary material in IJSEM Online.
Worldwide, increasing numbers of insects have evolved resistance to a wide range of pesticides, which hampers their control in the field and, therefore, threatens agriculture. Members of the ...carboxylesterase and cytochrome P450 monooxygenase superfamilies are prominent candidates to confer metabolic resistance to pyrethroid insecticides. Both carboxylesterases and P450 enzymes have been shown to be involved in pyrethroid resistance in Australian Helicoverpa armigera , the noctuid species possessing by far the most reported resistance cases worldwide. However, specific enzymes responsible for pyrethroid resistance in field populations of this species have not yet been identified. Here, we show that the resistance toward fenvalerate in an Australian strain of H. armigera is due to a unique P450 enzyme, CYP337B3, which arose from unequal crossing-over between two parental P450 genes, resulting in a chimeric enzyme. CYP337B3 is capable of metabolizing fenvalerate into 4′-hydroxyfenvalerate, which exhibits no toxic effect on susceptible larvae; enzymes from the parental P450 genes showed no detectable fenvalerate metabolism. Furthermore, a polymorphic H. armigera strain could be bred into a susceptible line possessing the parental genes CYP337B1 and CYP337B2 and a resistant line possessing only CYP337B3 . The exclusive presence of CYP337B3 in resistant insects of this strain confers a 42-fold resistance to fenvalerate. Thus, in addition to previously documented genetic mechanisms of resistance, recombination can also generate selectively advantageous variants, such as this chimeric P450 enzyme with an altered substrate specificity leading to a potent resistance mechanism.
Piriformospora indica, an endophytic root-colonizing fungus, efficiently promotes plant growth and induces resistance to abiotic stress and biotic diseases. P. indica fungal cell wall extract induces ...cytoplasmic calcium elevation in host plant roots. Here, we show that cellotriose (CT) is an elicitor-active cell wall moiety released by P. indica into the medium. CT induces a mild defense-like response, including the production of reactive oxygen species, changes in membrane potential, and the expression of genes involved in growth regulation and root development. CT-based cytoplasmic calcium elevation in Arabidopsis (Arabidopsis thaliana) roots does not require the BAK1 coreceptor or the putative Ca²⁺ channels TPC1, GLR3.3, GLR2.4, and GLR2.5 and operates synergistically with the elicitor chitin. We identified an ethyl methanesulfonate-induced mutant (cytoplasmic calcium elevation mutant) impaired in the response to CT and various other cellooligomers (n = 2–7), but not to chitooligomers (n = 4–8), in roots. The mutant contains a single nucleotide exchange in the gene encoding a poly(A) ribonuclease (AtPARN; At1g55870) that degrades the poly(A) tails of specific mRNAs. The wild-type PARN cDNA, expressed under the control of a 35S promoter, complements the mutant phenotype. Our identification of cellotriose as a novel chemical mediator casts light on the complex P. indica-plant mutualistic relationship.
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