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Janzen-Connell effects state that the accumulation of host-specific natural enemies near parent plants can negatively affect their offspring. Negative plant-soil feedbacks can produce patterns of seedling performance predicted by Janzen-Connell effects and influence plant populations, but their relevance in field conditions remains unclear. Here, using spatial point-pattern analysis, we examine the spatial distribution of Jacobaea vulgaris to assess whether distance- and density-dependent predictions of Janzen-Connell effects are evident in the field. We established 27 replicated 64 m2 plots at two grassland sites and mapped positions of rosette-bearing and flowering J. vulgaris plants within each plot. To investigate temporal distribution patterns, we tracked plant positions repeatedly in three plots during a single season. Additionally, we tested whether these patterns are soil-mediated. Soil samples were collected underneath flowering plants and at a distance of 0.5 meter, and used to compare seed germination, seedling survival, and growth under controlled conditions. Furthermore, we measured J. vulgaris growth in soil from patches with high J. vulgaris densities and in soil from areas outside these patches. The density of rosette-bearing plants was lower at close distances from flowering plants than expected from null models, suggesting negative distance-dependent plant recruitment. The degree of clustering decreased over time from rosette-bearing to flowering plants, indicating density-dependent self-thinning. Seed germination was higher in soil further away from flowering J. vulgaris plants than in soil underneath plants at one site, but soil distance was not an overall significant factor in explaining seed germination. However, seedling mortality and biomass did not differ between soils collected at the two distances, and plants produced similar biomass in soil collected from inside and outside J. vulgaris patches. Our study demonstrates conspecific distance- and density-dependent plant recruitment in J. vulgaris in the field, but we found no evidence this depends on belowground natural enemies. [ABSTRACT FROM AUTHOR]

The overall effect of a live soil inoculum collected from nature on plant biomass is often negative. One hypothesis to explain this phenomenon is that the overall net pathogenic effect of soil microbial communities reduces plant performance. Induced plant defenses triggered by the application of the plant hormones jasmonic acid (JA) and salicylic acid (SA) may help to mitigate this pathogenic effect of live soil. However, little is known about how such hormonal application to the plant affects the soil and how this, in turn, impacts plant growth. We grew four plant species in sterilized and inoculated live soil and exposed their leaves to two hormonal treatments (JA and SA). Two species (Jacobaea vulgaris and Cirsium vulgare) were negatively affected by soil inoculation. In these two species foliar application of SA increased biomass in live soil but not in sterilized soil. Two other species (Trifolium repens and Daucus carota) were not affected by soil inoculum and for these two species foliar application of SA reduced plant biomass in both the sterilized and live soil. Application of JA reduced plant biomass in both soils for all species. We subsequently carried out a multiple generation experiment for one of the plant species, J. vulgaris. In each generation, the live soil was a mixture of 10% soil from the previous generation and 90% sterilized soil and the same hormonal treatments were applied. The negative effects of live soil on plant biomass were similar in all four generations, and this negative effect was mitigated by the application of SA. Our research suggests that the application of SA can mitigate the negative effects of live soil on plant growth. Although the inoculum of soil containing a natural live soil microbial community had a strong negative effect on the growth of J. vulgaris , we found no evidence for an increase or decrease in negative plant-soil feedback in either the control or the SA treated plants. Also plant performance did not decrease consistently with succeeding generations. [ABSTRACT FROM AUTHOR]

Specialist herbivores feed on a restricted number of related plant species and may suffer food shortage if overexploitation leads to periodic defoliation of their food plants. The density, size and quality of food plants are important factors that determine the host plant choice of specialist herbivores. To explore how all these factors influence their oviposition behaviour, we used the cinnabar moth Tyria jacobaeae and the hybrids of a cross between Jacobaea vulgaris and J. aquatica as a study system. While defoliation by the cinnabar moth is common in the coastal area of The Netherlands, it is relatively rare in inland ragwort population. Ragworts contain pyrrolizidine alkaloids (PAs) and those that are found in coastal areas are rich in jacobine-like PAs while those that occur inland are rich in erucifoline-like PAs. We tested how the oviposition preference was influenced by plant size, nitrogen and water content and PA composition. We used cinnabar moth populations from a regularly defoliated area, Meijendel, and Bertogne, a rarely defoliated area. Our results revealed no effects of nitrogen or water content on oviposition preference. Moths from both populations laid larger egg batches on the plants rich in jacobine-like PAs. Moths from Meijendel preferred larger plants and spread their eggs over more egg batches that were, on average, smaller than those of Bertogne moths. These results suggest that Meijendel moths adopted a oviposition strategy to cope with potential defoliation. [ABSTRACT FROM AUTHOR]

Abstract: The cinnabar moth (Tyria jacobaeae, Arctiidae) normally feeds on Senecio jacobaea in the field. For the first time, naturally occurring populations of T. jacobaeae have been found thriving on Senecio adonidifolius, even though the moth''s preferred host, S. jacobaea, is available within 50–400 m. In the laboratory, the cinnabar moth has been shown to feed on and develop on S. adonidifolius despite its different leaf morphology, pyrrolizidine alkaloid (PA) profile and a large taxonomic distance to S. jacobaea. Here I examined whether T. jacobaeae has adapted to this new host in the field using adult oviposition behavior and plant acquired defense chemistry in pupae as criteria. Choice tests indicated local adaptation to this newly recorded host. T. jacobaeae reared on S. adonidifolius hosts laid more egg batches and total eggs on it than T. jacobaeae from S. jacobaea. The egg batches were smaller on S. adonidifolius possibly due to highly pinnate thread-like structure of its leaves. The bouquet of plant acquired PAs and the insect metabolized callimorphine in pupae differed widely between pupae collected from the two hosts. T. jacobaeae pupae taken from S. adonidifolius hosts contained more of the insect metabolized callimorphine than pupae taken from S. jacobaea hosts, but they did not differ in total PA concentration. Pupae taken from S. jacobaea hosts contained more unmetabolized plant PA''s than pupae from S. adonidifolius hosts. Additionally, 10% of T. jacobaeae larvae taken from S. adonidifolius in Biausse were infested with Carcelia dubia, a parasitic and rare tachinid fly that typically attacks arctiid moths. [Copyright &y& Elsevier]

We studied the variation in pyrrolizidine alkaloid (PA) patterns of lab-grown vegetative plants of 11 European Senecio jacobaea populations. Plants were classified as jacobine, erucifoline, mixed or senecionine chemotypes based on presence and absence of the PAs jacobine or erucifoline. Due to the presence of jacobine, total PA concentration in jacobine chemotypes was higher than in erucifoline chemotypes. Both relative and absolute concentrations of individual PAs differed between half-sib and clonal families, which showed that variation in PA patterns had a genetic basis. Within most populations relative abundance of PAs varied considerably between individual plants. Most populations consisted either of the jacobine chemotype or of the erucifoline chemotype, sometimes in combination with mixed or senecionine chemotypes. [Copyright &y& Elsevier]

This is the first study showing that alkaloids are present on the leaf surface of plants. A concentration of 30–230 pmol/cm2 pyrrolizidine alkaloids (PA''s) was detected in 8 different samples taken from Senecio jacobaea. PA concentration on the leaves was marginally correlated with PA concentration of the total leaf tissues. The PA spectrum on the leaf differed from the PA spectrum of the total leaf. [Copyright &y& Elsevier]

Plant specialised metabolites (SMs) are very diverse in terms of both their number and chemical structures with more than 200,000 estimated compounds. This chemical diversity occurs not only among different groups of compounds but also within the groups themselves. In the context of plant-insect interactions, the chemical diversity within a class of structurally related metabolites is generally also related to their bioactivity. In this study, we tested firstly whether individual SMs within the group of pyrrolizidine alkaloids (PAs) differ in their effects on insect herbivores (western flower thrips, Frankliniella occidentalis ). Secondly, we tested combinations of PA N -oxides to determine whether they are more active than their individual components. We also evaluated the bioactivity of six PA free bases and their corresponding N -oxides. At concentrations similar to that in plants, several PAs reduced thrip's survival but the effect also differed strongly among PAs. In general, PA free bases caused a lower survival than their corresponding N -oxides. Among the tested PA free bases, we found jacobine and retrorsine to be the most active against second instar larvae of thrips, followed by erucifoline and seneciphylline, while senecionine and monocrotaline did not exhibit significant dose-dependent effects on thrip's survival. In the case of PA N -oxides, we found that only senecionine N -oxide and jacobine N -oxide reduced thrip's survival, although the effect of senecionine N -oxide was weak. Combinations of PA N -oxides showed no synergistic effects. These findings indicate the differences observed in the effect of structurally related SMs on insect herbivores. It is of limited value to study the bioactivity of combined groups, such as PAs, without taking their composition into account. [ABSTRACT FROM AUTHOR]

Persian poppy (Papaver bracteatum Lindl.) is a perennial medicinal plant belonging to the Papaveraceae family that is endemic to the mountainous areas in Northern Iran. It is known for high amounts of the valuable benzylisoquinoline alkaloid thebaine. The effects of induced polyploidy as well as the effect of methyl Jasmonate (MeJA) elicitation on the root production of thebaine and on the expression of five alkaloid biosynthesis related genes were studied. The in vitro tetraploidy induction caused a significant increased expression of norcoclaurine synthase (NCS) and salutaridinol (SAT), and a significant decreased expression of berberine bridge enzyme (BBE) in the leaves. In the root tissues, the BBE , NCS , and SAT showed an increased expression in tetraploid plants, while codeinone reductase (COR) showed a decreased expression. A similar alteration pattern was found in mixoploid plants when compared to their diploid counterparts. MeJA at concentrations of 0.1 and 0.5 mM caused a remarkable increase in the thebaine content in the roots of treated plants, where the highest thebaine content was identified in plants elicited with 0.5 mM MeJA. Elicitation treatment caused a substantial increase in the expression of NCS and SAT in the leaves, while it had no major effect on BBE , codeine 3-O-demethylase (CODM) and COR. Expression analysis in the roots showed that MeJA caused a significant increase in the expression of only BBE and NCS , while expression of other studied genes remained unchanged. Our results may be exploited for improved thebaine production and the processing of Persian poppy. • Gene expression levels of NCS and SAT increased, while that of BBE decreased in the leaves of polyploidized plants. • In the roots, COR and CODM , both downstream of thebaine in the alkaloid pathway, are downregulated in tetraploids. • MeJA elicitation increased expression of NCS and SAT in the leaves, while that of BBE , CODM , and COR remained unchanged. • The elicitation-induced increase in root thebaine content was influenced mainly by the concentration of the elicitor. [ABSTRACT FROM AUTHOR]

Abstract: Secondary metabolites such as pyrrolizidine alkaloids (PAs) play a crucial part in plant defense. PAs can occur in plants in two forms: tertiary amine (free base) and N-oxide. PA extraction and detection are of great importance for the understanding of the role of PAs as plant defense compounds, as the tertiary PA form is known for its stronger influence on several generalist insects, whereas the N-oxide form is claimed to be less deterrent. We measured PA N-oxides and their reduced tertiary amines by liquid chromatography–tandem mass spectrometry (LC-MS/MS). We show that the occurrence of tertiary PAs is not an artifact of the extraction and detection method. We found up to 50% of tertiary PAs in shoots of Jacobine – chemotype plants of Jacobaea vulgaris. Jacobine and its derivatives (jacoline, jaconine, jacozine and dehydrojaconine) may occur for more than 20% in reduced form in the shoots and more than 10% in the roots. For 22 PAs detected in F2 hybrids (J. vulgaris × Jacobaea aquatica), we calculate the tertiary amine percentage (TA%=the tertiary amine concentration/(tertiary amine concentration+the corresponding N-oxide concentration)×100). We found that the TA% for various PAs was genotype-dependent. Furthermore, TA% for the different PAs were correlated and the highest correlations occurred between PAs which share high structural similarity. [ABSTRACT FROM AUTHOR]

Abstract: The pyrrolizidine alkaloid (PA) content of flowers, leaves, and roots of Anchusa strigosa (Boraginaceae) was analysed by ESI–LC–MS. Six PAs, including two new natural compounds, were detected, characterized by NMR spectroscopy, and quantified in each plant organ. The results indicated that the highest total concentration of PAs was in the leaves (23.63mg/g of dried part), followed by the flowers (19.77mg/g), and finally by the roots (1.80mg/g). All PAs isolated were subjected to Spodoptera exigua and Pieris brassicae larvae. Feeding activity by both herbivore species using a bioassay was inhibited up to circa 75% depending on PA and applied concentration. [Copyright &y& Elsevier]

Abstract: Pyrrolizidine alkaloids (PAs) of the macrocyclic senecionine type are secondary metabolites characteristic for most species of the genus Senecio (Asteraceae). These PAs are deterrent and toxic to most vertebrates and insects and provide plants with a chemical defense against herbivores. We studied the PA composition of 24 out of 26 species of Senecio section Jacobaea using GC-MS. The PA profiles of eight of these species have not been studied before and additional PAs were identified for most other species that were included in previous studies. With one exception (senecivernine) all 26 PAs identified in sect. Jacobaea can be regarded as derivatives of the biosynthetic backbone structure senecionine. Based on the PA profiles of the species of sect. Jacobaea and the results of previous tracer studies, we constructed two hypothetical biosynthetic scenarios of senecionine diversification. Both scenarios contain two major reactions: the conversion of the necine base moiety retronecine into the otonecine moiety and site-specific epoxidations within the necic acid moiety. Further reactions are site-specific hydroxylations, sometimes followed by O-acetylations, site-specific dehydrogenations, E, Z-isomerizations, and epoxide hydrolysis and chlorolysis. The GC-MS data and both biosynthetic scenarios were subsequently used to study the evolution of PA formation in sect. Jacobaea by reconstructing the evolutionary history of qualitative PA variation in this section. This was achieved by optimizing additive presence/absence data of PAs and types of enzymatic conversions on a maximum parsimony cladogram of section Jacobaea inferred from DNA sequence and morphological data. Besides showing large intra- and interspecific variation, PA distribution appears to be largely incidental within the whole clade. These results together with the finding that all but one of the PAs identified in sect. Jacobaea are also present in species of other sections of Senecio indicate that differences in PA profiles in Senecio can not be explained by the gain and loss of PA specific genes, but rather by a transient switch-off and switch-on of the expression of genes encoding PA pathway-specific enzymes. [Copyright &y& Elsevier]