Chemodiversity in flowers of Tanacetum vulgare has consequences on a florivorous beetle.

The chemical composition of plant individuals can vary, leading to high intraspecific chemodiversity. Diversity of floral chemistry may impact the responses of flower-feeding insects. Tanacetum vulgare plants vary significantly in their leaf terpenoid composition, forming distinct chemotypes. We investigated the composition of terpenoids and nutrients of flower heads and pollen in plants belonging to three chemotypes - dominated either by ß-thujone (BThu), artemisia ketone (Keto) or a mixture of (Z)-myroxide, santolina triene, and artemisyl acetate (Myrox) - using different analytical platforms. We tested the effects of these differences on preferences, weight gain and performance of adults of the shining flower beetle, Olibrus aeneus. The terpenoid composition and diversity of flower heads and pollen significantly differed among individuals belonging to the above chemotypes, while total concentrations of pollen terpenoids, sugars, amino acids, and lipids did not differ. Beetles preferred BThu over the Myrox chemotype in both olfactory and contact choice assays, while the Keto chemotype was marginally repellent according to olfactory assays. The beetles gained the least weight within 48 h and their initial mortality was highest when feeding exclusively on floral tissues of the Myrox chemotype. Short-term weight gain and long-term performance were highest when feeding on the BThu chemotype. In conclusion, the beetles showed chemotype-specific responses towards different T. vulgare chemotypes, which may be attributed to the terpenoid composition in flower heads and pollen rather than to differences in nutrient profiles. Both richness and overall diversity are important factors when determining chemodiversity of individual plants and their consequences on interacting insects.

Draft Genome of Tanacetum Coccineum: Genomic Comparison of Closely Related Tanacetum-Family Plants.

The plant Tanacetum coccineum (painted daisy) is closely related to Tanacetum cinerariifolium (pyrethrum daisy). However, T. cinerariifolium produces large amounts of pyrethrins, a class of natural insecticides, whereas T. coccineum produces much smaller amounts of these compounds. Thus, comparative genomic analysis is expected to contribute a great deal to investigating the differences in biological defense systems, including pyrethrin biosynthesis. Here, we elucidated the 9.4 Gb draft genome of T. coccineum, consisting of 2,836,647 scaffolds and 103,680 genes. Comparative analyses of the draft genome of T. coccineum and that of T. cinerariifolium, generated in our previous study, revealed distinct features of T. coccineum genes. While the T. coccineum genome contains more numerous ribosome-inactivating protein (RIP)-encoding genes, the number of higher-toxicity type-II RIP-encoding genes is larger in T. cinerariifolium. Furthermore, the number of histidine kinases encoded by the T. coccineum genome is smaller than that of T. cinerariifolium, suggesting a biological correlation with pyrethrin biosynthesis. Moreover, the flanking regions of pyrethrin biosynthesis-related genes are also distinct between these two plants. These results provide clues to the elucidation of species-specific biodefense systems, including the regulatory mechanisms underlying pyrethrin production.

Under fire-simultaneous volatilome and transcriptome analysis unravels fine-scale responses of tansy chemotypes to dual herbivore attack.

BACKGROUND: Tansy plants (Tanacetum vulgare L.) are known for their high intraspecific chemical variation, especially of volatile organic compounds (VOC) from the terpenoid compound group. These VOCs are closely involved in plant-insect interactions and, when profiled, can be used to classify plants into groups known as chemotypes. Tansy chemotypes have been shown to influence plant-aphid interactions, however, to date no information is available on the response of different tansy chemotypes to simultaneous herbivory by more than one insect species. RESULTS: Using a multi-cuvette system, we investigated the responses of five tansy chemotypes to feeding by sucking and/or chewing herbivores (aphids and caterpillars; Metopeurum fuscoviride Stroyan and Spodoptera littoralis Boisduval). Herbivory by caterpillars following aphid infestation led to a plant chemotype-specific change in the patterns of terpenoids stored in trichome hairs and in VOC emissions. The transcriptomic analysis of a plant chemotype represents the first de novo assembly of a transcriptome in tansy and demonstrates priming effects of aphids on a subsequent herbivory. Overall, we show that the five chemotypes do not react in the same way to the two herbivores. As expected, we found that caterpillar feeding increased VOC emissions, however, a priori aphid infestation only led to a further increase in VOC emissions for some chemotypes. CONCLUSIONS: We were able to show that different chemotypes respond to the double herbivore attack in different ways, and that pre-treatment with aphids had a priming effect on plants when they were subsequently exposed to a chewing herbivore. If neighbouring chemotypes in a field population react differently to herbivory/dual herbivory, this could possibly have effects from the individual level to the group level. Individuals of some chemotypes may respond more efficiently to herbivory stress than others, and in a group environment these "louder" chemotypes may affect the local insect community, including the natural enemies of herbivores, and other neighbouring plants.

Impact of municipal and industrial waste incinerators on PCBs content in the environment.

Polychlorinated biphenyls (PCBs) have been withdrawn from the market due to their toxicity, bioaccumulation capacity, and persistence. PCBs have been observed to potentially form in combustion processes under appropriate conditions and in the presence of precursors containing chlorine. The study covered a municipal waste incineration plant and an industrial waste incineration plant. The objective of the study was to assess the effect of these objects on PCB accumulation in soil and plants taking into account the distance from the emission object and wind direction. Soil samples were collected from layers: 0-5, 5-10, 10-20, and 20-30 cm. Test plants were collected from the same areas as the soil samples. The highest accumulation of PCBs was found in plants with large leaf area. Around the municipal waste incineration plant, these were Tanacetum vulgare leaves (12.45 ng/g), and around the industrial waste incineration plant-grasses (4.3 ng/g). In the case of soils, the accumulation of PCBs for both kind waste incinerators was similar, reaching approximately 3 ng/g. As the distance from the municipal waste incinerator and industrial waste incinerator increased, the accumulation of PCBs in the soil decreased. For municipal waste incinerator, no effect of wind direction on PCB accumulation in the soil was observed. In the majority of cases, the accumulation of PCBs in soils taken from the leeward side of the industrial waste incinerator was higher than that in soils from the windward side. In soils around the municipal waste incinerator, PCB compounds moved deep into the soil and reached the highest accumulation in the soil layer of 10-20 cm or 20-30 cm. In soils around the industrial waste incinerator, the highest accumulation of PCBs occurred in the soil layer of 0-5 cm.

Pyrethrin Biosynthesis: The Cytochrome P450 Oxidoreductase CYP82Q3 Converts Jasmolone To Pyrethrolone.

The plant pyrethrum (Tanacetum cinerariifolium) synthesizes highly effective natural pesticides known as pyrethrins. Pyrethrins are esters consisting of an irregular monoterpenoid acid and an alcohol derived from jasmonic acid (JA). These alcohols, referred to as rethrolones, can be jasmolone, pyrethrolone, or cinerolone. We recently showed that jasmolone is synthesized from jasmone, a degradation product of JA, in a single hydroxylation step catalyzed by jasmone hydroxylase (TcJMH). TcJMH belongs to the CYP71 clade of the cytochrome P450 oxidoreductase family. Here, we used coexpression analysis, heterologous gene expression, and in vitro biochemical assays to identify the enzyme responsible for conversion of jasmolone to pyrethrolone. A further T cinerariifolium cytochrome P450 family member, CYP82Q3 (designated Pyrethrolone Synthase; TcPYS), appeared to catalyze the direct desaturation of the C1-C2 bond in the pentyl side chain of jasmolone to produce pyrethrolone. TcPYS is highly expressed in the trichomes of the ovaries in pyrethrum flowers, similar to TcJMH and other T cinerariifolium genes involved in JA biosynthesis. Thus, as previously shown for biosynthesis of the monoterpenoid acid moiety of pyrethrins, rethrolones are synthesized in the trichomes. However, the final assembly of pyrethrins occurs in the developing achenes. Our data provide further insight into pyrethrin biosynthesis, which could ultimately be harnessed to produce this natural pesticide in a heterologous system.

Volatile, stored and phloem exudate-located compounds represent different appearance levels affecting aphid niche choice.

Intraspecific and intra-individual differences in emitted volatile compounds and in surface and phloem sap-related metabolites do not only affect host plant choice of monophagous aphids but may also guide them to the plant part that provides their ideal niche by maximising their fitness. However, little is known about the variation at these different plant appearance levels. We investigated the preferences of the monophagous aphid species Macrosiphoniella tancetaria and Uroleucon tanaceti for different plant parts (inflorescence stems, young and old leaves) of Tanacetum vulgare plants from two chemotypes, testing their reactions towards volatile, surface and phloem sap-related cues. Furthermore, we studied the variation in leaf glandular trichome density as well as in the composition of volatile, stored and phloem exudate-located specialised (secondary) plant compounds from the different plant parts of these chemotypes. Aphid species showed differences in their preferences. Aphids of M. tanacetaria had to assess the entire plant to choose the stem, whereas U. tanaceti only needed volatile cues to locate the old leaves, which are the plant parts representing their respective niches. Volatiles and stored metabolites varied in their composition and concentration between chemotypes. Stored metabolites additionally differed among plant parts, which was reflected in distinct trichome densities. The composition of phloem exudate-located specialised compounds mostly varied among plant parts. These pronounced differences in plant chemistry on multiple levels provide distinct perception levels for aphids probably driving their niche choice. This study demonstrates the importance to consider these multiple levels to elucidate plant-herbivore interactions with high resolution.

Metabotype variation in a field population of tansy plants influences aphid host selection.

It is well known that plant volatiles influence herbivores in their selection of a host plant; however, less is known about how the nonvolatile metabolome affects herbivore host selection. Metabolic diversity between intraspecific plants can be characterized using non-targeted mass spectrometry that gives us a snapshot overview of all metabolic processes occurring within a plant at a particular time. Here, we show that non-targeted metabolomics can be used to reveal links between intraspecific chemical diversity and ecological processes in tansy (Tanacetum vulgare). First, we show that tansy plants can be categorized into five subgroups based up on their metabolic profiles, and that these "metabotypes" influenced natural aphid colonization in the field. Second, this grouping was not due to induced metabolomic changes within the plant due to aphid feeding but rather resulted from constitutive differences in chemical diversity between plants. These findings highlight the importance of intraspecific chemical diversity within one plant population and provide the first report of a non-targeted metabolomic field study in chemical ecology.

Environmental assessment of the effects of a municipal landfill on the content and distribution of heavy metals in Tanacetum vulgare L.

Heavy metal pollution is an important concern because of its potential to affect human health. This study was conducted to analyze plants growing on a landfill body and in its surroundings to determine their potential for heavy metal accumulation. In addition, the enrichment coefficient (EC) for the plant/soil system was used for determining the environmental contamination from a landfill in terms of heavy metal accumulation. The samples were taken in 2013-2014. Of the analyzed metals, iron achieved the highest values in the samples, i.e. - stalk (103.4-6564.6 mg/kg DM), roots (6563.6-33,036.6 mg/kg DM), leaf (535.1-11,275 mg/kg DM) and soil (12,389-39,381.9 mg/kg DM). The highest concentrations were determined in 2013 for Fe, Mn and Zn. Iron achieved the highest concentrations in the years 2013-2014. Next, EC values were then calculated, with the highest noted for Cd. Cd, as well as Cr, Ni and Zn are accumulated mostly in the leaves, whereas Co, Cu, Fe, Hg, Mn and Pb are accumulated mainly in the roots of T. vulgare.

Accumulation, selection and covariation of amino acids in sieve tube sap of tansy (Tanacetum vulgare) and castor bean (Ricinus communis): evidence for the function of a basic amino acid transporter and the absence of a γ-amino butyric acid transporter.

Sieve tube sap was obtained from Tanacetum by aphid stylectomy and from Ricinus after apical bud decapitation. The amino acids in sieve tube sap were analyzed and compared with those from leaves. Arginine and lysine accumulated in the sieve tube sap of Tanacetum more than 10-fold compared to the leaf extracts and they were, together with asparagine and serine, preferably selected into the sieve tube sap, whereas glycine, methionine/tryptophan and γ-amino butyric acid were partially or completely excluded. The two basic amino acids also showed a close covariation in sieve tube sap. The acidic amino acids also grouped together, but antagonistic to the other amino acids. The accumulation ratios between sieve tube sap and leaf extracts were smaller in Ricinus than in Tanacetum. Arginine, histidine, lysine and glutamine were enriched and preferentially loaded into the phloem, together with isoleucine and valine. In contrast, glycine and methionine/tryptophan were partially and γ-amino butyric acid almost completely excluded from sieve tube sap. The covariation analysis grouped arginine together with several neutral amino acids. The acidic amino acids were loaded under competition with neutral amino acids. It is concluded from comparison with the substrate specificities of already characterized plant amino acid transporters, that an AtCAT1-like transporter functions in phloem loading of basic amino acids, whereas a transporter like AtGAT1 is absent in phloem. Although Tanacetum and Ricinus have different minor vein architecture, their phloem loading specificities for amino acids are relatively similar.

Differences in shoot and root terpenoid profiles and plant responses to fertilisation in Tanacetum vulgare.

Intraspecific chemical diversity is a common phenomenon especially found in shoots of essential oil-accumulating plant species. Abiotic factors can influence the concentration of essential oils, but the effects are inconsistent and little is known in how far these may vary within an individual and within species between chemotypes. Tanacetum vulgare L. occurs in various chemotypes that differ in the composition of mono- and sesquiterpenoids in their shoot tissues. We investigated how far shoot chemotype grouping is mirrored in root terpenoid profiles. Furthermore, we studied whether different fertilisation amounts influence the plant growth and morphological traits as well as the constitutive terpenoid concentration of leaves and roots of three chemotypes, trans-carvyl acetate, ß-thujone, and camphor, to different degrees. Shoot terpenoids were dominated by monoterpenoids, while the roots contained mainly sesquiterpenoids. The clear grouping in three chemotypes based on leaf chemistry was weakly mirrored in the root terpenoid composition. Furthermore, the leaf C/N ratio and the stem height differed between chemotypes. All plants responded to increased nutrient availability with increased total biomass and specific leaf area but decreased C/N and root/shoot ratios. Leaf terpenoid concentrations decreased with increasing fertiliser supply, independent of chemotype. In contrast to the leaves, the terpenoid concentrations of the roots were unaffected by fertilisation. Our results demonstrate that aboveground and belowground organs within a species can be under different selection pressures.

Tanacetum vulgare as a bioindicator of trace-metal contamination: a study of a naturally colonized open-pit lignite mine.

We investigated the possibility of use of Tanacetum vulgare (tansy) as an ecological indicator of metal concentration in a naturally colonized open-pit lignite mine in Belchatów (Poland). Tanacetum vulgare is the only species growing abundantly and spontaneously in the lignite mine waste dumps. Metal concentrations in roots, stems, leaves, flowers, and soil were measured in dump sites differing in type and time of reclamation and therefore differing in pollution levels. Tanacetum vulgare appeared to be an accumulator of chromium and iron in roots, whereas highest concentrations of manganese and zinc were found in leaves. A high bioaccumulation factor for cadmium (Cd) was observed in dumps and control sites, indicating that even small amounts of Cd in the environment may result in significant uptake by the plant. The lowest concentrations of metals were found in plants from sites situated on dumps reclaimed with argillaceous limestone.

Diurnal changes in assimilate concentrations and fluxes in the phloem of castor bean (Ricinus communis L.) and tansy (Tanacetum vulgare L.).

Reports about diurnal changes of assimilates in phloem sap are controversial. We determined the diurnal changes of sucrose and amino acid concentrations and fluxes in exudates from cut aphid stylets on tansy leaves (Tanacetum vulgare), and sucrose, amino acid and K(+) concentrations and fluxes in bleeding sap of castor bean pedicel (Ricinus communis). Approximately half of the tansy sieve tubes exhibited a diurnal cycle of sucrose concentrations and fluxes in phloem sap. Data from many tansy plants indicated an increased sucrose flux in the phloem during daytime in case of low N-nutrition, not at high N-nutrition. The sucrose concentration in phloem sap of young Ricinus plants changed marginally between day and night, whereas the sucrose flux increased 1.5-fold during daytime (but not in old Ricinus plants). The amino acid concentrations and fluxes in tansy sieve tubes exhibited a similar diurnal cycle as the sucrose concentrations and fluxes, including their dependence on N-nutrition. The amino acid fluxes, but not the concentrations, in phloem sap of Ricinus were higher at daytime. The sucrose/amino acid ratio showed no diurnal cycle neither in tansy nor in Ricinus. The K(+)-concentrations in phloem sap of Ricinus, but not the K(+) fluxes, decreased slightly during daytime and the sucrose/K(+)-ratio increased. In conclusion, a diurnal cycle was observed in sucrose, amino acid and K(+) fluxes, but not necessarily in concentrations of these assimilates. Because of the large variations between different sieve tubes and different plants, the nutrient delivery to sink tissues is not homeostatic over time.

How aphids decide what is good for them: experiments to test aphid feeding behaviour on Tanacetum vulgare (L.) using different nitrogen regimes.

Leaf-chewing herbivores select food with a protein/carbohydrate ratio of 0.8-1.5, whereas phloem sap, which aphids feed on, has a ratio of approximately 0.1. Enhanced N fertilization increases the amino acid concentration in phloem sap and elevates the N/C ratio. The study examines: (1) whether aphids select between plants of different N nutrition, (2) whether feeding time correlates with the amino acid composition of phloem sap, and (3) at which stage of probing aphids identify the quality of the plant. Uroleucon tanaceti (Mordvilko) and Macrosiphoniella tanacetaria (Kaltenbach), specialist aphids feeding on tansy (Tanacetum vulgare L.), were reared on this host plant grown essentially hydroponically (in Vermiculite) in the greenhouse on 1, 3, 6, or 12 mM NH(4)NO(3). One and 3 mM NH(4)NO(3) corresponds to the situation found in natural tansy stands. Aphid stylet penetration was monitored by electrical penetration graphs whilst phloem sap was sampled by stylectomy. Both aphid species settled 2-3 times more frequently on plants fertilized with 6 or 12 mM NH(4)NO(3). The phloem sap of these plants contained up to threefold higher amino acid concentrations, without a change in the proportion of essential amino acids. No time differences were observed before stylet penetration of plant tissue. After the first symplast contact, most aphids penetrated further, except M. tanacetaria on low-N plants, where 50% withdrew the stylet after the first probing. The duration of phloem feeding was 2-3 times longer in N-rich plants and the time spent in individual sieve tubes was up to tenfold longer. Aphids identified the nutritional quality of the host plant mainly by the amino acid concentration of phloem sap, not by leaf surface cues nor the proportion of essential amino acids. However, U. tanaceti infestation increased the percentage of methionine plus tryptophan in phloem tenfold, thus manipulating the plants nutritional quality, and causing premature leaf senescence.

Harvest regimen optimization and essential oil production in five tansy (Tanacetum vulgare L.) genotypes under a northern climate.

Tansy (Tanacetum vulgare L.) was cultivated at the Norwegian Crop Research Institute at the Apelsvoll Research Centre, Division Kise, in the period from 2000 to 2001. The study focused on different harvesting regimens for high biomass production and essential oil (EO) yield and quality. Two tansy genotypes from Canada (Richters and Goldsticks) and three Norwegian genotypes (Steinvikholmen, Alvdal, and Brumunddal) were studied. The Canadian genotypes reached a height of 130-145 cm and showed a higher dry weight of aerial plant parts compared to the Norwegian plants in 2000. Similar oil yields could be observed for the Canadian types and genotype Steinvikholmen in the range of 30.8-34.6 L/ha when the plants were harvested twice during budding and before flowering after regrowth (year 2001). In contrast, single harvesting at the full bloom stage resulted in higher oil yields, between 42.1 and 44.5 L/ha (Canadian genotypes), whereas 21.0-38.4 L/ha was obtained from the Norwegian types. Tansy genotypes could be grouped into the following chemotypes: the mixed chemotypes Steinvikholmen (thujone-camphor), Alvdal (thujone-camphor-borneol), Goldsticks (thujone-camphor-chrysanthenyl type), and Brumunddal (thujone-camphor-1,8-cineole-bornyl acetate/borneol-alpha-terpineol) and the distinct chemotype Richters, with average concentrations of (E)-chrysanthenyl acetate >40% in both leaf and flower EO.

Biosynthesis of the irregular monoterpene artemisia ketone, the sesquiterpene germacrene D and other isoprenoids in Tanacetum vulgare L. (Asteraceae).

The incorporation of [1-13C]-labeled glucose into the irregular monoterpene artemisia ketone, the regular monoterpenes camphor and beta-thujone, the sesquiterpene germacrene D, the diterpene trans-phytol and beta-sitosterol and isofucosterol has been studied in axenic cultures of Tanacetum vulgare L. (Asteraceae). Quantitative 13C NMR spectroscopic analysis of the resulting labeling patterns showed that the isoprene units of the monoterpenes and the diterpene are formed via the methylerythritol phosphate (MEP) pathway, whereas the isoprene building blocks of the sesquiterpene and the sterols originate from the mevalonic acid (MVA) pathway.