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Thrips are one of the most challenging pests in agricultural crops, including Chrysanthemum . In this study we tested via two plant assays whether solutions containing sticky rice germ oil (RGO) droplets could effectively trap thrips and lower thrips damage on Chrysanthemum . In the first assay, we additionally assessed the metabolomic effects of these RGO droplet sprays and thrips presence on plant chemistry via 1 H NMR and headspace GC-MS on multiple timepoints to investigate which plant metabolites were affected by spraying and their potential relation to plant resistance against thrips. In the second assay, we tested the individual RGO solution constituents against thrips. Our results suggested that the adhesive RGO droplets were not effective as a physical trap as only three out of 600 adult thrips were caught at the achieved coverage. However, average thrips damage was still reduced up to 50% and no negative effects on plant growth were observed up to 25 days. Results from the second plant assay indicated that the individual constituents of the solution containing RGO droplets may have direct effects against thrips. Metabolomics analysis of sprayed leaves via headspace GC-MS and 1 H NMR indicated that fatty acids and several volatile compounds such as 4(10)-thujene (sabinene), eucalyptol, cis -4-thujanol, and isocaryophyllene were highest on day 10, while sucrose, malic acid, o -Cymene, and 3-Methyl-2-butenoic acid were highest on day 25. Plants with thrips showed higher flavonoid, carbohydrate and glutamine acetic acid levels, and lower fatty acids and malic acid levels. RGO application increased the levels of fatty acids and alcohols present on top of and inside the Chrysanthemum leaves, while decreasing the concentrations of volatile compounds such as eucalyptol, chrysanthenone and eugenol in the Chrysanthemum leaves. Most interestingly, the thrips effect on the plant metabolome was no longer visible in RGO treated plants at the later harvesttime, suggesting that RGO application may overrule or prevent the metabolomic effects of thrips infestation. In conclusion, our study provides new information on how the application of a new plant-based plant protection product affects insect herbivores and alters crop phytochemistry for improved herbivore resistance., Competing Interests: A patent for the method to fabricate solutions with adhesive plant-derived oil droplets has been filed with the European Patent Office, application no. 22202752.6; EP4356732A1, by Wageningen University. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2025 Bierman, Fernandes, Choi, Seo, Vrieling, Macel, Knegt, Kodger, van Zwieten, Klinkhamer and Bezemer.)

With a continuous increase in world population and food production, chemical pesticide use is growing accordingly, yet unsustainably. As chemical pesticides are harmful to the environment and developmental resistance in pests is increasing, a sustainable and effective pesticide alternative is needed. Inspired by nature, we mimic one defense strategy of plants, glandular trichomes, to shift away from using chemical pesticides by moving toward a physical immobilization strategy via adhesive particles. Through controlled oxidation of a biobased starting material, triglyceride oils, an adhesive material is created while monitoring the reactive intermediates. After being milled into particles, nanoindentation shows these particles to be adhesive even at low contact forces. A suspension of particles is then sprayed and found to be effective at immobilizing a target pest, thrips, Frankliniella occidentalis . Small arthropod pests, like thrips, can cause crop damage through virus transfer, which is prevented by their immobilization. We show that through a scalable fabrication process, biosourced materials can be used to create an effective, sustainable physical pesticide., Competing Interests: Competing interests statement:A patent pertaining to this research has been filed with the European Patent Office Application No. EP22202752.

The defense mechanisms of plants evolve as they develop. Previous research has identified chemical defenses against Western flower thrips (WFT) in Gladiolus ( Gladiolus hybridus L.). Consequently, our study aimed to explore the consistency of these defense variations against WFT across the various developmental stages of Gladiolus grown under different conditions. Thrips bioassays were conducted on whole plants at three developmental stages, using the Charming Beauty and Robinetta varieties as examples of susceptible and resistant varieties, respectively. Metabolomic profiles of the leaves, buds and flowers before thrips infestation were analyzed. The thrips damage in Charming Beauty was more than 500-fold higher than the damage in Robinetta at all plant development stages. Relative concentrations of triterpenoid saponins and amino acids that were associated with resistance were higher in Robinetta at all plant stages. In Charming Beauty, the leaves exhibited greater damage compared to buds and flowers. The relative concentrations of alanine, valine and threonine were higher in buds and flowers than in leaves. The Metabolomic profiles of the leaves did not change significantly during plant development. In addition, we cultivated plants under different environmental conditions, ensuring consistency in the performance of the two varieties across different growing conditions. In conclusion, the chemical thrips resistance markers, based on the analysis of vegetative plants grown in climate rooms, were consistent over the plant's lifetime and for plants grown under field conditions.

Background: Bulk segregant analysis (BSA) can help identify quantitative trait loci (QTLs), but this may result in substantial bycatch of functionally irrelevant genes., Results: Here we develop a Gene Ontology-mediated approach to zoom in on specific genes located inside QTLs identified by BSA as implicated in a continuous trait. We apply this to a novel experimental system: flowering time in the giant woody Jersey kale, which we phenotyped in four bulks of flowering onset. Our inferred QTLs yielded tens of thousands of candidate genes. We reduced this by two orders of magnitude by focusing on genes annotated with terms contained within relevant subgraphs of the Gene Ontology. A pathway enrichment test then led to the circadian rhythm pathway. The genes that enriched this pathway are attested from previous research as regulating flowering time. Within that pathway, the genes CCA1, FT, and TSF were identified as having functionally significant variation compared to Arabidopsis. We validated and confirmed our ontology-mediated results through genome sequencing and homology-based SNP analysis. However, our ontology-mediated approach produced additional genes of putative importance, showing that the approach aids in exploration and discovery., Conclusions: Our method is potentially applicable to the study of other complex traits and we therefore make our workflows available as open-source code and a reusable Docker container., (© 2022. The Author(s).)

Herbivore population dynamics are strongly influenced by the interactions established through their shared host. Such plant-mediated interactions can occur between different herbivore species and different life developmental stages of the same herbivore. However, whether these interactions occur between leaf-feeding herbivores and their soil-dwelling pupae is unknown. We studied whether tomato (Solanum lycopersicum) leaf herbivory by the American serpentine leafminer Liriomyza trifolii affects the performance of conspecific pupae exposed to the soil headspace of the plant. To gain mechanistic insights, we performed insect bioassays with the jasmonate-deficient tomato mutant def-1 and its wild-type, along with phytohormones, gene expression and root volatiles analyses. Belowground volatiles accelerated leafminer metamorphosis when wild-type plants were attacked aboveground by conspecifics. The opposite pattern was observed for def-1 plants, in which aboveground herbivory slowed metamorphosis. Leafminer attack induced jasmonate and abscisic acid accumulation and modulated volatile production in tomato roots in a def-1-dependent manner. Our results demonstrate that aboveground herbivory triggers changes in root defence signalling and expression, which can directly or indirectly via changes in soil or microbial volatiles, alter pupal development time. This finding expands the repertoire of plant-herbivore interactions to herbivory-induced modulation of metamorphosis, with potential consequences for plant and herbivore community dynamics., (© 2022 The Authors. New Phytologist © 2022 New Phytologist Foundation.)

Concentration of plant secondary metabolites (SMs) show seasonal variations. However, it is still not well understood how these abiotic and biotic factors influence the seasonal variations of SMs. In addition, it is of interest to know if and how SMs are reallocated to the different plant organs, in particular whether SMs are reallocated to the remaining tissues when biomass is lost, e.g., during winter. Here we used Jacobaea vulgaris, Jacobaea aquatica, two F1 and four F2 hybrids that differed in their pyrrolizidine alkaloids (PAs) bouquet as a study system. A series of clones of these genotypes were investigated during their vegetative stage spanning 14 months in a semi-natural environment. We found that the total PA concentration in roots and shoots showed a gradual increase until the spring of the second year, whereafter it dropped substantially in shoots. The variation in PA composition due to seasonal changes was significant but relatively small. Senecionine-like PAs were the dominant PAs in roots, while jacobine-/erucifoline-like PAs were dominant in shoots. The variation of PA concentration was significantly correlated with temperature, day length, and plant age. A correlation analysis showed that PAs were not reallocated when biomass was lost in winter. Overall, our study showed that PA composition of each genotype changed over seasons in a different manner but seasonal variation did not overrule the differences in PA composition among genotypes., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Despite the extensive studies on latex, some fundamental questions on their chemical specialization and the factors influencing this specialization have yet to be investigated. To address this issue, latexes and their bearing tissues from diverse species were profiled by 1 HNMR and GC-MS. Additionally, the antiherbivory activity of these materials was tested against thrips ( Frankliniella occidentalis Pergande, 1895). The multivariate data analysis showed a clear separation between latexes and leaves from the same species. Conversely, the chemical profiles of latexes from different species were highly similar, that is, they displayed much less metabolic species-specificity. These shared chemical profiles of latexes were reflected in their overall higher mortality index (80.4% ± 7.5) against thrips compared with their bearing tissues (55.5% ± 14.9). The metabolites correlated to the antiherbivory activity of latexes were triterpenoids and steroids. However, the activity could not be attributed to any single terpenoid. This discrepancy and the reduction of the latex activity after fractionation suggested a complementary effect of the compounds when in a mixture as represented by the latex. Additionally, aqueous fractions of several latexes were found to possess simple spectra, even with only 1 metabolite. These metabolites were determined to be organic acids that might be involved in the modulation of the rate of latex coagulation, potentially increasing the sealing and trapping effects of the latex., Competing Interests: The authors declare that they have no conflict of interest., (Thieme. All rights reserved.)

It is increasingly evident that plants interact with their outside world through the production of volatile organic compounds, 1-5 but whether the volatiles have evolved to serve in plant defense is still a topic of considerable debate. 3 , 6-8 Unharmed leaves constitutively release small amounts of volatiles, but when the leaves are damaged by herbivorous arthropods, they emit substantially more volatiles. These herbivore-induced plant volatiles (HIPVs) attract parasitoids and predators that kill insect herbivores, 9-12 and this can benefit the plants. 13 , 14 As yet, however, there is no tangible evolutionary evidence that this tritrophic interplay contributes to the selection forces that have shaped the volatile emissions of plants. 2 , 3 , 5-8 , 15 With this in mind, we investigated the evolutionary changes in volatile emissions in invasive common ragwort and the respective defensive roles of its constitutive and inducible volatiles. This Eurasian plant has invaded other continents, where it evolved for many generations in the absence of specialized herbivores and their natural enemies. We found that, compared to native ragworts, invasive plants release higher levels of constitutive volatiles but considerably lower levels of herbivore-induced volatiles. As a consequence, invasive ragwort is more attractive to a specialist moth but avoided by an unadapted generalist moth. Importantly, conforming to the indirect defense hypothesis, a specialist parasitoid was much more attracted to caterpillar-damaged native ragwort, which was reflected in higher parasitism rates in a field trial. The evolution of foliar volatile emissions appears to be indeed driven by their direct and indirect roles in defenses against insects., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

Understanding the mechanisms involved in host plant resistance opens the way for improved resistance breeding programs by using the traits involved as markers. Pest management is a major problem in cultivation of ornamentals. Gladiolus ( Gladiolus hybridus L.) is an economically important ornamental in the Netherlands. Gladiolus is especially sensitive to attack by western flower thrips ( Frankliniella occidentalis (Pergande) (Thysanoptera:Thripidae)). The objective of this study was, therefore, to investigate morphological and chemical markers for resistance breeding to western flower thrips in Gladiolus varieties. We measured thrips damage of 14 Gladiolus varieties in a whole-plant thrips bioassay and related this to morphological traits with a focus on papillae density. Moreover, we studied chemical host plant resistance to using an eco-metabolomic approach comparing the 1 H NMR profiles of thrips resistant and susceptible varieties representing a broad range of papillae densities. Thrips damage varied strongly among varieties: the most susceptible variety showed 130 times more damage than the most resistant one. Varieties with low thrips damage had shorter mesophylls and epidermal cells, as well as a higher density of epicuticular papillae. All three traits related to thrips damage were highly correlated with each other. We observed a number of metabolites related to resistance against thrips: two unidentified triterpenoid saponins and the amino acids alanine and threonine. All these compounds were highly correlated amongst each other as well as to the density of papillae. These correlations suggest that papillae are involved in resistance to thrips by producing and/or storing compounds causing thrips resistance. Although it is not possible to distinguish the individual effects of morphological and chemical traits statistically, our results show that papillae density is an easy marker in Gladiolus-breeding programs targeted at increased resistance to thrips.

Based on the hypothesis that the variation of the metabolomes of latex is a response to selective pressure and should thus be affected differently from other organs, their variation could provide an insight into the defensive chemical selection of plants. Metabolic profiling was used to compare tissues of three Euphorbia species collected in diverse regions. The metabolic variation of latexes was much more limited than that of other organs. In all the species, the levels of polyisoprenes and terpenes were found to be much higher in latexes than in leaves and roots of the corresponding plants. Polyisoprenes were observed to physically delay the contact of pathogens with plant tissues and their growth. A secondary barrier composed of terpenes in latex and in particular, 24-methylenecycloartanol, exhibited antifungal activity. These results added to the well-known role of enzymes also present in latexes, show that these are part of a cooperative defense system comprising biochemical and physical elements.

Fungal endophytes isolated from two latex bearing species were chosen as models to show their potential to expand their host plant chemical diversity. Thirty-three strains were isolated from Alstonia scholaris (Apocynaceae) and Euphorbia myrsinites (Euphorbiaceae). High performance thin layer chromatography (HPTLC) was used to metabolically profile samples. The selected strains were well clustered in three major groups by hierarchical clustering analysis (HCA) of the HPTLC data, and the chemical profiles were strongly correlated with the strains' colony size. This correlation was confirmed by orthogonal partial least squares (OPLS) modeling using colony size as "Y" variable. Based on the multivariate data analysis of the HPTLC data, the fastest growing strains of each cluster were selected and used for subsequent experiments: co-culturing to investigate interactions between endophytes-phytopathogens, and biotransformation of plant metabolites by endophytes. The strains exhibited a high capacity to fight against fungal pathogens. Moreover, there was an increase in the antifungal activity after being fed with host-plant metabolites. These results suggest that endophytes play a role in plant defense mechanisms either directly or by biotransformation/induction of metabolites. Regarding HPTLC-based metabolomics, it has proved to be a robust approach to monitor the interactions among fungal endophytes, the host plant and potential phytopathogens.

Phytohormone applications are used to mimic herbivory and can induce plant defences. This study investigated (i) metabolomic changes in leaf tissues of Jacobaea vulgaris and J. aquatica after methyl jasmonate (MeJA) and salicylic acid (SA) applications and (ii) the effects on a leaf-chewing, a leaf-mining and a piercing-sucking herbivore. MeJA treated leaves showed clearly different metabolomic profiles than control leaves, while the differences in metabolomic profiles between SA treated leaves and control leaves were less clear. More NMR peaks increased than decreased after MeJA treatment while this pattern was reversed after SA treatment. The leaf-chewing (Mamestra brassicae) and the leaf-mining herbivores (Liriomyza trifolii) fed less on MeJA-treated leaves compared to control and SA-treated leaves while they fed equally on the latter two. In J. aquatica but not in J. vulgaris, SA treatment reduced feeding damage by the piercing-sucking herbivore (Frankliniella occidentalis). Based on the herbivory and metabolomic data after phytohormone application, we made speculations as follows: For all three herbivore species, plants with high levels of threonine and citric acid showed less herbivory while plants with high levels of glucose showed more herbivory. Herbivory by thrips was lower on plants with high levels of alanine while it was higher on plants with high levels of 3,5-dicaffeoylquinic acid. The plant compounds that related to feeding of piercing-sucking herbivore were further verified with previous independent experiments., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Western flower thrips (WFT), Frankliniella occidentalis, is a serious insect pest of Chrysanthemum [Chrysanthemum × morifolium Ramat. (Asteraceae)]. Here we have investigated whether genotypic variation in constitutive and inducible resistance to WFT correlates with phenotypic differences in leaf trichome density and the activity of the defense-related enzyme polyphenol oxidase (PPO) in chrysanthemum. Non-glandular and glandular leaf trichome densities significantly varied among ninety-five chrysanthemum cultivars. Additional analyses in a subset of these cultivars, differing in leaf trichome density, revealed significant variation in PPO activities and resistance to WFT as well. Constitutive levels of trichome densities and PPO activity, however, did not correlate with chrysanthemum resistance to WFT. Further tests showed that exogenous application of the phytohormone jasmonic acid (JA) increased non-glandular trichome densities, PPO activity and chrysanthemum resistance to WFT, and that these effects were cultivar dependent. In addition, no tradeoff between constitutive and inducible resistance to WFT was observed. JA-mediated induction of WFT resistance, however, did not correlate with changes in leaf trichome densities nor PPO activity levels. Taken together, our results suggest that chrysanthemum can display both high levels of constitutive and inducible resistance to WFT, and that leaf trichome density and PPO activity may not play a relevant role in chrysanthemum defenses against WFT.

Auxins are commonly used for commercial propagation of chrysanthemums by stem cuttings. Recent studies imply that these root-promoting hormones also affect plant defense responses. The underlying motive of this study stems from the serendipitous observation that water dipping of auxin-coated cuttings beneficially affected thrips herbivory. Therefore, the primary objective of this investigation was to explore the role of indole-3-butyric acid (IBA) in relation to herbivore susceptibility in chrysanthemum. We observed contrasting findings concerning the physical presence of IBA and it's role in promoting susceptibility of cuttings to thrips, which may in part be explained by the phenotypical variations of cuttings generated from mother plants. Nonetheless, we repeatedly demonstrated considerable protection, in some experiments up to 37%, against thrips and leaf miner upon water dipping of IBA-coated cuttings. Assessment of polyphenol oxidase activity (PPO), 14 days after dipping treatment, suggests that neither direct induction nor priming of plant defenses are involved. Future experiments aimed at understanding the early signaling events may help to explain the underlying mechanisms involved in conferring herbivore protection. We propose a dual role for auxins in early integrated pest management strategies to maximize plant development and minimize herbivory through feasible, cost-effective water dipping treatments.

Induction of defenses is one of the most widely accepted eco-friendly approaches for management of pests and diseases. Seeds are receptive to resistance-inducing chemicals and could offer broad-spectrum protection at the early stages of development. However, seed treatment with elicitors has previously been shown to differentially influence induced defense responses among cultivars and thus, could hamper commercial exploitation. In this context, the objective of the present study was to evaluate the genotype-dependent ability of jasmonic acid (JA) to induce resistance against western flower thrips (WFT) at the seed stage. We examined the variation in inducibility of resistance in eight commercial tomato cultivars. Causal factors accounting for discrepancies in JA-induced responses at the seed stage were phenotypically and biochemically evaluated. Seed receptivity to exogenous JA appeared to be cultivar dependent. Thrips associated silver damage was only reduced in JA seed-treated plants of cultivar Carousel. Enhancement of resistance, was not associated with activation of defense-related traits such as polyphenol oxidase activity (PPO), trichomes or volatiles. Sulfuric acid scarification, prior to JA seed incubation, significantly augmented the embryonic responsiveness to JA in cv. Moneymaker without an adverse effect on growth. Hence, these results support the hypothesis that seed coat permeability is a key factor for successfully inducing JA mediated thrips defenses. The outcome of our study is of translational value as it creates opportunities for the seed industry to perform pre-treatments on non-responsive cultivars as well as for tomato breeding programs to select for genetic traits that affect seed permeability., (Copyright © 2020 Mouden, Kappers, Klinkhamer and Leiss.)

Abiotic and biotic properties of soil can influence growth and chemical composition of plants. Although it is well-known that soil microbial composition can vary greatly spatially, how this variation affects plant chemical composition is poorly understood. We grew genetically identical Jacobaea vulgaris in sterilized soil inoculated with live soil collected from four natural grasslands and in 100% sterilized soil. Within each grassland we sampled eight plots, totalling 32 different inocula. Two samples per plot were collected, leading to three levels of spatial variation: within plot, between and within grasslands. The leaf metabolome was analysed with 1 H Nuclear magnetic resonance spectroscopy (NMR) to investigate if inoculation altered the metabolome of plants and how this varied between and within grasslands. Inoculation led to changes in metabolomics profiles of J. vulgaris in two out of four sites. Plants grown in sterilized and inoculated soils differed in concentrations of malic acid, tyrosine, trehalose and two pyrrolizidine alkaloids (PA). Metabolomes of plants grown in inoculated soils from different sites varied in glucose, malic acid, trehalose, tyrosine and in one PA. The metabolome of plants grown in soils with inocula from the same site was more similar than with inocula from distant sites. We show that soil influences leaf metabolomes. Performance of aboveground insects often depends on chemical composition of plants. Hence our results imply that soil microbial communities, via affecting aboveground plant metabolomes, can impact aboveground plant-insect food chains but that it is difficult to make general predictions due to spatial variation in soil microbiomes.

The effect of environmental factors on the chemical composition of plants eventually resulting in plant growth regulation is an age-old issue in plant biology. Nowadays, the acceleration in changes in environmental conditions (e.g. global warming) can act as an incentive to investigate their correlation with metabolic changes. In this study, Cistus monspeliensis plants grown on the island of Sardinia (Italy) were used to explore the geographical-mediated metabolic variation and its repercussion on plant-fungus interactions. Samples of different ecotypes of C. monspeliensis were collected and chemically profiled by 1 H NMR and HPTLC-based metabolomics and the relationship between the variations of biological activity was examined by multivariate data analysis. The ecotypes, collected from different geographical zones and altitudes, exhibited clearly distinguishable chemical profiles, particularly in their terpene and phenolic contents. In particular, multivariate data analysis revealed several diterpenes of the labdane and clerodane series among the terpenes and methoxyflavonoids to be responsible for the differentiation. The antifungal activity of the plants was used to explore the correlation between chemical variation and biological activity. Results showed that there was a strong correlation between the metabolic profiles and the antifungal activity, revealing terpenes and methoxylated flavonoids as the main involved metabolites. This demonstrated that environmental factors can influence the chemical variation of plant ecotypes, resulting in the generation of chemotypes that are potentially adapted to their niche conditions including the plant-fungal interactions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Collectively, plants produce a huge variety of secondary metabolites (SMs) which are involved in the adaptation of plants to biotic and abiotic stresses. The most characteristic feature of SMs is their striking inter- and intraspecific chemical diversity. Cytochrome P450 monooxygenases (CYPs) often play an important role in the biosynthesis of SMs and thus in the evolution of chemical diversity. Here we studied the diversity and evolution of CYPs of two Jacobaea species which contain a characteristic group of SMs namely the pyrrolizidine alkaloids (PAs)., Results: We retrieved CYPs from RNA-seq data of J. vulgaris and J. aquatica, resulting in 221 and 157 full-length CYP genes, respectively. The analyses of conserved motifs confirmed that Jacobaea CYP proteins share conserved motifs including the heme-binding signature, the PERF motif, the K-helix and the I-helix. KEGG annotation revealed that the CYPs assigned as being SM metabolic pathway genes were all from the CYP71 clan but no CYPs were assigned as being involved in alkaloid pathways. Phylogenetic analyses of full-length CYPs were conducted for the six largest CYP families of Jacobaea (CYP71, CYP76, CYP706, CYP82, CYP93 and CYP72) and were compared with CYPs of two other members of the Asteraceae, Helianthus annuus and Lactuca sativa, and with Arabidopsis thaliana. The phylogenetic trees showed strong lineage specific diversification of CYPs, implying that the evolution of CYPs has been very fast even within the Asteraceae family. Only in the closely related species J. vulgaris and J. aquatica, CYPs were found often in pairs, confirming a close relationship in the evolutionary history., Conclusions: This study discovered 378 full-length CYPs in Jacobaea species, which can be used for future exploration of their functions, including possible involvement in PA biosynthesis and PA diversity.

The evolution of increased competitive ability (EICA) hypothesis and the shifting defense hypothesis (SDH) predict that evolutionary changes occur in a suite of traits related to defense and growth in invasive plant species as result of the absence of specialist herbivores. We tested how this suite of traits changed due to the absence of specialist herbivores in multiple invasive regions that differ in climatic conditions with native and invasive Jacobaea vulgaris in a controlled environment. We hypothesized that invasive J. vulgaris in all invasive regions have i) a higher plant growth and underlying traits, such as photosynthetic capacity, ii) lower regrowth-related traits, such as carbohydrate storage, and iii) an increased plant qualitative defense, such as pyrrolizidine alkaloids (PAs). Our results show that invasive J. vulgaris genotypes have evolved a higher photosynthetic rate and total PA concentration but a lower investment in root carbohydrates, which supports the SDH hypothesis. All the traits changed consistently and significantly in the same direction in all four invasive regions, indicative of a parallel evolution. Climatic and soil variables did differ between ranges but explained only a very small part of the variation in trait values. The latter suggests that climate and soil changes were not the main selective forces on these traits.

Historically, latex-bearing plants have been regarded as important medicinal resources in many countries due to their characteristic latex ingredients. They have also often been endowed with a social or cultural significance in religious or cult rituals or for hunting. Initial chemical studies focused on the protein or peptide content but recently the interest extended to smaller molecules. Latex has been found to contain a broad range of specialized metabolites such as terpenoids, cardenolides, alkaloids, and phenolics, which are partly responsible for their antibacterial, antifungal, anthelmintic, cytotoxic, and insect-repellent activities. The diversity in biology and chemistry of latexes is supposedly associated to their ecological roles in interactions with exogenous factors. Latexes contain unique compounds that are different to those found in their bearing plants. Exploring the feasibility of plant latex as a new type of bioactive chemical resource, this review paper covers the chemical characterization of plant latexes, extending this to various other plant exudates. Also, the factors influencing this chemical differentiation and the production, transportation, and chemistry of the latex exudates are described, based on ecological and biochemical mechanisms. We also proposed a latex coagulation model involving 4 general conserved steps. Therefore, the inherent defensive origin of latexes is recognized as their most valuable character and encourages one to pay attention to these materials as alternative sources to discover metabolites with insecticidal or antimicrobial activity., Competing Interests: The authors declare that they have no conflict of interest., (Georg Thieme Verlag KG Stuttgart · New York.)

Western flower thrips (WFT) are a major pest on many crops, including tomato. Thrips cause yield losses, not only through feeding damage, but also by the transmission of viruses of which the Tomato Spotted Wilt Virus is the most important one. In cultivated tomato, genetic diversity is extremely low, and all commercial lines are susceptible to WFT. Several wild relatives are WFT resistant and these resistances are based on glandular trichome-derived traits. Introgression of these traits in cultivated lines did not lead to WFT resistant commercial varieties so far. In this study, we investigated WFT resistance in cultivated tomato using a F2 population derived from a cross between a WFT susceptible and a WFT resistant cultivated tomato line. We discovered that this WFT resistance is independent of glandular trichome density or trichome-derived volatile profiles and is associated with three QTLs on chromosomes 4, 5 and 10. Foliar metabolic profiles of F3 families with low and high WFT feeding damage were clearly different. We identified α-tomatine and a phenolic compound as potential defensive compounds. Their causality and interaction need further investigation. Because this study is based on cultivated tomato lines, our findings can directly be used in nowadays breeding programs., (� The Author(s) 2019. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Introduction: To date, most studies of natural variation and metabolite quantitative trait loci (mQTL) in tomato have focused on fruit metabolism, leaving aside the identification of genomic regions involved in the regulation of leaf metabolism., Objective: This study was conducted to identify leaf mQTL in tomato and to assess the association of leaf metabolites and physiological traits with the metabolite levels from other tissues., Methods: The analysis of components of leaf metabolism was performed by phenotypying 76 tomato ILs with chromosome segments of the wild species Solanum pennellii in the genetic background of a cultivated tomato (S. lycopersicum) variety M82. The plants were cultivated in two different environments in independent years and samples were harvested from mature leaves of non-flowering plants at the middle of the light period. The non-targeted metabolite profiling was obtained by gas chromatography time-of-flight mass spectrometry (GC-TOF-MS). With the data set obtained in this study and already published metabolomics data from seed and fruit, we performed QTL mapping, heritability and correlation analyses., Results: Changes in metabolite contents were evident in the ILs that are potentially important with respect to stress responses and plant physiology. By analyzing the obtained data, we identified 42 positive and 76 negative mQTL involved in carbon and nitrogen metabolism., Conclusions: Overall, these findings allowed the identification of S. lycopersicum genome regions involved in the regulation of leaf primary carbon and nitrogen metabolism, as well as the association of leaf metabolites with metabolites from seeds and fruits.

Plants produce an extremely diverse array of metabolites that mediate many aspects of plant-environment interactions. In the context of plant-herbivore interactions, it is as yet poorly understood how natural backgrounds shape the bioactivity of individual metabolites. We tested the effects of a methanol extract of Jacobaea plants and five fractions derived from this extract, on survival of western flower thrips (WFT). When added to an artificial diet, the five fractions all resulted in a higher WFT survival rate than the methanol extract. In addition, their expected combined effect on survival, assuming no interaction between them, was lower than that of the methanol extract. The bioactivity was restored when the fractions were combined again in their original proportion. These results strongly suggest synergistic interactions among the fractions on WFT survival rates. We then tested the effects of two pyrrolizidine alkaloids (PAs), free base retrorsine and retrorsine N-oxide, alone and in combination with the five shoot fractions on WFT survival. The magnitude of the effects of the two PAs depended on the fraction to which they were added. In general, free base retrorsine was more potent than retrorsine N-oxide, but this was contingent on the fraction to which these compounds were added. Our results support the commonly held, though seldom tested, notion that the efficacy of plant metabolites with respect to plant defence is dependent on their phytochemical background. It also shows that the assessment of bioactivity cannot be decoupled from the natural chemical background in which these metabolites occur.

In this study we investigated the effect of methyl jasmonate (MeJA) application on pyrrolizidine alkaloid (PA) concentration and composition of two closely related Jacobaea species. In addition, we examined whether MeJA application affected herbivory of the polyphagous leaf feeding herbivore Spodoptera exigua. A range of concentrations of MeJA was added to the medium of Jacobaea vulgaris and J. aquatica tissue culture plants grown under axenic conditions. PA concentrations were measured in roots and shoots using LC-MS/MS. In neither species MeJA application did affect the total PA concentration at the whole plant level. In J. vulgaris the total PA concentration decreased in roots but increased in shoots. In J. aquatica a similar non-significant trend was observed. In both Jacobaea species MeJA application induced a strong shift from senecionine- to erucifoline-like PAs, while the jacobine- and otosenine-like PAs remained largely unaffected. The results show that MeJA application does not necessarily elicits de novo synthesis, but rather leads to PA conversion combined with reallocation of certain PAs from roots to shoots. S. exigua preferred feeding on control leaves of J. aquatica over MeJA treated leaves, while for J. vulgaris both the control and MeJA treated leaves were hardly eaten. This suggests that the MeJA-induced increase of erucifoline-like PAs can play a role in resistance of J. aquatica to S. exigua. In J. vulgaris resistance to S. exigua may already be high due to the presence of jacobine-like PAs or other resistance factors.

Ultraviolet radiation (UV) is an important modulator of plant defenses against biotic stresses. We have recently described that different supplemental UV exposure times and irradiance intensities enhanced tomato (Solanum lycopersicum) resistance to Western flower thrips (Frankliniella occidentalis). UV increased jasmonic acid-isoleucine (JA-Ile) and salicylic acid (SA) levels, as well as the expression of JA- and SA-responsive genes, before thrips herbivory. Here we report how UV affects tomato defense responses upon thrips infestation, and resistance to pathogens that are susceptible to the activation of SA-associated defenses. Our experiments reveal that, at 7 days after thrips infestation, UV did not enhance the levels of jasmonates, auxin or abscisic acid. UV also did not affect the expression of JA-responsive genes in the cultivar Moneymaker, the jasmonate deficient mutant def-1, the type-VI trichome deficient mutant od-2, or their wild-type Castlemart. However, UV strongly activated SA-associated defense responses in def-1 after thrips infestation. Further bioassays showed that UV increased def-1 resistance to the hemi-biotrophic bacterial pathogen Pseudomonas syringae pv. tomato DC3000, which is susceptible to SA-mediated defenses. Our results suggest that UV might enhance tomato resistance to this pathogen in the JA deficient genotype through the activation of SA defenses.

Ultraviolet (UV) radiation can modulate plant defenses against herbivorous arthropods. We investigated how different UV exposure times and irradiance intensities affected tomato (Solanum lycopersicum) resistance to thrips (Frankliniella occidentalis) by assessing UV effects on thrips-associated damage and host-selection, selected metabolite and phytohormone contents, expression of defense-related genes, and trichome density and chemistry, the latter having dual roles in defense and UV protection. Short UV daily exposure times increased thrips resistance in the cultivar 'Moneymaker' but this could not be explained by changes in the contents of selected leaf polyphenols or terpenes, nor by trichome-associated defenses. UV irradiance intensity also affected resistance to thrips. Further analyses using the tomato mutants def-1, impaired in jasmonic acid (JA) biosynthesis, od-2, defective in the production of functional type-VI trichomes, and their wild-type, 'Castlemart', showed that UV enhanced thrips resistance in Moneymaker and od-2, but not in def-1 and Castlemart. UV increased salicylic acid (SA) and JA-isoleucine concentrations, and increased expression of SA- and JA-associated genes in Moneymaker, while inducing expression of JA-defensive genes in od-2. Our results demonstrate that UV-mediated enhancement of tomato resistance to thrips is probably associated with the activation of JA-associated signaling, but not with plant secondary metabolism or trichome-related traits.

In cultivated tomato (Solanum lycopersicum), increases in photosynthetically active radiation (PAR) induce type VI leaf glandular trichomes, which are important defensive structures against arthropod herbivores. Yet, how PAR affects the type VI trichome-associated leaf chemistry and its biological significance with respect to other photomorphogenic responses in this agronomically important plant species is unknown. We used the type VI trichome-deficient tomato mutant odorless-2 (od-2) and its wild type to investigate the influence of PAR on trichome-associated chemical defenses against thrips (Frankliniella occidentalis). High PAR increased thrips resistance in wild-type plants, but not in od-2. Furthermore, under high PAR, thrips preferred od-2 over the wild type. Both genotypes increased type VI trichome densities under high PAR. Wild-type plants, however, produced more trichome-associated allelochemicals, i.e. terpenes and phenolics, these being undetectable or barely altered in od-2. High PAR increased leaf number and thickness, and induced profound but similar metabolomic changes in wild-type and od-2 leaves. Enhanced PAR also increased levels of ABA in wild-type and od-2 plants, and of auxin in od-2, while the salicylic acid and jasmonate concentrations were unaltered. However, in both genotypes, high PAR induced the expression of jasmonic acid-responsive defense-related genes. Taken together, our results demonstrate that high PAR-mediated induction of trichome-associated chemical defenses plays a prominent role in tomato-thrips interactions.

Variation in the induction of plant defenses along the plant canopy can determine distribution and colonization of arthropod herbivores within the plant. In tomato, type VI glandular trichomes, which are epidermal defensive structures, and their derived volatiles are induced by the phytohormone jasmonic acid (JA). How JA-mediated induction of these trichome-associated chemical defenses depends on the leaf developmental stage and correlates with resistance against herbivory is unknown. We showed that application of JA reduced thrips-associated damage, however the amplitude of this response was reduced in the fully developed leaves compared to those still developing. Although JA increased type-VI trichome densities in all leaf developmental stages, as well as JA-inducible defensive proteins, these increases were stronger in developing leaves. Remarkably, the concentration of trichome-derived volatiles was induced by JA to a larger degree in developing leaves than in fully developed leaves. In fully developed leaves, the increase in trichome-derived volatiles was explained by an enhanced production per trichome, while in developing leaves this was mainly caused by increases in type-VI trichome densities. Together, we showed that JA-mediated induction of trichome density and chemistry depends on leaf development stage, and it might explain the degree of thrips-associated leaf damage in tomato., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Resins are one of the first sites of interaction between plants and biotic and abiotic factors. Despite their evident morphological and chemical differentiation from other plant organs, the detailed correlation between resins and biological or environmental factors is not yet clear. In this study, 1 H nuclear magnetic resonance (NMR), gas chromatography coupled with mass spectrometry (GC-MS) and high-performance thin-layer chromatography (HPTLC)-based profiling techniques were applied to the metabolic characterisation of plant resins of different species and season of collection, using samples from five different species that were collected during early and late spring. The 1 H NMR analysis confirmed the main metabolic groups in the resins to be terpenoids and further GC-MS analysis revealed a notable chemical variation between the species and collection periods. Abies grandis displayed a significant differentiation from the other species, showing a higher number of monoterpenes. The HPTLC-based profiling method hyphenated with multivariate data analysis (MVDA) also showed a clear separation confirming the GC-MS terpenoidal profiling results. Additionally, the unknown compounds were obtained by preparative TLC for identification. Based on the results of the three analytical platforms, it was concluded that the major difference in chemical composition of pine species was between species rather than the collection period. Nonetheless, the chemical profiles of resins from different species and collection periods can be well discriminated and correlated to mono- and sesquiterpenes in the case of species and diterpenes for the collection periods., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

We hypothesized that due to the absence of specialist herbivores in introduced ranges, invasive plants have evolved decreased allocation to carbohydrate storage for regrowth ability and as a consequence allocate more to growth. In this study, we compared plant growth, carbohydrate storage and regrowth ability of invasive and native Jacobaea vulgaris in response to complete shoot defoliation. We used invasive J. vulgaris genotypes from three geographically and climactically distinct regions and compared these with native genotypes from Europe. We found that invasive genotypes initially grew larger while native genotypes regrew larger after defoliation. Before defoliation, the carbohydrate storage in roots of invasive genotypes was 38% lower than native genotypes. Biomass after regrowth increased with root carbohydrate storage while it decreased with structural root mass, showing that it is crucial to study root storage and structural components separately in order to investigate plant regrowth. All studied traits of invasive populations from the three geographically and climatologically distinct regions changed in the same expected direction suggesting that the shifts in herbivore guild were causal to the observed change in growth and regrowth ability rather than environmental factors.

Western flower thrips (WFT) Frankliniella occidentalis (Pergande) is a key agricultural pest of cultivated tomatoes. Induced host plant resistance by activating jasmonic acid (JA) signaling pathway constitutes a promising method for WFT control. The phytotoxin coronatine (COR), produced by Pseudomonas syringae pv. tomato DC3000 ( Pst ), mimics the plant hormone JA-Isoleucine and can promote resistance against herbivorous arthropods. Here we determined the effect of Pst and COR on tomato resistance against WFT, induction of JA and salicylic acid (SA) associated defenses, and plant chemistry. Additionally, we investigated the presence of other components in Pst -derived and filtered culture medium, and their interactive effect with COR on tomato resistance to WFT. Our results showed that infiltration of COR or Pst reduced WFT feeding damage in tomato plants. COR and Pst induced the expression of JA-associated gene and protein marker. COR also induced expression of a SA-related responsive gene, although at much less magnitude. Activation of JA defenses in COR and Pst infiltrated plants did not affect density of type VI leaf trichomes, which are defenses reported to be induced by JA. An untargeted metabolomic analysis showed that both treatments induced strong changes in infiltrated leaves, but leaf responses to COR or Pst slightly differed. Application of the Pst -derived and filtered culture medium, containing COR but not viable Pst , also increased tomato resistance against WFT confirming that the induction of tomato defenses does not require a living Pst population to be present in the plant. Infiltration of tomato plants with low concentrations of COR in diluted Pst -derived and filtered culture medium reduced WFT feeding damage in a greater magnitude than infiltration with an equivalent amount of pure COR indicating that other elicitors are present in the medium. This was confirmed by the fact that the medium from a COR-mutant of Pst also strongly reduced silver damage. In conclusion, our results indicate that induction of JA defenses by COR, Pst infection, the medium of Pst and the medium of a Pst COR - mutant increased resistance against WFT. This was not mediated by the reinforcement of leaf trichome densities, but rather the induction of chemical defenses.

Plants have developed diverse defence mechanisms to ward off herbivorous pests. However, agriculture still faces estimated crop yield losses ranging from 25% to 40% annually. These losses arise not only because of direct feeding damage, but also because many pests serve as vectors of plant viruses. Herbivorous thrips (Thysanoptera) are important pests of vegetable and ornamental crops worldwide, and encompass virtually all general problems of pests: they are highly polyphagous, hard to control because of their complex lifestyle, and they are vectors of destructive viruses. Currently, control management of thrips mainly relies on the use of chemical pesticides. However, thrips rapidly develop resistance to these pesticides. With the rising demand for more sustainable, safer, and healthier food production systems, we urgently need to pinpoint the gaps in knowledge of plant defences against thrips to enable the future development of novel control methods. In this review, we summarize the current, rather scarce, knowledge of thrips-induced plant responses and the role of phytohormonal signalling and chemical defences in these responses. We describe concrete opportunities for breeding resistance against pests such as thrips as a prototype approach for next-generation resistance breeding.

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., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The high structural diversity of plant metabolites suggests that interactions among them should be common. We investigated the effects of single metabolites and combinations of plant metabolites on insect herbivores. In particular we studied the interacting effects of pyrrolizidine alkaloid (PAs), and chlorogenic acid (CGA), on a generalist herbivore, Frankliniella occidentalis. We studied both the predominantly occurring PA N -oxides and the less frequent PA free bases. We found antagonistic effects between CGA and PA free bases on thrips mortality. In contrast PA N -oxides showed synergistic interactions with CGA. PA free bases caused a higher thrips mortality than PA N -oxides while the reverse was through for PAs in combination with CGA. Our results provide an explanation for the predominate storage of PA N -oxides in plants. We propose that antagonistic interactions represent a constraint on the accumulation of plant metabolites, as we found here for Jacobaea vulgaris . The results show that the bioactivity of a given metabolite is not merely dependent upon the amount and chemical structure of that metabolite, but also on the co-occurrence metabolites in, e.g., plant cells, tissues and organs. The significance of this study is beyond the concerns of the two specific groups tested here. The current study is one of the few studies so far that experimentally support the general conception that the interactions among plant metabolites are of great importance to plant-environment interactions.

Plant defenses inducible by herbivorous arthropods can determine performance of subsequent feeding herbivores. We investigated how infestation of tomato (Solanum lycopersicum) plants with the Western flower thrips (Frankliniella occidentalis) alters host plant suitability and foraging decisions of their conspecifics. We explored the role of delayed-induced jasmonic acid (JA)-mediated plant defense responses in thrips preference by using the tomato mutant def-1, impaired in JA biosynthesis. In particular, we investigated the effect of thrips infestation on trichome-associated tomato defenses. The results showed that when offered a choice, thrips preferred non-infested plants over infested wild-type plants, while no differences were observed in def-1. Exogenous application of methyl jasmonate restored the repellency effect in def-1. Gene expression analysis showed induction of the JA defense signaling pathway in wild-type plants, while activating the ethylene signaling pathway in both genotypes. Activation of JA defenses led to increases in type-VI leaf glandular trichome densities in the wild type, augmenting the production of trichome-associated volatiles, i.e. terpenes. Our study revealed that plant-mediated intraspecific interactions between thrips are determined by JA-mediated defenses in tomato. We report that insects can alter not only trichome densities but also the allelochemicals produced therein, and that this response might depend on the magnitude and/or type of the induction., (© The Author 2017. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists.)

With mounting concerns over health and environmental effects of pesticides, the search for environmentally acceptable substitutes has amplified. Plant secondary metabolites appear in the horizon as an attractive solution for green crop protection. This paper reviews the need for changes in the techniques and compounds that, until recently, have been the mainstay for dealing with pest insects. Here we describe and discuss main strategies for selecting plant-derived metabolites as candidates for sustainable agriculture. The second part surveys ten important insecticidal compounds, with special emphasis on those involved in human health. Many of these insecticidal metabolites, however, are crystalline solids with limited solubility which might potentially hamper commercial formulation. As such, we introduce the concept of natural deep eutectic solvents for enhancing solubility and stability of such compounds. The concept, principles and examples of green pest control discussed here offer a new suite of environmental-friendly tools designed to promote and adopt sustainable agriculture.

Hybridization can lead to novel qualitative or quantitative variation of secondary metabolite (SM) expression that can have ecological and evolutionary consequences. We measured pyrrolizidine alkaloid (PA) expression in the shoots and roots of a family including one Jacobaea vulgaris genotype and one Jacobaea aquatica genotype (parental genotypes), two F(1) hybrid genotypes, and 102 F(2) hybrid genotypes using liquid chromatography-tandem mass spectrometry (LC-MS/MS). We detected 37 PAs in the roots and shoots of J. vulgaris, J. aquatica and the hybrids. PA concentrations and compositions differed between genotypes, and between roots and shoots. Three otosenine-like PAs that only occurred in the shoots of parental genotypes were present in the roots of F(2) hybrids; PA compositions were sometimes novel in F(2) hybrids compared with parental genotypes, and in some cases transgressive PA expression occurred. We also found that PAs from within structural groups covaried both in the roots and in the shoots, and that PA expression was correlated between shoots and roots. Considerable and novel variation present among F(2) hybrids indicates that hybridization has a potential role in the evolution of PA diversity in the genus Jacobaea, and this hybrid system is useful for studying the genetic control of PA expression., (© 2011 Institute of Biology Leiden. New Phytologist © 2011 New Phytologist Trust.)

Sex allocation theory explains how size-related variations in male and female fitness may favor the evolution of size-dependent sex allocation in hermaphrodites. Although empirical studies show that sex allocation changes gradually with size in many species, theoretical studies tend to predict an abrupt sex reversal from one sex to the other, that is, single-sexed sequential hermaphrodites. We show that this discrepancy between data and theory collapses if one takes into account that size affects male and female fitness through distinct routes. Using the classification of budget (larger individuals spend a greater budget on reproduction) and direct (e.g., larger plants are taller and may disperse pollen more efficiently) effects of size suggested by Klinkhamer et al., we propose a simple general framework appropriately incorporating these two categories of size effects in male and female fitness expressions. Analytical and numerical results show that a gradual sex change is evolutionarily stable for a large set of parameter values. Sex reversal is selected only in the absence of budget effects of size. We provide further predictions on size-dependent sex allocation and assess the relative importance of budget and direct effect for creating different patterns.

We measured the relationship between selfing rates and flower number in an experimental population of bumblebee pollinated Cynoglossum officinale, with plants differing in flower number. Results were compared with the prediction of a model based on pollen dynamics and pollinator behaviour. The selfing rate, as measured by multilocus oligonucleotide DNA fingerprinting, increased with flower number and ranged from 0% to 70%. Flowers on large plants received an equal number of visits from bumblebees as flowers on small plants. On large plants more flowers in a row were visited, inducing geitonogamy. The overall relationship between selfing rate and number of flowers can be explained by pollen dynamics and pollinator behaviour without invoking postpollination processes such as differential pollen tube growth and abortion., (Blackwell Science Ltd.)

In a study of Cynoglossum officinale L. it was found that the number of seeds per flower increased with plant weight, and that flower production was inversely related to the number of seeds per flower. As a result, the number of seeds and total seed weight increased proportionally with plant weight. Seed size was not related to plant weight. Differences in seed size within the individual were, in part, related to the position of the seed on the cyme and not related to the number of seeds within a flower. The relationship, between plant size and female fitness found in this study is discussed in relation to life-history theory.