Multiple indole glucosinolates and myrosinases defend Arabidopsis against Tetranychus urticae herbivory

Arabidopsis (Arabidopsis thaliana) defenses against herbivores are regulated by the jasmonate (JA) hormonal signaling pathway, which leads to the production of a plethora of defense compounds. Arabidopsis defense compounds include tryptophan-derived metabolites, which limit Arabidopsis infestation by the generalist herbivore two-spotted spider mite, Tetranychus urticae. However, the phytochemicals responsible for Arabidopsis protection against T. urticae are unknown. Here, we used Arabidopsis mutants disrupted in the synthesis of tryptophan-derived secondary metabolites to identify phytochemicals involved in the defense against T. urticae. We show that of the three tryptophan-dependent pathways found in Arabidopsis, the indole glucosinolate (IG) pathway is necessary and sufficient to assure tryptophan-mediated defense against T. urticae. We demonstrate that all three IGs can limit T. urticae herbivory, but that they must be processed by myrosinases to hinder T. urticae oviposition. Putative IG breakdown products were detected in mite-infested leaves, suggesting in planta processing by myrosinases. Finally, we demonstrate that besides IGs, there are additional JA-regulated defenses that control T. urticae herbivory. Together, our results reveal the complexity of Arabidopsis defenses against T. urticae that rely on multiple IGs, specific myrosinases, and additional JA-dependent defenses.

Three IGs and specific myrosinases help protect Arabidopsis thaliana against herbivory by the two-spotted spider mite T. urticae.     

Concentration of glucosinolates in relation to habitat and insect herbivory for the native crucifer Cardamine cordifolia

Understanding plant-insect interactions requires further data on herbivory in relation to the variation in concentration of characteristic secondary compounds. We report here analyses of the glucosinolate contents for a native perennial, montane crucifer Cardamine cordifolia in relation to: (a) plant characteristics; (b) insect herbivory; and (c) habitat. The only pattern of variation of glucosinolate content with leaf characteristics found was an inverse correlaton between leaf weight and total isothiocyanate-yielding glucosinolates (IYG) in shaded plants. There was a highly significant, negative relationship between total IYG and leaf damage by insects, particularly in typical shaded habitats. Higher insect-caused damage on denser, smaller leaves of plants from the driest soils was observed. Additionally, plants occurring in sun-exposed habitats from the beginning of the growing season, both naturally and experimentally, had similar (or lower) concentrations of total IYG, and were significantly more damaged by insects, than those in the more usual shaded habitats. The experimental removal of shade cover in mid-season resulted in significantly elevated quantities of total IYG in the first year, with a relaxation of that stress-induced response in the second year. We suggest that the insect herbivore guild on Cardamine cordifolia responds to concentration and composition of glucosinolates and exerts its greatest pressure on plants with lower concentrations. Differential herbivory, consumption mediated in part by glucosinolate concentration, appears to contribute to microhabitat occurrence of C. cordifolia.     

The role of isoprene in insect herbivory

Several hypotheses have previously been proposed to explain the function of isoprene in plants, including its ability to protect the leaf metabolic machinery from transient high temperature^1,2 and from oxidative stress.³ Isoprene may also serve as a metabolic overflow mechanism for carbon or photosynthetic energy^4–6 and may promote flowering in neighbouring plants.⁷ We have reported recently that isoprene can be detected by a herbivore, Manduca sexta, and that it directly deters them from feeding, with an isoprene emission threshold level of <6 nmol m⁻² s⁻¹.⁸ We demonstrated this using both in vivo experiments, using isoprene-emitting transgenic tobacco plants (Nicotiana tabacum cv. Samsun) and non-emitting azygous control plants, and in vitro experiments, using an artificial (isoprene-emitting and non-emitting control) diet. Here we discuss the potential role of isoprene in plant-herbivore interactions and the possibility that isoprene actually serves multiple purposes in plants.Key words: multiple functions, deterrence, signal, Manduca sextaIsoprene (C₅H₈; 2-methyl 1,3-butadiene) is a volatile organic compound that is produced in many, but not all, plant species.⁹ Because of its high volatility, once produced, isoprene is rapidly released from the leaf surface into the atmosphere, the chemical and physical properties of which can be altered due to the high chemical reactivity and large mass flux rate of isoprene.^10,11 While isoprene production in plants consumes considerable amounts of energy,¹² its role in plants has not yet been fully explained. Isoprene production and emission may confer significant benefits for plants that balance or outweigh its high production costs,¹³ but why only some plants produce the compound and why several beneficial effects have been demonstrated is not clear.Isoprene has recently been reported to directly deter M. sexta caterpillars from feeding. This is supported both by in vivo and in vitro studies and we propose that isoprene may confer competitive advantage to plant against herbivory.⁸ The reason why isoprene functions as a deterrent is not clear. The very obvious explanation is that isoprene is a harmful substance acting, for example as a digestibility reducing substance or toxin, and that it might operate together with other types of plant defences. However, another possibility is that isoprene might be a harmless deterrent as, for example, in the case of the alkaloid gramine in grasses. A grasshopper (Locusta migratoria) normally avoids this compound, but when force fed, habituation occurs and chronic intake does not affect herbivore fitness.¹⁴ This type of substance may be a chemical mimic or be associated with other more toxic compounds.¹⁵ It is also possible that the avoidance behaviour towards harmless deterrents may not be associated at all with toxicity but rather with the avoidance of non-host plants,¹⁶ or avoidance of unsuitable food sources to reduce the risk of feeding on a toxic plant.¹⁷Using nutritional indices, it is possible to investigate the effect of a compound on food consumption and utilisation.¹⁸ We therefore investigated the effect of isoprene on food consumption and utilization by M. sexta using nutritional indices. In this experiment, third instar caterpillars were force fed on isoprene-emitting or non-emitting artificial diets for 24 hour. There were no significant differences (p > 0.05, n = 27) in relative growth rate, consumption index, approximate digestibility, efficiency of conversion of ingested food to body mass and efficiency of conversion of digested food to body mass between the caterpillars that fed on the isoprene emitting-diet and those that fed on the non-emitting diet (Fig. 1). This indicates that isoprene does not directly influence food uptake and utilization by the caterpillars, and that the observed avoidance behaviour⁸ may be related to avoidance of isoprene behaving as a ‘non-host plant’ signal, or isoprene might act in association with other defence mechanism in plants.Open in a separate windowFigure 1Quantification of food consumption and utilization of third instar M. sexta larvae. Larvae fed on isoprene-emitting (55 nmol m⁻² s⁻¹) or non-emitting artificial diet for 24 hours. Larval weight gained, frass produced and leaf weight consumed were measured and analysed according to Waldbauer nutritional indices (1968). ANOVA was used to analyse the data at 95% confidence and no significant confidence was found (n = 27). RGR, relative growth rate; CI, consumption index; AD, approximate digestibility; ECI, efficiency of conversion of ingested food to body mass; ECD, efficiency of conversion of digested food to body mass.It should be noted that this experiment was only performed over 24 hours and it therefore does not probe the longer term effect of isoprene on caterpillar fitness. The function of isoprene in plant-herbivore interactions should therefore be investigated further by extending the experimental period and measurement of caterpillar development, both by using in vitro (isoprene-emitting and non-emitting artificial diet) and in vivo (isoprene-emitting and non-emitting transgenic plants) model systems. The recent availability of transgenic plants, including (a) one that does not normally emit isoprene but is induced to produce isoprene^19,20 and (b) one that normally produces isoprene but has its isoprene emission suppressed,² can facilitate this research and provide insights into the function of isoprene in planta.Various adaptation strategies for reducing the production costs of terpenes have been proposed, including sharing biosynthetic enzymes among multiple pathways, minimizing enzyme turnover rate, using a single enzyme to generate multiple products and using products for more than one function.²¹ The possibility of isoprene being a compound with multiple functions has been hypothesized,⁸ as isoprene has been shown, by inhibitor studies and genetic manipulation, to be able to defend plants from high temperature episodes^1,2 and against ozone³ and now also to protect against herbivory.⁸ It has also been shown to promote flowering in neighboring plants.⁷ Further investigations are, however, required to determine if isoprene emission can deter herbivore insects from a wide range of plant species, or whether this and other effects are specific to plant species. The multiple functions of isoprene may explain how the benefits of isoprene production by plants can outweigh the costs of its production, and why some, but not all, plants produce this reactive and volatile compound.     

The Arabidopsis extracellular UNUSUAL SERINE PROTEASE INHIBITOR functions in resistance to necrotrophic fungi and insect herbivory

Protease inhibitors (PIs) function in the precise regulation of proteases, and are thus involved in diverse biological processes in many organisms. Here, we studied the functions of the Arabidopsis UNUSUAL SERINE PROTEASE INHIBITOR (UPI) gene, which encodes an 8.8 kDa protein of atypical sequence relative to other PIs. Plants harboring a loss‐of‐function UPI allele displayed enhanced susceptibility to the necrotrophic fungi Botrytis cinerea and Alternaria brassicicola as well as the generalist herbivore Trichoplusia ni. Further, ectopic expression conferred increased resistance to B. cinerea and T. ni. In contrast, the mutant has wild‐type responses to virulent, avirulent and non‐pathogenic strains of Pseudomonas syringae, thus limiting the defense function of UPI to necrotrophic fungal infection and insect herbivory. Expression of UPI is significantly induced by jasmonate, salicylic acid and abscisic acid, but is repressed by ethylene, indicating complex phytohormone regulation of UPI expression. The upi mutant also shows significantly delayed flowering, associated with decreased SOC1 expression and elevated levels of MAF1, two regulators of floral transition. Recombinant UPI strongly inhibits the serine protease chymotrypsin but also weakly blocks the cysteine protease papain. Interestingly, jasmonate induces intra‐ and extracellular UPI accumulation, suggesting a possible role in intercellular or extracellular functions. Overall, our results show that UPI is a dual‐specificity PI that functions in plant growth and defense, probably through the regulation of endogenous proteases and/or those of biotic invaders.     

Piecing together the transport pathway of aliphatic glucosinolates

Glucosinolates are sulfur-rich secondary metabolites characteristic of the Brassicales order. Transport of glucosinolates was suggested more than 30 years ago through a number of studies which indicated that glucosinolates are produced in maternal tissue and subsequently transported to the seed. These observations laid the foundation for numerous studies on glucosinolate transport which have provided a wealth of information on biochemical properties of glucosinolate transport, source–sink relationships between organs and on the transport routes of glucosinolates. However, most of the conclusions and hypotheses proposed in these studies have not been discussed in context of each other to provide a complete overview of the current state of knowledge on glucosinolate transport. In this review, we are thus piecing together the glucosinolate pathway by presenting and critically analyzing all data on glucosinolate research. Furthermore, the data on glucosinolate transport is considered in the light of the newest findings on glucosinolate synthesis and distribution. The aim is to provide a comprehensive and updated set of hypotheses which may prove useful in directing future research on glucosinolate transport.     

Pronounced effects of slug herbivory on seedling recruitment of Brassica cultivars and accessions,especially those with low levels of aliphatic glucosinolates

In demographic studies on Brassica cultivars and accessions we observed large genotypic variation in the ability of seedlings to establish. Here we quantify the role of slugs during establishment by including and excluding slug herbivory.Slug exclusion during the first 10 days after germination led to 26.5 times more surviving seedlings in modern canola varieties of Brassica napus, 4.3 times more in feral B. napus and 1.9 times more in wild Brassica rapa. While seed and seedlings of canola were low in aliphatic glucosinolates (AGS), feral B. napus and wild B. rapa had much higher AGS levels. Consequently, we find a positive correlation between the survival of young seedlings and AGS concentration. Concentrations of indole glucosinolates (IGS) in seeds were much lower than those of AGS and did not correlate significantly with survival. Subsequently, we exposed 10-day-old seedlings to herbivory. In the 4 days following exposure, slug herbivory was negatively correlated to AGS concentration but not to IGS.In choice experiments in the lab, the preference of the slug Arion lusitanicus also correlated negatively with AGS content. A. lusitanicus preferred to feed on B. napus rather than on wild B. rapa, but had no significant preference when presented with plants similar in AGS content.Slugs can be a limiting factor for seedling recruitment in populations of B. napus, especially for modern canola cultivars with a low AGS content.     

Comparative analysis of quantitative trait loci controlling glucosinolates,myrosinase and insect resistance in Arabidopsis thaliana

Evolutionary interactions among insect herbivores and plant chemical defenses have generated systems where plant compounds have opposing fitness consequences for host plants, depending on attack by various insect herbivores. This interplay complicates understanding of fitness costs and benefits of plant chemical defenses. We are studying the role of the glucosinolate-myrosinase chemical defense system in protecting Arabidopsis thaliana from specialist and generalist insect herbivory. We used two Arabidopsis recombinant inbred populations in which we had previously mapped QTL controlling variation in the glucosinolate-myrosinase system. In this study we mapped QTL controlling resistance to specialist (Plutella xylostella) and generalist (Trichoplusia ni) herbivores. We identified a number of QTL that are specific to one herbivore or the other, as well as a single QTL that controls resistance to both insects. Comparison of QTL for herbivory, glucosinolates, and myrosinase showed that T. ni herbivory is strongly deterred by higher glucosinolate levels, faster breakdown rates, and specific chemical structures. In contrast, P. xylostella herbivory is uncorrelated with variation in the glucosinolate-myrosinase system. This agrees with evolutionary theory stating that specialist insects may overcome host plant chemical defenses, whereas generalists will be sensitive to these same defenses.     

Reducing the impact of insect herbivory in eucalypt plantations through management of extrinsic influences on tree vigour

In this paper it is argued that concepts developed in ecologically derived insect–plant interaction models can contribute directly to the management of insect herbivory in eucalypt plantations. Common to most species of commercially planted eucalypt is their genetic potential for early rapid growth. Several plant defence theories predict that intrinsically fast growing plants are able to tolerate relatively high levels of herbivory. The risk of this strategy failing increases when plants are exposed to external stressful factors that reduce canopy growth and vigour. Results from a young Eucalyptus camaldulensis plantation stressed by moisture deficit and two young Eucalyptus dunnii plantations, stressed by flooding and weed competition, respectively, are summarized. In all three cases, the stress‐inducing agents reduced canopy growth rates and architecture so that the proportion of leaf tissue damaged by insects increased and the tree’s ability to tolerate this damage decreased. Therefore, alleviating tree stress through improved silvicultural practices or improved site selection techniques may indirectly reduce the impact of insect herbivory. In resource‐limiting environments, an alternative approach may be to plant eucalypt species that although slower growing, are predicted to have better defended foliage. Manipulation of these natural antiherbivore plant strategies are not exclusive of other management approaches, such as the need for routine surveillance of key pest insects or the genetic selection of natural insect resistance and selective chemical control techniques, but should be viewed as an overarching concept for plantation health.     

CYP79F1 and CYP79F2 have distinct functions in the biosynthesis of aliphatic glucosinolates in Arabidopsis

Cytochromes P450 of the CYP79 family catalyze the conversion of amino acids to oximes in the biosynthesis of glucosinolates, a group of natural plant products known to be involved in plant defense and as a source of flavor compounds, cancer-preventing agents and bioherbicides. We report a detailed biochemical analysis of the substrate specificity and kinetics of CYP79F1 and CYP79F2, two cytochromes P450 involved in the biosynthesis of aliphatic glucosinolates in Arabidopsis thaliana. Using recombinant CYP79F1 and CYP79F2 expressed in Escherichia coli and Saccharomyces cerevisiae, respectively, we show that CYP79F1 metabolizes mono- to hexahomomethionine, resulting in both short- and long-chain aliphatic glucosinolates. In contrast, CYP79F2 exclusively metabolizes long-chain elongated penta- and hexahomomethionines. CYP79F1 and CYP79F2 are spatially and developmentally regulated, with different gene expression patterns. CYP79F2 is highly expressed in hypocotyl and roots, whereas CYP79F1 is strongly expressed in cotyledons, rosette leaves, stems, and siliques. A transposon-tagged CYP79F1 knockout mutant completely lacks short-chain aliphatic glucosinolates, but has an increased level of long-chain aliphatic glucosinolates, especially in leaves and seeds. The level of long-chain aliphatic glucosinolates in a transposon-tagged CYP79F2 knockout mutant is substantially reduced, whereas the level of short-chain aliphatic glucosinolates is not affected. Biochemical characterization of CYP79F1 and CYP79F2, and gene expression analysis, combined with glucosinolate profiling of knockout mutants demonstrate the functional role of these enzymes. This provides valuable insights into the metabolic network leading to the biosynthesis of aliphatic glucosinolates, and into metabolic engineering of altered aliphatic glucosinolate profiles to improve nutritional value and pest resistance.     

Long-distance phloem transport of glucosinolates in Arabidopsis

Glucosinolates are a large group of plant secondary metabolites found mainly in the order Capparales, which includes a large number of economically important Brassica crops and the model plant Arabidopsis. In the present study, several lines of evidence are provided for phloem transport of glucosinolates in Arabidopsis. When radiolabeled p-hydroxybenzylglucosinolate (p-OHBG) and sucrose were co-applied to the tip of detached leaves, both tracers were collected in the phloem exudates at the petioles. Long-distance transport of [(14)C]p-OHBG was investigated in wild-type and transgenic 35S::CYP79A1 plants, synthesizing high amounts of p-OHBG, which is not a natural constituent of wild-type Arabidopsis. In both wild-type and 35S::CYP79A1 plants, radiolabeled p-OHBG was rapidly transported from the application site into the whole plant and intact p-OHBG was recovered from different tissues. The pattern of distribution of the radioactivity corresponded to that expected for transport of photoassimilates such as sucrose, and was consistent with translocation in phloem following the source-sink relationship. Radiolabeled p-OHBG was shown to accumulate in the seeds of wild-type and 35S::CYP79A1 plants, where p-OHBG had been either exogenously applied or endogenously synthesized from Tyr in the leaves. p-OHBG was found in phloem exudates collected from cut petioles of leaves from both wild-type and 35S::CYP79A1 plants. Phloem exudates were shown to contain intact glucosinolates, and not desulphoglucosinolates, as the transport form. It is concluded that intact glucosinolates are readily loaded into and transported by the phloem.     

植物竞争和昆虫取食调节入侵植物对土壤细菌群落和功能的影响

以入侵植物空心莲子草(Alternanthera philoxeroides(Mart.)Griseb)、本土近缘种莲子草(Alternan-thera sessilis(L.)DC)、生防昆虫莲草直胸跳甲(Agasicles hygrophila(Selman&Vogt))和本地昆虫虾钳菜披龟甲(Cassida pi...     

Molecular mapping of seed aliphatic glucosinolates in Brassica juncea.

An RFLP genomic map with 316 loci was used to study the inheritance of aliphatic glucosinolates in Brassica juncea using doubled-haploid (DH) populations developed from a cross between RLM-514, an agronomically superior non-canola quality B. juncea (high erucic acid and high glucosinolates), and an agronomically poor canola quality B. juncea breeding line. Two QTLs (GSL-A2a and GSL-A2b) associated with 3-butenyl were consistent across years and locations, and explained 75% of the phenotypic variance in the population. Three QTLs (GSL-A2a, GSL-F, GSL-B3) affected 2-propenyl and explained 78% of the phenotypic variance in the population. For total aliphatic glucosinolates, five QTLs explained 30% to 45% of the total phenotypic variance in the population in different environments. Several QTLs (GSL-A7 and GSL-A3) were highly inconsistent in different environments. Major QTLs (GSL-A2a and GSL-A2b) associated with individual glucosinolates were non-significant for total aliphatic glucosinolates. A marker-assisted selection strategy based on QTLs associated with individual glucosinolates rather than total aliphatic glucosinolates is proposed for B. juncea.     

昆虫植食对植物有性生殖的影响

植物在个体发育的各个阶段都与不同的群落成员相互作用,如竞争的植物、有益的传粉者和敌对的植食动物。昆虫植食在各类生态系统中普遍存在,并可能对植物有性生殖产生各种影响。植食昆虫可通过对植物有性生殖结构的消耗直接对植物生殖产生影响,也可通过影响植物资源分配和花性状等改变传粉者服务,从而间接对植物有性生殖带来正面、负面或中性的影响。同一植物的植食昆虫和传粉者往往对植物的吸引性状 （如花大小、气味、颜色等）有相同的偏好,因此植食者与传粉者均能对植物有性生殖性状施加选择压力。本文从昆虫植食对植物有性生殖的直接影响、间接影响以及植食昆虫对植物有性生殖性状选择的影响三个方面进行综述,以期为昆虫植食和生物资源多样性保护相关研究提供参考。     

Bus, a bushy Arabidopsis CYP79F1 knockout mutant with abolished synthesis of short-chain aliphatic glucosinolates

A new mutant of Arabidopsis designated bus1-1 (for bushy), which exhibited a bushy phenotype with crinkled leaves and retarded vascularization, was characterized. The phenotype was caused by an En-1 insertion in the gene CYP79F1. The deduced protein belongs to the cytochrome P450 superfamily. Because members of the CYP79 subfamily are believed to catalyze the oxidation of amino acids to aldoximes, the initial step in glucosinolate biosynthesis, we analyzed the level of glucosinolates in a CYP79F1 null mutant (bus1-1f) and in an overexpressing plant. Short-chain glucosinolates derived from methionine were completely lacking in the null mutant and showed increased levels in the overexpressing plant, indicating that CYP79F1 uses short-chain methionine derivatives as substrates. In addition, the concentrations of indole-3-ylmethyl-glucosinolate and the content of the auxin indole-3-acetic acid and its precursor indole-3-acetonitrile were increased in the bus1-1f mutant. Our results demonstrate for the first time that the formation of glucosinolates derived from methionine is mediated by CYP79F1 and that knocking out this cytochrome P450 has profound effects on plant growth and development.     

Survey of aliphatic glucosinolates in Sicilian wild and cultivated Brassicaceae

In the frame of the activities carried out to exploit Sicilian local cultivars of brassicas, we focused our attention on some of the potential health compounds of various local cruciferous crops. These compounds are of interest to improve the quality of the produce with the aim to develop new cultivars capable of providing functional foods able to prevent disease. In this context, we surveyed for the presence of specific glucosinolates in local cultivars of broccoli, cauliflower, kale, and in some wild species widespread in Sicily, using as control various commercial cultivars. Glucosinolate composition varied extensively among species and crops of the same species, such as cauliflower, broccoli and kale. Cultivar variation for glucosinolate profile was also observed for some crops. For example, Sicilian cultivars of cauliflower possessing colored curds displayed a high content of glucosinolates, glucoraphanin in particular, compared to white curd commercial cultivars. Also some wild species had a high content of other glucosinolates.     

Genetic diversity increases insect herbivory on oak saplings

A growing body of evidence from community genetics studies suggests that ecosystem functions supported by plant species richness can also be provided by genetic diversity within plant species. This is not yet true for the diversity-resistance relationship as it is still unclear whether damage by insect herbivores responds to genetic diversity in host plant populations. We developed a manipulative field experiment based on a synthetic community approach, with 15 mixtures of one to four oak (Quercus robur) half-sib families. We quantified genetic diversity at the plot level by genotyping all oak saplings and assessed overall damage caused by ectophagous and endophagous herbivores along a gradient of increasing genetic diversity. Damage due to ectophagous herbivores increased with the genetic diversity in oak sapling populations as a result of higher levels of damage in mixtures than in monocultures for all families (complementarity effect) rather than because of the presence of more susceptible oak genotypes in mixtures (selection effect). Assemblages of different oak genotypes would benefit polyphagous herbivores via improved host patch location, spill over among neighbouring saplings and diet mixing. By contrast, genetic diversity was a poor predictor of the abundance of endophagous herbivores, which increased with individual sapling apparency. Plant genetic diversity may not provide sufficient functional contrast to prevent tree sapling colonization by specialist herbivores while enhancing the foraging of generalist herbivores. Long term studies are nevertheless required to test whether the effect of genetic diversity on herbivory change with the ontogeny of trees and local adaptation of specialist herbivores.     

Composition and content of glucosinolates in developing Arabidopsis thaliana

The glucosinolate composition and content in various tissues of Arabidopsis thaliana (L.) Heynh. ecotype Columbia during development from seeds to bolting plants were determined in detail by high-performance liquid chromatography. Comparison of the glucosinolate profiles of leaves, roots and stems from mature plants with those of green siliques and mature seeds indicated that a majority of the seed glucosinolates were synthesized de novo in the silique. A comparison of the glucosinolate profile of mature seeds with that of cotyledons indicated that a major part of seed glucosinolates was retained in the cotyledons. Turnover of glucosinolates was studied by germination of seeds containing radiolabelled p-hydroxybenzylglucosinolate (p-OHBG). Approximately 70% of the content of [14C]p-OHBG in the seeds was detected in seedlings at the cotyledon stage and [14C]p-OHBG was barely detectable in young plants with rosettes of six to eight leaves. The turn-over of p-OHBG was found to coincide with the expression of the glucosinolate-degrading enzyme myrosinase, which was detectable at very low levels in seedlings at the cotyledon stage, but which dramatically increased in leaves from plants at later developmental stages. This indicates that there is a continuous turnover of glucosinolates during development and not only upon tissue disruption.     

The ecology of predispersal insect herbivory on tree reproductive structures in natural forest ecosystems

Plant-insect interactions are key model systems to assess how some species affect the distribution, the abundance, and the evolution of others. Tree reproductive structures represent a critical resource for many insect species, which can be likely drivers of demography, spatial distribution, and trait diversification of plants. In this review, we present the ecological implications of predispersal herbivory on tree reproductive structures by insects (PIHR) in forest ecosystems. Both insect's and tree's perspectives are addressed with an emphasis on how spatiotemporal variation and unpredictability in seed availability can shape such particular plant-animal interactions. Reproductive structure insects show strong trophic specialization and guild diversification. Insects evolved host selection and spatiotemporal dispersal strategies in response to variable and unpredictable abundance of reproductive structures in both space and time. If PIHR patterns have been well documented in numerous systems, evidences of the subsequent demographic and evolutionary impacts on tree populations are still constrained by time-scale challenges of experimenting on such long-lived organisms, and modeling approaches of tree dynamics rarely consider PIHR when including biotic interactions in their processes. We suggest that spatially explicit and mechanistic approaches of the interactions between individual tree fecundity and in sect dynamics will clarify predictions of the demogenetic implications of PIHR in tree populations. In a global change context, further experimental and theoretical contributions to the likelihood of life-cycle disruptions between plants and their specialized herbivores, and to how these changes may gen erate novel dynamic patterns in each partner of the interaction are increasingly critical.