No simple sum: seasonal variation in tannin phenotypes and leaf-miners in hybrid oaks

Parent-hybrid asymmetries in the biosynthesis of defense chemistry are believed to affect the distribution of herbivorous insects within plant hybrid zones. While tannins are often produced as complex, dynamic mixtures, the ecological effects of biological and ontogenetic variation in tannin metabolism within hybrid zones are poorly understood. Here, we examine correlations between the seasonal variation of absolute and relative concentrations of individual ellagitannins and total proanthocyanidins in the Quercus gambelii × Q. grisea hybrid complex and a community of leaf-mining Lepidoptera (Gracillariidae, Heliozelidae, Nepticulidae, Tischeriidae), and examine the interactive effects between tannins on leaf-miner abundance. We found strong seasonal variation in oak tannin phenotypes, but two general phenotypes are characteristic of oaks of the Q. gambelii × Q. grisea complex, based upon biosynthetic variation in high molecular weight ellagitannins. Leaf-miner community structure was significantly correlated with the dominant tannin phenotypes in the Q. gambelii × Q. grisea hybrid zone in mid-summer (July, August). Additionally, leaf-miner density was significantly negatively correlated with the ratio of vescalagin-derivatives and total proanthocyanidins in August and September. Overall, hybridization affects biological and ontogenetic variation in ellagitannin and proanthocyanidin biosynthesis within the Q. gambelii × Q. grisea hybrid complex and this variation correlates with herbivore distribution and abundance. Future studies of phenolic chemistry in plant-herbivore interactions in hybrid zones should include a more rigorous quantification of tannin phenotypes as complex, temporally dynamic mixtures.     

Lignoid chemical defenses in the freshwater macrophyte Saururus cernuus

Summary. Chemical defense against herbivores has rarely been investigated for freshwater plants, possibly due to the common misconception that herbivory on aquatic macrophytes is low and would not select for chemical defenses. In previous work, the freshwater angiosperm Saururus cernuus was shown to be a low preference food for omnivorous crayfish despite its high nutrient value and relatively soft texture. We used feeding by the crayfish Procambarus clarkii to guide fractionation of the deterrent lipid-soluble extract of this plant, leading to the identification of seven deterrent lignoid metabolites, (–)-licarin A, (+)-saucernetin, (–)-dihydroguaiaretic acid, (–)-sauriols A and B, (–)-saucerneol, and (–)-saucerneol methyl ether. Lignans have been implicated in terrestrial plant chemical defenses as insect growth inhibitors, insect toxins, nematocides, antibacterial, and antifungal agents. However, these activities have rarely been demonstrated using ecologically relevant methodologies in terrestrial systems, and never before in freshwater systems. The widespread nature of lignans amongst very distantly related plants, along with their rich diversity of molecular structure, suggests that they could play a large role in mediating plant-herbivore interactions. In addition to the lignoid compounds we identified, there were other compounds present in low concentration or unstable compounds that were deterrent, that did not appear to be lignans, but that we were unable to identify. This plant thus appears to be defended by a complex mixture of natural products. Received 6 June 2000; revised 23 August 2000; accepted 2 September 2000     

Differential responses in American (<Emphasis Type="Italic">Castanea dentata</Emphasis> Marshall) and Chinese (<Emphasis Type="Italic">C. mollissima</Emphasis> Blume) chestnut (Falales: Fagaceae) to foliar application of jasmonic acid

Summary.  Tannins are plant defense compounds that exhibit antibiotic (e.g. toxic) and antixenotic (e.g. repellent) effects against an array of plant pests. They are broadly divided into two major groups, hydrolysable tannins (gallotannins and ellagitannins) and proanthocyanidins, each with an undetermined number of compounds. We investigated constitutive levels of hydrolysable tannins and proanthocyanidins in the leaves and stems of American (Castanea dentata Marshall) (Fagales: Fagaceae) and Chinese (C. mollissima Blume) chestnut. American chestnut contained more proanthocyanidins in leaves and stems than Chinese chestnut, but Chinese chestnut contained more foliar hydrolysable tannins. Regardless of these differences, gypsy moth (Lymantria dispar L, Lepidoptera: Lymantriidae) performance did not differ when fed American and Chinese chestnuts. We also investigated the effects of jasmonic acid (JA) treatment on differential tannin induction in American and Chinese chestnut leaves and stems. JA treatment increased proanthocyanidins in American chestnut stems and hydrolysable tannins in both tissue types of American chestnut, but did not influence tannin concentrations in Chinese chestnut leaves or stems. HPLC-ESI-MS analysis of pooled samples suggested that hydrolysable tannins in each tissue were qualitatively comprised primarily of ellagitannins, and JA generally increased the number of hydrolysable tannins that could be detected by ESI-MS. Third, we investigated the performance of gypsy moths on JA treated and untreated American and Chinese chestnut. Caterpillar relative growth was not influenced by JA treatment on Chinese chestnut, but decreased in response to JA application on American chestnut. Our results indicate that JA-dependent defenses differ between these chestnut species. This study improves our understanding of ecologically important differences in tannin induction and herbivore susceptibility in Castanea, and has important implications in efforts for American chestnut restoration and commercial chestnut production.     

Maternal provisioning of sequestered defensive steroids by the Asian snake <Emphasis Type="Italic">Rhabdophis tigrinus</Emphasis>

Summary.   Rhabdophis tigrinus obtains defensive steroids (bufadienolides) from its diet and sequesters those compounds in specialized structures on its neck known as nuchal glands. Hatchling snakes lacking these steroids must acquire them from toads consumed as prey. Here we show that females provision bufadienolides to their offspring in amounts correlated to the quantity in their own nuchal glands; thus, chemically protected mothers produce defended offspring. Bufadienolides can be provisioned to embryos via deposition in yolk and by transfer across the egg membranes within the oviducts. Maternally provisioned bufadienolides persist in the nuchal glands of juvenile snakes from the time of hatching in late summer until the following spring, when toads of ingestible size become abundant. Therefore, maternal provisioning may provide chemical protection from predators for young R. tigrinus in the absence of dietary sources of bufadienolides.     

Cardenolide content,emetic potency,and thin-layer chromatography profiles of monarch butterflies,Danaus plexippus,and their larval host-plant milkweed,Asclepias humistrata,in Florida

Summary This paper is the fourth in a series on cardenolide fingerprints of monarch butterflies (Danaus plexippus, Danainae) and their host-plant milkweeds (Asclepiadaceae) in the eastern United States. Cardenolide concentrations ofAsclepias humistrata plants from north central Florida ranged from 71 to 710 µg/0.1 g dry weight, with a mean of 417 µg/0.1 g. Monarchs reared individually on these plants contained cardenolide concentrations ranging from 243 to 575 µg/0.1 g dry weight, with a mean of 385 µg/0.1 g. Cardenolide uptake by butterflies was independent of plant concentration, suggesting that sequestration saturation occurs in monarchs fed cardenolide-rich host plants. Thinlayer chromatography resolved 19 cardenolides in the plants and 15 in the butterflies. In addition to humistratin,A. humistrata plants contained several relatively non-polar cardenolides of the calotropagenin series which are metabolized to more polar derivatives in the butterflies. These produced a butterfly cardenolide fingerprint clearly distinct from those previously established for monarchs reared on otherAsclepias species. In emetic assays with the blue jay,Cyanocitta cristata, the 50% emetic dose (ED₅₀) per jay was 57.1 µg, and the average number of ED₅₀ units per butterfly was 13.8, establishing that this important south eastern milkweed produces highly emetic, chemically defended monarchs. Our data provide further support for the use of cardenolide fingerprints of wild-caught monarchs to make ecological predictions concerning defence against natural enemies, seasonal movement and larval host-plant utilization by monarch butterflies during their annual cycle of migration, breeding and overwintering.     

Characterization of bioactive plant ellagitannins by chromatographic, spectroscopic and mass spectrometric methods

Ellagitannins are widely distributed plant polyphenols showing potent anti-herbivore activities, but rather complex chemical structures. Here we show how ellagitannins with different structures can be efficiently characterized from plant extracts by utilizing the information obtained from four universal methods used for ellagitannin analysis: Sephadex LH-20 gel chromatography, high-performance liquid chromatography, ultraviolet spectroscopy, and mass spectrometry. We show that by combining the information obtained from the ellagitannins’ chromatographic elution order, specific shapes of the ultraviolet spectra, molecular masses and characteristic mass spectral fragmentations, it is possible to classify these polyphenols into different ellagitannin sub-groups, and even characterize the structures within the sub-groups. These findings aid in the fast screening of complex plant samples for their ellagitannin composition without having to isolate compounds for individual characterization.     

Intraguild predation by <Emphasis Type="Italic">Harmonia axyridis</Emphasis> on coccinellids revealed by exogenous alkaloid sequestration

Summary.  Under laboratory conditions, the multicolored Asian lady beetle, Harmonia axyridis is well known as an intraguild predator of other ladybirds. However the real impact of this exotic species on native species was poorly investigated in the field. Because many ladybird species produce alkaloids as defensive compounds, we propose here a new method of intraguild predation monitoring in coccinellids based on alkaloid quantification by GC-MS. In laboratory experiments, adaline was unambiguously detected in fourth instar larvae of H. axyridis having ingested one egg or one first instar larva of Adalia bipunctata. Although prey alkaloids in the predator decreased with time, traces were still detected in pupae, exuviae and imagines of H. axyridis having ingested one prey when they were fourth instar larvae. Analysis of H. axyridis larvae collected in two potato fields shows for the first time in Europe the presence of exogenous alkaloids in 9 out of 28 individuals tested. This new method of intraguild predation detection could be used more widely to follow the interactions between predators and potential chemically defended insect preys.     

Thermal effects on swimming activity and habitat choice in juvenile Pacific cod (Gadus macrocephalus)

The behavioral responses of fishes to temperature variation have received less attention than physiological responses, despite their direct implications for predator–prey dynamics in aquatic ecosystems. In this paper, we describe the temperature dependence of swimming performance and behavioral characteristics of juvenile Pacific cod (Gadus macrocephalus; 75–125 mm total length). Maximum swimming speeds increased with temperature and body size. Routine swimming speeds of Pacific cod in small groups of similarly sized fish (N = 6) increased with body size and were 34 % faster at 9 °C than at 2 °C. The response to temperature was opposite that previously described for juvenile walleye pollock (Theragra chalcogramma), reflecting species-specific differences in behavioral responses. In a separate experiment, we demonstrated the effect of temperature on habitat selection of juvenile Pacific cod: Use of an artificial eelgrass patch in a 5-m-long laboratory tank was significantly greater at 9 °C than at 2 °C. These results illustrate that temperature affects a range of behavioral traits that play important roles in determining the frequency and outcomes of predator–prey interactions.     

Some polyphagous Homoptera gain protection from their natural enemies by feeding on the toxic plantsSpartium junceum andErythrina corallodendrum (Leguminosae)

Summary Interactions were studied among alkaloid-containing legumes (Erythrina corallodendrum andSpartium junceum) and non-toxic plants (Citrus sinensis, Cucurbita moschata andEuphorbia tirucalli), several polyphagous homopterans,Aphis craccivora (Aphididae),Icerya purchasi, I. aegyptiaca (Margarodidae),Lepidosaphes ulmi (Diaspididae) andPlanococcus citri (Pseudococcidae), and some major natural enemies of these homopterans. Significant reductions in survival due to negative effects of alkaloid containing as compared with non-alkaloidal plants were recorded for the predatorsRodolia cardinalis andChilocorus bipustulatus, but not forCryptolaemus montrouzieri (Coleoptera: Coccinellidae),Chrysoperla carnea (Neuroptera: Chrysopidae) andSympherobius sanctus (Neuroptera: Sympherobiidae). The development time of the larvae or pupae ofR. cardinalis, C. carnea andS. sanctus was longer on the toxic plants than on the non-toxic ones. The percentage of parasitism ofA. craccivora collected from the non-alkaloidal plantsVicia palaestina andMelilotus albus was much higher than that onS. junceum. The parasitoid complexes ofA. craccivora differed between both plant groups. The nutritive value of honeydew ofI. purchasi andA. craccivora, as expressed by the life span ofEncyrtus infelix (Hymenoptera: Encyrtidae) adults, was also investigated. Life spans were significantly longer when the wasps fed on honeydew produced on non-alkaloidal plants (C. sinensis andPittosporum tobira) than on alkaloid containing plants whenI. purchasi — but notA. craccivora — was the producer. It is suggested that the chemical defense ofE. corallodendrum andS. junceum is exploited by polyphagous phytophages to reduce predation. In nature, population growth and density of four of the investigated homopterans are conspicuously high when they developed on the alkaloid containing plant species, and very low on non-alkaloid plants. The efficiency of their natural enemies may be reduced by sequestration of alkaloids (or other toxic plant compounds) or their transfer into excreted honeydew. Therefore it is assumed that a generalist phytophagous homopteran may be protected from its natural enemies, although at different rates of efficiency, if it can safely sequester the host allelochemical when it develops on toxic species within its host range.     

Alterations in antioxidant defense system of workers chronically exposed to arsenic,cadmium and mercury from coal flying ash

Humans are exposed to different stress factors that are responsible for over-production of reactive oxygen species. Exposure to heavy metals is one of these factors. The aim of the study was to analyze the effect of chronic exposure to heavy metals through coal flying ash on the efficiency of antioxidative defensive mechanisms, represented by the activity of superoxide dismutase, glutathione peroxidase and ascorbic acid. Nonessential elements such as arsenic and mercury levels showed a significant increase (p > 0.001) in the power plant workers rather than in the control subjects. There were no significant differences of blood cadmium between power plant workers and control subjects. We found a significant positive correlation (p < 0.05) between BAs/SZn (r = 0.211), BAs/BSe (r = 0.287), BCd/SCu (r = 0.32) and BHg/BSe (r = 0.263) in the plant workers. Red blood cell antioxidant enzymes and plasma ascorbic acid were significantly lower in power plants workers than in the control group (p < 0.002). We can conclude that levels of mercury, arsenic and cadmium in blood, despite their concentration within the reference values, significantly affect plasma ascorbic acid concentration, superoxide dismutase and glutathione peroxidase activity, which are able to increase the risk of oxidative stress.

     

Identification and biological activity of sex pheromone components from females of the plum moth Illiberis rotundata Jordan (Lepidoptera: Zygaenidae: Procridinae)

The plum moth, Illiberis rotundata Jordan (Lepidoptera: Zygaenidae: Procridinae), is a pest of orchards in Japan and China. Few chemical ecological studies have been directed towards the Zygaenidae and particularly the Procridinae. To investigate the sex pheromone of this species, extracts of pheromone glands from adult female I. rotundata were analyzed by coupled gas chromatography-electroantennography (GC-EAG) and coupled gas chromatography-mass spectrometry (GC-MS). Whilst GC-EAG on male moths showed an active peak, identified as 2-butyl (7Z)-dodecenoate, GC-MS also revealed the presence of the homologue 2-butyl (9Z)-tetradecenoate. Electroantennographic investigations, as well as field tests, strongly suggested the natural compounds to have the (R)-configuration at the stereogenic centre. Field results demonstrate 0.2 mg of a 1:1-mixture of (2R)-butyl (7Z)-dodecenoate and (2R)-butyl (9Z)-tetradecenoate to be a powerful lure that may be used in pest control measures against I. rotundata. The chemical structures of the new pheromone components show the same features as those of other zygaenid species: unsaturated fatty acids esterified with a short chain chiral alcohol. This is the first example of a two-component blend constituting the pheromone of a procridinid species.     

Defensive chemistry of lycid beetles and of mimetic cerambycid beetles that feed on them

Summary.  Beetles of the family Lycidae have long been known to be chemically protected. We present evidence that North American species of the lycid genera Calopteron and Lycus are rejected by thrushes, wolf spiders, and orb-weaving spiders, and that they contain a systemic compound that could account, at least in part, for this unacceptability. This compound, a novel acetylenic acid that we named lycidic acid, proved actively deterrent in feeding tests with wolf spiders and coccinellid beetles. Species of Lycuscommonly figure as models of mimetic associations. Among their mimics are species of the cerambycid beetle genus Elytroleptus, remarkable because they prey upon the model lycids. We postulated that by doing so Elytroleptus might incorporate the lycidic acid from their prey for their own defense. However, judging from analytical data, the beetles practice no such sequestration, explaining why they remain relatively palatable (in tests with wolf spiders) even after having fed on lycids. Chemical analyses also showed the lycids to contain pyrazines, such as were already known from other Lycidae, potent odorants that could serve in an aposematic capacity to forestall predatory attacks. David Utterback: Deceased     

A laboratory study of behavioral interactions of the Antarctic keystone sea star Odontaster validus with three sympatric predatory sea stars

The circumpolar sea star Odontaster validus is ubiquitous in the nearshore marine benthos of Antarctica. Despite its ecological importance, little is known of its behavioral interactions with other common sympatric sea stars. To examine these interactions we employed time-lapse video analyses conducted in a large laboratory tank (1.8 m diameter circular tank, 1,629 L). In each experimental trial, 34 adult O. validus were placed in a tight circular grouping on one side of the tank, and one adult individual of one of three common sympatric species of predatory sea star (Labidiaster annulatus, Diplasterias brandti, or Perknaster aurorae) was placed on the opposite side of the tank. Digital images of sea star movements were then captured at one min intervals over a 24 h period and aspects of sea star movements subsequently analyzed. Each 24 h treatment was replicated three times, as was a control treatment consisting only of O. validus. O. validus had significantly elevated levels of activity in the presence of P. aurorae when compared with the other two sea stars (potential chemically mediated response), and displayed a distinct “flight response” (change in direction and twofold to sixfold increase of speed) upon tactile contact with this species. Moreover, an “alarm response” was detected when individuals of O. validus that encountered a fleeing conspecific also fled the vicinity. In contrast, our results indicated that O. validus displays virtually no chemical or tactile behavioral responses to the large multi-armed L. annulatus and only weak tactile responses to D. brandti.     

Pyrrolizidine alkaloids on three trophic levels – evidence for toxic and deterrent effects on phytophages and predators

Summary. Pyrrolizidine alkaloids (PAs) present a model system in the investigation of tritrophic interactions mediated by plant secondary compounds. However, their toxicity for insect herbivores has never been experimentally proven. Here, we demonstrate the toxic effects of a PA on growth and survival of the eri silk moth Philosamia ricini. In a feeding experiment, larvae of this generalist herbivore fed with an artificial PA diet gained weight significantly slower than control animals, and died as pupae. We suggest that derivatives of the ingested PA N-oxide damage developmental functions during metamorphosis. A tracer test with [¹⁴C]senecionine N-oxide revealed that the caterpillars lack adaptations that would prevent conversion of the chemical into the pro-toxic free base. In contrast, the PA adapted leaf beetle Longitarsus anchusae accumulates PAs as N-oxides. We tested the purpose of sequestration in this species as defence against predators. Through a series of prey choice experiments with three carabid predator species, chemically non-protected bark beetle pupae were chosen almost uniformly over L. anchusae pupae. In a following choice test with one of these predators, artificially PA-treated mealworm segments deterred the predator from feeding. Overall the study corroborates the immediate toxic effect of PAs on non-adapted herbivores and the protective effect that adapted insects may gain by sequestering them. It thereby underlines the potential for PAs to play a central role in multitrophic interactions between plants, phytophages and their predators.     

Effects of salinity stress on neurotransmission, energy metabolism, and anti-oxidant biomarkers of Carcinus maenas from two estuaries of the NW Iberian Peninsula

This study investigated the effects of salinity on biomarkers of oxidative stress, energy metabolism, and neurotransmission of Carcinus maenas from an estuary low impacted by pollution and from an estuary under chemical stress in the NW Iberian Peninsula. Crabs were collected in the field and, following an acclimation period, they were exposed for 7 days to five salinity levels ranging from 4 to 45 psu. At the end of the exposure period, stress biomarkers were determined in samples of muscle and digestive gland. The biomarkers assessed in the muscle were the activities of the enzymes cholinesterases (ChE), of which acetylcholinesterase is involved in neurotransmission, and lactate dehydrogenase (LDH) and isocitrate dehydrogenase (IDH) that are involved in energy metabolism. The biomarkers assessed in the digestive gland were (1) the activities of the enzymes glutathione S-transferases (GST), glutathione reductase (GR), and glutathione peroxidase (GPx), involved in phase II biotransformation and the anti-oxidant defence system; (2) the levels of total glutathiones (TG), also belonging to the anti-oxidant system; and (3) the levels of lipid peroxidation as a measure of oxidative damage. The results showed a significant influence of salinity on neurotransmission, energy metabolism, anti-oxidant status, and oxidative damage of C. maenas. For some biomarkers, this influence was dependent on whether the crabs were collected at the low-polluted estuary or at the contaminated estuary. In particular, crabs collected at the low-polluted estuary showed altered neurotransmission and anti-oxidant defences (GR). Crabs collected at the impacted estuary showed alterations in neurotransmission, energy metabolism (IDH and LDH), biotransformation, and anti-oxidant defences (GST, GR, GPx, and TG), as well as in oxidative damage, indicating that salinity change superimposes higher stress on these organisms. For ChE, IDH, and TG, altered responses were induced by both hypo- and hypersalinity.     

Effect of unfavorable trophic scenarios on amylase and protease activity of <Emphasis Type="Italic">Nephrops norvegicus</Emphasis> (L.) larvae during their first vertical migration: a laboratory approach

In Portuguese waters, Nephrops norvegicus larvae hatch at 400–800 m depth and need to perform a vertical migration to food-rich shallower waters to find suitable prey. The effect of suboptimal feeding on digestive enzymes activity of N. norvegicus larvae during this early period of their larval life remains unknown. Protease and amylase activities were investigated ex situ using flurometry in laboratory-hatched larvae exposed to different feeding and/or starving scenarios in the 24 h following hatching, the period during which they typically accomplish their upward vertical migration. Amylase activity was very low in comparison with protease activity, indicating that carbohydrates are not a primary energy reserve. Larvae starved for 12 h and subsequently fed displayed no increase in amylase activity, which suggests that feeding may be required before 12 h post-hatch to trigger amylase activity. Protease activity was high under all feeding conditions, and the increase in protease activity under sustained starvation indicated the catabolism of protein reserves. The ability of first-stage N. norvegicus larvae to metabolize protein reserves may play a decisive role for their survival during their first vertical migration, as it enables them to overcome the deleterious effects of short-term starvation and/or suboptimal feeding.     

Development of sol–gel formulations for slow release of pheromones

A new type of dispenser for slow-release of sex pheromones and other semiochemicals was developed based on sol–gel polymers that can be useful for monitoring, mass trapping, and mating disruption in integrated pest management (IPM). Sol–gel matrices exhibit glass characteristics and allow control of the degree of cross-linking during the polymerization process in order to provide an optimal release rate for a particular pheromone. The advantages of sol–gel (silica) matrixes include keeping the added molecules chemically stable and enabling the sol–gel material to be applied in any desired thickness and pheromone quantity, and thereby readily modify release rates. In addition, sol–gels are primarily silica and water that are common in the environment and therefore safe for field dispensing. We developed a method for the entrapment of pheromones in sol–gel matrices that allowed release at an almost constant rate over many days in the field. For example, 2.5 mg (E)-5-decenyl acetate pheromone of peach twig borer, Anarsia lineatella, entrapped in various sol–gel formulations released 14–45 μg/day for up to 28 days. The codling moth (Cydia pomonella) pheromone in sol–gels was used in field tests to capture more codling moth males than unbaited control traps. We describe how the method may be modified to entrap other types of pheromones by making sol–gels with different pore sizes.     

Sex and life-history stage alter herbivore responses to a chemically defended red alga

Intraspecific variation in resistance to herbivory among genders and life-history phases of primary producers can significantly alter the ecological and evolutionary consequences of plant-herbivore interactions. Seaweeds (macroalgae) with complex life histories have multiple distinct phases with associated variation in traits that can potentially lead to differences in resistance to consumers and provide a unique system in which to simultaneously test the effects of sex and life-history stage on herbivory. We tested the susceptibility to grazing of the three life-history stages and separate sexes of the chemically defended red alga Asparagopsis armata against the sea hare Aplysia parvula, and we related this to the plant quality traits of different stages and genders. Differences in nutrient content and halogenated secondary metabolites between life-history phases were highly sex dependent. Male gametophytes had a low concentration of secondary metabolites and the highest nutrient content. The highest secondary metabolite content was found within the female gametophyte, in the wall of the reproductive structures (cystocarps) that contain the microscopic carposporophyte phase. Feeding choices by A. parvula were consistent with differences in algal quality and defense and resulted in the haploid male gametophytes being the most preferred food type. The diploid carposporophyte found inside the chemically rich cystocarps was the least consumed life-history stage. Selective herbivory of male gametophytes by A. parvula is consistent with an observed shift in gametophyte sex ratio in the field from unity at the beginning of the reproductive season to female bias at the end. The variation in susceptibility to herbivory found between sex and life-history stages of A. armata represents the first example of sex-biased consumption in seaweeds and may contribute to the maintenance of complex life histories such as those found in red algae.     

Bacterial colonization of the phyllosphere of nineteen plant species and antimicrobial activity of their leaf secondary metabolites against leaf associated bacteria

Summary. The scope of this work was to examine whether leaf constitutive secondary metabolites play a role in determining bacterial colonization of the phyllosphere. To this aim, we surveyed nineteen native or cultivated plant species that share a common bacterial pool in a North Mediterranean area, and estimated the size of total and ice nucleation active (INA) bacterial populations on their leaves. Large differences in the colonization of their phyllosphere were found; the population size of epiphytic bacteria ranged from 7.5 × 10² to 1 × 10⁶ CFU/g fresh weight, in eucalypt and celery, respectively. Species native in Mediterranean-type climate areas, particularly those belonging to the group of aromatic plants, are characterized by scarce presence of INA bacteria. The antibacterial activity of essential oils, surface phenolics and leaf tissue extracts was also estimated against the INA strains P. syringae and E. herbicola, isolated from two of these plant species. E. herbicola proved more sensitive than P. syringae. Of the species examined, oregano [Origanum vulgare L. subsp. hirtum (Link.) Ietswaart], an aromatic plant, had the highest antimicrobial activity, whereas six species showed no activity at all. Further experiments were performed with oregano and bean (Phaseolus vulgaris L.) that represent two extremes in their secondary metabolite content. Both plants were inoculated with P. syringae. By the end of incubation, the bacterial population on bean plants was about 100 times higher than that on oregano leaves. Scanning electron micrographs showed that bacterial growth on oregano leaves was confined to sites away from glandular hairs. Results from the bacterial colonization survey together with those from the toxicity tests showed that all species rich in antibacterial secondary metabolites harbored low leaf bacterial populations. These results provide substantial evidence that leaf secondary metabolites function as constitutive defense chemicals against microbial invasions. However, the fact that species with non- or moderately active leaf secondary metabolites are not always highly colonized suggests mediation of other unknown factors, the contribution of which requires further investigation.     

Intraspecific variation of sesquiterpene lactones associated to a latitudinal gradient in <Emphasis Type="Italic">Smallanthus macroscyphus</Emphasis> (Heliantheae: Asteraceae)

According to theory, variation in plant secondary metabolism against herbivores is driven by variation in biotic and abiotic conditions interacting with plants genotype to determine the expression of resistance traits. Particularly, it has been long postulated that plants growing along latitudinal gradients experience changes in biotic and abiotic interactions, specifically leading to a decrease of plant toxicity towards the poles. We tested this hypothesis using the asteraceous species Smallanthus macroscyphus. Smallanthus species are known to contain sesquiterpene lactones (STLs), bitter compounds with a broad spectrum of biological activities, including deterrence to herbivores. S. macroscyphus showed a decrease in chemical diversity of STLs when investigating populations growing from the tropical regions to less tropical ones. Populations from lower latitudes were found to be more chemically diverse with enhydrin, uvedalin and fluctuanin as main components, while populations southward were chemically fairly uniform, with polymatin A as the main and largely dominant STL. The STL chemistry of S. macroscyphus is in agreement with the hypothesis that plants of tropical forests have a greater diversity of secondary metabolites when compared to their temperate counterparts.