Eucalypt responses to fertilization and reduced herbivory

Summary We manipulated soil fertility and insect attack for two species of Eucalyptus in natural stands of subalpine woodland on shallow, infertile granitic soils. E. pauciflora and E. stellulata responded in similar ways to simultaneous insecticide and fertilizer treatments. Eliminating herbivorous insects produced the largest changes — improved plant growth, increased leaf N and P, and reduced leaf specific density. Fertilizer regime modified some leaf properties, but had little effect on tree growth. E. stellulata trees were initially shorter than E. pauciflora, but grew faster without herbivores; by the end of the experiment both species were the same size when herbivores were removed. Foliage N and P levels increased most in trees with the most balanced fertilizer addition (NPK), and increased in all trees protected from insects, regardless of fertilizer regime. In this system, herbivorous insects exacerbated the effects of nutrientpoor soils, and may affect dominance of Eucalyptus species in mature forests.     

Induced responses in Ipomoea hederacea: simulated mammalian herbivory induces resistance and susceptibility to insect herbivores

Multispecies interactions between plants and natural enemies are ubiquitous, and often lead to diffuse interactions between plants and their herbivores. Non-specific induced responses, where responses induced by one species affect other species, are one potential mechanism generating diffuse interactions. Using 57 inbred lines of the Ivyleaf morning glory, Ipomoea hederacea, in a greenhouse experiment, we examined whether simulated mammalian herbivory induced responses that could affect plant resistance to the generalist insect herbivore, Spodoptera exigua. Inbred lines were highly variable for induced responses, ranging from induced resistance to induced susceptibility, with the rank-order for resistance in inbred lines changing between clipping and control treatments. We failed to detect significant genetic correlations between induced responses and trichome density, or that clipping modified the negative relationship between trichome density and Spodoptera exigua consumption and biomass. Our results suggest that non-specific induced responses can mediate the diffuse evolutionary relationship between I. hederacea and its herbivores, and that genetic variation in induced responses are an important component of this interaction. Handling Error: Heikki Hokkanen     

Plasticity and overcompensation in grass responses to herbivory

Several hypotheses predict defoliation-induced increases in individual plant fitness. In this paper we examine three such hypotheses: the Herbivore Optimization Hypothesis (HOH); the Continuum of Responses Hypothesis (CRH); and the Growth Rate Model (GRM). All three have in common predictions based on responses of defoliated individuals with the objective of explaining community and higher level phenomena. The latter two extend theory by specifying conditions for overcompensatory responses. They differ in whether overcompensation is sensitive to conditions external (CRH) or internal (GRM) to the plant. We tested these hypotheses with field experiments in a grassland system in which two native, perennial grass species replace each other along a short topographic/resource gradient. We detected positive, neutral, and negative changes in plant mass in response to partial defoliation. Patterns of responses to the edaphic and competitive environment combinations were unique to each species and neither the CRH nor the GRM were able to consistently predict responses in these grasses. Predictions of the HOH were fully supported only by the species naturally limited to lower-resource environments: overcompensation occurred in natural environments and it occurred at herbivory levels these plants experience naturally. Thus, the overcompensatory response can be important for the maintenance of local plant population distributions. However, new mechanistic theory is needed to account for the trend common to both species: overcompensatory responses to herbivory were greater in the edaphic environment in which each species was naturally most abundant.     

Herbivory pattern and induced responses to simulated herbivory inQuercus mongolica var.grosseserrata

Seasonal changes in leaf traits and the herbivory pattern ofQuercus mongolica var.grosseserrata were studied, and simulated herbivory experiments were carried out in order to evaluate leaf trait responses. Leaves ofQ. mongolica emerged simultaneously in spring and most were retained until autumn. Nitrogen concentration was highest when leaves first emerged and decreased rapidly with leaf age. Leaf mass per area (LMA) increased with leaf age. Herbivore attack was concentrated in the first 20 days after bud-break, which corresponded to the high nutritional value of the leaves for herbivores at this time. Simulated herbivory experiments indicated that LMA increased with artificial leaf damage, suggesting an increase in leaf toughness, and that nitrogen concentration decreased later in the season in comparison with intact leaves. As a result, herbivore attack following artificial leaf damage decreased with increasing initial leaf damage. However, leaf longevity was not affected by initial leaf damage. These responses were considered to be a strategy to disperse herbivory damage among leaves.     

Induced defense mechanisms in an aquatic angiosperm to insect herbivory

In terrestrial angiosperms, defense and resistance mechanisms against herbivores have been studied extensively; yet this topic is poorly understood in aquatic angiosperms. We investigated induced response mechanisms in Myriophyllum spicatum to the generalist insect herbivore Acentria ephemerella in three independent experiments. Various morphological and chemical response variables were examined in grazed apical shoots and compared to undamaged controls. We further estimated plant palatability of induced and non-induced apices in choice assays, and assessed the growth response of Acentria larvae in no-choice feeding assays. Leaves of induced apices were splayed out horizontally and changed in color from green to red. The dry matter content and thus plant toughness increased by up to 19 %, but silica levels stayed constant. Induced apices exhibited a decline in chlorophyll content of up to 34 %, reflected also by a 10 % decrease in nitrogen levels, while nitrogen increased by 14 % in lower parts of grazed shoots. Also, herbivore-deterring total phenolic compounds increased by up to 20 % in apices. In choice trials, Acentria larvae strongly avoided grazed tips, and growth was reduced by 25 % on induced apices. In total, we observed five different induced resistance and defensive traits in grazed apices: changes in appearance, increased plant toughness, delocalization of N-containing metabolites, increased polyphenols, and reduced nutritional value. The observed changes prevent herbivore damage and loss of apical tissue, which are most valuable for plant fitness. Our study presents the first evidence of multiple, parallel defense strategies including constitutive and induced defense mechanisms in a freshwater angiosperm.     

Induced responses in three tropical dry forest plant species – direct and indirect effects on herbivory

Induced changes in plant quality are hypothesized to reduce herbivore numbers and subsequent damage to the plant. The resultant decrease in herbivory may be due to direct negative impacts on herbivores, through the reduction in foliage quality as food, or due to indirect effects of plant-induced traits interacting with the third trophic level, increasing predation and parasitism rates on herbivores. The relative importance of induced responses as direct and/or indirect defenses has not been evaluated in natural systems. Moreover, few studies have evaluated the influence of early-season damage on late-season herbivory in natural systems, particularly in the tropics. The presence of induced responses and subsequent impact on folivory as a consequence of early-season damage were evaluated in three plant species ( Croton pseudoniveus , Bursera instabilis and Piper stipulaceum ) in a tropical dry forest in Mexico. A two-factorial experiment was applied to determine if induced responses influenced subsequent herbivory directly, by reducing foliage quality, or indirectly, through their interaction with parasitoids and predatory arthropods. Plants from all three species with reduced early-season damage had higher herbivory rates through the rest of the growing season, compared to plants that were damaged during leaf expansion. Chemical analyses showed that early-season damage induced the production of total phenolics and condensed tannins for C. pseudoniveus and B. instabilis , respectively. The mechanism by which these compounds affected subsequent herbivory was most likely by directly reducing foliage quality as food for herbivores, given that predatory arthropods and parasitoids had no effects on herbivory in this study. I conclude that early-season damage in these three species influenced later-season herbivory through the induction of plant responses that may act to reduce plant quality as food for herbivores.     

The alkaloidal responses of wild tobacco to real and simulated herbivory

Summary I compared the induced alkaloidal response in undamaged leaves of plants subjected to herbivory by the larvae of Manduca sexta and to different simulations of this herbivory; all herbivory treatments removed similar amounts of leaf mass. Although larval feeding induced a significant increase (2.2x) in alkaloid concentrations compared to undamaged plants, the alkaloid responses to larval feeding were significantly lower than the responses to an herbivory simulation (4x controls) which involved removing the same amount of leaf area from the same positions on the leaf, over a similar time period. Moreover, another herbivory simulation, identical in amount of leaf mass removed and duration of damage to the larval feeding, but without regard to spatial array of leaf damage, resulted in an alkaloidal response (5.5x controls) higher still than the previous herbivory simulation. In a second experiment the importance of leaf vein damage on the induced alkaloidal response was examined. Here, leaf removal that involved cutting leaf tissues from between secondary veins before removing the midrib, resulted in alkaloidal responses that were significantly lower (1.7x controls) than responses from leaf removal that involved cutting both veins and midribs along with the intervein tissues (2.6x controls). Vein damage alone did not produce a significant response. These results indicate that herbivory is difficult to simulate: that how a leaf is damaged can be as important as the magnitude of leaf damage in determining a plant's response to damage.     

Immunosuppression by human gangliosides: I. Relationship of carbohydrate structure to the inhibition of T cell responses.

Causes of cellular immunodeficiency frequently associated with cancer remain poorly understood. One possible mechanism is tumor cell membrane shedding of immunosuppressive molecules, such as the sialic acid-containing glycosphingolipids, gangliosides. To explore this interesting hypothesis and establish structure-activity relationships, we examined the effects of a series of highly purified human gangliosides on T cell function. In all, ten individual molecular species of two major biosynthetic pathways were compared for their ability to inhibit human T cell proliferative responses. They include GM1, GD1a, GD1b, and GT1b (the predominant normal brain species), and GM4, GM3, GM2, GD3, GD2 and GQ1b. Strikingly, each HPLC-purified molecule, from the simplest monosialoganglioside to the most complex polysialoganglioside, had potent inhibitory activity; even the ganglioside with the most elemental carbohydrate structure (GM4, one sialic acid linked to a monosaccharide) strongly inhibits T cell proliferative responses to tetanus toxoid (ID90 = 1.5 microM). The data also reveal a complex interplay between elements of oligosaccharide structure in determining immunosuppressive activity. Sialic acid is critical to maximal activity, and (i) immunosuppression is most potent in gangliosides containing a terminal sialic acid. (ii) Total desialylation almost abolishes activity and (iii) partial alteration (lactone formation) reduces activity. (iv) Activity is generally but not always higher with higher numbers of sialic acid residues/molecule, and (v) some larger neutral glycosphingolipids retain measurable immunosuppressive activity. Overall, the potent inhibition by gangliosides supports the hypothesis that shedding of these molecules by tumors creates a highly immunosuppressive microenvironment around the tumor, thereby inhibiting the function of infiltrating host leukocytes and contributing to diminished T cell responses in cancer.     

Induced Resistance to 6-Methylpurine and Cycloheximide in Tetrahymena. I. Germ Line Mutants of T. THERMOPHILA

Two new mutant lines of Tetrahymena thermophila ( T. pyriformis, syngen 1), each conferring resistance to a different agent, are described. Resistance to cycloheximide and 6-methylpurine are each determined by dominant genes, ChxA2 and Mpr; the traits show phenotypic assortment. The method used to select these mutations, the critical importance of backcrossing to wild type following mutagenesis, and the utility of these marker genes in further mutagenic selection schemes and studies of the sexual cycle of Tetrahymena are noted.     

Growth,reproduction and defence in nettles: responses to herbivory modified by competition and fertilization

We studied the effects of environmental factors on the ability to compensate for simulated herbivory in an annual (Urtica urens) and two perennial (U. dioica subspp. dioica and sondenii) nettle species in order to test the compensatory continuum hypothesis. Further, we studied the expression of costs of structural defence in different environmental conditions. Herbivory interacted significantly with density and fertilization for height growth, branching and reproductive traits. The negative effects of simulated herbivory on relative height growth were less detrimental for plants grown in low density and high fertilization level, which supports the compensatory continuum hypothesis. However, with respect to other traits measured, our results do not support the idea of compensatory continuum. The negative effects of apical excision on inflorescence biomass were relatively more prominent in plants growing at low density and receiving more fertilization than on those growing in worse conditions. In addition, branching was reduced by apical excision regardless of resource levels. The lack of compensation for herbivory is explained by the role which the damaged or removed tissue plays in plant development and function. In the perennial U. dioica, defensive responses to herbivory, measured as changes in trichome density, were stronger than in the annual U. urens. In the southern subspecies dioica, trichome density increased in newly emerged leaves after leaf clipping. In the northern subspecies sondenii, trichome density increased after apical excision. The differences in the defensive responses between the two subspecies of U. dioica may be due to differential natural herbivory pressures on subspecies inhabiting different geographical regions. We observed negative phenotypic correlations between structural defence and other plant traits, which suggests the existence of costs of defence. In addition to differences among the species studied, the expression of costs of defence was dependent on the resource levels.     

Early physiological responses of Abies alba and Rubus fruticosus to ungulate herbivory

For the last 50 years, Chamois (Rupicapra rupicapra) population exponentially increased in the French Alps. This herbivore faces a food shortage in the winter; its diet mainly includes woody species (Abies alba Mill.) and brambles (Rubus fruticosus). The physiological response following simulated herbivory of these two plants was assessed. More precisely, the effects of leaf clipping and ungulate saliva application on the epidermal flavonoids and chlorophyll contents were considered as an induced structural response. The chlorophyll fluorescence (Fv/Fm) was also measured to provide information on the photosynthesis status as an induced functional response. The non-invasive techniques used in this experiment highlighted an induced response, emerging during the days following the simulated herbivory. R. fruticosus was drastically affected, photosynthesis was disturbed (decreased Fv/Fm) and mortality began on day 5 reaching 100 % less than 1 week later. Physiological parameters were also affected in A. alba (slight decrease of Fv/Fm and lower content of flavonoids), but this species recovered and survived no matter what the stressing treatment was. These results did not point out an efficient repulsive-induced response of A. alba and R. fruticosus to herbivory. The existence of constitutive defences such as prickles (R. fruticosus) or low-digestible tannins (A. alba) is no more efficient to escape from ungulates consumption. Nevertheless, in spite of the increase of the ungulates exponential demography, these two plants do not face rarefaction problem in the French Alps. Thus, survival of A. alba stands is probably linked to the recovery ability of saplings, when R. fruticosus stands maintain themselves thanks to the strong resprouting capacity of this species.     

Plant responses to selective grazing by bison: interactions between light,herbivory and water stress

Two abundant tallgrass prairie forb species, Ambrosia psilostachya and Vernonia baldwinii, are commonly found intact in patches where the grasses have been selectively grazed by bison. Microclimatic patterns and physiological responses of these forbs were measured in grazed and ungrazed patches. These experiments demonstrated that bison herbivory indirectly enhanced water availability and productivity of forbs growing in grazed patches. This was due primarily to the reduction in transpiring grass leaf area in grazed patches and an increase in light availability. In grazed patches, incident light at forb mid-canopy height was 53% greater than ungrazed sites at midseason and soil temperatures were always warmer (e.g., 10°C at 5 cm), perhaps enabling forbs to initiate growth earlier in the spring. Enhanced leaf xylem pressure potential and stomatal conductance in plants in grazed areas were most evident when water availability was low (i.e., late in the growing season and over short-term dry periods characteristic of the tallgrass prairie environment). Relative to individuals in ungrazed areas, end-of-season biomass of A. psilostachya was 40% greater and reproductive biomass and head number of V. baldwinii was 45% and 40% greater, respectively, in plants in grazed patches. A favorable growing environment maintained in grazed patches during periods of water limitation enhances carbon gain in forbs leading to increased biomass and potential fitness.     

T cell responses to alloantigens. I. Studies of in vivo and in vitro immunologic memory and suppression by limit dilution analysis

A limit dilution technique was used to study the frequency of cytotoxic T cell precursors (f pre Tc) in the peripheral blood lymphocytes (pbl) of naive and alloimunized mice. It was found that alloimmune mice showed a 2- to 5-fold specific increase in the f pre Tc reactive to the immunizing alloantigens. This technique was further adapted for use as a sensitive in vitro assay for alloantigen-specific suppressor T cells. It was found that nonimmunosuppressed B6AF1 mice bearing surviving B10.BR cardiac allografts had circulating alloantigen-specific suppressor cells. In vitro it was shown that the culture of heat-inactivated B10.D2 spleen cells with B6AF1 spleen cells gave rise to alloantigen-specific B6AF1 suppressor cells.