Diterpenoid Alkaloid Concentration in Tall Larkspur Plants Damaged by Larkspur Mirid

Tall larkspur (Delphinium barbeyi) is a serious poisonous plant threat to cattle on mountain rangelands. The larkspur mirid [Hopplomachus affiguratus] has been proposed as a biological tool to damage tall larkspur in an effort to deter grazing by cattle and thus prevent poisoning. Preliminary data suggested that it may also reduce toxic alkaloid levels. The objective of this study was to determine if damage caused by the larkspur mirid would reduce toxic alkaloid concentration. Larkspur mirids were collected in the field in 1992 and placed on potted plants in the greenhouse. The resulting mirid-damaged leaves were lower in toxic alkaloids than leaves from uninfested plants. In the 1995 field study, toxic and total norditerpenoid alkaloid concentrations were measured in two larkspur populations having established mirid populations and in two newly infested larkspur populations. In the 1996 field study, three widely separated larkspur populations infested with mirids were sampled. Mirid-damaged leaves were lower in toxic alkaloids in both years, but there were no differences in flowering heads. However, only at Yampa, Colorado, did mirids reduce toxic alkaloids to levels that would not pose a threat to cattle. There was no difference in toxic or total alkaloid concentration between larkspur populations with long-term mirid infestations compared to newly infested plants. The plant-to-plant variability in alkaloid concentration was greater than differences due to mirids.     

A Functional Explanation for Patterns of Norditerpenoid Alkaloid Levels in Tall Larkspur (Delphinium barbeyi)

Concentrations of norditerpenoid alkaloids vary among larkspur (Delphinium) species, locations, and years, but environmental stresses seem to have little effect on alkaloid levels. There is a need for a functional hypothesis of alkaloid synthesis and metabolism to explain the observed trends in concentration and to predict the toxicity of larkspur populations. This study was replicated at two locations over two years in the mountains of central Utah. Ten tall larkspur (D. barbeyi) plants were marked at each location, and a single stalk was harvested from each plant at weekly intervals throughout the growing season. Concentrations of toxic and total alkaloids were measured by Fourier-transformed infrared spectroscopy (FTIR), and alkaloid pools were calculated by multiplying the alkaloid concentration by the dry weight of the plant to determine the amount of alkaloids in the stalk. Alkaloid pools in the stalks increased for the first three weeks, leveled off, and then declined to low levels as the plants began to senesce. Concentrations of alkaloids declined through the season, as the alkaloids were diluted in the increasing biomass as the plants grew. These patterns will be used to predict potential toxicity of larkspur populations.     

Influence of alkaloid concentration on acceptability of tall larkspur (Delphinium spp.) to cattle and sheep

Tall larkspur (Delphinium spp.) is a serious toxic plant problem on western U.S. ranges. The major toxins in tall larkspur are methyllycaconitine (MLA) and 14-deacetylnudicauline (14-DAN); the sum of both is termed the toxic alkaloid concentration. Toxic alkaloids comprise about 20–50% of the total alkaloid concentration in tall larkspur. Toxic and total alkaloid concentration generally declines with maturity, whereas cattle and sheep consumption of larkspur typically increases with plant maturity. We hypothesized that cattle and sheep consumption of tall larkspur was negatively related to higher concentrations of total or toxic alkaloid. We compared consumption of several collections of dried, ground larkspur and fresh larkspur in a series of trials. In another trial, a crude alkaloid fraction was extracted with ethanol, added to alfalfa hay, and consumption compared to untreated alfalfa hay, alcohol-treated hay, and the essentially alkaloid-free plant residue. In all cases we correlated amounts eaten with total and toxic alkaloid concentration. A grazing trial was also conducted to relate larkspur consumption over time to alkaloid concentrations. Total alkaloid concentrations in dried, whole-plant collections ranged from 9.3 to 38.8 mg/g of dry weight, whereas toxic alkaloid concentrations varied from 0.0 to 7.1 mg/g. In one pen trial, cattle preferred a larkspur collection (P<0.01) that contained no toxic alkaloids but had a high total alkaloid concentration (39 mg/g). There was no correlation (P>0.05), however, between concentrations of total or toxic alkaloids and amount of dry plant consumed in this or any other trial. Conversely, sheep consumption tended to be negatively influenced by total and toxic alkaloid concentration (P0.08). In the trials with extract, cattle preferred the alcohol-treated hay and rejected the alkaloid-free residue (P<0.01), whereas the alkaloid-treated hay was of intermediate acceptability. Cattle preferred the alkaloid-treated hay over the alkaloid-free residue, indicating that alkaloids did not deter consumption. Conversely, the alkaloid-treated hay was less preferred than either untreated or alcohol-treated hay, suggesting a negative effect on acceptability. There was no correlation between alkaloid concentration and amount of treated feed eaten. In field trials, the amount of composited, fresh leaves or flowers eaten by cattle was influenced by plant part (P=0.04), but was not related (P>0.05) to alkaloid concentration. Cattle preferred leaves over flowers when offered individual plants differing in phenological stage and/or amount of shade, but alkaloid concentration was not related to consumption. We conclude that knowledge of the concentration of total or toxic alkaloid in tall larkspur will give little or no indication of plant acceptability to cattle. Even though accurate predictions can be made about the potential toxicity of larkspur based on the concentration of toxic alkaloids, predictions about consumption must be based primarily on plant phenology.     

Influence of Light and Photosynthesis on Alkaloid Concentration in Larkspur

Concentrations of toxic norditerpenoid alkaloids vary greatly in tall larkspur (Delphinium barbeyi) and may be influenced by environmental stress. We evaluated the effect of shade, darkness, and inhibition of photosynthesis on toxic alkaloid concentration. In plants treated with metribuzin to inhibit photosynthesis, alkaloid concentration increased, but dry weight of the plants decreased as growth ceased, leaving absolute alkaloid content similar to that of control plants. Short-term shade (70% reduction in sunlight for three days), dark treatments from leaves collected at night, and aluminum foil covered leaves all increased alkaloid concentration in comparison to untreated control plants. It appears that absolute amounts of alkaloids remained the same, but the mass of stressed plants declined as nonstructural carbohydrates were depleted, thus increasing the relative concentration of alkaloids. We conclude that norditerpenoid alkaloids in larkspur do not respond to short-term light stress. Alkaloid concentration was lower in larkspur plants growing beneath forest canopy and in potted plants in a long-term shade study (70% reduction in sun light for 21 days) than plants growing in open sunlight. Long-term shade may have reduced synthesis of norditerpenoid alkaloids, particularly in the earlier developmental stages of the plant. Shade stress or photosynthesis inhibition apparently did not increase norditerpenoid alkaloid synthesis, which contrasts with the carbon/nutrient balance theory of plant defense.     

Ingestion of tall larkspur by cattle

Tall larkspur (Delphinium spp.) is a palatable but toxic poisonous plant in the western United States. The toxins in tall larkspur are diterpenoid alkaloids. We examined the influences of food flavor and postingestive consequences on consumption of a 33% larkspur pellet during 30-min feeding periods for five days using esophageally fistulated cattle that were sham-fed larkspur pellets. Consumption by the sham-fed group was compared to a control group fed alfalfa pellets, and a larkspur group fed only larkspur pellets. Sham-fed cattle did not decrease (P > 0.1) feed consumption compared to controls, indicating no significant difference in food flavor. The larkspur group decreased (P < 0.05) feed consumption by 41% relative to controls and by 31% relative to sham-fed animals (P = 0.08). This reduction in feed consumption indicates the adverse postingestive consequences of tall larkspur ingestion, as the larkspur group apparently developed a conditioned taste aversion to the larkspur pellet. Even though these animals were averted to the pellets, they showed none of the classical signs of intoxication from ingestion of tall larkspur.This research was supported in part by the Utah Agricultural Experiment Station, Utah State University, Logan, Utah 84322. Approved as journal article No. 3846.     

Distribution of Norditerpene Alkaloids in Tall Larkspur Plant Parts Through the Growing Season

Previous research showed that toxic and total alkaloid pools in tall larkspur (Delphinium barbeyi) increased during early growth, then declined precipitously during the late flower and pod stage of growth. The objective of this study was to measure the concentration and pools of toxic and total alkaloids in tall larkspur plant parts, including roots, and to evaluate the changes in these pools over the growing season as an estimate of diterpenoid alkaloid kinetics in tall larkspur. Twenty entire plants were harvested at each phenological stage: beginning of growth in the spring, early flower, early pod, late pod, and senescence. The plants were separated into their respective parts, freeze-dried, extracted, and analyzed for toxic and total alkaloid concentration, and alkaloid pools were calculated. Concentration of toxic and total alkaloids in leaves and stems declined as the plants matured, while concentration in flowers and pods increased (P < 0.004). Concentration of alkaloids in the root declined in the early growth, then increased at the end of the season (P = 0.002). Alkaloid pools in the root decreased during early growth, with a corresponding increase of pools in foliar parts. In the late flower and pod stage, alkaloid pools in the leaves and stems declined rapidly, while the pool in the crown and roots tended to increase.     

Alkaloidal responses to damage inNicotiana native to North America

We performed field tests of alkaloid induction inNicotiana attenuata plants growing in southwestern Utah with mimicry of the two major types of damage inflicted by invertebrate and vertebrate herbivores: leaf damage and stalk removal, respectively. In undamaged plants, seasonal increases in leaf nicotine content occurred at a rate of 0.046% leaf dry mass/day. Leaf damage doubled the accumulation rate to 0.086–0.138% leaf dry mass/day, while stalk removal resulted in a quadrupling of the accumulation rate to 0.206% leaf dry mass/day. These damage-induced increases in nicotine accumulation are significantly larger than between-plant and phenological variations. Leaf damage to the nornicotine-(N. repanda andN. trigonophylla) and anabasine-accumulating (N. glauca)Nicotiana species native to North America resulted in 1.5- to 5-fold increases in their principal leaf alkaloid pools. We conclude that alkaloid induction is not limited to nicotine-accumulatingNicotiana species and that herbivores feeding on previously damaged plants are likely to encounter tissues with alkaloid titers significantly higher than those of undamaged plants.     

The Biogeographical Distribution of Duncecap Larkspur (Delphinium occidentale) Chemotypes and Their Potential Toxicity

Larkspurs (Delphinium spp.) are poisonous plants found on rangelands in western North America. Larkspur’s toxicity has been attributed to the norditerpenoid alkaloids, which are divided into two main structural groups: the highly toxic (N-methylsuccinimido) anthranoyllycoctonine type (MSAL type) and the less toxic 7,8-methylenedioxylycoctonine type (MDL type). Plants high in the MSAL-type alkaloids are thought to be the most toxic to cattle, and the concentrations of these alkaloids have been used as a predictor of plant toxicity. Duncecap larkspur, Delphinium occidentale, occurs throughout much of the Intermountain West and Northwestern United States. Specimens from field collections and herbaria deposits were evaluated taxonomically and chemically. Two distinct alkaloid profiles were identified: one that contains the MSAL-type alkaloids and one that contains little, if any, MSAL-type alkaloids. Thus, plants with these two alkaloid profiles should differ in their toxic potential. Each profile was unique in its geographical distribution. These findings have important implications in grazing management decisions on D. occidentale-infested rangelands, and they demonstrate that botanical classification alone is not a good indicator to determine the toxic risk of D. occidentale.     

Tall Larkspur Ingestion: Can Cattle Regulate Intake Below Toxic Levels?

Tall larkspur (Delphinium barbeyi) is a toxic forb often consumed by cattle on mountain rangelands, with annual fatalities averaging about 5%. This study examined the relationship between food ingestion and toxicity in cattle. Two grazing studies suggested that larkspur consumption above 25–30% of cattle diets for one or two days led to reduced larkspur consumption on subsequent days. We subsequently hypothesized that cattle can generally limit intake of larkspur to sublethal levels. This hypothesis was tested by feeding a 27% larkspur pellet in experiment 1. Cattle given a 27% larkspur pellet ad libitum showed distinct cyclic patterns of intake, where increased larkspur consumption on one or two days was followed by reduced (P < 0.025) consumption on the following day. The amount of larkspur (mean 2007 g/day; 17.8 mg toxic alkaloid/kg body wt) consumed was just below a level that would produce overt signs of toxicity. Experiment 2 was conducted to examine cattle response to a toxin dose that varied with food intake. Lithium chloride (LiCl) paired with corn ingestion was used as a model toxin, and we hypothesized that if increased (decreased) consumption was followed by a stronger (weaker) dose of LiCl, cattle would show a transient reduction (increase) in corn intake. There was no difference (P > 0.05) between controls and treatment animals at the 20 or 40 mg LiCl/kg dose in the percentage of corn consumed, but the 80 mg LiCl/kg dose induced a cyclic response (mean 46%) compared to intake by controls (mean 96%) (P < 0.001). At the 80 mg/kg dose, LiCl induced an aversion to corn; when corn intake decreased on subsequent days and LiCl dose also decreased, cattle responded by increasing corn intake and apparently extinguishing the transient food aversion. Experiment 3 was similar to the LiCl trial, except that tall larkspur was the toxin. Cattle responded to oral gavage of ground larkspur with distinct cycles; days of higher corn consumption were followed by one to three days of reduced consumption. Corn intake for controls was higher (P < 0.01) than for larkspur-treated animals (means 84 and 52%, respectively; day × treatment interaction P < 0.01). The threshold for toxic effects on corn intake was 14 mg toxic alkaloid/kg body weight. In conclusion, cattle apparently limit ingestion of some toxins so that periods of high consumption are followed by periods of reduced consumption to allow for detoxification. Cyclic consumption generally enables cattle to regulate tall larkspur consumption below a toxic threshold and allows cattle the opportunity to safely use an otherwise nutritious, but toxic, plant.     

Neotyphodium coenophialum Mycelial Protein and Herbage Mass Effects on Ergot Alkaloid Concentration in Tall Fescue

Livestock grazing endophyte-infected tall fescue (Festuca arundinacea Schreb.) pastures often suffer from ergot poisoning. The endophyte,Neotyphodium coenophialum (Morgan-Jones and Gams) Glenn, Bacon, and Hanlin, comb. nov., also provides drought-tolerant, insect-resistant, and disease-resistant qualities to the plant. Therefore, producers are faced with a biological dilemma of risking loss of pasture by using endophyte-free tall fescue pasture or animal losses with endophyte-infected tall fescue pasture. One potential solution is to breed endophyte-infected tall fescue with lower levels of alkaloids. However, breeding could select for plants that are antagonistic to the endophyte, resulting in reduced plant vigor as a consequence of disruption of the mutualistic association between the organisms. The objectives of this study were to determine the relationship between plant mass and endophyte mycelial proteins versus ergot alkaloid concentration. To examine the endophyte effect on mycelial mass, tissue culture regenerants from tall fescue genotype PDN2 were infected with endophyte isolates EDN11, EDN12, and EDN2 to eliminate confounding effects of multiple plant genotypes. Crosses with PDN11 as the maternal parent and plant genotypes DN2, DN12, and DN15 as paternal parents were used to produce a population of different plant genotypes, all containing the same endophyte. Fungal mycelial protein was extracted from lyophilized tall fescue leaf sheath tissue and immunochemically quantified with monoclonal antibodies specific toN. coenophialum proteins. Ergot alkaloid concentration was also immunochemically measured. Quantities of mycelial protein and ergot alkaloids were calculated by regressing experimental values against standards of each. There was no correlation between herbage mass and alkaloid concentration or fungal protein quantity and alkaloid concentration when different fungal isolates were inserted into the same plant genotype. Coefficients of determination (r ²) were low (0.31 and 0.17) between leaf sheath and leaf blade alkaloid concentrations and endophyte protein when different plant genotypes contained the same endophyte isolate. Likewise,r ² values were low between leaf sheath and leaf blade alkaloid concentrations and herbage mass. These data suggest that little or no antagonism occurred to the endophyte among plants low in alkaloid concentration.     

Temporal and Spatial Variation in Alkaloid Levels in Achnatherum robustum, a Native Grass Infected with the Endophyte Neotyphodium

The native North American perennial grass Achnatherum robustum (Vasey) Barkworth [= Stipa robusta (Vasey) Scribn.] or sleepygrass is toxic and narcotic to livestock. The causative agents are alkaloidal mycotoxins produced from infections by a systemic and asexual Neotyphodium endophyte. Recent studies suggest that toxicity is limited across the range of sleepygrass in the Southwest USA. We sampled 17 populations of sleepygrass with varying distance from one focal population known for its high toxicity levels near Cloudcroft, NM, USA. For some, we sampled individual plants twice within the same growing season and over successive years (2001–2004). We also determined infection levels in each population. In general, all populations were highly infected, but infection levels were more variable near the focal population. Only infected plants within populations near the Cloudcroft area produced alkaloids. The ergot alkaloid, ergonovine, comprised the bulk of the alkaloids, with lesser amounts of lysergic and isolysergic acid amides and ergonovinine alkaloids. Levels of all alkaloids were positively correlated among individual plants within and between growing seasons. Infected plants that produced no alkaloids in 1 yr did not produce any alkaloids within the same growing season or in other years. Levels of alkaloids in sleepygrass populations declined with distance from the Cloudcroft population, although infection levels increased. Infected plants in populations in northern New Mexico and southern Colorado produced no alkaloids at all despite 100% infectivity. Our results suggest that only specific Neotyphodium haplotypes or specific Neotyphodium–grass combinations produce ergot alkaloids in sleepygrass. The Neotyphodium haplotype or host–endophyte combination that produces toxic levels of alkaloids appears restricted to one locality across the range of sleepygrass. Because of the wide variation in alkaloid levels among populations, interactions between the endophyte and host, and consequences for herbivores, competitors, and pathogens and other components of the community, are likely to vary widely across the geographic range of this native grass.     

Influence of Defoliation on Toxic Alkaloid Concentration and Alkaloid Pools in Tall Larkspur

This study was replicated at two locations in the mountains of central Utah. In 1997, ten uniform plants of tall larkspur (Delphinium barbeyi) in the early bud stage (40 cm in height) were selected at each site and clipped at 5 cm above soil level. In 1998, one stalk from each plant was harvested on a weekly basis; in 1999, one stalk was harvested at four times during its phenological development. Toxic and total alkaloid concentrations were measured and alkaloid pools in the entire stalk were calculated. Clipping reduced stalk height to less than 50 cm in 1998 and 65 cm in 1999, compared to over 100 cm in unclipped control plants. Alkaloid concentration was similar to control plants, but toxic alkaloid pools were 70% lower than control plants, because of the reduction in biomass of the stalks. Clipping reduced subsequent vigor and the amount of toxic and total alkaloids in tall larkspur.     

Elevational Trends in Defense Chemistry, Vegetation, and Reproduction in Sanguinaria canadensis

Evaluation of biotic interactions along geographic gradients reveals that pressure on plant populations by herbivores and pathogens increases as latitude decreases, and is accompanied by a parallel increase in the number and toxicity of alkaloid-bearing plants. We compared rhizome alkaloid content with plant reproductive and vegetative characters in Sanguinaria canadensis (Papaveraceae) along an elevational gradient over two growing seasons to ascertain 1) if alkaloid production in bloodroot varies among populations and systematically with elevation, and 2) if there exists a correlation between isoquinoline alkaloid, vegetative and reproductive production. In general, alkaloid content in bloodroot rhizomes declines with elevation, increases with rhizome water content, varies by site, and fluctuates seasonally with plant growth and reproduction. Alkaloid content was positively correlated with vegetative and reproductive effort with few exceptions. Analysis of total protopine and benzophenanthridine alkaloid concentrations revealed generally similar patterns as those of individual alkaloid concentrations, although significant differences did appear between individual alkaloid concentrations.     

Use of NMR for predicting protein concentration in soybean seeds based on oil measurements

Increasing the level of protein in soybean seeds has been a major target for soybean [Glycine max (L.) Merr.] breeders. The objective of this study was to examine the potential of predicting soybean seed protein based on oil values as determined by NMR. Seed protein and oil concentrations were determined in an F₂ population generated from the cross between a G. max (NK, S08-80), and a G. soja (PI 458536) cultivar. The protein concentration in the population ranged from 40.4 to 52.6%. Protein-oil regression analysis was used to generate an equation for predicting seed protein concentration based on oil readings. The regression equation Protein=62.3–1.3 [Oil] (R ²=0.46) was developed, with a corresponding correlation of −0.69 between the traits. With this equation, the mean protein concentration of the selected 25% of the population (a simulated breeding pressure) was greater than the mean of the unselected population (46.1%, SE=0.13) by about 1.9%. Individual F₂ plants that exceeded the mean protein value of the population constituted 86.4% of the selected samples. Selection based on oil concentration, however, failed to include 27.1% of the plants that were among the top 25% for protein concentration. Selection of high-protein plants based on NMR oil measurement was reasonably effective in the test population and might offer a new and rapid method of selecting high-protein individuals in soybean populations derived from the wild soybean progenitor, G. soja. If further tested on other populations and samples, it might be used as an analytical alternative for an indirect measurement of protein concentration based on NMR measurements of the oil.     

Courtship pheromone production and body size as correlates of larval diet in males of the arctiid moth,Utetheisa ornatrix

Hydroxydanaidal, the corematal courtship pheromone of maleUtetheisa ornatrix, shows pronounced quantitative variation in natural populations of the moth. Males that, as larvae, fed on seed-bearing rather than immature food plants (Crotalaria spectabilis orC. mucronata) produce higher levels of hydroxydanaidal. Such males also have higher systemic loads of pyrrolizidine alkaloid, the known metabolic precursor of hydroxydanaidal, whichUtetheisa sequester from their larval diet and which is concentrated in the seeds ofCrotalaria. Males raised on seed-bearing plants also achieve higher adult weight. In the context of sexual selection, therefore, femaleUtetheisa could, through assessment of male hydroxydanaidal levels, gauge both the alkaloid content and body weight of their suitors.     

Antifungal leaf-surface metabolites correlate with fungal abundance in sagebrush populations

A central component in understanding plant–enemy interactions is to determine whether plant enemies, such as herbivores and pathogens, mediate the evolution of plant secondary metabolites. Using 26 populations of a broadly distributed plant species, sagebrush (Artemisia tridentata), we examined whether sagebrush populations in habitats with a greater prevalence of fungi contained antifungal secondary metabolites on leaf surfaces that were more active and diverse than sagebrush populations in habitats less favorable to fungi. Because moisture and temperature play a key role in the epidemiology of most plant–pathogen interactions, we also examined the relationship between the antifungal activity of secondary metabolites and the climate of a site. We evaluated the antifungal activity of sagebrush secondary metabolites against two fungi, a wild Penicillium sp. and a laboratory yeast, Saccharomyces cerevisiae, using a filter-paper disk assay and bioautography. Comparing the 26 sagebrush populations, we found that fungal abundance was a good predictor of both the activity (r ² = 0.36 for Saccharomyces, r ² = 0.37 for Penicillium) and number (r ² = 0.34 for Saccharomyces) of antifungal secondary metabolites. This suggests that selection imposed by fungal pathogens has led to more effective antifungal secondary metabolites. We found that the antifungal activity of sagebrush secondary metabolites was negatively related to average vapor pressure deficit of the habitat (r ² = 0.60 for Saccharomyces, r ² = 0.61 for Penicillium). Differences in antifungal activity among populations were not due to the amount of secondary metabolites, but rather to qualitative differences in the composition of antifungal compounds. Although all populations in habitats with high fungal prevalence had secondary metabolites with high antifungal activity, different suites of compounds were responsible for this activity, suggesting independent outcomes of selection on plants by fungal pathogens. The location of antifungal secondary metabolites on the leaf surface is consistent with their putative defense role, and we found no evidence supporting other functions, such as protection from ultraviolet light or oxidation. That the antifungal activity of sagebrush secondary metabolites was similar for two different fungi provides support for broad antifungal defenses. The incidence and severity of fungal disease in the field (caused by Puccinia tanaceti) were similar in moist and dry habitats, possibly reflecting an equilibrium between plant defense and fungal attack, as sites with greater fungal abundance compensated with more effective secondary metabolites. The geographic correlation between fungal abundance and antifungal secondary metabolites of sagebrush, coupled with our other data showing heritable variation in these metabolites, suggests that pathogenic fungi have selected for antifungal secondary metabolites in sagebrush.     

Sequestration, Maintenance, and Tissue Distribution of Pyrrolizidine Alkaloid N-Oxides in Larvae of Two Oreina Species

Oreina cacaliae and O. speciosissima are leaf beetles that, as larvae and adults, sequester pyrrolizidine alkaloid N-oxides (PAs) as defensive compounds from their host plants Adenostyles alliariae and Senecio nemorensis. As in most Oreina species, O. speciosissima is also defended by autogenously produced cardenolides (mixed defensive strategy), whereas O. cacaliae does not synthesize cardenolides and is exclusively dependent on host-plant-acquired PAs (host-derived defense). Adults of the two Oreina species were found to have the same PA storage capacity. The larvae, however, differ; larvae of O. speciosissima possess a significantly lower capability to store PAs than O. cacaliae. The ability of Oreina larvae to sequester PAs was studied by using tracer techniques with ¹⁴C-labeled senecionine N-oxide. Larvae of the two species efficiently take up [¹⁴C]senecionine N-oxide from their food plants and store the alkaloid as N-oxide. In O. cacaliae, there is a slow but continuous loss of labeled senecionine N-oxide. This effect may reflect the equilibrium between continuous PA uptake and excretion, resulting in a time-dependent tracer dilution. No noticeable loss of labeled alkaloid is associated with molting. Senecionine N-oxide is detectable in all tissues. The hemolymph is, with ca. 50–60% of total PAs, the major storage compartment, followed by the integument, with ca 30%. The alkaloid concentration in the hemolymph is approximately sixfold higher than in the solid tissues. The selectivity of PA sequestration in larvae is comparable to PA sequestration in the bodies of adult beetles.     

Development of an Insect Herbivore and its Pupal Parasitoid Reflect Differences in Direct Plant Defense

In nature, plants defend themselves by production of allelochemicals that are toxic to herbivores. There may be considerable genetic variation in the expression of chemical defenses because of various selection pressures. In this study, we examined the development of the small cabbage butterfly, Pieris rapae, and its gregarious pupal ectoparasitoid, Pteromalus puparum, when reared on three wild populations (Kimmeridge, Old Harry, Winspit) of cabbage, Brassica oleracea, and a Brussels sprout cultivar. Wild plant populations were obtained from seeds of plants that grow naturally along the south coast of Dorset, England. Significant differences in concentrations of allelochemicals (glucosinolates) were found in leaves of plants damaged by P. rapae. Total glucosinolate concentrations in Winspit plants, the population with the highest total glucosinolate concentration, were approximately four times higher than in the cultivar, the strain with the lowest total glucosinolate concentration. Pupal mass of P. rapae and adult body mass of Pt. puparum were highest when reared on the cultivar and lowest when developing on Kimmeridge plants, the wild strain with the lowest total glucosinolate concentration. Development of male parasitoids was also more negatively affected than female parasitoids. Our results reveal that plant quality, at least for the development of ‘adapted’ oligophagous herbivores, such as P. rapae, is not based on total glucosinolate content. The only glucosinolate compound that corresponded with the performance of P. rapae was the indole glucosinolate, neoglucobrassicin. Our results show that performance of ectoparasitoids may closely reflect constraints on the development of the host.     

Ergovaline and peramine production in endophyte-infected tall fescue: Independent regulation and effects of plant and endophyte genotype

Peramine and ergovaline have ecological and economical significance because they possess insect and/or mammalian toxicity properties. The relationship between these endophytically derived alkaloids in tall fescue (Festuca arundinacea Schreb.) is unknown. Seasonal and plant tissue effects on the concentration of peramine and ergovaline was investigated in field and greenhouse experiments. The relationship between the alkaloids and the regulatory effects of endophyte and plant on their content were investigated among progeny of reciprocal crosses between high- and low-ergovaline and peramine plant-endophyte combinations. Variation in peramine concentration ranged from 750 to 1742 µg/kg in greenhouse-grown plants, and there was no seasonal trend in peramine content. There was a correlation (r=0.69) between peramine and ergovaline content among leaf tissues of field-grown plants, but there was no correlation between the alkaloids in the culm (r=0.20) or panicle (r=0.17) tissues. Mean leaf ergovaline concentration of progeny derived from the low-ergovaline parent (163 µg/kg) was less than the midparent value (228 µg/kg), but mean of progeny from the high-ergovaline parent was not different from the mid-parent value. Ranges within each progeny set were approximately double their mean. Mean leaf peramine concentrations of the progeny sets were similar to the mid-parent value (3354 µg/kg) but ranges were from 1716 to 8753 µg/kg. There was no correlation between leaf peramine and ergovaline (r=0.01). These data suggest that endophytically produced alkaloids are independently regulated and are controlled by both plant and endophyte genotype.     

Transfer of pyrrolizidine and quinolizidine alkaloids toCastilleja (Scrophulariaceae) hemiparasites from composite and legume host plants

Castilleja (Scrophulariaceae) species of the western United States contain pyrrolizidine and quinolizidine alkaloids. TheCastilleja obtain the alkaloids by root parasitism on host plants, withSenecio atratus andS. triangularis (Asteraceae) furnishing the pyrrolizidines, and quinolizidines being obtained fromLupinus species andThermopsis montana (Leguminosae). Individual plants within a givenCastilleja species population may be devoid of alkaloids while others have high alkaloid content. No populations have been found which are made up of both pyrrolizidine- and quinolizidine-containing individuals. These results have important implications forCastilleja ecology, for the study of insect herbivores which areCastilleja specialists, and in the development of systems for the investigation of the role of alkaloids as plant defenses.Paper 10 in the series Chemistry of the Scrophulariaceae. Paper 9: D.R. Gardner, J. Narum, D. Zook, and F.R. Stermitz.J. Nat. Prod. 50: In press.