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.     

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.     

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.     

Wheat Gene Expression is Differentially Affected by a Virulent Russian Wheat Aphid Biotype

An improved understanding of the complex interactions between plants and aphids is emerging. Recognition of aphid feeding in plant tissues involves production of several defense response signaling pathways and downstream production of defense and detoxification compounds. Feeding by Russian wheat aphid, Diuraphis noxia (Kurdjumov), a serious pest of cereal crops worldwide, induces foliar deformity and chlorophyll loss during compatible wheat-D. noxia interactions. Experiments described here revealed significant differences in level and pattern of gene expression in defense response signaling and metabolic pathways between compatible and incompatible D. noxia-wheat interactions. The jasmonate (JA)-signaling genes LOX, AOS, and AOC were significantly more upregulated (~3- to 7 fold) in incompatible interactions than in compatible interactions (~2.5 to 3.5 fold) as early as 1 h post D. noxia infestation (hpi). Cellulose synthase, responsible for strengthening plant cell walls via cellulose production, was also more upregulated in incompatible interactions (4 to 7 fold) than in compatible interactions (1 to 3.5 fold). In contrast, glycolysis and citric acid cycle genes were significantly downregulated (~1.5 to 2 fold) in incompatible interactions and upregulated or less downregulated in compatible interactions from 6 to 72 hpi. Differences in expression of JA-signaling genes between feeding site tissues and non-feeding site tissues suggest that D. noxia defense response signals in wheat are restricted primarily to aphid feeding sites in the initial 6 hpi. This is the first report of differential upregulation of plant genes at 1 hpi in incompatible interactions involving aphid herbivory. Early wheat plant defense responses in incompatible D. noxia interactions at 1, 3, and 6 hpi appear to be important aspects of D. noxia resistance in wheat.     

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.     

Predicting toxicity of tall larkspur (Delphinium barbeyi): measurement of the variation in alkaloid concentration among plants and among years

Tall larkspur (Delphinium barbeyi) is the principal mountain larkspur responsible for the majority of cattle deaths on mountain rangelands in western Colorado and central and southern Utah in the United States. Ten plants in each of two tall larkspur populations in the mountains near Ferron and Salina, Utah, were marked, and single stalks were harvested periodically through the growing season for 4 yr. Toxic alkaloid concentration [alkaloids containing the N-(methylsuccimimido)-anthranilik ester group] was determined by Fourier transform infrared (FTIR) spectroscopy. Individual larkspur plants varied in alkaloid concentrations, especially in early growth (14–38 mg/g). As the concentration declined over the growing season, variation among plants also declined. There were yearly differences in alkaloid concentration among individual plants (P < 0.01) and populations (P < 0.001), even after accounting for differences in phenological growth between years. Variables such as precipitation, temperature, days since snow melt, growing degree days (sum of mean temperature each day from snow melt), and plant height and weight were all considered in a Mallows Cp multiple regression selection procedure to predict alkaloid concentration. The mixed model procedure in SAS adjusted the regression equation for locations and years. Growing degree days was the best single predictor of alkaloid levels: ln y = (3.581 – 0.00423 GDD), R ² = 0.85. Internal validation of this equation within individual years and locations from which the equation was developed, produced correlations between observed versus predicted values ranging from r = 0.73 to 0.93. External validations on nine other larkspur populations produced correlations ranging from r = 0.76 to 0.99. This predictive equation can provide a tool for ranchers and land managers to make management decisions of when to graze cattle in larkspur areas.     

Further Alkaloids Common to Ants and Frogs: Decahydroquinolines and a Quinolizidine

Three alkaloids—two minor decahydroquinolines (DHQs) and a major quinolizidine—were detected in an extract of a Brazilian myrmicine ant (Solenopsis (Diplorhoptrum) sp. picea group). One DHQ (3) was identical to a known frog-skin alkaloid, cis-195A (cis-5-methyl-2-propyldecahydroquinoline), while the second DHQ, an isomer of 3, designated 195J, was assigned a tentative cis-2-methyl-5-propyldecahydroquinoline structure (2) based on mass and infrared spectra. The third alkaloid proved identical to the frog-skin alkaloid 195C, for which a structure had not been previously proposed. Mass and infrared spectral analysis, including chemical ionization tandem mass spectrometry, indicated a 4-methyl-6-propylquinolizidine structure (1) for 195C. The four possible diastereomers were synthesized and the (6Z,10E)-4-methyl-6-propylquinolizidine diastereomer (1b) was identical to the natural alkaloid. Skin extracts of a population of a Madagascan mantelline frog contained, among other alkaloids, minor amounts of the same alkaloid triad 1–3 with 1 again predominating. The common occurrence of alkaloids 1–3 in both ant and frog supports the hypothesis that ants are a likely dietary source for sequestered frog-skin alkaloids and brings to six, the alkaloid classes common to ant and frog.     

Seasonal and spatial variations of alkaloids in Merendera montana in relation to chemical defense and phenology

Merendera montana (Liliaceae) is an endemic plant broadly, but exclusively, occurring in the Iberian Peninsula where it is one of the most abundant species in rangelands grazed by sheep and on the routes used by migrating sheep herds. In spite of showing a peculiar phenology adapted to Mediterranean regions (leaves remain green from fall to spring and are absent in summer), this species has also successfully colonized high altitude environments (up to 2300 m), and abundant populations can be found in montane and subalpine pastures. Chemical analysis has shown a high forage quality as well as the pre- sence of tropolone alkaloids (similar to other species of the genus Merendera), mainly colchicine, 3-demethylcolchicine, and colchicoside. In general terms, the alkaloid content was significantly lower in the subterranean corms than in the leaves. No remarkable seasonal variations in total alkaloids was found, although an increase of colchicoside in the corms and a decrease of colchicine was noted in summer. Both chemical deterrents and phenology could be the cause of the widespread ecological adaptation of Merendera montana. The alkaloid content would prevent intake by wild herbivores during winter, while the absence of aerial organs in summer, coinciding exactly with the highest domestic herbivore pressure, would permit this species to colonize rich soil habitats where there is low plant competition.     

Detrimental effects ofCinchona leaf alkaloids on larvae of the polyphagous insectSpodoptera exigua

YoungCinchona ledgeriana plants contain two types of alkaloid: indole alkaloids in the leaves and quinoline alkaloids in the root. FromCinchona leaves, a crude alkaloid extract was made, containing the cinchophylline type of indole alkaloids and a small amount of 5-methoxytryptamine. The leaf alkaloid extract exerted a strong detrimental effect on the growth of larvae of the polyphagous beet armyworm,Spodoptera exigua (Lepidoptera). Feeding of larvae on an artificial diet containing the leaf alkaloids at the same concentrations as those found in the plant resulted in significant growth reduction, retardation in development, and mortality of the larvae. Cinchophyllines are composed of 5-methoxytryptamine coupled to a corynantheal unit. When incorporated into the artificial diet, 5-methoxytryptamine alone had no effect on the 5.exigua larvae. Corynantheal, however, had a strong detrimental effect on growth of the larvae, its effect being comparable to that of the leaf alkaloid extract. In contrast to the indole-type leaf alkaloids, the quinolinetypeCinchona root alkaloids did not affect growth and development of the larvae. These results suggest that the indole-type alkaloids, which inCinchona plants are present at the highest concentrations in the young, vulnerable leaflets, are involved in the chemical defense of the plant against herbivorous insects.     

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.     

Honeydew analysis for detecting phloem transport of plant natural products

Analysis of honeydew excreted by various sap-sucking insects indicated the presence of certain plant secondary constituents in the phloem of their host plants. Honeydew excreted by mealybugs (Pseudococcus longispinus), living onCastanospermum australe, contained the indolizidine alkaloid castanospermine, a potent -glucosidase inhibitor. Similarly, honeydew from green peach aphids (Myzus persicae), feeding onSenecio vulgaris flower buds, contained the pyrrolizidine alkaloid senecionine, itsN-oxide, and hydrolytic products including retronecine. Cardenolides were detected in the honeydew of oleander aphids (Aphis nerii) feeding on oleander (Nerium oleander), indicating that these compounds are translocated in the phloem. On the other hand, honeydew from greenbugs (Schizaphis graminum), feeding on barley, lacked gramine or related indole metabolites. Similarly MBOA, the breakdown product of DIMBOA, was not detected in the honeydew of greenbugs living on DIMBOA-containing wheat.     

Influence of Phenological Stage on Swainsonine and Endophyte Concentrations in Oxytropis sericea

Locoweeds are defined as Astragalus and Oxytropis species that cause intoxication due to the alkaloid swainsonine. Swainsonine concentrations in Oxytropis sericea were influenced by location, plant part, and the developmental stage of the plant. Concentrations followed similar trends at each location, generally increasing over the growing season in above-ground parts until the plant reaches maturity with no change in concentration in the crowns. At the onset of senescence, swainsonine decreased in floral parts to less than half of the peak concentration. Similar to swainsonine concentrations, endophyte amounts were influenced by location, plant part, and the developmental stage of the plant. Likewise, endophyte amounts generally increased over the growing season in above ground parts and remained static in the crowns at all four locations. Swainsonine in Oxytropis sericea was positively associated with the endophyte Undifilum, which is responsible for swainsonine biosynthesis.     

Full-scale study on dynamic state membrane bio-reactor with modified intermittent aeration

In order to evaluate the performance of a newly developed dynamic state membrane bio-reactor (MBR) with MIA, a full-scale plant with the capacity of 210 m³/d was operated for treating sewage. Due to the MIA, the effect of residual oxygen could be completely removed during the non-aeration period and recycle ratio could be reduced to 1 recycle-to-influent ratio. Even though the plant was operated in winter season, stable performance could be achieved (BOD, CODcr, SS, TN, TP and E. coli removal efficiency; 98.2, 95.2, 99.8, 72.7, 71.4 and 99.9%, respectively). Therefore newly developed dynamic state MBR with the MIA can be one of the useful biological nutrient removal (BNR) processes for stable nutrient removal.     

Molecular Identification of Endophytic Bacteria in Leucojum aestivum In Vitro Culture,NMR-Based Metabolomics Study and LC-MS Analysis Leading to Potential Amaryllidaceae Alkaloid Production

In this study, endophytic bacteria belonging to the Bacillus genus were isolated from in vitro bulblets of Leucojum aestivum and their ability to produce Amaryllidaceae alkaloids was studied. Proton Nuclear Magnetic Resonance (¹H NMR)-based metabolomics combined with multivariate data analysis was chosen to compare the metabolism of this plant (in vivo bulbs, in vitro bulblets) with those of the endophytic bacteria community. Primary metabolites were quantified by quantitative ¹H NMR (qNMR) method. The results showed that tyrosine, one precursor of the Amaryllidaceae alkaloid biosynthesis pathway, was higher in endophytic extract compared to plant extract. In total, 22 compounds were identified including five molecules common to plant and endophyte extracts (tyrosine, isoleucine, valine, fatty acids and tyramine). In addition, endophytic extracts were analyzed using Liquid Chromatography-Mass Spectrometry (LC-MS) and Gas Chromatography-Mass Spectrometry (GC-MS) for the identification of compounds in very low concentrations. Five Amaryllidaceae alkaloids were detected in the extracts of endophytic bacteria. Lycorine, previously detected by ¹H NMR, was confirmed with LC-MS analysis. Tazettine, pseudolycorine, acetylpseudolycorine, 1,2-dihydro-chlidanthine were also identified by LC-MS using the positive ionization mode or by GC-MS. In addition, 11 primary metabolites were identified in the endophytic extracts such as tyramine, which was obtained by decarboxylation of tyrosine. Thus, Bacillus sp. isolated from L. aestivum bulblets synthesized some primary and specialized metabolites in common with the L. aestivum plant. These endophytic bacteria are an interesting new approach for producing the Amaryllidaceae alkaloid such as lycorine.     

Sequestration and metabolism of protoxic pyrrolizidine alkaloids by larvae of the leaf beetle Platyphora boucardi and their transfer via pupae into defensive secretions of adults

Several neotropical leaf-beetles of the genus Platyphora ingest and specifically metabolize plant acquired pyrrolizidine alkaloids (PAs) of the lycopsamine type (e.g., rinderine or intermedine) and enrich the processed alkaloids in their exocrine defensive secretions. In contrast to the related palaearctic leaf beetles of the genus Oreina, which absorb and store only the non-toxic alkaloid N-oxides, Platyphora sequesters PAs exclusively as protoxic tertiary amines. In this study, the ability of P. boucardi larvae to accumulate PAs was investigated. Tracer studies with [¹⁴C]rinderine and its N-oxide revealed that P. boucardi larvae, like adult beetles, utilize the two alkaloidal forms with the same efficiency, but accumulate the alkaloid as a tertiary amine exclusively. Ingested rinderine is rapidly epimerized to intermedine, which is localized in the hemolymph and all other tissues; it is also detected on the larval surface. Like adults, larvae are able to synthesize their own alkaloid esters (beetle PAs) from orally administered [¹⁴C]retronecine and endogenous aliphatic 2-hydroxy acids. These retronecine esters show the same tissue distribution as intermedine. A long-term feeding experiment lasting for almost four months revealed that retronecine esters synthesized from [¹⁴C]retronecine in the larvae are transferred from larvae via pupae into the exocrine glands of adult beetles. Pupae contain ca. 45% of the labeled retronecine originally ingested, metabolized, and stored by larvae; ca. 12% of larval radioactivity could be recovered from the defensive secretions of adults sampled successively over two and a half months. Almost all of this radioactivity is found in the insect-made retronecine esters that are highly enriched in the defensive secretions, i.e., more than 200-fold higher concentration compared to pupae.     

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.     

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.     

Individual and Geographic Variation of Skin Alkaloids in Three Species of Madagascan Poison Frogs (Mantella)

Alkaloid profiles for 81 individual mantellid frogs, Mantella baroni (Boulenger 1988) (N = 19), M. bernhardi (N = 51), and M. madagascariensis (Grandidier 1877) (N = 11), from six different populations from Madagascar were examined. Marked individual differences in alkaloid composition (number, type, and amount) were observed between different species and between populations of the same species. Disjunct populations of each of the three species differed significantly in alkaloid composition. Sympatric populations of M. baroni and M. madagascariensis also differed significantly in alkaloid composition. In M. bernhardi, differences in alkaloid composition were marginally associated with different sexes. A total of 111 alkaloids, including isomers, were detected in analysis of the individuals from the three species. The majority (47%) appear likely to be obtained from dietary mites, whereas many of the others (18%) are presumed to be from ants, and a few (4%) are from millipedes. Putative dietary sources for the remaining alkaloids are generally unknown, but beetles are probably the source of at least some of the tricyclic alkaloids (6%). In addition, alkaloid compositions from extracts of groups of individuals from five additional populations of M. baroni and from one population of M. bernhardi (Vences et al. 1994) and one population of M. cowanii (Boulenger 1882) were examined. An additional 50 alkaloids, including isomers, were detected in the combined samples, bringing the total number of alkaloids identified from these four species of mantellid frogs to 161. Alkaloid compositions in mantellid poison frogs are diverse and highly dependent on geographic location that appear to be largely determined by the nature and availability of alkaloid-containing prey items. Electronic supplementary material The online version of this article (doi: ) contains supplementary material, which is available to authorized users.     

Environmental and Genotypic Influences on Isoquinoline Alkaloid Content in Sanguinaria canadensis

In a common garden, we investigated genetic and environmental influences on alkaloid production using Sanguinaria canadensis as a model. Nutrient and shade regimes were applied to replicated clones over one growing season, and induction of alkaloid production in bloodroot was tested on a whole-plant basis using jasmonic acid as an elicitor. Alkaloid concentrations increased with decreasing light intensity and fertilizer levels. Induction was not achieved by foliar application of jasmonic acid. Genetic influences represented by clone effects may be indicated by variation in alkaloid concentration by clone, but this experimental design did not allow us to distinguish genetic from pre-experiment environmental influences on the rhizomes.     

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.