The alkaloid profiles of Sophora nuttalliana and Sophora stenophylla

Sophora is a diverse genus in the family Fabaceae, comprised of herbs, shrubs, and trees that occurs throughout the world, primarily in the northern hemisphere. Species of Sophora are known to contain quinolizidine alkaloids that are toxic and potentially teratogenic. Two perennial herbaceous species occur in North America, Sophora stenophylla and Sophora nuttalliana. The quinolizidine alkaloid composition of these two species was investigated throughout their geographical distribution using field collections and herbarium specimens. Both species contain quinolizidine alkaloids, and S. nuttalliana contains the teratogen anagyrine. Lastly, neither species contains the neurotoxin swainsonine as implied by the common name “white loco” for S. nuttalliana.     

Plant alkaloids that cause developmental defects through the disruption of cholinergic neurotransmission

The exposure of a developing embryo or fetus to alkaloids from plants, plant products, or plant extracts has the potential to cause developmental defects in humans and animals. These defects may have multiple causes, but those induced by piperidine and quinolizidine alkaloids arise from the inhibition of fetal movement and are generally referred to as multiple congenital contracture‐type deformities. These skeletal deformities include arthrogyrposis, kyposis, lordosis, scoliosis, and torticollis, associated secondary defects, and cleft palate. Structure‐function studies have shown that plant alkaloids with a piperidine ring and a minimum of a three‐carbon side‐chain α to the piperidine nitrogen are teratogenic. Further studies determined that an unsaturation in the piperidine ring, as occurs in gamma coniceine, or anabaseine, enhances the toxic and teratogenic activity, whereas the N‐methyl derivatives are less potent. Enantiomers of the piperidine teratogens, coniine, ammodendrine, and anabasine, also exhibit differences in biological activity, as shown in cell culture studies, suggesting variability in the activity due to the optical rotation at the chiral center of these stereoisomers. In this article, we review the molecular mechanism at the nicotinic pharmacophore and biological activities, as it is currently understood, of a group of piperidine and quinolizidine alkaloid teratogens that impart a series of flexure‐type skeletal defects and cleft palate in animals. Birth Defects Research (Part C) 99:235–246, 2013. Published 2013 Wiley Priodicals, Inc.     

Support for the removal of Cyclolobium from tribe Millettieae (Leguminosae) from its quinolizidine alkaloid status

Fruits of Cyclolobium brasiliense Benth. (Leguminosae; Papilionoideae) were found to contain quinolizidine alkaloids. Several tetracyclic sparteine-type alkaloids, the bipiperidyl alkaloid ammodendrine and the α-pyridone alkaloid N-methylcytisine were identified. The presence of quinolizidine alkaloids in this monotypic genus supports a relationship with tribe Brongniartieae and genistoid tribes rather than its current placement in tribe Millettieae.     

Alkaloid profiles of Dermatophyllum arizonicum,Dermatophyllum gypsophilum,Dermatophyllum secundiflorum,Styphnolobium affine,and Styphnolobium japonicum previously classified as Sophora species

Sophora arizonica, Sophora gypsophila, Sophora secundiflora, Sophora affinis, and Sophora japonica were recently reclassified as Dermatophyllum arizonicum, Dermatophyllum gypsophilum, Dermatophyllum secundiflorum, Styphnolobium affine, and Styphnolobium japonicum, respectively. Some legumes of the sub family Papilionoideae including Sophora species are reported to contain a variety of quinolizidine alkaloids. The quinolizidine alkaloid profiles of D. arizonicum, D. gypsophilum, D. secundiflorum, S. affine, and S. japonicum were investigated qualitatively and quantitatively using field collections and herbarium specimens throughout their range of geographical distribution for the native species. This is the first report of the alkaloid profiles of D. arizonicum and D. gypsophilum. Alkaloid profiles of the other species were compared to previous reports. The Dermatophyllum species contain quinolizidine alkaloids, and the teratogen anagyrine (11), while the Styphnolobium species do not contain quinolizidine alkaloids. The chemotaxonomic data are consistent with the reclassification of each species.     

(+)-2,3-dehydro-10-oxo-alpha-isosparteine in Uresiphita reversalis larvae fed on Cytisus monspessulanus leaves

Quinolizidine alkaloids, found in the leaves of Cytisus monspessulanus L. (Leguminosae), were characterized in the cuticle of larvae of the pyralid moth Uresiphita reversalis (Lepidoptera: Pyralidae) when the latter were fed on this weed. By GC-MS analysis of the methanolic extracts of the cuticle, four quinolizidine alkaloids, N-methylcytisine, cytisine, aphylline and anagyrine, were identified as possible defense substances. In addition, the quinolizidine alkaloid, (+)-2,3-dehydro-10-oxo-alpha-isosparteine was characterized in both the insect and host plant.     

Alkaloids of Bolusanthus speciosus

Investigation of the alkaloids of Bolusanthus speciosus afforded a new quinolizidine alkaloid, 6β-hydroxylupanine. The structure of this alkaloid was assigned on the basis of spectroscopic methods and by chemical transformations. The other alkaloids isolated were cytisine, N-methylcytisine, 11α-allylcytisine, anagyrine, 13-hydroxyanagyrine, 5,6-dehydrolupanine, lupanine, sparteine and β-isoparteine. The biosynthetic significance of these findings is discussed briefly.     

Localization of the Enzymes of Quinolizidine Alkaloid Biosynthesis in Leaf Chloroplasts of Lupinus polyphyllus

Studies with purified chloroplasts of Lupinus polyphyllus LINDL. leaflets indicate that the first two enzymes of quinolizidine alkaloid biosynthesis, lysine decarboxylase and 17-oxosparteine synthase, are localized in the chloroplast stroma. Thus, both enzymes share the same subcellular compartment as the biosynthetic pathway of lysine, the precursor of quinolizidine alkaloids. The activity of diaminopimelate decarboxylase, the final enzyme in lysine biosynthesis, is about two to three orders of magnitude higher than that of the enzymes of alkaloid formation.     

Inhibition of seed germination by quinolizidine alkaloids

Germination of Lactuca sativa L. was inhibited by mixtures of quinolizidine alkaloids. The alkaloid esters resulted in the strongest inhibition: 6 mM 13-tigloyloxylupanine inhibited germination by 100%, whereas the other lupin alkaloids, such as lupanine and sparteine, gave a 45 and 20% inhibition, respectively. Seedlings of Lupinus albus L., which are not affected by quinolizidine alkaloids, excrete lupanine and 13-tigloyloxylupanine into the surrounding medium by their roots. It is assumed that lupin alkaloids are potential compounds of plant-plant interaction (i.e. allelopathy) besides their role in plant-herbivore interrelations.     

Concomitant occurrence of pyrrolizidine and quinolizidine alkaloids in the hemiparasite Osyris alba L. (Santalaceae)

The investigation of the alkaloid extracts of the hemiparasitic plant Osyris alba, collected from three different localities in southern France, revealed the concomitant presence of both pyrrolizidine (PA) and quinolizidine (QA) alkaloids in the samples from two of these localities. The sample from the third locality contained only PAs. The eight QAs identified were sparteine, N-methylcytisine, cytisine, methyl-12-cytisine acetate, hydroxy-N-methylcytisine, N-acetylcytisine, lupanine, and anagyrine. Of the eleven detected PAs, eight were identified as chysin A, chysin B, 1-carboxypyrrolizidine-7-olide, senecionine, integerrimine, retrorsine, senecivernine and a new alkaloid janfestine (7R-hydroxychysin A or 1R-carbomethoxy-7R-hydroxypyrrolizidine). PAs were mainly present as their N-oxides This is, to our knowledge, the first report demonstrating the simultaneous presence of two classes of alkaloids, quinolizidine and pyrrolizidine alkaloids, in a single parasitic plant. As these alkaloids do not occur in the same host plant, the results indicate that Osyris must have tapped more than one host plant concomitantly. Since both quinolizidine and pyrrolizidine alkaloids serve as defence compounds against herbivores, affecting different molecular targets, the simultaneous acquisition of the two types of alkaloids by a single plant could provide a novel mode of defence of hemiparasites against herbivores.     

Iridoids and alkaloids from Castilleja host plants for platyptilia pica. Rhexifoline content of P. pica

Iridoid and alkaloid analyses were conducted on Castilleja sulphurea, C. occidentalis, C. rhexifolia and C. hispida (Scrophulariaceae). Pyrrolizidine alkaloids were found in C. rhexifolia and some C. sulphurea populations, but other C. sulphurea populations had quinolizidine alkaloids or none at all. No C. occidentalis populations contained alkaloids. C. hispida was found to contain lamprolobine and the quinolizidine alkaloid anagyrine. All taxa contained the pyridinemonoterpene rhexifoline. The iridoid content of the Castilleja species were all qualitatively similar. Major iridoids were aucubin, catalpol, penstemonoside and shanzhiside methyl ester, with traces of 8-epiloganin and gardoside methyl ester. Larvae of Platyptilia pica (Pterophoridae) hosted by Castilleja were found to excrete and not sequester iridoids. The adult moths contained rhexifoline alkaloid, but at a low concentration level. Systematic implications of the results for Castilleja are discussed.     

Chemotaxonomy of Portuguese Ulex: quinolizidine alkaloids as taxonomical markers

Six species of Portuguese Ulex L. in a total of nineteen populations were studied by GC-EIMS as to their content in quinolizidine alkaloids. Sparteine, beta-isosparteine, jussiaeiine A, N-methylcytisine, cytisine, 5,6-dehydrolupanine, rhombifoline, lupanine, jussiaeiine B, N-formylcytisine, N-acetylcytisine, anagyrine, jussiaeiine C, jussiaeiine D, pohakuline, baptifoline, and epibaptifoline were detected. Analysis of the chromatograms showed that the chemical profile of all species was mainly composed of N-methylcytisine, cytisine, anagyrine, and jussiaeiines A, B, C and D. Therefore a quantification study of these alkaloids in all the populations studied was done by GC. These data were then submitted to cluster analysis and principal component analysis, which allowed the definition of five chemotypes and the recognition of hybrids. N-methylcytisine, cytisine, and jussiaeiines A, C and D are recognized as markers of this genus in Portugal.     

Turnover and transport of quinolizidine alkaloids. Diurnal fluctuations of lupanine in the phloem sap,leaves and fruits of Lupinus albus L.

Quinolizidine alkaloids formed in the leaves of Lupinus albus L. are translocated via the phloem to the other plant organs, especially the maturing fruits. Compared with amino-acid transport in the phloem, the alkaloids contribute about 8% to the overall nitrogen being exported from the leaf. Since it is likely that the alkaloids are subsequently degraded in the target tissues a minor role of quinolizidine alkaloids might be nitrogen transport. A marked diurnal fluctuation of alkaloids was observed in the leaves, the phloem sap, the roots and the fruits with an increase during the day and an amplitude of several hundred percent thus providing evidence for a rapid turnover of endogenous alkaloids.Abbreviations QA quinolizidine alkaloids - GLC gas-liquid chromatography     

Intraspecific variation in alkaloid profile of four lupine species with implications for the pea aphid probing behaviour

The alkaloid profile of white lupine Lupinus albus L. cv. ‘Boros’ and ‘Butan’, narrow-leaf lupine L. angustifolius L. cv. ‘Bojar’, ‘Graf’, ‘Karo’, ‘Mirela’, and ‘Sonet’, yellow lupine L. luteus L. cv. ‘Dukat’, ‘Parys’, ‘Perkoz’, and ‘Talar’, and wild species big-leaf lupine L. polyphyllus Lindl. was studied. The pea aphid Acyrthosiphon pisum Harris probing behaviour was monitored using the electrical penetration graph (EPG) technique.Four types of feeding behaviour were recognized. First, on highly acceptable L. luteus cv. ‘Dukat’, the total and mean duration of probing, time to reach phloem phase, and the duration of the first phloem phase were comparable to those in aphids on control plant Pisum sativum. Second, on partially acceptable L. luteus cv. ‘Talar’, pathway activities were slightly impeded, and the probes were more numerous and slightly shorter than on control plants. The phloem phase occurred sporadically, and feeding was terminated early after a brief period of ingestion. Third, on unpalatable but acceptable L. angustifolius cv. ‘Bojar’ and ‘Sonet’, and L. luteus cv. ‘Parys’ and ‘Perkoz’, the probes were numerous and usually epidermal. The phloem phase occurred rarely, and when it did, it was short and consisted mainly of watery salivation. Finally, on unacceptable L. albus cv. ‘Boros’ and ‘Butan’, L. angustifolius cv. ‘Graf’, ‘Karo’, ‘Mirela’, and L. polyphyllus, total probing time and individual probes were short and phloem phase did not occur.Eighteen alkaloids were identified: one piperidine alkaloid (ammodendrine), one indole (gramine), ten quinolizidine alkaloids (one tricyclic and nine tetracyclic compounds), and six esters. All lupine varieties that contained lupanine, its derivatives and especially their esters appeared to be unacceptable to the pea aphid, independent of the total concentration of any specific lupanine alkaloid. In contrast, sparteine and its derivatives did not seem to affect aphid probing significantly.     

Activation of chloroplast-localized enzymes of quinolizidine alkaloid biosynthesis by reduced thioredoxin

Previous studies from the laboratory of the authors have shown that the tetracyclic quinolizidine alkaloids are synthesized in leaf chloroplasts of Lupinus polyphyllus. Additionally, alkaloid formation reveals a light dependent diurnal rhythm in vivo. The present study shows that the principal biosynthetic enzymes, lysine decarboxylase and 17-oxosparteine synthase, assayed in acetone powder extracts and isolated chloroplasts of L. polyphyllus, were activated by reduced E. coli thioredoxin. Since both enzymes display optimal activity at pH 8 and were rather inactive at pH 7, both thioredoxin and the light mediated shift in the hydrogen ion concentration of the chloroplast stroma from pH 7 to pH 8 may be involved in the light controlled alkaloid formation.Abbreviations DTE dithioerythritol     

Alkaloid profile of leaves and seeds of Lupinus hintonii C. P. Smith

L. hintonii C. P. Smith grows in the Central Highland forests of Mexico at altitudes between 2800 m to 3200 m above see level. Members of the genus Lupinus produce quinolizidine alkaloids as main chemical defensive compounds against herbivores. Surprisingly alkaloid profiles are rather constant within this species, while substantial variation was found when compared to morphologically closely related other taxa. As part of a phytochemical project on Mexican wild lupins, we report on the alkaloid profiles of seeds and leaves of L. hintonii. 19 alkaloids could be identified by capillary GLC-MS. Six major alkaloids occurred in leaves and seeds: 13-hydroxylupanine (28% and 45% respectively), tetrahydrorhombifoline (31% and 23% respectively), angustifoline (2% and 4% respectively), lupanine (7% and 5% respectively), 13alpha-tigloyloxylupanine (19% and 5% respectively) and 4alpha-angeloyl-3beta-hydroxylupanine (9% and 2%). This chemical pattern resembles that of the North American lupin L. floribundus.     

Quinolizidine alkaloid profiles of two taxa of Teline maderensis

The alkaloid composition of the aerial parts of two taxa of Teline maderensis was studied by capillary GLC and GLC-MS. N-Methylcytisine was the major alkaloid found in both plants. Contents of cytisine and lupanine were higher in T. maderensis var. paivae while anagyrine content was more pronounced in T. maderensis var. maderensis. The alkaloids dehydrocytisine, N-acetylcytisine and epibaptifoline appeared only in T. maderensis var. maderensis and N-formylcytisine was identified as a minor constituent in T. maderensis var. paivae, and detected only in trace amounts in the other variety of the plant.     

Determination of quinolizidine alkaloids in Sophora tonkinensis by HPCE

A simple, rapid and reliable high-performance capillary electrophoresis method has been developed to determine quantitatively the alkaloid content of Sophora tonkinensis, a Chinese herb commonly known as shan-dou-gen. A total of seven quinolizidine alkaloids (cytisine, sophocarpine, matrine, lehmannine, sophoranol, oxymatrine and oxysophocarpine) could be readily separated within 15 min. The running buffer was 50 mM phosphate buffer (pH 2.5) containing 1% hydroxypropyl-beta-cyclodextrin and 3.3% isopropanol in water. The applied voltage was 25 kV, the capillary temperature was 25 degrees C, the detection wavelength was 200 nm and scopolamine butylbromide was used as internal standard. The method was used to analyse the chemical constituents of two commercial alternatives to shan-dou-gen. The alkaloid constituents of authentic shan-dou-gen gave a specific HPCE electropherogram that could be used to distinguish the drug from potential substitutes. Furthermore, the content of oxymatrine and the total content of the seven quinolizidine alkaloids could be used as quantitative markers in order to assess the quality of S. tonkinensis.     

Post-damage alkaloid concentration in sweet and bitter lupin varieties and its effect on subsequent herbivory

Abstract:  While most lupin species possess quinolizidine alkaloids, sweet (low alkaloid) varieties are more palatable but at the same time more susceptible to herbivory. Nevertheless, as they are not totally devoid of alkaloids, it may be possible that their alkaloid levels increase after damage. The aim of this study was to compare inductive responses to herbivory in sweet and bitter varieties of Lupinus albus (L.) and Lupinus angustifolius (L.), and to assess if these responses were effective to stop subsequent herbivory. Two experiments were carried out; in the first, Anticarsia gemmatalis (Hübner; Lep., Noctuidae) caterpillars were introduced in field-growing lupin plants and allowed to feed for 72 h, after which leaves were collected and analysed for alkaloid content and composition. The second experiment was a bioassay, in which leaves collected from experiment 1, from treated and control plants, were offered to another set of Anticarsia caterpillars, and consumption was recorded after 24 h. We found that both L. albus varieties (sweet and bitter) had an increase in their alkaloid concentration after damage, while none of the L. angustifolius varieties had. The sweet L. albus variety, Rumbo, had a greater inductive response than the bitter variety. When leaves were offered to caterpillars (bioassay), this variety showed the greatest difference between consumption of controls and previously eaten leaves, implying that alkaloid levels reached after damage were effective to deter subsequent herbivores as a result of plants probably overcoming a 'palatability threshold'.     

Cellular localization of quinolizidine alkaloids by laser desorption mass spectrometry (LAMMA 1000)

Stem sections of Lupinus polyphyllus and Cytisus scoparius have been analyzed for the distribution of quinolizidine alkaloids by laser desorption mass spectrometry, employing a LAMMA 1000 instrument. Sparteine and lupanine could be recorded and were found to be restricted to the epidermis and probably also to the neighbouring 1 or 2 subepidermal cell layers.Abbreviations QA quinolizidine alkaloids - GLC gas liquid chromatography - MS mass spectrometry     

Phytochemical differences between Calia secundiflora (Leguminosae) growing at two sites in Mexico

The ecology and quinolizidine alkaloid chemistry of Calia secundiflora (Ortega) Yakovlev growing at two sites in Mexico were compared. At one site (Hidalgo) the vegetation was dominated by Flourensia resinosa and C. secundiflora, at the other site (Queretaro) C. secundiflora and Dodanaea viscosa were dominant. The Hidalgo site had shallower soils with less organic matter, N, P, and CaCO3. Seeds of C. secundiflora from each site accumulated a similar range of quinolizidine alkaloids, but the profile of alkaloids in the leaves and roots were different. The leaves and roots of plants at Hidalgo accumulated a similar range of alkaloids to the seeds with cytisine and/or N-methylcytisine being most abundant, whereas at Queretaro the leaves and roots accumulated lupinine, with other alkaloids being relatively minor constituents. The latter profile has not been reported previously for C. secundiflora.