Anthraquinone,anthrone and phenylpyrone components of Aloe nyeriensis var. kedongensis leaf exudate

The leaf exudate of Aloe nyeriensis var. kedongensis yielded six compounds which were identified on the basis of spectral data and inter-conversions as two groups of three allied compounds. These were (a) 4-methoxy-6(2′,4′-dihydroxy-6′- methylphenyl)-pyran-2-one, its 2′-O-β-D-glucopyranoside (aloenin) and the 2″-O-p-coumaroyl ester of aloenin, (b) the anthracene derivatives l,2,8-trihydroxy- 6-methylanthraquinone (nataloe-emodin), its 2-O-β-D-glucopyranosyl ester and the corresponding 10-C -β-D-glucopyranoside nataloin.     

The chemotaxonomic significance of the phenyl pyrone aloenin in the genus Aloe

The phenyl pyrone, aloenin was positively identified in 16 species in a greater chemotaxonomic study on 380 species of Aloe. A large number of species have previously been suggested to be related on the basis of their macromorphological characters. The leaf exudate composition of the 16 species are presented together with a summary of the salient morphological characters. The possible taxonomic relationships between aloenin producing species, not previously thought to be associated with one another are discussed and illustrates the need to explore additional characters of taxonomic value in this large genus of ca. 420 species where no natural classification system exists.     

The uncoupling activity of substituted anthracene derivatives on isolated mitochondria from Phaseolus aureus

Unsubstituted anthraquinones and anthrones have a low uncoupling activity on isolated mitochondria. The presence of phenolic groups is a prerequisite for a marked effect. The number of phenolic groups is of minor importance but their substitution pattern is important. The methylene group of the anthrones increases the activity.     

Cape aloes—A review of the phytochemistry,pharmacology and commercialisation of Aloe ferox

Aloe ferox Mill. (= A. candelabrum A. Berger), commonly known as the bitter aloe or Cape aloe, is a polymorphic species indigenous to South Africa. The plant has been used since ancient times as a generic chemopreventive and anti-tumour remedy in folk medicine and it has a well-documented history of use as a laxative. In addition to the plethora of traditional medicinal uses, A. ferox has recently gained popularity as an ingredient in cosmetic formulations and food supplements. Anti-oxidant, antimicrobial, anti-inflammatory, anticancer and antimalarial activities, etc. have been reported. In addition, the ability of Cape aloes to enhance the transport of poorly permeable drugs has enjoyed recent research interest. Due to its medicinal and commercial importance it has been a popular research topic for natural product scientists who have isolated several chromones and anthrones from the leaf exudate and finished product (bitters). A summary of the historical and modern day uses, commercialisation, chemical composition and biological properties of this coveted ethnomedicinally and commercially important species is presented.     

Studies upon the copper fungicides. IX. Investigations with the exudate from fungus spores

Determinations have been made of the copper dissolved from pure copper compounds by water, by a standardized solution of the exudate of Neurospora sitophila spores, by standard solutions of malic and succinic acids and their sodium salts, and by glycine and mannitol.
The results compared with the known fungicidal performances of the copper compounds show that the amounts of copper yielded to water and to the standardized spore extract or glycine solution provide an indication of fungicidal properties.
Although soluble copper in excess of that dissolved by water can only appear from alkaline Bordeaux deposit by complex ion formation, results show that this is not necessarily the only means by which copper can dissolve from other copper compounds.
In a chemical investigation of the exudate of N. sitophila spores, mannitol, succinate and fumarate have been isolated. None of these materials can dissolve an appreciable amount of copper from alkaline Bordeaux deposit. Malate, found by other workers to be a constituent of the exudate of N. sitophila spores cultured on potato dextrose agar, has not been detected in such a solution from spores grown on malt agar. The quantity of mannitol in spore exudate has been shown to depend on the nutrient medium on which the fungus is grown.     

Vibrational, 1H-NMR spectroscopic,and thermal characterization of gladiolus root exudates in relation to Fusarium oxysporum f. sp. gladioli resistance

Fourier transform Raman (FT Raman) and IR (FTIR) and (1)H-NMR spectroscopies coupled with differential scanning calorimetry (DSC) were applied to the characterization of root exudates from two cultivars of gladiolus (Spic Span and White Prosperity) with different degrees of resistance and susceptibility to Fusarium oxysporum gladioli, the main pathogen of gladiolus. This work was aimed at correlating the composition of root exudates with the varietal resistance to the pathogen. Spectroscopic analysis showed that White Prosperity root exudate differs from Spic Span root exudate by a higher relative amount of the aromatic-phenolic and sugarlike components and a lower relative amount of carbonylic and aliphatic compounds. DSC analysis confirmed the spectroscopic results and showed that White Prosperity root exudate is characterized by an aromatic component that is present in a higher amount than in the Spic Span root exudate. The results are discussed in relation to the spore germination tests showing that White Prosperity, which is characterized by a remarkable resistance toward F. oxysporum gladioli, exudes substances having a negative influence on microconidial germination of the pathogen; root exudates from Spic Span, one of the most susceptible cultivars to F. oxysporum gladioli, proved to have no effect. White Prosperity's ability to inhibit conidial germination of F. oxysporum gladioli can be mainly related to the presence of a higher relative amount of aromatic-phenolic compounds.     

Therapeutic and prophylactic uses of invertebrates in contemporary Spanish ethnoveterinary medicine

Zootherapeutic practices in ethnoveterinary medicine are important in many socio-cultural environments around the world, particularly in developing countries, and they have recently started to be inventoried and studied in Europe. In light of this, the purpose of this review is to describe the local knowledge and folk remedies based on the use of invertebrates and their derivative products in contemporary Spanish ethnoveterinary medicine. An overview in the fields of ethnozoology, ethnoveterinary medicine and folklore was made. Automated searches in the most important databases were performed. All related works were examined thoroughly and use-reports were obtained from 53 documentary sources. The traditional use of 18 invertebrate species and five ethnotaxa and a total of 86 empirical remedies based on the use of a single species was recorded. The two most relevant zoological groups were found to be insects and molluscs. A broad diversity of body parts or derivative products have been and are used to treat or prevent ca. 50 animal diseases or conditions, in particular diseases of the skin and subcutaneous tissue, different infectious livestock diseases, and disorders of the eye and adnexa. Cattle, sheep and equines form the group of domestic animals in which the greatest number of remedies are mentioned. In addition, seven magical remedies and practices are documented. In comparison with other culturally related areas, this is a rich heritage. The use-reports included here will help in the search for new and low-cost drugs for treating livestock and alternative materials for pharmaceutical purposes, future research addressing the validation of the effects and the development of organic farming.     

The occurrence and taxonomic distribution of the anthrones aloin, aloinoside and microdontin in Aloe

A chemotaxonomic survey of 380 species of Aloe indicated the presence of the anthrone isomers aloin A and B together with the aloinoside isomers and microdontin A and B in 36 (10%) species of Aloe. This group, referred to as the microdontin chemotype, is thus characterised by a combination of exudate compounds and not merely a single phytochemical marker, implying taxonomic significance of leaf exudate compounds. The 36 representatives of the group occupy disparate taxonomic positions in the largely artificial hierarchy of the present classification system. Although many of the species have previously been considered as related (based on macromorphology only), a large number of species have not been associated with one another before. The chemical profiles and leaf exudate compositions of the species are presented, followed by a brief summary of the morphological diversity. Whilst conceding the possibility of convergent evolution, the geographical distribution of the species and thoughts on possible relationships between the taxa are discussed.     

THE STIGMATIC SECRETION OF THE SWEETPOTATO

The stigmatic exudate of sweetpotato, when removed with organic solvents, consisted chiefly of lipid and phenolic compounds. Only traces of sugar were obtained. The esterified lipids were similar in chain length to capric and lauric acids. The two prinicpal phenolic compounds have UV absorption peaks, and bathochromic shifts on ionization with NaOH similar to those of esters of caffeic acid. On acid hydrolysis, caffeic acid was obtained from these and from three minor phenolic compounds. From two of the minor phenolic compounds, glucose was released by hydrolysis. The phenolic content of stigmas increased up to 20–24 hr before anthesis, and then gradually decreased. Neither compatible nor incompatible pollination affects the amount or composition of the stigmatic exudate.     

The Composition of Phloem Exudate and Xylem Sap from Tree Tobacco (Nicotiana glauca Grah.)

The composition of xylem sap and exudate from stem incisionsof Nicotiana glauca Grah. was compared in detail. Exudationfrom stem incisions occurred over a 5 min period in certainplants, enabling collection of 5–30 µl of sap. Therate of exudation showed an exponential decline. Exudate hada high dry matter content (170–196 mg ml^–1) andhigh sugar (sucrose) levels. Xylem sap had a low pH (5.8) andexudate a pH of 7.9. Glutamine dominated the amino compoundsin xylem sap and exudate, and K⁺ was the major cation. Totalamino compounds in stem exudate reached 10.8 mg ml^–1 whereasxylem sap contained much lower levels (0.28 mg ml^–1).All mineral elements and amino compounds with the exceptionof calcium were more concentrated in stem exudate than in xylemsap. Sucrose was labelled heavily in stem exudate following pulsingof an adjacent leaf with ¹⁴CO₂. A concentration gradient ofsugar (2.1 bar m^–1) was recorded for stems. Levels ofsucrose, amino compounds and K⁺ ions in stem exudate showeda diurnal periodicity. Each commodity reached maximum concentrationat or near noon and minimum concentration about dawn. The evidencesuggests that exudate from stem incisions of N. glauca is arepresentative sample of solutes translocated in the phloem. Nicotiana glauca Grah., phloem sap, xylem sap, sucrose, amino compounds, mineral ions     

Anthraquinones as defensive compounds in eggs of Galerucini leaf beetles: Biosynthesis by the beetles?

Eggs of leaf beetles of the tribe Galerucini, subfamily Galerucinae, contain polyketides that are unusual in insects: 1,8-dihydroxylated anthraquinones (chrysazin, chrysophanol) and anthrones (dithranol, chrysarobin) deterring predators. The host plants do not contain these compounds. In the present study, we tested the hypothesis that the beetles, but not bacterial or fungal microorganisms living as endosymbionts within the beetles, produce the anthraquinones. The tansy leaf beetle Galeruca tanaceti was used as Galerucini model organism. It was treated with antimicrobial substances to eradicate the microorganisms and inhibit the hypothesised endosymbiotic anthraquinone production. Despite treatment, female G. tanaceti laid eggs containing anthraquinones. Although broad spectrum antimicrobials were used, it cannot be excluded that the potential endosymbiotic microorganisms are resistant. Given that the hypothesised endosymbionts are transferred via the eggs from one generation to the next, bacterial or fungal DNA was expected to be present in the eggs. With the exception of Wolbachia pipientis, however, no further 16S rDNA from bacteria responsible for anthraquinone biosynthesis was detected in eggs of untreated beetles. Because Wolbachia were also found in closely related anthraquinone-free insects, we exclude these bacteria as producers of the defensive polyketides. Nor was any 18S rDNA from fungi with anthraquinone biosynthetic abilities detected. Our results indicate that anthraquinones and anthrones in eggs of Galerucini are produced by beetle enzymes and not by endosymbiotic microorganisms within the eggs.     

Biochemical composition of stigmatic exudate of Eichhornia crassipes (Mart.) solms

The stigmatic exudate of Eichhornia crassipes (Mart.) Solms is moderately soluble in water, but is readily soluble in 80% ethanol. Soluble sugars, fructose, sucrose, total phenol, hydroxy phenol, free amino acids, soluble proteins and free fatty acids were found in the exudate. The exudate is devoid of organic acids and alkaloids, while proteins, soluble sugars, free fatty acids and free amino acids were the principal constituents. The stigma possesses detectable quantities of these compounds and shows a progressive increase in the amount of these substances up to the stage just prior to anthesis. The ultraviolet absorption profile of the stigmatic exudate in ethanol shows characteristic absorption peaks and bathochromatic shifts on addition of NaOH. The peaks probably correspond to those of simple phenolic compounds.     

Quantification by ELISA of Cytokinins in Root-Pressure Exudates of Urtica dioica Plants Grown under Different Nitrogen Levels

ELISAs for [9R]iP, [9R]Z and (diH) [9R]Z have been worked out which, in combination with HPLC., allow the quantitative determination of cytokinins of the isopentenyladenin-, zeatin- and dihydrozeatin-type in samples of plant material. The three ELISAs are optimally applicable in the range from 0.5 to 20 pmoles of cytokinins per assay but reliable measurements can still be performed down to 0.05 pmoles. The assay has been applied to root pressure exudates obtained from Urtica dioica plants grown under controlled conditions and at different nitrogen (nitrate) levels. The predominant cytokinins of the xylem exudate were Z, Z-nucleotide, iP, [9R]iP and iP-nucleotide, while (diH)Z was present to a minor extent. The concentrations of these compounds were found to range between 0.2 and 3 pmoles per ml exudate. Interestingly, no effect of the different nitrogen nutrition on the pattern and concentration of cytokinins in the xylem exudate could be established.     

Ethnoveterinary medicines used for horses in Trinidad and in British Columbia, Canada

This paper investigates the commonalities in ethnoveterinary medicine used for horses between Trinidad (West Indies) and British Columbia (Canada). These research areas are part of a common market in pharmaceuticals and are both involved in the North American racing circuit. There has been very little research conducted on medicinal plants used for horses although their use is widespread. The data on ethnoveterinary medicines used for horses was obtained through key informant interviews with horse owners, trainers, breeders, jockeys, grooms and animal care specialists in two research areas: Trinidad and British Columbia (BC). A participatory validation workshop was held in BC. An extensive literature review and botanical identification of the plants was also done. In all, 20 plants were found to be used in treating racehorses in Trinidad and 97 in BC. Of these the most-evidently effective plants 19 of the plants used in Trinidad and 66 of those used in BC are described and evaluated in this paper. Aloe vera, Curcuma longaand Ricinus communisare used in both research areas. More research is needed in Trinidad to identify plants that respondents claimed were used in the past. Far more studies have been conducted on the temperate and Chinese medicinal plants used in BC and therefore these ethnoveterinary remedies reflect stronger evidence of efficacy.     

Secretory tissues in the East African shrubby aloes

Examination of Aloe leaf sections revealed the presence of three types of cells at the phloem pole of the vascular bundles, aloin cells, outer bundle sheath cells and fibres. Three species contain fibres alone and produce a sparse exudate with few components staining purple with Fast Blue B on thin-layer chromatograms. The majority of species have aloin cells of various sizes and in the tetraploid species these produce a copious exudate containing anthraquinone and chromone derivatives. It is suggested that the aloin cells act as storage tissue and that the compounds are synthesized in the surrounding layer of cells of smaller diameter, many of which can be seen to contain globules of unknown constitution. Anatomical observations support the idea that the shrubby tetraploid Aloe species are derived from a form similar to the diploid Aloe morijensis. Two forms of this species have been described, one with fibres only and few exudate components and another with both fibres and aloin cells and an exudate containing compounds of the tetraploid species. A line of diploid plants similar in anatomy and chemistry to the first form is represented by A. fibrosa and A. babatiensis while a line of tetraploids with affinities to the second form is represented by A. nyeriensis, A. cheranganiensis, A. elgonica, A. dawei and A.yavellana.     

Bradyrhizobium japonicum nodD1 can be specifically induced by soybean flavonoids that do not induce the nodYABCSUIJ operon.

Besides genistein and daidzein, which are active inducers of the nodYABCSUIJ operon in Bradyrhizobium japonicum, soybean seeds also excrete compounds that are not inducers of the nodYABCSUIJ genes but enhance induction of this operon in the presence of a suboptimal genistein concentration. This synergism was studied in detail, and specific compounds were identified in seed exudate which specifically induce the nodD1 gene but not the nodYABCSUIJ operon. Therefore, our current hypothesis is that the observed synergism is caused by a specific induction of nodD1. The specific nodD1 inducers from soybean seed extract have been purified and characterized chemically. They appear to be derivatives of genistein, glycitein, and daidzein with glucose, malonyl, and acetyl groups attached. Both root and seed exudate appear to contain these compounds, with the seed being the major source. No hydrolysis of these compounds to their aglycone forms was detected in the presence of B. japonicum. A model for nod gene induction in B. japonicum is discussed.     

Induction of the nodA promoter of Rhizobium leguminosarum Sym plasmid pRL1JI by plant flavanones and flavones.

An expression vector containing the Rhizobium leguminosarum nodA promoter cloned in front of the Escherichia coli lacZ gene was used to characterize the properties of the R. leguminosarum nodA gene-inducing compound(s) present in sterile root exudate of the host plant Vicia sativa L. subsp. nigra (L.). The major inducing compound was flavonoid in nature, most likely a flavanone. The commercially available flavonoids naringenin (5,7,4'-trihydroxyflavanone), eriodictyol (5,7,3'4'-tetrahydroxyflavanone), apigenin (5,7,4'-trihydroxyflavone), and luteolin (5,7,3',4'-tetrahydroxyflavone) induced the nodA promoter to the same level as the root exudate. On the basis of chromatographic properties, it was concluded that none of these compounds is identical to the inducer that is present in root exudate. The induction of the nodA promoter by root exudate and by the most effective inducer naringenin was very similar, as judged from the genetic requirements and the kinetics of induction.     

Extracellular organics from specific cultures of Heterosigma akashiwo (Raphidophyceae) irreversibly alter respiratory activity in mammalian cells

Raphidophyte blooms have been well documented in several coastal areas around the world. Centring raphidophyte-bloom research has been a focus evolving around issues of ichthyotoxicity, allelopathy and anti-predatory activity. However, the details of these phenomena such as the identity of the compounds and the mechanisms underlying these processes are poorly understood. One such raphidophyte, Heterosigma akashiwo (Hada) Hara et Chihara, has historically received much attention with regard to its ichthyotoxic and allelopathic properties. In this study, we collected extracellular organic compounds from cultures of nine H. akashiwo isolates and tested those exudates on two mammalian cell lines: rat osteoblastic sarcoma (UMR-106) and human embryonic kidney (HEK-293). A tetrazolium colourimetric assay was used to determine the activity of mitochondrial dehydrogenases. Exposure of the mammalian cell lines to exudates collected from cultures of H. akashiwo (strain 764) significantly increased activity in a concentration- and time-dependent manner. Exudate concentrations of as little as 0.3 mg ml⁻¹ elicited a stimulatory response in the mammalian cells. This is comparable to the range of exudate concentrations that were originally in the algal cultures (>0.1 mg ml⁻¹). Significant increases in activity were observed 12–24 h following continuous or 1 h (transient) exposure to the exudate. Production of the stimulatory bioactive exudate was not altered by nutrient-stressed H. akashiwo cultures (reduced iron, phosphate or nitrate). Collectively, these bioactive compound(s) consistently increased cellular activity 3–15-fold. Interestingly, of the nine isolates tested, four of them produced the stimulatory exudate, whereas four others did not produce the stimulatory compound(s) and isolate 560R produced a compound(s) that was inhibitory in nature. Thus, we have shown that cultures of H. akashiwo produce organic compounds that can alter the metabolic activity of mammalian cells. Future isolation and characterization of these bioactive compounds may determine them to have ecological relevance, potentially involved in the ichthyotoxic, allelopathic and/or anti-predatory behaviour of this alga.     

A contribution to the phytochemistry of the East African tetraploid shrubby aloes and their diploid allies

Compounds in the leaf exudates of a group of shrubby aloes from East Africa have been re-examined by two-dimensional thin-layer chromatography using Fast Blue B as a revealing reagent. Exudates from the most abundant taxa, the two subspecies of Aloe nyeriensis , were found to contain substances giving four distinct chromatographic patterns. One was distinguished by the presence of the anthrone C-glycoside barbaloin while two others contained instead homonataloin with or without aloenin. The fourth group contained neither of these but a similar compound of unknown constitution. Exudate patterns from the neighbouring but localized tetraploid species A. cheranganiensis , the more remote A. dawei and A. elonica , together with the diploid A. yavellana , related to one or other of the groupings of A. nyeriensis. Some plants of a localized diploid species, A. morijensis , also yielded a similar pattern but other plants of rather more slender form contained quite different compounds resembling those in the diploids A. fibrosa and A. babatiensis. This supports the hypothesis that A. morijensis is close to the putative ancestor of the tetraploid East African shrubby aloes. The other widespread shrubby species in this locality, the diploid A. rabaiensis , is unrelated taxonomically and chemically.