An examination of phytochemical variation in Aloe elgonica Bullock

Compounds in the leaf exudates from 43 individuals of A. elgonica collected on Mount Elgon on the Kenya-Uganda border and raised at Kew were examined by two-dimensional thin-layer chromatography. The compounds reacting with Fast Blue B to give coloured zones showed patterns of variation throughout the species. These patterns could not be correlated with the considerable variations in chromosome form and leaf surface anatomy. The two major compounds in the leaf exudates were the anthrone C-glycosides, barbaloin and homonataloin, which were never observed together in the same plant. Aloenin occurs throughout the species in every plant examined, varying only in amount as detected by staining.     

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.     

Observations on the phytochemistry of the Aloë leaf-exudate compounds

Exudates from the cut surfaces of Aloë leaves contain compounds, many of which can be recognized by their colour reaction with fast blue B salt after separation on thin-layer chromatograms. About 90 chromatographic zones were observed from c. 240 Aloë species. These included 24 orange-staining zones, 35 purple-staining zones and a variety of zones staining shades of grey, green, blue and brown. A few of these substances have been identified as known compounds. Each of the remaining compounds is given a code according to its colour reaction and each is referred to a standard plant source in which it is prominent. Thus a convenient basis for phytochemical discussions is provided. The distribution of the known compounds in the genus as recorded in the literature is compared with present findings. Chemical relationships often followed accepted taxonomic groupings. In particular, correlations were found among shrubby African species and a group of species from Somalia.     

The phylogenetic significance of colour patterns in marine teleost larvae

Ichthyologists, natural‐history artists, and tropical‐fish aquarists have described, illustrated, or photographed colour patterns in adult marine fishes for centuries, but colour patterns in marine fish larvae have largely been neglected. Yet the pelagic larval stages of many marine fishes exhibit subtle to striking, ephemeral patterns of chromatophores that warrant investigation into their potential taxonomic and phylogenetic significance. Colour patterns in larvae of over 200 species of marine teleosts, primarily from the western Caribbean, were examined from digital colour photographs, and their potential utility in elucidating evolutionary relationships at various taxonomic levels was assessed. Larvae of relatively few basal marine teleosts exhibit erythrophores, xanthophores, or iridophores (i.e. nonmelanistic chromatophores), but one or more of those types of chromatophores are visible in larvae of many basal marine neoteleosts and nearly all marine percomorphs. Whether or not the presence of nonmelanistic chromatophores in pelagic marine larvae diagnoses any major teleost taxonomic group cannot be determined based on the preliminary survey conducted, but there is a trend toward increased colour from elopomorphs to percomorphs. Within percomorphs, patterns of nonmelanistic chromatophores may help resolve or contribute evidence to existing hypotheses of relationships at multiple levels of classification. Mugilid and some beloniform larvae share a unique ontogenetic transformation of colour pattern that lends support to the hypothesis of a close relationship between them. Larvae of some tetraodontiforms and lophiiforms are strikingly similar in having the trunk enclosed in an inflated sac covered with xanthophores, a character that may help resolve the relationships of these enigmatic taxa. Colour patterns in percomorph larvae also appear to diagnose certain groups at the interfamilial, familial, intergeneric, and generic levels. Slight differences in generic colour patterns, including whether the pattern comprises xanthophores or erythrophores, often distinguish species. The homology, ontogeny, and possible functional significance of colour patterns in larvae are discussed. Considerably more investigation of larval colour patterns in marine teleosts is needed to assess fully their value in phylogenetic reconstruction. © 2013 The Authors. Zoological Journal of the Linnean Society published by John Wiley & Sons Ltd on behalf of The Linnean Society of London     

Chromones and anthrones from Aloe marlothii and Aloe rupestris

A phytochemical investigation of the leaf exudate of Aloe marlothii has resulted in the isolation of a new chromone (7-O-methylaloeresin A) and a new anthrone (5-hydroxyaloin A 6'-O-acetate). Furthermore 7-O-methylaloesin was isolated as a natural product for the first time from the leaf exudate of Aloe rupestris. The structure elucidation of these compounds was based on spectral data including 2D NMR. The chemotaxonomic value of 7-O-methylaloesin in Aloe series Asperifoliae and section Pachydendron is discussed.     

Comparative chromatographic patterns of leaf exudate components from Aloe Section Pachydendron Haw

Leaf exudates from individuals of 29 species included in Aloe Section Pachydendron have been separated by TLC and HPLC to reveal their phenolic components. All the zones are described by their chromatographic behaviour and UV absorption properties, but not all can be identified as known compounds so are distinguished by an arbitrary code. Section Pachydendron has been shown to be chemically heterogeneous akhough there are some correlations between species said to be taxonomically related. Without attaching taxonomic significance four chemical groupings can be discerned: (1) species in which chromones are prominent; (2) species in which andirone and anthraquinone glycosides are prominent; (3) species containing mainly purple-staining phenolic compounds; and (4) species with few leaf exudate phenolic compounds. This survey emphasizes the uncertain taxonomy of the Section and the need for more extensive collection and analysis.     

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.     

The ecology of multiple colour defences

Individuals of many species are considered to rely on a single type of external appearance to escape predation but there are many notable exceptions. To develop an ecological framework to explain why some individuals employ different colour patterns as part of their defensive repertoire, we collate examples of colour change that are associated with living in different environments and microhabitats, examples of age-related colour change, colour defences tailored to different predators, and startle displays, where hidden conspicuous colour patterns are suddenly revealed. The over-arching commonality to all these examples is that the use of more than one defense-related external appearance is associated with a spatial or temporal change in predation risk. For example, coarse scale temporal changes in an animal’s background frequently select for gradual colour changes, while fine-scale spatial heterogeneity selects for more rapid colour changes and we provide a graphical framework for this. Irrespective of the mechanisms underlying colour change, using more than one colour defence appears driven by variation in predation risk rather than by idiosyncratic abilities to alter external appearances as is commonly believed, although physiological and energetic factors will play some role.     

Effect of marker choice and thermal cycling protocol on zooplankton DNA metabarcoding studies

DNA metabarcoding is a promising approach for rapidly surveying biodiversity and is likely to become an important tool for measuring ecosystem responses to environmental change. Metabarcoding markers need sufficient taxonomic coverage to detect groups of interest, sufficient sequence divergence to resolve species, and will ideally indicate relative abundance of taxa present. We characterized zooplankton assemblages with three different metabarcoding markers (nuclear 18S rDNA, mitochondrial COI, and mitochondrial 16S rDNA) to compare their performance in terms of taxonomic coverage, taxonomic resolution, and correspondence between morphology‐ and DNA‐based identification. COI amplicons sequenced on separate runs showed that operational taxonomic units representing >0.1% of reads per sample were highly reproducible, although slightly more taxa were detected using a lower annealing temperature. Mitochondrial COI and nuclear 18S showed similar taxonomic coverage across zooplankton phyla. However, mitochondrial COI resolved up to threefold more taxa to species compared to 18S. All markers revealed similar patterns of beta‐diversity, although different taxa were identified as the greatest contributors to these patterns for 18S. For calanoid copepod families, all markers displayed a positive relationship between biomass and sequence reads, although the relationship was typically strongest for 18S. The use of COI for metabarcoding has been questioned due to lack of conserved primer‐binding sites. However, our results show the taxonomic coverage and resolution provided by degenerate COI primers, combined with a comparatively well‐developed reference sequence database, make them valuable metabarcoding markers for biodiversity assessment.     

Genetic divergence in the superspecies Manacus

The bearded manakins in the genus Manacus are lekking, neotropical passerines. Male plumage colour varies with geographical location and classification is based solely on these plumage patterns. It has recently been suggested that in this group of birds, plumage patterns may be a misleading taxonomic character. In this study we used microsatellite variation in a collection of museum samples to establish the amount of genetic divergence between the previously described bearded manakin species/subspecies. We found substantial genetic substructuring between species/subspecies and that plumage patterns indeed may be a misleading taxonomic character because the presence of yellow in male nuptial plumage is found in most divergent forms. We did not detect a significant isolation by distance relationship although the P -value was close to significance. Physical barriers such as rivers and mountains may affect gene flow and play a role in shaping genetic structure of the genus Manacus . Accordingly, boundaries between species/subspecies often coincide with large rivers, mountains and seas.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 81 , 439–447.     

Full replacement of the function of the secondary electron acceptor phylloquinone(= vitamin K1) by non-quinone carbonyl compounds in green plant photosystem I photosynthetic reaction centers

One-carbonyl quinonoid compounds, fluorenone (fluoren-9-one), anthrone, and their derivatives are introduced into spinach photosystem (PS) I reaction centers in place of the intrinsic secondary electron acceptor phylloquinone (= vitamin K1). Anthrone and 2-nitrofluorenone fully mediated the electron-transfer reaction between the reduced primary electron acceptor chlorophyll A0- and the tertiary electron acceptor iron-sulfur centers. It is concluded that the PS I phylloquinone-binding site has a structure that enables various compounds with different molecular structures to function as the secondary acceptor and that the reactions of incorporated compounds are mainly determined by their redox properties rather than by their molecular structure. Carbonyl groups increase the binding affinity of the quinone/quinonoid compounds but do not seem to be essential to their function. The quinonoid compounds as well as quinones incorporated into the PS I phylloquinone-binding sites are estimated to function at redox potentials more negative than in organic solvents.     

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.     

A colour reaction for the differentiation of ascomycetous and hemibasidiomycetous yeasts

Seventy yeast strains, representative of twenty-six ascogenous genera, four saprobic hemibasidiomycetous genera and thirteen genera of the Cryptococcales were tested for their reaction with the stabilized aromatic diazonium compound, Diazonium Blue B salt. An aqueous, buffered solution of this compound gave a characteristic red colouration with the colonies of the hemibasidiomycetous species and those Cryptococcales characterized by the hemibasidiomycetous cell-wall type. The characteristic colour reaction was not observed with colonies of either the ascomycetous yeasts or those Cryptococcales characterized by the ascomycetous cell-wall type.The possible taxonomic use of the colour reaction with Diazonium Blue B salt as an affinitive characteristic is discussed.     

Chromosome number and flavonoid synthesis in Briza L. (Gramineae)

Following a recent reidentification of the apigenin C-glycosides in diploid Briza media, the luteolin C-glycosides were reexamined and three acylated derivatives of a luteolin C-glycoside were found. In an attempt to identify the chromosome or group of chromosomes responsible for the change in flavonoid synthesis from 4'-hydroxy- in diploid plants to 3',4'-dihydroxyflavone C-glycosides in autotetraploid plants, leaf flavonoids of artificially produced aneuploids of B. media were examined. Among these plants three different leaf flavonoid profiles were recognized: diploid, tetraploid, and a "modified" tetraploid pattern. All the aneuploids with the normal or "modified" tetraploid pattern were trisomic for one of the small acrocentric chromosomes. Induced polyploids of other Briza species were usually found to have similar flavonoid patterns in the two chromosome races. Flavonoid sulfates were found in three South American species but are absent from all the European species.     

Searching for biodiversity indicators in running waters: do bryophytes, macroinvertebrates, and fish show congruent diversity patterns?

The degree to which different taxonomic groups show congruence in diversity patterns has attracted increased attention, yet such studies on stream biota are lacking. We examined environmental correlates of and congruence in the species richness patterns of bryophytes, macroinvertebrates, and fish in 101 boreal streams in Finland. Congruence in species richness among the taxonomic groups was generally low, mainly because of their differing responses to major environmental gradients. Bryophytes and macroinvertebrates showed the strongest degree of congruence, but even this relationship had a relatively weak predictive power. Bryophyte diversity showed the strongest relationship with water colour, followed by habitat stability, and stream size. Macroinvertebrate diversity increased with stream size, and further variation was accounted for by water colour and acidity. Fish species richness showed a weak and complex relationship with geographical location, stream size, and in-stream habitat characteristics. The regression models explained 23, 45, and 26% of the variation in species richness of bryophytes, macroinvertebrates, and fish, respectively. Our results suggest that indicator taxa may be of limited value in stream biodiversity inventories. Habitat-based approaches are suggested as an alternative surrogate measure in the conservation evaluation of lotic biodiversity.     

Species boundaries, populations and colour morphs in the coral reef three-spot damselfish (Dascyllus trimaculatus) species complex

Coloration patterns of tropical reef fishes is commonly used for taxonomic purposes, yet few studies have focused on the relationship between species boundaries and coloration types. The three-spot damselfish (Dascyllus trimaculatus) species complex comprises four species that vary both in geographical ranges and colour patterns making them an ideal model to study these relationships. We analysed the mitochondrial control region of 122 individuals from all four species collected from 13 localities. Individuals from two species (Dascyllus albisella and D. strasburgi) grouped into monophyletic clades, while the two other species (D. trimaculatus and D. auripinnis) were found to be paraphyletic. Coloration patterns were therefore not found to be good predictors of genetic isolation. In contrast, geographical origin was always consistent with the observed genetic pattern.     

Interference by azide in the estimation of carbohydrates with anthrone

The anthrone reaction for the determination of carbohydrates, hexoses, pentoses, and polysaccharides is a general method which is very widely used. In this method a bacteriostatic agent, usually sodium azide, benzoic acid, or other bacteriostatic agents are added to the solution. These compounds, especially azide, appear to interfere with the development of the specific colour, introducing a serious error in the determination. A detailed study of the effect of azide and other interfering agents on a number of various carbohydrates have been made with some suggestions as to how errors may be avoided.     

Characteristics of a New Sterol-nonrequiring Mycoplasma

Two Mycoplasma strains recovered from tissue culture environments were found to grow in complex media devoid of serum or serum fractions containing cholesterol and in a cholesterol-free synthetic medium. Neither strain was capable of synthesizing pigmented carotenoids, although these compounds are present in, and characteristic of, other sterol-nonrequiring mycoplasmas. Serological tests and an analysis of their cell protein patterns obtained by gel electrophoresis indicated that the isolates were similar to each other but distinct from other sterol-nonrequiring serotypes, Mycoplasma laidlawii and M. granularum, as well as from sterol-requiring species. The existence of Mycoplasma other than M. laidlawii and M. granularum without sterol requirements suggested the need for some taxonomic changes in this group of organisms.