Balkans’ Asteraceae Species as a Source of Biologically Active Compounds for the Pharmaceutical and Food Industry

Herbal drugs are a useful source of different bioactive compounds. Asteraceae species, as the most widespread vascular plants, can be used both as food and as medicine due to the great diversity of recorded chemical components – different phenolic compounds, terpenes, carotenoids, vitamins, alkaloids, etc. The Balkan Peninsula is characterized by great diversity of plants from Asteraceae family, including presence of rare and endemic species. In this review, results of the survey of chemical composition and biological activity, mainly focusing on antioxidant, antimicrobial and anticancer effects of selected Balkans’ Asteraceae species were provided. In addition, information on edible plants from Asteraceae family is presented, due to growing interest for the so‐called ‘healthy diet’ and possible application of Balkans’ Asteraceae species as food of high nutritional value or as a source of functional food ingredients.     

Evolutionary origin of the Asteraceae capitulum: Insights from Calyceraceae

? Premise of the study: Phylogenies based on molecular data are revealing that generalizations about complex morphological structures often obscure variation and developmental patterns important for understanding the evolution of forms, as is the case for inflorescence morphology within the well-supported MGCA clade (Menyanthaceae + Goodeniaceae + Calyceraceae + Asteraceae). While the basal families share a basic thyrsic/thyrsoid structure of their inflorescences, Asteraceae possesses a capitulum that is widely interpreted as a racemose, condensed inflorescence. Elucidating the poorly known inflorescence structure of Calyceraceae, sister to Asteraceae, should help clarify how the Asteraceae capitulum evolved from thyrsic/thyrsoid inflorescences. ? Methods: The early development and structure of the inflorescence of eight species (five genera) of Calyceraceae were studied by SEM, and patterns of evolutionary change were interpreted via phylogenetic character mapping. ? Key results: The basic inflorescence structure of Calyceraceae is a cephalioid (a very condensed botryoid/thyrsoid). Optimization of inflorescence characters on a DNA sequence-derived tree suggests that the Asteraceae capitulum derives from a simple cephalioid through two morphological changes: loss of the terminal flower and suppression of the cymose branching pattern in the peripheral branches. ? Conclusions: Widely understood as a condensed raceme, the Asteraceae capitulum is the evolutionary result of a very reduced, condensed thyrsoid. Starting from that point, evolution worked separately only on the racemose developmental control/pattern within Asteraceae and mainly on the cymose developmental control/pattern within Calyceraceae, producing head-like inflorescences in both groups but with very different diversification potential. We also discuss possible remnants of the ancestral cephalioid structure in some Asteraceae.     

A comprehensive generic-level phylogeny of the sunflower family: Implications for the systematics of Chinese Asteraceae

The sunflower family (Asteraceae) is the largest and the most diverse flowering plant family, comprising 24 000–30 000 species and 1600–1700 genera. In China, Asteraceae are also the largest family, with approximately 2336 indigenous species in 248 genera. In the past two decades, molecular phylogenetic analyses has contributed greatly to our understanding of the systematics of Asteraceae. Nevertheless, the large-scale analyses and knowledge about the relationships of Chinese Asteraceae at the generic level as a whole are far from complete due to difficulties in sampling. In this study, we presented a three-marker (rbcL, ndhF, and matK) phylogeny of Asteraceae, including 506 genera (i.e., approximately one-third of Asteraceae genera). The study sampled 200 Chinese genera (i.e., approximately 80% of Chinese Asteraceae genera). The backbones of the new phylogeny were largely congruent with earlier studies, with 13 subfamilies and 45 tribes recognized. Chinese Asteraceae were distributed in 7 subfamilies (Mutisioideae, Wunderlichioideae, Carduoideae, Pertyoideae, Gymnarrhenoideae, Cichorioideae, and Asteroideae) and 22 tribes (Mutiseae, Hyalideae, Cardueae, Pertyeae, Gymnarrheneae, Vernonieae, Cichorieae, Doroniceae, Senecioneae, Astereae, Anthemideae, Gnaphalieae, Calenduleae, Inuleae, Athroismeae, Helenieae, Coreopsideae, Neurolaeneae, Tageteae, Millieae, Eupatorieae, and Heliantheae). Chinese Asteraceae lacked 6 basal subfamilies and 23 tribes. Several previously ambiguous relationships were clarified. Our analyses also resolved some unplaced genera within Chinese Asteraceae. Finally, our phylogenetic tree was used to revise the classification for all genera of Chinese Asteraceae. In total, 255 genera, 22 tribes, and 7 subfamilies in China are recognized.     

Characterization of strigolactones exuded by Asteraceae plants

Strigolactones (SLs), originally characterized as germination stimulants for root parasitic weeds, are now recognized as hyphal branching factors for symbiotic arbuscular mycorrhizal fungi and as a novel class of plant hormones inhibiting shoot branching. In the present study, SLs in root exudates of 13 Asteraceae plants including crops, a weed, and ornamental plants were characterized. High performance liquid chromatography/tandem mass spectrometry (LC–MS/MS) analyses revealed that all the Asteraceae plants examined exuded known SLs and, except for sunflower (Helianthus annuus), high germination stimulant activities at retention times corresponding to these SLs were confirmed. The two major SLs exuded by these Asteraceae plants were orobanchyl acetate and orobanchol. 5-Deoxystrigol and 7-hydroxyorobanchyl acetate were detected in root exudates from several Asteraceae species examined in this study.     

Development of novel low-copy nuclear markers for Hieraciinae (Asteraceae) and their perspective for other tribes

? Premise of the study: The development of three low-copy nuclear markers for low taxonomic level phylogenies in Asteraceae with emphasis on the subtribe Hieraciinae is reported. ? Methods and Results: Marker candidates were selected by comparing a Lactuca complementary DNA (cDNA) library with public DNA sequence databases. Interspecific variation and phylogenetic signal of the selected genes were investigated for diploid taxa from the subtribe Hieraciinae and compared to a reference phylogeny. Their ability to cross-amplify was assessed for other Asteraceae tribes. All three markers had higher variation (2.1-4.5 times) than the internal transcribed spacer (ITS) in Hieraciinae. Cross-amplification was successful in at least seven other tribes of the Asteraceae. Only three cases indicating the presence of paralogs or pseudogenes were detected. ? Conclusions: The results demonstrate the potential of these markers for phylogeny reconstruction in the Hieraciinae as well as in other Asteraceae tribes, especially for very closely related species.     

Two chloroplast DNA inversions originated simultaneously during the early evolution of the sunflower family (Asteraceae)

The chloroplast DNA (cpDNA) inversion in the Asteraceae has been cited as a classic example of using genomic rearrangements for defining major lineages of plants. We further characterize cpDNA inversions in the Asteraceae using extensive sequence comparisons among 56 species, including representatives of all major clades of the family and related families. We determine the boundaries of the 22-kb (now known as 22.8 kb) inversion that defines a major split within the Asteraceae, and in the process, we characterize the second and a new, smaller 3.3-kb inversion that occurs at one end of the larger inversion. One end point of the smaller inversion is upstream of the trnE-UUC gene, and the other end point is located between the trnC-GCA and rpoB genes. Although a diverse sampling of Asteraceae experienced substantial length variation and base substitution during the long evolutionary history subsequent to the inversion events, the precise locations of the inversion end points are identified using comparative sequence alignments in the inversion regions. The phylogenetic distribution of two inversions is identical among the members of Asteraceae, suggesting that the inversion events likely occurred simultaneously or within a short time period shortly after the origin of the family. Estimates of divergence times based on ndhF and rbcL sequences suggest that two inversions originated during the late Eocene (38-42 MYA). The divergence time estimates also suggest that the Asteraceae originated in the mid Eocene (42-47 MYA).     

Assessment of airborne Asteraceae pollen in Brisbane, Australia

In the southern hemisphere, the airborne prevalence and seasonal distribution of Asteraceae pollen are poorly characterized. In this study, we assessed the longitudinal distribution of Asteraceae pollen using an historic aerobiological dataset. The influence of weather parameters on pollen derived from Asteraceae species was additionally explored. Asteraceae pollen was collected using a static Burkard 7-day spore trap during a 5-year (June 1994–May 1999) aerometric surveillance program in Brisbane, Australia. Two discrete seasonal intervals were identified each sampling year. The first interval (September and October) consisted of small quantities of unidentifiable Asteraceae pollen. The second interval (March and April) was characterized by the highest concentrations of Asteraceae pollen that were predominantly derived from Baccharis halimifolia. Associations between daily Asteraceae pollen concentrations and temperature were observed during each sampling year. The results of the study demonstrated that pollen derived from B. halimifolia, an invasive weed species introduced in the late nineteenth century, continue to be in the environment of Brisbane. Given the public health burden associated with personal Asteraceae pollen exposure in other geographic locations, future Australian-based clinical studies should characterize the contribution of Asteraceae pollen, in particular B. halimifolia, to allergic sensitization and respiratory morbidity.     

Seed Inputs to Microsite Patch Recovery on Two Tropandean Landslides in Ecuador

To understand landslide regeneration and provide information necessary for restoration, we sampled seed rain, seed pool, and plant cover on two Ecuadorian landslides. We trapped 1304 seeds and found that, while most seeds were in the family Asteraceae, there was substantial variation in seed rain among plant families. Four hundred and seventy-five seedlings emerged from soil samples, including nonvascular and vascular families; again, species in Asteraceae dominated, with species in Piperaceae also very common. Plant cover, consisting of members of four fern families and 20 vascular plant families—with species in Asteraceae, Melastomataceae and Poaceae most common—was scored as a percentage of the total plot area. Principal components analysis (PCA) showed that, for all three of these plant life stages (seed rain, seed-propagule pool, plant cover), spatial variation was dominated by differences between the two landslides rather than within-landslide plot differences. PCA also showed that plots separated best on axes defined by the families Cecropiaceae, Urticaceae, Melastomataceae, Papilionaceae, Asteraceae, and Araceae with clumping of families in PCA space suggesting common successional strategies. Another multivariate technique, canonical correspondence analysis (CCA), showed that the combined seed rain and seed pool data could predict the percent cover of the family Verbenaceae and that the current plant cover families could predict Asteraceae seeds and seedlings. Finally, we use our past and present landslide data, along with multivariate modeling results, to suggest strategies for successful landslide restoration.     

Evolution of host-parasite relationships of Golovinomyces (Ascomycete: Erysiphaceae) inferred from nuclear rDNA sequences

To understand the evolution of host-parasite relationships in the genus Golovinomyces (Ascomycete: Erysiphaceae), which are obligate parasitic fungi of plants, we investigated the phylogenetic relationships of the genus based on 60 internal transcribed spacer (ITS) and 41 28S rDNA sequences. Five major groups, each represented by isolates from a single tribe of the Asteraceae, were identified in the taxa analyzed in this study. Host plants of four groups were strictly restricted to the Asteraceae. The fifth group, the Lactuceae group, is a large group composed of isolates collected from the tribe Lactuceae of the Asteraceae and all other plant families, which suggests a close affinity between Golovinomyces and the Asteraceae in the early stages of their evolution. Tree topology comparisons between the asteraceous hosts and their parasites suggest that Golovinomyces diverged along with the phylogeny of host tribes Carsueae, Astereae, Heliantheae, and Lactuceae of the Asteraceae. However, a conflict of branching order between the tribe Anthemideae and their parasites suggests that host-jumping has occurred in the tribe Anthemideae. Consequently, we suggest that there are two different phases in the evolutionary history of the host-parasite relationships of Golovinomyces. One phase is divergence in accord with the phylogeny of their hosts, which occurred within the Asteraceae. The another phase is host-jumping, which occurred from the Asteraceae to other families and within the Asteraceae.     

The value of sampling anomalous taxa in phylogenetic studies: major clades of the Asteraceae revealed

The largest family of flowering plants Asteraceae (Compositae) is found to contain 12 major lineages rather than five as previously suggested. Five of these lineages heretofore had been circumscribed in tribe Mutisieae (Cichorioideae), a taxon shown by earlier molecular studies to be paraphyletic and to include some of the deepest divergences of the family. Combined analyses of 10 chloroplast DNA loci by different phylogenetic methods yielded highly congruent well-resolved trees with 95% of the branches receiving moderate to strong statistical support. Our strategy of sampling genera identified by morphological studies as anomalous, supported by broader character sampling than previous studies, resulted in identification of several novel clades. The generic compositions of subfamilies Carduoideae, Gochnatioideae, Hecastocleidoideae, Mutisioideae, Pertyoideae, Stifftioideae, and Wunderlichioideae are novel in Asteraceae systematics and the taxonomy of the family has been revised to reflect only monophyletic groups. Our results contradict earlier hypotheses that early divergences in the family took place on and spread from the Guayana Highlands (Pantepui Province of northern South America) and raise new hypotheses about how Asteraceae dispersed out of the continent of their origin. Several nodes of this new phylogeny illustrate the vast differential in success of sister lineages suggesting focal points for future study of species diversification. Our results also provide a backbone exemplar of Asteraceae for supertree construction.     

云南菊科入侵植物入侵机制及其利用研究进展

植物入侵已对生物多样性、社会经济和人类健康造成了严重的威胁。云南由于地形复杂、气候和生态系统类型多样而成为植物入侵的重灾区。菊科是入侵植物中最多的一个科;但目前尚缺乏有关云南菊科入侵植物入侵机制和利用的报道。云南菊科入侵植物约有86种,入侵的主要机制:云南地形复杂,气候和生态系统类型多样,为入侵植物的生存提供了适宜的环境;同时,这些菊科入侵植物具有较强的繁殖能力和竞争能力、多样化的生态对策及丰富的化感物质,且其天敌缺乏等。这些因素的综合作用使很多菊科外来植物成功入侵云南。综合利用是治理入侵植物的一种有效措施。云南菊科入侵植物资源丰富,产量高,在医药、饲料、工业原料等方面具有一定的开发利用价值。     

Sesquiterpene lactone-based classification of three Asteraceae tribes: a study based on self-organizing neural networks applied to chemosystematics

This work describes an application of artificial neural networks on a small data set of sesquiterpene lactones (STLs) of three tribes of the family Asteraceae. Structurally different types of representative STLs from seven subtribes of the tribes Eupatorieae, Heliantheae and Vernonieae were selected as input data for self-organizing neural networks. Encoding the 3D molecular structures of STLs and their projection onto Kohonen maps allowed the classification of Asteraceae into tribes and subtribes. This approach allowed the evaluation of structural similarities among different sets of 3D structures of sesquiterpene lactones and their correlation with the current taxonomic classification of the family. Predictions of the occurrence of STLs from a plant species according to the taxa they belong to were also performed by the networks. The methodology used in this work can be applied to chemosystematic or chemotaxonomic studies of Asteraceae.     

Studies in the Goodeniaceae and the Brunoniaceae with a discussion of their relationship to Asteraceae and Calyceraceae

The relationships between Asteraceae, Calyceraceae, Goodeniaceae, and Brunoniaceae are discussed, and some SEM-observations of microcharacters within the two latter families are presented. Based on morphological evidence neither Goodeniaceae s.str. nor Goodeniaceae s.lat. (i.e., incl. Brunonia) appear to be the putative sister taxon of Asteraceae. A cladistic analysis demonstrates that Brunonia australis may constitute a separate, monotypic family or alternatively be deeply nested within Goodeniaceae s.lat Brunonia shares a number of potential synapomorphies with Asteraceae. Of special interest are the peculiar hairs and feathery pappus bristles which very much resemble those found within Asteraceae subfam. Barnadesioideae. However, studies with SEM disclose that these structures rather constitute autapomorphies for the taxa in question. In conclusion, Calyceraceae remain to be the best sister candidate of the sunflower family.     

Comparative analysis of NBS domain sequences of NBS-LRR disease resistance genes from sunflower, lettuce, and chicory

Plant resistance to many types of pathogens and pests can be achieved by the presence of disease resistance (R) genes. The nucleotide binding site-leucine rich repeat (NBS-LRR) class of R-genes is the most commonly isolated class of R-genes and makes up a super-family, which is often arranged in the genome as large multi-gene clusters. The NBS domain of these genes can be targeted by polymerase chain reaction (PCR) amplification using degenerate primers. Previous studies have used PCR derived NBS sequences to investigate both ancient R-gene evolution and recent evolution within specific plant families. However, comparative studies with the Asteraceae family have largely been ignored. In this study, we address recent evolution of NBS sequences within the Asteraceae and extend the comparison to the Arabidopsis thaliana genome. Using multiple sets of primers, NBS fragments were amplified from genomic DNA of three species from the family Asteraceae: Helianthus annuus (sunflower), Lactuca sativa (lettuce), and Cichorium intybus (chicory). Analysis suggests that Asteraceae species share distinct families of R-genes, composed of genes related to both coiled-coil (CC) and toll-interleukin-receptor homology (TIR) domain containing NBS-LRR R-genes. Between the most closely related species, (lettuce and chicory) a striking similarity of CC subfamily composition was identified, while sunflower showed less similarity in structure. These sequences were also compared to the A. thaliana genome. Asteraceae NBS gene subfamilies appear to be distinct from Arabidopsis gene clades. These data suggest that NBS families in the Asteraceae family are ancient, but also that gene duplication and gene loss events occur and change the composition of these gene subfamilies over time.     

Polyploidy in Asteraceae of the xerophytic scrub of the Ecological Reserve of the Pedregal of San Angel,Mexico City

Asteraceae species diversity is high in the xerophytic scrub of the Ecological Reserve of the Pedregal of San Angel (REPSA), located in the southern part of the Basin of the Valley of Mexico. Here we determined whether the frequency of polyploidy is high in the reserve, given the enhanced ability of polyploids to colonize new habitats. In addition, we compared the frequency of polyploidy in Asteraceae in the reserve with the frequency in three oceanic archipelagos and two continental areas in Mexico. This was done to see how the ‘virtual’ island of the open lava flow in the reserve compares with volcanic islands at different distances from source areas. Chromosome numbers for 75 species of Asteraceae were obtained from published literature. Based on the possession of three or more basic chromosome sets in a nucleus, 33% were polyploids. If taxa with haploid chromosome numbers of n ≥ 14 or n ≥ 11 were considered to be polyploids, the proportion of polyploids rose to 57 and 75%, respectively. When using a phylogenetic approach, the highest percentage of polyploids (84%) was obtained and it could be inferred whether they are palaeo‐ or neopolyploids; thus, we consider that this criterion better reflects the events of polyploidy in Asteraceae. A high frequency of polyploid species in Asteraceae in REPSA suggests that polyploids may have contributed to the species diversity and the vegetation structure of the xerophytic scrub of this reserve. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 211–229.     

Biochemical Conservation and Evolution of Germacrene A Oxidase in Asteraceae

Sesquiterpene lactones are characteristic natural products in Asteraceae, which constitutes ∼8% of all plant species. Despite their physiological and pharmaceutical importance, the biochemistry and evolution of sesquiterpene lactones remain unexplored. Here we show that germacrene A oxidase (GAO), evolutionarily conserved in all major subfamilies of Asteraceae, catalyzes three consecutive oxidations of germacrene A to yield germacrene A acid. Furthermore, it is also capable of oxidizing non-natural substrate amorphadiene. Co-expression of lettuce GAO with germacrene synthase in engineered yeast synthesized aberrant products, costic acids and ilicic acid, in an acidic condition. However, cultivation in a neutral condition allowed the de novo synthesis of a single novel compound that was identified as germacrene A acid by gas and liquid chromatography and NMR analyses. To trace the evolutionary lineage of GAO in Asteraceae, homologous genes were further isolated from the representative species of three major subfamilies of Asteraceae (sunflower, chicory, and costus from Asteroideae, Cichorioideae, and Carduoideae, respectively) and also from the phylogenetically basal species, Barnadesia spinosa, from Barnadesioideae. The recombinant GAOs from these genes clearly showed germacrene A oxidase activities, suggesting that GAO activity is widely conserved in Asteraceae including the basal lineage. All GAOs could catalyze the three-step oxidation of non-natural substrate amorphadiene to artemisinic acid, whereas amorphadiene oxidase diverged from GAO displayed negligible activity for germacrene A oxidation. The observed amorphadiene oxidase activity in GAOs suggests that the catalytic plasticity is embedded in ancestral GAO enzymes that may contribute to the chemical and catalytic diversity in nature.     

Geographic space,relief, and soils predict plant community patterns of Asteraceae in rupestrian grasslands,Brazil

We investigated the effects of geographic distance and environmental heterogeneity on Asteraceae communities in the rupestrian grasslands of the Espinhaço Range, southeastern Brazil. Asteraceae species composition and relative abundance were sampled in 21–1 ha plots distributed across park conservation units, comprising an environmental gradient in soils and relief (elevation and slope). We found high levels of heterogeneity overall in assemblages of herbaceous and woody Asteraceae species at the intermediate spatial scale of this study (10–100 km). We found that both relief and soil properties (texture and total exchangeable bases) explained significant proportions of variation in community composition, suggesting that definitions or expansions of conservation units should span the highest possible variability in observed relief and soil values. We also found that Asteraceae distribution exhibited strong spatial structure independent of environmental variation, reinforcing the need to consider conservation across larger areas. Considering the dystrophic character of the soils found in the Espinhaço Range and the few differences in the fertility observed between the sampling units, our results also support the hypothesis that plant distributions in the tropics are controlled by a hierarchy of environmental gradients, where variables such as altitude and exchangeable bases and Al texture are important predictors in low‐fertility soils. We suggest that Asteraceae could be a model family for conservation planning in other countries and regions because it is particularly well represented in open and montane areas of the Neotropics in general. Abstract in Portuguese is available with online material.     

Identification and validation of Asteraceae miRNAs by the expressed sequence tag analysis

MicroRNAs (miRNAs) are small non-coding RNA molecules that play a vital role in the regulation of gene expression. Despite their identification in hundreds of plant species, few miRNAs have been identified in the Asteraceae, a large family that comprises approximately one tenth of all flowering plants. In this study, we used the expressed sequence tag (EST) analysis to identify potential conserved miRNAs and their putative target genes in the Asteraceae. We applied quantitative Real-Time PCR (qRT-PCR) to confirm the expression of eight potential miRNAs in Carthamus tinctorius and Helianthus annuus. We also performed qRT-PCR analysis to investigate the differential expression pattern of five newly identified miRNAs during five different cotyledon growth stages in safflower. Using these methods, we successfully identified and characterized 151 potentially conserved miRNAs, belonging to 26 miRNA families, in 11 genus of Asteraceae. EST analysis predicted that the newly identified conserved Asteraceae miRNAs target 130 total protein-coding ESTs in sunflower and safflower, as well as 433 additional target genes in other plant species. We experimentally confirmed the existence of seven predicted miRNAs, (miR156, miR159, miR160, miR162, miR166, miR396, and miR398) in safflower and sunflower seedlings. We also observed that five out of eight miRNAs are differentially expressed during cotyledon development. Our results indicate that miRNAs may be involved in the regulation of gene expression during seed germination and the formation of the cotyledons in the Asteraceae. The findings of this study might ultimately help in the understanding of miRNA-mediated gene regulation in important crop species.