Accelerated diversification correlated with functional traits shapes extant diversity of the early divergent angiosperm family Annonaceae.

A major goal of phylogenetic systematics is to understand both the patterns of diversification and the processes by which these patterns are formed. Few studies have focused on the ancient, species-rich Magnoliales clade and its diversification pattern. Within Magnoliales, the pantropically distributed Annonaceae are by far the most genus-rich and species-rich family-level clade, with c. 110 genera and c. 2,400 species. We investigated the diversification patterns across Annonaceae and identified traits that show varied associations with diversification rates using a time-calibrated phylogeny of 835 species (34.6% sampling) and 11,211 aligned bases from eight regions of the plastid genome (rbcL, matK, ndhF, psbA-trnH, trnL-F, atpB-rbcL, trnS-G, and ycf1). Twelve rate shifts were identified using BAMM: in Annona, Artabotrys, Asimina, Drepananthus, Duguetia, Goniothalamus, Guatteria, Uvaria, Xylopia, the tribes Miliuseae and Malmeeae, and the Desmos-Dasymaschalon-Friesodielsia-Monanthotaxis clade. TurboMEDUSA and method-of-moments estimator analyses showed largely congruent results. A positive relationship between species richness and diversification rate is revealed using PGLS. Our results show that the high species richness in Annonaceae is likely the result of recent increased diversification rather than the steady accumulation of species via the 'museum model'. We further explore the possible role of selected traits (habit, pollinator trapping, floral sex expression, pollen dispersal unit, anther septation, and seed dispersal unit) in shaping diversification patterns, based on inferences of BiSSE, MuSSE, HiSSE, and FiSSE analyses. Our results suggest that the liana habit, the presence of circadian pollinator trapping, androdioecy, and the dispersal of seeds as single-seeded monocarp fragments are closely correlated with higher diversification rates; pollen aggregation and anther septation, in contrast, are associated with lower diversification rates.

A jungle tale: Molecular phylogeny and divergence time estimates of the Desmopsis-Stenanona clade (Annonaceae) in Mesoamerica.

The predominantly Asian tribe Miliuseae (Annonaceae) includes over 37 Neotropical species that are mainly distributed across Mesoamerica, from southern Mexico to northern Colombia. The tremendous ecological and morphological diversity of this clade, including ramiflory, cauliflory, flagelliflory, and clonality, suggests adaptive radiation. Despite the spectacular phenotypic divergence of this clade, little is known about its phylogenetic and evolutionary history. In this study we used a nuclear DNA marker and seven chloroplast markers, and maximum parsimony, maximum likelihood and Bayesian inference methods to reconstruct a comprehensive time-calibrated phylogeny of tribe Miliuseae, especially focusing on the Desmopsis-Stenanona clade. We also perform ancestral area reconstructions to infer the biogeographic history of this group. Finally, we use ecological niche modeling, lineage distribution models, and niche overlap tests to assess whether geographic isolation and ecological specialization influenced the diversification of lineages within this clade. We reconstructed a monophyletic Miliuseae that is divided into two strongly supported clades: (i) a Sapranthus-Tridimeris clade and (ii) a Desmopsis-Stenanona clade. The colonization of the Neotropics and subsequent diversification of Neotropical Miliuseae seems to have been associated with the expansion of the boreotropical forests during the late Eocene and their subsequent fragmentation and southern displacement. Further speciation within Neotropical Miliuseae out of the Maya block seems to have occurred during the last 15 million years. Lastly, the geographic structuring of major lineages of the Desmopsis-Stenanona clade seems to have followed a climatic gradient, supporting the hypothesis that morphological differentiation between closely related species resulted from both long-term isolation between geographic ranges and adaptation to environmental conditions.

Phylogenomics and evolution of floral traits in the Neotropical tribe Malmeeae (Annonaceae).

Androdioecy is the rarest sexual system among plants. The majority of androdioecious species are herbaceous plants that have evolved from dioecious ancestors. Nevertheless, some woody and androdioecious plants have hermaphrodite ancestors, as in the Annonaceae, where androdioecious genera have arisen several times in different lineages. The majority of androdioecious species of Annonaceae belong to the Neotropical tribe Malmeeae. In addition to these species, Pseudoxandra spiritus-sancti was recently confirmed to be androdioecious. Here, we describe the morphology of male and bisexual flowers of Pseudoxandra spiritus-sancti, and investigate the evolution of androdioecy in Malmeeae. The phylogeny of tribe Malmeeae was reconstructed using Bayesian inference, maximum parsimony and maximum likelihood of 32 taxa, using DNA sequences of 66 molecular markers of the chloroplast genome, sequenced by next generation sequencing. The reconstruction of ancestral states was performed for characters associated with sexual systems and floral morphology. The phylogenetic analyses reconstructed three main groups in Malmeeae, (Malmea (Cremastosperma, Pseudoxandra)) sister to the rest of the tribe, and (Unonopsis (Bocageopsis, Onychopetalum)) sister to (Mosannona, Ephedranthus, Klarobelia, Oxandra, Pseudephedranthus fragrans, Pseudomalmea, Ruizodendron ovale). Hermaphroditism is plesiomorphic in the tribe, with four independent evolutions of androdieocy, which represents a synapomorphy of two groups, one that includes three genera and 14 species, the other with a single genus of seven species. Male flowers are unisexual from inception and bisexual flowers possess staminodes and functional stamens. Pseudoxandra spiritus-sancti is structurally androdioecious.

A mega-phylogeny of the Annonaceae: taxonomic placement of five enigmatic genera and support for a new tribe, Phoenicantheae.

The Annonaceae, the largest family in the early-divergent order Magnoliales, comprises 107 genera and c. 2,400 species. Previous molecular phylogenetic studies targeting different taxa have generated large quantities of partially overlapping DNA sequence data for many species, although a large-scale phylogeny based on the maximum number of representatives has never been reconstructed. We use a supermatrix of eight chloroplast markers (rbcL, matK, ndhF, psbA-trnH, trnL-F, atpB-rbcL, trnS-G and ycf1) to reconstruct the most comprehensive tree to date, including 705 species (29%) from 105 genera (98%). This provides novel insights into the relationships of five enigmatic genera (Bocagea, Boutiquea, Cardiopetalum, Duckeanthus and Phoenicanthus). Fifteen main clades are retrieved in subfamilies Annonoideae and Malmeoideae collectively, 14 of which correspond with currently recognised tribes. Phoenicanthus cannot be accommodated in any existing tribe, however: it is retrieved as sister to a clade comprising the tribes Dendrokingstonieae, Monocarpieae and Miliuseae, and we therefore validate a new tribe, Phoenicantheae. Our results provide strong support for many previously recognised groups, but also indicate non-monophyly of several genera (Desmopsis, Friesodielsia, Klarobelia, Oxandra, Piptostigma and Stenanona). The relationships of these non-monophyletic genera-and two other genera (Froesiodendron and Melodorum) not yet sampled-are discussed, with recommendations for future research.

Correlated evolutionary rates across genomic compartments in Annonaceae.

The molecular clock hypothesis is an important concept in biology. Deviations from a constant rate of nucleotide substitution have been found widely among lineages, genomes, genes and individual sites. Phylogenetic research can accommodate for these differences in applying specific models of evolution. Lineage-specific rate heterogeneity however can generate bi- or multimodal distributions of substitution rates across the branches of a tree and this may mislead phylogenetic inferences with currently available models. The plant family Annonaceae is an excellent case to study lineage-specific rate heterogeneity. The two major sister subfamilies, Annonoideae and Malmeoideae, have shown great discrepancies in branch lengths. We used high-throughput sequencing data of 72 genes, 99 spacers and 16 introns from 24 chloroplast genomes and nuclear ribosomal DNA of 23 species to study the molecular rate of evolution in Annonaceae. In all analyses, longer branch lengths and/or higher substitution rates were found for the Annonoideae compared to the Malmeoideae. The Annonaceae had wide variability in chloroplast length, ranging from minimal 175,684bp to 201,723 for Annonoideae and minimal 152,357 to 170,985bp in Malmeoideae, mostly reflecting variation in inverted-repeat length. The Annonoideae showed a higher GC-content in the conserved parts of the chloroplast genome and higher omega (dN/dS)-ratios than the Malmeoideae, which could indicate less stringent purifying selection, a pattern that has been found in groups with small population sizes. This study generates new insights into the processes causing lineage-specific rate heterogeneity, which could lead to improved phylogenetic methods.

Cell cycle arrest and apoptosis induction by methanolic leaves extracts of four Annonaceae plants.

BACKGROUND: Uvaria longipes (Craib) L.L.Zhou, Y.C.F.Su & R.M.K.Saunders, Artabotrys burmanicus A.DC, Marsypopetalum modestum (Pierre) B.Xue & R.M.K.Saunders and Dasymaschalon sp. have been used for traditional medicine to treat cancer-like symptoms in some ethnic groups of Thailand and Laos. METHODS: We evaluated the anti-cancer activity of these Annonaceae plants against several human cancer cell lines. The apoptosis induction was detected by Annexin/propidium iodide (PI) staining. Phytochemical screening was tested by standard protocols and bioactive compounds were determined by HPLC. RESULTS: The crude extracts from leaves of U. longipes, Dasymaschalon sp., A. burmanicus, and M. modestum showed particular effects that were found to vary depending on the cancer cell line, suggesting that the effect was in a cell-type specific manner. Interestingly, the induction of apoptotic cell death was prominent by the leaves-derived crude extract of M. modestum. This crude was, therefore, subjected to cell cycle analysis by PI staining. Results showed that this crude extract arrested cell cycle and increased the percentage of cells in the SubG1 phase in some cancer cell lines. The phytochemical screening tests indicated that all crude extracts contained tannins and flavonoids. HPLC of flavonoids using standards identified rutin as an active compound in U. longipes and Dasymaschalon sp., whereas quercetin was found in U. longipes and M. modestum. CONCLUSIONS: These crude extracts provide a new source for rutin and quercetin, which might be capable of inducing cancer cell apoptotic death in a cell-type specific manner. This suggests, by analyzing the major bioactive compounds, the potential use of these crudes for chemotherapy in the future.

Alphonsea glandulosa (Annonaceae), a New Species from Yunnan, China.

Alphonsea glandulosa sp. nov. is described from Yunnan Province in south-west China. It is easily distinguished from all previously described Alphonsea species by the possession of glandular tissue at the base of the adaxial surface of the inner petals. Nectar was observed throughout the flowering period, including the pistillate phase and subsequent staminate phase. Small curculionid beetles were observed as floral visitors and are inferred to be effective pollinators since they carry pollen grains. A phylogenetic analysis was conducted to confirm the placement of this new species within Alphonsea and the evolution of the inner petal glands and specialized pollinator reward tissues throughout the family.

Chemical Composition and Bactericidal Activity of the Essential Oils of Four Species of Annonaceae Growing in Brazilian Amazon.

A study was conducted to determine the chemical composition and the antibacterial activity of essential oils from Bocageopsis multiflora (Mart.) R.E. Fr., Ephedranthus amazonicus R.E. Fr., Guatteria blepharophylla Mart. and Xylopia aromatica (Lam.) Mart . The essential oils were obtained from the leaves by hydrodistillation and analyzed by GC-FID and GC-MS. The chromatograms showed the predominance of sesquiterpenoids. Spathulenol was the major constituent of oils of B. inultiflora (20.3%), X aromatica (21.5%) and E. amazonicus (16.9%); followed by caryophyllene oxide of G. blepharophylla (55.7%). Other constituents with significant percentages were the sesquiterpenes P-bisabolene (11.9%) in the samples oil of B. multiflora, humulene epoxide 11 (16.3%) -of E. amazonicus, the monoterpenes trans-pinocarveol (10.2%) and dihydrocarveol (11.6%) of X aromatica. The essential oils of X. aromatica and G. blepharophylla showed strong activity against Gram-positive bacteria Streptococcus sanguinis by microdilution method. The antimicrobial activity against Staphylococcus aureus also was detected by G. blepharophylla essential oil.

Acetogenins from Annonaceae.

In recent decades, annonaceous acetogenins have become highly studied plant secondary metabolites in terms of their isolation, structure elucidation, synthesis, biological evaluation, mechanism of action, and toxicity. The aim of the present contribution is to summarize chemical and biological reports published since 1997 on annonaceous acetogenins and synthetic acetogenin mimics. The compounds are considered biologically in terms of their cytotoxicity for cancer cell lines, neurotoxicity, pesticidal effects, and miscellaneous activities.

Alkaloids of the Annonaceae: occurrence and a compilation of their biological activities.

This chapter presents an overview of the chemistry and pharmacology of the alkaloids found in species of the Annonaceae family. The occurrence of alkaloids from Annonaceae species, as well as their chemical structures and pharmacological activities are summarized in informative and easy-to-understand tables. Within the Annonaceae family, the genera Annona, Duguetia, and Guatteria have led to many important publications. Valuable and comprehensive information about the structure of these alkaloids is provided. The alkaloids of the aporphine type represent the predominant group in this family. Many of the isolated alkaloids exhibit unique structures. In addition to the chemical structures, the pharmacological activities of some alkaloids are also presented in this chapter. Thus, the leishmanicidal, antimicrobial, antitumor, cytotoxic, and antimalarial activities observed for these alkaloids are highlighted. The chapter is presented as a contribution for the scientific community, mainly to enable the search for alkaloids in species belonging to the Annonaceae family.

A plastid DNA phylogeny of tribe Miliuseae: insights into relationships and character evolution in one of the most recalcitrant major clades of Annonaceae.

PREMISE OF THE STUDY: Tribe Miliuseae (∼25 genera and ∼510 species) includes a substantial part of the species and generic diversity in the pantropical flowering-plant family Annonaceae (∼108 genera and ∼2400 species). Previous molecular phylogenetic analyses have failed to resolve the backbone phylogeny of the tribe, impeding biogeographical and evolutionary studies. We use a dense generic taxon sample (∼89% of generic diversity in Miliuseae) and plastid DNA sequence data (∼7 kb) to clarify the phylogenetic relationships of and within the tribe. METHODS: Parsimony and Bayesian phylogenetic reconstructions and ancestral character-state reconstructions of several reproductive characters were performed. KEY RESULTS: Dendrokingstoniae, Monocarpieae, and Miliuseae are recovered in a strongly supported clade, and each tribe is strongly supported as monophyletic. Miliuseae are characterized by a synapomorphic cryptoaperturate/disulculate pollen apertural system. Stenanona is shown to be nested within the paraphyletic genus Desmopsis. The only Neotropical clade (Sapranthus, Tridimeris, Desmopsis, and Stenanona) in the predominantly Asian Miliuseae is shown to be closely related to an undescribed genus from continental Southeast Asia and the Indo-Malayan and Austral-Pacific genus Meiogyne. Ancestral character-state reconstructions of several reproductive characters that are diagnostically important at the generic level indicate a considerable degree of homoplasy. CONCLUSIONS: The results improve our understanding of the relationships of and within Miliuseae, but parts of the backbone of the phylogeny remain poorly supported. Additional data from variable nuclear markers or reduced-genome-representation approaches seem to be required to further resolve relationships within this recalcitrant clade.

Chemical constituents from Glyphaea brevis and Monodora myristica: chemotaxonomic significance.

A chemical investigation of the Glyphaea brevis leaves and of the Monodora myristica fruits led to the identification of thirteen compounds, seven linear long-chain aliphatic compounds, 1, 2, 4, 6, and 9-11, three steroids, 3a, 3b, and 7, two triterpenes, 5a and 5b, and one polyol, 8. The compounds 2 and 8, previously mentioned in the literature, are here characterized by their complete (1)H- and (13)C-NMR assignments. This is the first report of a full NMR assignment for linear fatty acid esters of aliphatic primary alcohols and for meso-erythritol. Compound 5b and 8 were isolated for the first time from plant extracts of the Tiliaceae family, and compounds 9-11 from the Annonaceae plant family. Our results constitute the basis for further chemotaxonomic studies on the two species.

First fossil record of Alphonsea Hk. f. & T. (Annonaceae) from the Late Oligocene sediments of Assam, India and comments on its phytogeography.

A new fossil leaf impression of Alphonsea Hk. f. & T. of the family Annonaceae is described from the Late Oligocene sediments of Makum Coalfield, Assam, India. This is the first authentic record of the fossil of Alphonsea from the Tertiary rocks of South Asia. The Late Oligocene was the time of the last significant globally warm climate and the fossil locality was at 10°-15°N palaeolatitude. The known palaeoflora and sedimentological studies indicate a fluvio-marine deltaic environment with a mosaic of mangrove, fluvial, mire and lacustrine depositional environments. During the depositional period the suturing between the Indian and Eurasian plates was not complete to facilitate the plant migration. The suturing was over by the end of the Late Oligocene/beginning of Early Miocene resulting in the migration of the genus to Southeast Asia where it is growing profusely at present. The present study is in congruence with the earlier published palaeofloral and molecular phylogenetic data. The study also suggests that the Indian plate was not only a biotic ferry during its northward voyage from Gondwana to Asia but also a place for the origin of several plant taxa.

The leaf essential oils of five Vietnamese Desmos species (Annonaceae).

The leaf essential oils of five Desmos species from Vietnam have been extracted by steam distillation and subjected to GC and GC-MS analyses. The plant samples were Desmos cochinchinensis Lour., D. penduculosus (A. DC.) Ban, D. penducolosus var. tonkinensis Ban, D. chinensis Lour., and D. dumosus (Roxb.) Saff. The oils were rich in sesquiterpene hydrocarbons (65.9%-88.9%) and monoterpene hydrocarbons (6.3%-30.9%). The oxygenated counterparts were less common. The quantitatively significant constituents of the oils were alpha-pinene (2.4%-12.1%), beta-elemene (2.2-39.5%), beta-caryophyllene (13.9-26.3%), germacrene D (9.9-15.5%), bicyclogermacrene (2.0-11.4%) and alpha-humulene (3.8-7.5%). The studied oils could be classified into two chemical forms: oils with abundance of beta-caryophyllene, germacrene D and alpha-pinene (D. cochinchinensis, D. penducolosus var. tonkinensis, D. chinensis and D. Dumosus) and oil with high amounts of beta-elemene, beta-caryophyllene and germacrene D (D. penduculosus).

Rust fungi on Annonaceae: the genus Sphaerophragmium.

Seven species of the rust genus Sphaerophragmium occur on members of the tropical plant family Annonaceae. Uropyxis gerstneri is recombined to S. gerstneri. A new species, S. xylopiae, is described from Xylopia acutiflora. The host plant of S. boanense is identified as Mitrella sp. Sphaerophragmium pulchrum is transferred to Dicheirinia. The anatomy of telia with teliospores and parasitizing mycelium is described and illustrated in detail. A new type of M-haustorium, which emanates laterally from intracellular hypha, is detected in S. monodorae. An identification key is given.

7,7-Dimethylaporphine alkaloids from the stem of Guatteriopsis friesiana.

Phytochemical investigation of a methanolic extract of the stem of Guatteriopsis friesiana afforded two new 7,7-dimethylaporphine alkaloids, 6,6a-dihydrodemethoxyguadiscine (1) and guatteriopsiscine (3), together with demethoxyguadiscine (2), liriodenine (4), corypalmine (5), and coreximine (6). Their structures were elucidated on the basis of spectroscopic methods (UV, IR, EIMS, HRESIMS, 1D/2D NMR). The absolute configurations of 1 and 3 were determined from the circular dichroism curves. The presence of 7,7-dimethylaporphine alkaloids in this species is important for the chemotaxonomy of Guatteriopsis. Antimicrobial activity of compounds 1-5 was investigated, and 4 showed activity against Rhodococcus equi, with a MIC value of 10 microg x mL(-1).

Larvicidal and IGR activity of extract of Tanzanian plants against malaria vector mosquitoes.

BACKGROUND & OBJECTIVES: This paper reports the larvicidal activity of seventeen Tanzanian plant species against the malaria vector, Anopheles gambiae s.s. Giles larvae. Some of the plants are used traditionally as sources of insecticidal materials. METHODS: The crude extracts from the leaves, stem and root barks of the investigated plants were obtained by solvent extraction and then bio-assayed following WHO protocols showed LC50 values 10 to 400 ppm after 24 h exposure. The structures were determined on interpretation of spectroscopic data. RESULTS: The most active extracts were those from the stem and root barks of Annona squamosa, Uvaria faulknerae, U. kirkii and Uvariodendron pycnophyllum, all of which had LC50 values between 10 and 100 ppm. Long-term exposure beyond 24 h also showed more susceptibility of the larvae to the extracts. Larvae deformities by forming tail-like structures were observed for the methanol extracts of Tessmannia martiniana var pauloi. INTERPRETATION & CONCLUSION: The results suggest that the investigated plant extracts are promising as larvicides against An. gambiae s.s. Giles mosquitoes and could be useful leads in the search for new and biodegradable plant derived larvicide products.

Phylogeny of the basal angiosperm genus Pseuduvaria (Annonaceae) inferred from five chloroplast DNA regions, with interpretation of morphological character evolution.

Phylogenetic relationships within the magnoliid basal angiosperm genus Pseuduvaria (Annonaceae) are investigated using chloroplast DNA sequences from five regions: psbA-trnH spacer, trnL-F, matK, rbcL, and atpB-rbcL spacer. Over 4000 nucleotides from 51 species (of the total 53) were sequenced. The five cpDNA datasets were analyzed separately and in combination using maximum parsimony (MP), maximum likelihood (ML), and Bayesian methods. The phylogenetic trees constructed using all three phylogenetic methods, based on the combined data, strongly support the monophyly of Pseuduvaria following the inclusion of Craibella phuyensis. The trees generated using MP were less well resolved, but relationships are similar to those obtained using the other methods. ML and Bayesian analyses recovered trees with short branch lengths, showing five main clades. This study highlights the evolutionary changes in seven selected morphological characters (floral sex, stamen and carpel numbers, inner petal color, presence of inner petal glands, flowering peduncle length, and monocarp size). Although floral unisexuality is ancestral within the genus, several evolutionary lineages reveal reversal to bisexuality. Other phylogenetic transitions include the evolution of sapromyophily, and fruit-bat frugivory and seed dispersal, thus allowing a wide range of adaptations for species survival.

Evolution of syncarpy and other morphological characters in African Annonaceae: a posterior mapping approach.

The congenital fusion of carpels, or syncarpy, is considered a key innovation as it is found in more than 80% of angiosperms. Within the magnoliids however, syncarpy has rarely evolved. Two alternative evolutionary origins of syncarpy were suggested in order to explain the evolution of this feature: multiplication of a single carpel vs. fusion of a moderate number of carpels. The magnoliid family Annonaceae provides an ideal situation to test these hypotheses as two African genera, Isolona and Monodora, are syncarpous in an otherwise apocarpous family with multicarpellate and unicarpellate genera. In addition to syncarpy, the evolution of six other morphological characters was studied. Well-supported phylogenetic relationships of African Annonaceae and in particular those of Isolona and Monodora were reconstructed. Six plastid regions were sequenced and analyzed using maximum parsimony and Bayesian inference methods. The Bayesian posterior mapping approach to study character evolution was used as it accounts for both mapping and phylogenetic uncertainty, and also allows multiple state changes along the branches. Our phylogenetic analyses recovered a fully resolved clade comprising twelve genera endemic to Africa, including Isolona and Monodora, which was nested within the so-called long-branch clade. This is the largest and most species-rich clade of African genera identified to date within Annonaceae. The two syncarpous genera were inferred with maximum support to be sister to a clade characterized by genera with multicarpellate apocarpous gynoecia, supporting the hypothesis that syncarpy arose by fusion of a moderate number of carpels. This hypothesis was also favoured when studying the floral anatomy of both genera. Annonaceae provide the only case of a clear evolution of syncarpy within an otherwise apocarpous magnoliid family. The results presented here offer a better understanding of the evolution of syncarpy in Annonaceae and within angiosperms in general.

[Progress in study of flavonoids from Annonaceae and biological activities of these compounds].

More than 50 new flavonoids derived from Annonaceae are reported in the last two decades. Many genuses in Annonaceae contain flavonoids having structural novelty and broad pharmacological activities. Due to the pharmacological interest of some of these compounds, chemical investigations on this topic have grown considerably in the decades. Here the biological activities of some of these flavonoids are also briefly discussed.