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Many major branches in the Tree of Life are marked by stereotyped body plans that have been maintained over long periods of time. One possible explanation for this stasis is that there are genetic or developmental constraints that restrict the origin of novel body plans. An alternative is that basic body plans are potentially quite labile, but are actively maintained by natural selection. We present evidence that the conserved floral morphology of a species-rich flowering plant clade, Malpighiaceae, has been actively maintained for tens of millions of years via stabilizing selection imposed by their specialist New World oil-bee pollinators. Nine clades that have lost their primary oil-bee pollinators show major evolutionary shifts in specific floral traits associated with oil-bee pollination, demonstrating that developmental constraint is not the primary cause of morphological stasis in Malpighiaceae. Interestingly, Malpighiaceae show a burst in species diversification coinciding with the origin of this plant–pollinator mutualism. One hypothesis to account for radiation despite morphological stasis is that although selection on pollinator efficiency explains the origin of this unique and conserved floral morphology, tight pollinator specificity subsequently permitted greatly enhanced diversification in this system.

Lophopterys floribunda is a Neotropical species of Malpighiaceae endemic to Brazil, occurring in both the Amazon and Atlantic Forest. Instead of the typical bi-glandular sepals reported for Neotropical Malpighiaceae, this species presents a single, large gland on the lateral sepals. In addition, ant patrolling was observed at the apex of bracts and bracteoles during fieldwork. Thus, this work aimed to describe the sepalar gland of L. floribunda and other secretory structures in its flowers and inflorescence. Samples of bracts, bracteoles, sepals, petals, and anther were collected and submitted to usual anatomical techniques. Unexpected nectaries at the apex of bracts and bracteoles, not visible to the naked eye, were described and represent a new type of structure for the family due to both their position and size. Mutualistic ants consume the exudate produced by these tiny nectaries, and such structures enable a specific visitation pattern for Lophopterys. Typical epithelial elaiophores occur on the lateral sepals, formed by an invaginated epidermis, which predominantly produce lipid secretion. The petal marginal glands are anatomically similar to the standard type of colleter, which exude mucilaginous substances. The exudate produced by the petal marginal glands was considered to have an additional role of contributing to the maintenance of the closed bud during the beginning of development. The globose epidermal cells containing lipids, proteins, and polysaccharides observed in the connective may be responsible for the typical aroma emitted by these flowers. The diversity of secretory structures reported here has application in both systematic and ecological studies of Malpighiaceae.

Alicia anisopetala and Callaeum psilophyllum are two closely related species that belong to the christianelloid clade of the family Malpighiaceae. Both species are pollinated by oil-collecting bees and exhibit variations at specimen and population level in the number of elaiophores per flower. These floral glands that secrete non-volatile oils constitute an ancestral trait for the family. There is evidence that the observed variations in the number of elaiophores can be the result of processes of connation or reduction associated with differences in their vascularization. In order to identify which process occurs in each species, we conducted an anatomical study in natural populations of both species distributed along a wide range of their geographical distributions in Argentina. We collected flowers of different individuals, counted the number of elaiophores per flower, carried out exomorphological observations, and used conventional histological techniques to examine the vascularization of these glands. The floral anatomy of both species does not show any modifications in other whorls related to the fusion or reduction of elaiophores. Our results indicate that the process of loss of elaiophores in A. anisopetala is caused by incomplete connation and in C. psilophyllum by reduction, suggesting that the processes that lead to the loss of elaiophores in Malpighiaceae are homoplastic and would not reflect phylogenetic signals.

The stereotyped floral morphology of Neotropical Malpighiaceae species-zygomorphic and calyx with oil glands-is correlated with oil-bee pollination. In contrast, the floral trends of the Paleotropical lineages are actinomorphy and lack of calyx glands, probably due to the absence of oil-collecting bees. The Paleotropical genus Acridocarpus is an exception because of its zygomorphic, gland-bearing flowers. Glands throughout Acridocarpus inflorescences were morphoanatomically evaluated to verify the occurrence of patterns related to species and geographic distribution. Herbarium (25 species) and fresh samples of A. longifolius were processed according to standard anatomical techniques. To verify the presence of glucose and protein, strip tests were applied to the exudate. Fresh samples were fixed and submitted to histochemical tests. Based on the occurrence, number and placement of glands, three distribution patterns were recognized on the bracteole and ten on the calyx. Bract, bracteole and sepal glands have a typical nectary structure with a palisade-like epidermis and vascularized parenchyma. Glands were classified as short-stalked, sessile or immersed. Histochemical tests performed on bract and sepal glands of A. longifolius were positive for proteins, polysaccharides and phenolic compounds, and negative for oil compounds. Glucose and protein were detected in the exudate. These results allow us to recognize the glands in Acridocarpus inflorescences as nectaries. This comprehensive morphoanatomical study helps to clarify the correlation between patterns of floral morphology and secretion consumers, as well as to better understand floral evolution in Malpighiaceae across their dispersal events.

Background and AimsMalpighiales are one of the largest angiosperm orders and have undergone radical systematic restructuring based on molecular phylogenetic studies. The clade has been recalcitrant to molecular phylogenetic reconstruction, but has become much more resolved at the suprafamilial level. It now contains so many newly identified clades that there is an urgent need for comparative studies to understand their structure, biology and evolution. This is especially true because the order contains a disproportionally large diversity of rain forest species and includes numerous agriculturally important plants. This study is a first broad systematic step in this endeavour. It focuses on a comparative structural overview of the flowers across all recently identified suprafamilial clades of Malpighiales, and points towards areas that desperately need attention.MethodsThe phylogenetic comparative analysis of floral structure for the order is based on our previously published studies on four suprafamilial clades of Malpighiales, including also four related rosid orders (Celastrales, Crossosomatales, Cucurbitales, Oxalidales). In addition, the results are compiled from a survey of over 3000 publications on macrosystematics, floral structure and embryology across all orders of the core eudicots.Key ResultsMost new suprafamilial clades within Malpighiales are well supported by floral structural features. Inner morphological structures of the gynoecium (i.e. stigmatic lobes, inner shape of the locules, placentation, presence of obturators) and ovules (i.e. structure of the nucellus, thickness of the integuments, presence of vascular bundles in the integuments, presence of an endothelium in the inner integument) appear to be especially suitable for characterizing suprafamilial clades within Malpighiales.ConclusionsAlthough the current phylogenetic reconstruction of Malpighiales is much improved compared with earlier versions, it is incomplete, and further focused phylogenetic and morphological studies are needed. Once all major subclades of Malpighiales are elucidated, more in-depth studies on promising structural features can be conducted. In addition, once the phylogenetic tree of Malpighiales, including closely related orders, is more fully resolved, character optimization studies will be possible to reconstruct evolution of structural and biological features within the order.

The repeated origin of similar phenotypes is invaluable for studying the underlying genetics of adaptive traits; molecular evidence, however, is lacking for most examples of such similarity. The floral morphology of neotropical Malpighiaceae is distinctive and highly conserved, especially with regard to symmetry, and is thought to result from specialization on oil-bee pollinators. We recently demonstrated that CYCLOIDEA2-like genes (CYC2A and CYC2B) are associated with the development of the stereotypical floral zygomorphy that is critical to this plant-pollinator mutualism. Here, we build on this developmental framework to characterize floral symmetry in three clades of Malpighiaceae that have independently lost their oil bee association and experienced parallel shifts in their floral morphology, especially in regard to symmetry. We show that in each case these species exhibit a loss of CYC2B function, and a strikingly similar shift in the expression of CYC2A that is coincident with their shift in floral symmetry. These results indicate that similar floral phenotypes in this large angiosperm clade have evolved via parallel genetic changes from an otherwise highly conserved developmental program.

• Premise of the study: Few studies have analyzed the physiological performance of different life stages and the expression of ontogenetic niche shifts in lianas. Here, we analyzed the photosynthetic and morphological acclimation of seedlings of Stigmaphyllon lindenianum, Combretum fruticosum, and Bonamia trichantha to distinctive light conditions in a tropical dry forest and compared their response with the acclimation response of adult canopy lianas of the same species. We expected acclimation to occur faster through changes in leaf photochemistry relative to adaptation in morphology, consistent with the life history strategies of these lianas.• Methods: Seedlings were assigned to the following light treatments: high light (HH), low light (LL), sun to shade (HL), and shade to sun (LH) in a common garden. After 40 d, HL and LH seedlings were exposed to opposite light treatments. Light response curves, the maximum photosynthetic rate in the field (Amax), and biomass allocation were monitored for another 40 d on leaves expanded before transfer.• Key results: Photosynthetic responses, Amax, and biomass of Stigmaphyllon and Combretum varied with light availability. Physiological characters were affected by current light environment. The previous light environment (carryover effects) only influenced Amax. Morphological characters showed significant carryover effects. Stigmaphyllon showed high morphological and physiological plasticity. Sun-exposed seedlings of this liana increased stem biomass and switched from self-supporting to climbing forms.• Conclusions: Acclimation in seedlings of these lianas is consistent with the response of adult lianas in the canopy in direction, but not in magnitude. There was no evidence for ontogenetic niche shifts in the acclimation response.

This aim of this study was to evaluate the lignification in young stems of the Lophanthera lactescens Ducke plant grown in vitro L. lactescens (Malpighiaceae), a species endemic in the Brazilian Amazon that possesses both medicinal properties and could be used in the forest product industry. Plants grown in vitro condition in MS medium were analyzed using Infrared (IR) microspectroscopy in a diffuse reflectance mode, fluorescence microscopy. Moreover, histochemical tests such as the Wiesner and Maüle test were run to monitor the process of lignification in cell walls. The spectra of IR microscopy recorded using cross section tissue are representative of guaiacyl/syringyl lignin, based on the 1336 and 1246 cm-1 signal. Individuals presenting stem sprains, resulting from the marked development, produced gelatinous fibers with a clear cellulose layer. Initially, fluorescence microscopy demonstrated lignin deposition in the cell corner region having progressive deposition in the secondary wall of tracheary elements of the stem. Esta pesquisa teve como objetivo avaliar a lignificação em caules jovens de plantas de Lophanthera lactescens Ducke crescidas in vitro. L. lactescens (Malpighiaceae) é uma espécie endêmica da Amazônia Brasileira que, além de propriedades medicinais, apresenta potencial para a indústria de produtos florestais. As plantas foram crescidas in vitro, usando-se meio nutritivo MS. As análises foram realizadas por microespectrometria no infravermelho, no modo reflectância difusa. Microscopia com fluorescência e testes histoquímicos de Wiesner e Maüle foram realizados para o acompanhamento do processo de lignificação. Os espectros registrados a partir da microscopia no infravermelho mostraram sinais em 1336 e 1246 cm-1, característicos de ligninas siringílica e guaiacílica. Indivíduos que apresentaram entorses no caule, em decorrência do acentuado desenvolvimento, produziram fibras gelatinosas, com a camada de celulose evidente. A microscopia com fluorescência evidenciou deposição de lignina inicialmente nos ângulos das células, com depósito progressivo ocorrendo na parede secundária dos elementos traqueais do caule.

The aim of this study was to evaluate the effects of soy-based beverages manufactured with water-soluble soy extract, containing probiotic strains (Lactobacillus acidophilus LA-5 and Bifidobacterium longum BB-46) and/or acerola by-product (ABP) on pooled faecal microbiota obtained from lean and obese donors. Four fermented soy beverages (FSs) (“placebo” (FS-Pla), probiotic (FS-Pro), prebiotic (FS-Pre), and synbiotic (FS-Syn)) were subjected to in vitro digestion, followed by inoculation in the TIM-2 system, a dynamic in vitro model that mimics the conditions of the human colon. Short- and branched-chain fatty acids (SCFA and BCFA) and microbiota composition were determined. Upon colonic fermentation in the presence of the different FSs formulations, acetic and lactic acid production was higher than the control treatment for faecal microbiota from lean individuals (FMLI). Additionally, SCFA production by the FMLI was higher than for the faecal microbiota from obese individuals (FMOI). Bifidobacterium spp. and Lactobacillus spp. populations increased during simulated colonic fermentation in the presence of FS-Syn in the FMLI and FMOI. FS formulations also changed the composition of the FMOI, resulting in a profile more similar to the FMLI. The changes in the composition and the increase in SCFA production observed for the FMLI and FMOI during these in vitro fermentations suggest a potential modulation effect of these microbiotas by the consumption of functional FSs.Key points• Soy beverages increased Bifidobacterium abundance in microbiota from obese individuals.• The synbiotic beverage increased Bifidobacterium abundance in microbiota from lean individuals.• The synbiotic beverage changed the microbiota from obese individuals, approaching the lean profiles.

The flowers of the species of Malpighiaceae in the Neotropical Region are relatively uniform in their morphology due to their dependence on oil-collecting bees as their main pollinators. However, many species of the genus Galphimia seem to have acquired a different floral syndrome, lacking markedly zygomorphic flowers and developed elaiophores in the calyx. Likewise, these species present anthers with great development, probably in response to the selection of pollinators that collect pollen. Galphimia australis incorporated some of these traits but also retained some residual characteristics typical of species pollinated by oil bees. This leads to many questions on how these flowers ensure their pollination. Inquiring about the reduction or modification of these characteristics allows us to understand how G. australis achieves a different pollination syndrome. In this research, we carry out a detailed morphological and anatomical study of the flowers and pollen grain devolvement of G. australis and floral visitors were observed and captured. Results were analyzed in order to determine how this species changed from the oil-floral syndrome, typical of neotropical Malpighiaceae, to one syndrome with pollen as the main reward.