Arceuthobium M.Bieb. (Viscaceae) parasitizing an angiosperm: the unique case of A. azoricum Hawksw. & Wiens in the Azores islands.

Arceuthobium azoricum Wiens & Hawksw. is a rare Azorean endemic epiphytic hemiparasite (mistletoe), that typically parasitizes branches of the gymnosperm Juniperus brevifolia (Hochst. ex Seub.) Antoine (Cupressaceae), also an Azorean endemic. Here we describe one population of A. azoricum, on Pico Island, parasitizing Erica azorica Hochst. ex Seub (Ericaceae) also an Azores endemic. Our molecular analysis (using nuclear ribosomal ITS) showed no differences between individuals parasitizing Erica and Juniperus. Moreover, a preliminary analysis showed no differences in morphological characters between accessions sampled from the different hosts. Given that this is the first bona fide record of Arceuthobium parasitizing an angiosperm, this population represents a unique host shifting event and its conservation is important because it may allow new insights into host recognition mechanisms in mistletoes. Immediate attention should be given to characterize this Pico Island population using appropriate molecular methods and additional morphological analyses.

Vascular plant taxa occurrences in exotic woodland and in natural and production forests on the Islands of São Miguel, Terceira and Pico (Azores).

Background: The data presented here originated from field expeditions carried out between 2017 and 2018, within the framework of Forest-Eco2 project: "Towards an Ecological and Economic valorisation of the Azorean Forest". The project aimed to quantify the ecological value of the Azorean forests, including carbon accumulation and to design and propose measures that could further enhance forest sustainability. For that, 90 forest plots were sampled on three Azores islands - São Miguel, Terceira and Pico - equally distributed into natural forest, exotic woodland and production forest. The aim of this report is to further expand knowledge on biodiversity trends enclosed in the different forest types present in the Azores, by providing a list of the occurrences of the 105 different vascular plant taxa together with a brief characterisation of their origin and life-form. New information: We provide an inventory of indigenous and non-indigenous vascular plant taxa from 90 forest stands. A total of 105 taxa were identified and registered, belonging to 60 families, 91 genera, 101 species and four subspecies. A total of 35% of the taxa were endemic, 27% native and 38% non-indigenous, including 19% of invasive taxa. Endangered and vulnerable taxa were registered, including Elaphoglossumhirtum (Sw.) C.Chr., Lactucawatsoniana Trel. and others which were considered by the authors a priority for conservation (e.g. Arceuthobiumazoricum Wiens & Hawksw., Bellisazorica Hochst. ex Seub., Saniculaazorica Guthnick ex Seub., Platantheramicrantha (Hochst. ex Seub.) Schltr.). Our records provide detailed and updated knowledge of Azorean Forest flora and highlight the role of natural forests as indigenous plant diversity hotspots and exotic woodland as a source of invasive taxa within the Archipelago.

Genome size variation within Crithmum maritimum: Clues on the colonization of insular environments.

Angiosperms present an astonishing diversity of genome sizes that can vary intra- or interspecifically. The remarkable new cytogenomic data shed some light on our understanding of evolution, but few studies were performed with insular and mainland populations to test possible correlations with dispersal, speciation, and adaptations to insular environments. Here, patterns of cytogenomic diversity were assessed among geographic samples (ca. 114) of Crithmum maritimum (Apiaceae), collected across the Azores and Madeira archipelagos, as well as in adjacent continental areas of Portugal. Using flow cytometry, the results indicated a significant intraspecific genome size variation, spanning from reduced sizes in the insular populations to larger ones in the mainland populations. Moreover, there was a tendency for an increase in genome size along the mainland populations, associated with lower temperatures, higher precipitation, and lower precipitation seasonality. However, this gradient might be the result of historic phylogeographical events associated with previous dispersal and extinction of local populations. Overall, our findings provided evidence that smaller genome sizes might play a critical role in the colonization of islands, corroborating other studies that argue that organisms with smaller genomes use fewer resources, having a selective advantage under insular environments. Although further studies are needed to improve our understanding of the mechanisms underlying genome size evolution on islands, conservation strategies must be promoted to protect the rich cytogenomic diversity found among C. maritimum populations, which occur in coastal areas that are particularly threatened by human activity, pollution, invasive species, and climate changes.

Diversity and biological activities of medicinal plants of Santiago island (Cabo Verde).

Plants continue to constitute key elements of medical practice in West African countries. The Cabo Verde archipelago hosts a great diversity of medicinal plants and local markets are considered important sites for trading plants harvested by rural communities. This study has two main goals: (i) to assess the medicinal uses of native species in Santiago, the biggest island of the archipelago, and (ii) to evaluate the antioxidant, antimicrobial and antidiabetic/antihyperglycemic activities of two native trees (Tamarix senegalensis and Sideroxylon marginatum) used in traditional medicine and traded in local markets. Our results revealed that on Santiago Island, 24 native plants are used in traditional medicine. The main uses of these species (e.g., forage, timber, food and fibres), their medicinal applications, the plant parts used, their mode of administration and conservation status are presented here for the first time. Moreover, the pharmacological characterization of two native tree species revealed that hydroethanolic extracts were richer in phenolic compounds and more active than their aqueous counterparts. All the studied extracts revealed significant antioxidant properties (DPPH and FRAP assays) and were generally moderately active against Gram-positive bacteria. All the extracts inhibited the activities of the carbohydrate digestive enzymes α-glucosidase and α-amylase in a dose-dependent manner. For α-glucosidase, the detected inhibitory activity (IC50 values from 2.0 ± 0.2 µg/mL to 9.9 ± 1.2 µg/mL) was significantly higher than that of acarbose, suggesting that extracts of both species can delay glucose absorption, thereby assisting in slowing down the progression of diabetes. Our findings highlight the crucial importance that medicinal plants have for the Cabo Verdean population, while also raising awareness on the need for sustainable use and conservation of native flora, and of tree species traded in local markets in particular.

Antimalarial and Cytotoxic Activity of Native Plants Used in Cabo Verde Traditional Medicine.

Medicinal plants have historically been a source of drugs in multiple applications, including the treatment of malaria infections. The Cabo Verde archipelago harbors a rich diversity of native plants, most of which are used for medicinal purposes. The present study investigated the in vitro antiplasmodial activities of four native plants from Cabo Verde (i.e., Artemisia gorgonum, Lavandula rotundifolia, Sideroxylon marginatum, and Tamarix senegalensis). Traditional preparations of these medicinal plants, namely aqueous extracts (infusions) and ethanolic extracts, were tested against both chloroquine-sensitive (3D7) and chloroquine-resistant (Dd2) Plasmodium falciparum strains using the SYBR Green detection method. The in vitro cytotoxicity was evaluated in Caco-2 and PLP2 cells using a sulforhodamine B colorimetric assay. An ethanolic extract of A. gorgonum and infusions of T. senegalensis exhibited high antiplasmodial activities (EC50 < 5 µg/mL) without cytotoxicity (GI50 > 400 µg/mL). Extracts of L. rotundifolia and S. marginatum exhibited moderate activities, with EC50 values ranging from 10-30 µg/mL. The A. gorgonum ethanolic extract showed activity toward early ring stages, and parasites treated with the T. senegalensis infusions progressed to the early trophozoite stage, although did not develop further to the late trophozoite or schizont stages. Antimalarial activities and the lack of cytotoxicity of the extracts are reported in the present study and support previous claims by traditional practitioners for the use of these plants against malaria while suggesting their ethnopharmacological usefulness as future antimalarials.

Monitoring Yellow Mealworm (Tenebrio molitor) as a Potential Novel Allergenic Food: Effect of Food Processing and Matrix.

The consumption of insects has increased in western countries, raising concerns about their potential to induce food allergic reactions in sensitized/allergic individuals. This work intended to develop a real-time PCR approach for the detection/quantification of yellow mealworm (Tenebrio molitor) as a potential allergenic food in complex matrices. For this purpose, reference mixtures simulating the production of pork sausages and wheat biscuits containing known amounts of mealworm were used. Real-time PCR with TaqMan probe targeting the cytochrome b gene of T. molitor was able to detect up to 2 fg of insect DNA, and 1.0 and 0.1 mg/kg of mealworm flour in autoclaved sausages and baked biscuits, respectively. Generally, the method showed acceptable analytical performance parameters, confirming its suitability/applicability for a wide range of foods. However, real-time PCR data showed significant differences among food matrix and processing, highlighting the importance of using appropriate calibration models for quantitative analysis. Finally, the real-time PCR approach was successfully validated with blind mixtures and applied to commercial samples, demonstrating its efficacy and reliability in the quantification of mealworm in processed foodstuffs.

Authentication of incense (Pittosporum undulatum Vent.) honey from the Azores (Mel dos Açores) by a novel real-time PCR approach.

'Mel dos Açores' is a unique nectar honey produced from the exceptional and diverse flora of the Azores archipelago, categorised as incense honey ('mel de incenso') or multifloral honey ('mel multiflora'). Incense honey should contain over 30 % of pollen grains of Pittosporum undulatum Vent. In this work, a real-time PCR method targeting the ITS region was proposed for the first time to detect P. undulatum in the honey from the Azores. The approach exhibited high analytical performance, achieving a quantification limit of 0.01 pg of incense DNA. The method was successfully applied to 22 honey samples, from which incense was detected in all 9 monofloral incense honeys and in 5 out of 10 multifloral samples from the Azores. Generally, the quantitative results for incense DNA were in good agreement with the melissopalynological data. Therefore, a simple, cost-effective and reliable tool was herein proposed to authenticate and valorise the Azores honey.

Evaluation of the Polyphenolic Composition and Bioactivities of Three Native Cabo Verde Medicinal Plants.

The use of medicinal plants in a variety of health conditions remains essential for the discovery of new treatments. The present study aimed to investigate the bioactive properties of three native plants from Cabo Verde Islands, namely Artemisia gorgonum Webb, Sideroxylon marginatum (Decne. ex Webb) Cout., and Tamarix senegalensis DC., contributing to the characterization of less-known medicinal plants and their potential benefits for human health. Known compounds, such as kaempferol, quercetin, caffeyolquinic, and apigenin derivatives, among others, were detected in the plant species under study. Overall, all species demonstrated good antioxidant capacity, especially the ethanolic extracts of A. gorgonum (EC50 = 0.149 mg/mL) in TBARS assay. Moreover, the ethanolic extracts of the studied plants showed cytotoxic properties against tumor cells, and again the A. gorgonum extract proved to be the most effective in inhibiting tumor growth, mainly in the CaCO2 (GI50 = 17.3 µg/mL) and AGS (GI50 = 18.2 µg/mL) cell lines. Only the ethanolic extracts of T. senegalensis and S. marginatum demonstrated anti-inflammatory activity, albeit weak (EC50 = 35 and 43 µg/mL, respectively). The present study contributed to increased knowledge about the bioactive properties of these plants commonly used in traditional medicine, some of which was discussed for the first time, opening new perspectives for their use in a wider range of health conditions, especially in African countries, where access to modern health care is more limited.

Multiplexed shotgun genotyping (MSG) data resolve phylogenetic relationships within and among archipelagos in Macaronesian Tolpis.

PREMISE: Plants endemic to oceanic archipelagos are suitable for studying evolution, being isolated on substrates of different ages. Evolution has been recent, rendering traditionally employed sequences insufficiently variable for resolving relationships. This study includes sampling in the genus Tolpis (Asteraceae) from the Azores, Madeira, and Cape Verde, and expands upon an earlier study demonstrating the efficacy of multiplexed shotgun genotyping (MSG) for resolving relationships in Canarian Tolpis. METHODS: Genomic libraries for 90 accessions of Tolpis and two from the outgroup were generated for genotyping individuals using MSG. Loci were de novo assembled with iPyrad, which clusters MSG loci within and between samples. A maximum likelihood phylogeny was generated with RAxML. Ancestral area reconstruction was inferred using R package BioGeoBEARS. RESULTS: MSG data recovered a highly resolved phylogeny from population to inter-archipelago levels. Ancestral area reconstruction provided biogeographic hypotheses for the radiation of Macaronesian Tolpis. CONCLUSIONS: Four major clades were resolved. The Madeiran endemic T. macrorhiza is sister to other Tolpis. Species from the Canaries, Cape Verdes, and the continent are sister to T. succulenta from Madeira, which has a sister subclade of Azorean populations composed of T. succulenta and T. azorica. Population-level resolution suggests unrecognized taxa on several archipelagos. Ancestral reconstruction suggests initial dispersal from the continent to Madeira, with dispersal to the Azores, then dispersal from Madeira to the Canary Islands, with both subsequent dispersal to the Cape Verdes and back-dispersal to the continent. Single-island radiations and inter-island dispersal are implicated in divergence in Macaronesian Tolpis.

Taxonomic, structural diversity and carbon stocks in a gradient of island forests.

Assessment of forest ecosystems and their services is seen as a key action for the advancement of biodiversity objectives, and to inform the development and implementation of related policies and planning. The Azorean forest is important for timber production, the protection of soil and water resources, and for its recreational and aesthetic value. However, its role in carbon accumulation has not been fully addressed. We assessed plant diversity, forest structure and carbon stocks in a gradient of three forest types (Natural Forest-NF; Exotic Woodland-EW and Production Forest-PF) in three of the Azores islands. We used biodiversity indices and found that NF harbored the highest plant diversity levels and PF the lowest. Diversity levels were lower for structural than for taxonomic data, particularly for PF. The highest tree carbon stock was found at EW in one of the islands, while PF consistently exhibited relatively high tree carbon stocks in the three islands. The largest soil carbon stocks were found at EW, while leaf litter carbon stocks were higher at PF. We concluded that NF play a fundamental role as plant diversity hotspots but have lower relevance as carbon stocks what might be associated with montane environmental conditions. PFs provide economic assets and act as carbon sinks, while EWs play a major role as carbon sinks in soil, but also at tree level in the oldest forests.

Diversity and Cytogenomic Characterization of Wild Carrots in the Macaronesian Islands.

The Macaronesian islands constitute an enormous reservoir of genetic variation of wild carrots (subtribe Daucinae; Apiaceae), including 10 endemic species, but an accurate understanding of the diversification processes within these islands is still lacking. We conducted a review of the morphology, ecology, and conservation status of the Daucinae species and, on the basis of a comprehensive dataset, we estimated the genome size variation for 16 taxa (around 320 samples) occurring in different habitats across the Macaronesian islands in comparison to mainland specimens. Results showed that taxa with larger genomes (e.g., Daucus crinitus: 2.544 pg) were generally found in mainland regions, while the insular endemic taxa from Azores and Cabo Verde have smaller genomes. Melanoselinum decipiens and Monizia edulis, both endemic to Madeira Island, showed intermediate values. Positive correlations were found between mean genome size and some morphological traits (e.g., spiny or winged fruits) and also with habit (herbaceous or woody). Despite the great morphological variation found within the Cabo Verde endemic species, the 2C-values obtained were quite homogeneous between these taxa and the subspecies of Daucus carota, supporting the close relationship among these taxa. Overall, this study improved the global knowledge of DNA content for Macaronesian endemics and shed light into the mechanisms underpinning diversity patterns of wild carrots in the western Mediterranean region.

Exploring physicochemical and cytogenomic diversity of African cowpea and common bean.

In sub-Saharan Africa, grain legumes (pulses) are essential food sources and play an important role in sustainable agriculture. Among the major pulse crops, the native cowpea (Vigna unguiculata) and introduced common bean (Phaseolus vulgaris) stand out. This paper has two main goals. First, we provide a comprehensive view of the available genetic resources of these genera in Africa, including data on germplasm collections and mapping biodiversity-rich areas. Second, we investigate patterns of physicochemical and cytogenomic variation across Africa to explore the geographical structuring of variation between native and introduced beans. Our results revealed that 73 Vigna and 5 Phaseolus species occur in tropical regions of Africa, with 8 countries accounting for more than 20 native species. Conversely, germplasm collections are poorly represented when compared to the worldwide collections. Regarding the nuclear DNA content, on average, V. unguiculata presents significantly higher values than P. vulgaris. Also, V. unguiculata is enriched in B, Mg, S, and Zn, while P. vulgaris has more Fe, Ca, and Cu. Overall, our study suggests that the physicochemical and cytogenomic diversity of native Vigna species is higher than previously thought, representing valuable food resources to reduce food insecurity and hunger, particularly of people living in African developing countries.

Exploring glucosinolates diversity in Brassicaceae: a genomic and chemical assessment for deciphering abiotic stress tolerance.

Brassica is one of the most economically important genus of the Brassicaceae family, encompassing several key crops like Brassica napus (cabbage) and broccoli (Brassica oleraceae var. italica). This family is well known for their high content of characteristic secondary metabolites such as glucosinolates (GLS) compounds, recognize for their beneficial health properties and role in plants defense. In this work, we have looked through gene clusters involved in the biosynthesis of GLS, by combining genomic analysis with biochemical pathways and chemical diversity assessment. A total of 101 Brassicaceae genes involved in GLS biosynthesis were identified, using a multi-database approach. Through a UPGMA and PCA analysis on the 101 GLS genes recorded, revealed a separation between the genes mainly involved in GLS core structure synthesis and genes belonging to the CYP450s and MYBs gene families. After, a detailed phylogenetic analysis was conducted to better understand the disjunction of the aliphatic and indolic genes, by focusing on CYP79F1-F2 and CYP81F1-F4, respectively. Our results point to a recent diversification of the aliphatic CYP79F1 and F2 genes in Brassica crops, while for indolic genes an earliest diversification is observed for CYP81F1-F4 genes. Chemical diversity revealed that Brassica crops have distinct GLS chemo-profiles from other Brassicaceae genera; being highlighted the high contents of GLS found among the Diplotaxis species. Also, we have explored GLS-rich species as a new source of taxa with great agronomic potential, particularly in abiotic stress tolerance, namely Diplotaxis, the closest wild relatives of Brassica crops.

Immunoreactivity of Lupine and Soybean Allergens in Foods as Affected by Thermal Processing.

Lupine and soybean are important technological aids for the food industry. However, they are also capable of inducing severe allergic reactions in food-sensitized/allergic individuals. In this context, this work intended to study the combined effects of thermal processing and food matrix on the immunoreactivity of lupine and soybean proteins used as ingredients in bakery and meat products, respectively. For this purpose, the effects of baking, mild oven cooking, and autoclaving on the protein profiles were evaluated, using model mixtures simulating the production of lupine-containing breads and soybean-containing cooked hams/sausages, by native- and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and immunoblotting using specific antibodies. The results showed that lupine gamma-conglutin immunoreactivity was slightly decreased in wheat flour mixtures compared to rice, but it was more pronounced in baked products. In meat mixtures, substantial protein fragmentation was noted after autoclaving, with decreased immunoreactivity of soybean trypsin inhibitor. The analysis of 22 commercial products enabled the identification of lupine gamma-conglutin in four bakery samples and soybean trypsin-inhibitor in five sausages, and further differentiated autoclaved from other milder thermally treated products. Generally, the immunoreactivity of target proteins was reduced by all the tested thermal treatments, though at a higher extent after autoclaving, being slightly altered by the food matrix.

How rapidly do self-compatible populations evolve selfing? Mating system estimation within recently evolved self-compatible populations of Azorean Tolpis succulenta (Asteraceae).

Genome-wide genotyping and Bayesian inference method (BORICE) were employed to estimate outcrossing rates and paternity in two small plant populations of Tolpis succulenta (Asteraceae) on Graciosa island in the Azores. These two known extant populations of T. succulenta on Graciosa have recently evolved self-compatibility. Despite the expectation that selfing would occur at an appreciable rate (self-incompatible populations of the same species show low but nonzero selfing), high outcrossing was found in progeny arrays from maternal plants in both populations. This is inconsistent with an immediate transition to high selfing following the breakdown of a genetic incompatibility system. This finding is surprising given the small population sizes and the recent colonization of an island from self-incompatible colonists of T. succulenta from another island in the Azores, and a potential paucity of pollinators, all factors selecting for selfing through reproductive assurance. The self-compatible lineage(s) likely have high inbreeding depression (ID) that effectively halts the evolution of increased selfing, but this remains to be determined. Like their progeny, all maternal plants in both populations are fully outbred, which is consistent with but not proof of high ID. High multiple paternity was found in both populations, which may be due in part to the abundant pollinators observed during the flowering season.

Bioconcentrations of herbicides used in sugarcane crops in tilapia (Oreochromis niloticus) and the risk for human consumption.

The practice of intensive herbicide use in the sugarcane industry has a high risk of compromising the quality of the water and the organisms that live there due to losses through runoff, leaching and other processes. In this work, the dynamics of four herbicides present in three different mixtures were evaluated through their incorporation and elimination in the muscle tissue of tilapia (Oreochromis niloticus). The highest mean values of bioconcentration factors were 1.730 for ametryn, 0.891 for tebuthiuron, 0.322 for hexazinone and 4.783 for diuron. Diuron presented the highest risk regarding the consumption of tilapia fillets by the population. However, considering that the fish would reach maximum levels of diuron when exposed to extremely high concentrations, an individual weighing 70 kg would need to ingest approximately 1.5 kg of this food product to surpass the acceptable daily intake of 0.007 mg kg-1 body weight. It was concluded that the risk of injury to the population consuming tilapia fillets from fish exposed to herbicides in water arising from sugarcane activities is very low. According to the risk estimation performed in this work, which is substantiated by the assumptions of the World Health Organization and the International Life Sciences Institute, there is a low risk of injury to the population consuming tilapia fillets from fish exposed to water containing herbicides in concentrations arising from sugarcane activities. However, as the risk was estimated from laboratory conditions, caution should be taken where herbicide applications are carried out with high frequency near water bodies, as the consumption of fish from these areas is quite common.

Shortcomings of Phylogenetic Studies on Recent Radiated Insular Groups: A Meta-Analysis Using Cabo Verde Biodiversity.

Over the previous decades, numerous studies focused on how oceanic islands have contributed to determine the phylogenetic relationships and times of origin and diversification of different endemic lineages. The Macaronesian Islands (i.e., Azores, Madeira, Selvagens, Canaries, and Cabo Verde), harbour biotas with exceptionally high levels of endemism. Within the region, the vascular plants and reptiles constitute two of the most important radiations. In this study we compare relevant published phylogenetic data and diversification rates retrieved within Cabo Verde endemic lineages and discuss the importance of choosing appropriate phylogeny-based methods to investigate diversification dynamics on islands. From this selective literature-based review, we summarize the software packages used in Macaronesian studies and discuss their adequacy considering the published data to obtain well-supported phylogenies in the target groups. We further debate the importance of Next Generation Sequencing (NGS), to investigate the evolutionary processes of diversification in the Macaronesian Islands. Analysis of genomic data provides phylogenetic resolution for rapidly evolving species radiations, suggesting a great potential to improve the phylogenetic signal and divergence time estimates in insular lineages. The most important Macaronesian reptile radiations provide good case-studies to compare classical phylogenetic methods with new tools, such as phylogenomics, revealing a high value for research on this hotspot area.

Phylogeography of the Macaronesian Lettuce Species Lactuca watsoniana and L. palmensis (Asteraceae).

The phylogenetic relationships and phylogeography of two relatively rare Macaronesian Lactuca species, Lactuca watsoniana (Azores) and L. palmensis (Canary Islands), were, until this date, unclear. Karyological information of the Azorean species was also unknown. For this study, a chromosome count was performed and L. watsoniana showed 2n = 34. A phylogenetic approach was used to clarify the relationships of the Azorean endemic L. watsoniana and the La Palma endemic L. palmensis within the subtribe Lactucinae. Maximum parsimony, Maximum likelihood and Bayesian analysis of a combined molecular dataset (ITS and four chloroplast DNA regions) and molecular clock analyses were performed with the Macaronesian Lactuca species, as well as a TCS haplotype network. The analyses revealed that L. watsoniana and L. palmensis belong to different subclades of the Lactuca clade. Lactuca watsoniana showed a strongly supported phylogenetic relationship with North American species, while L. palmensis was closely related to L. tenerrima and L. inermis, from Europe and Africa. Lactuca watsoniana showed four single-island haplotypes. A divergence time estimation of the Macaronesian lineages was used to examine island colonization pathways. Results obtained with BEAST suggest a divergence of L. palmensis and L. watsoniana clades c. 11 million years ago, L. watsoniana diverged from its North American sister species c. 3.8 million years ago and L. palmensis diverged from its sister L. tenerrima, c. 1.3 million years ago, probably originating from an African ancestral lineage which colonized the Canary Islands. Divergence analyses with *BEAST indicate a more recent divergence of the L. watsoniana crown, c. 0.9 million years ago. In the Azores colonization, in a stepping stone, east-to-west dispersal pattern, associated with geological events might explain the current distribution range of L. watsoniana.

Phylogenetic relationships and phylogeography of relevant lineages within the complex Campanulaceae family in Macaronesia.

Macaronesia has long been recognized as a natural model for studying evolutionary processes in plant diversification. Several studies have attempted to focus on single lineages, and few have covered the diversification of a family across all the archipelagos. We used a comprehensive sample to clarify the phylogenetic relationships and the biogeographic history of the Macaronesian Campanulaceae. Hypotheses related to the colonization of these archipelagos will be used to examine the diversification patterns of different lineages. We sequenced the ITS region and six cpDNA markers (atpB, matK, petD, rbcL, trnL-F, and psbA-trnH) from 10 Campanulaceae species, including seven endemic species in Macaronesia. The phylogeny of these taxa was reconstructed using maximum parsimony, maximum likelihood, and Bayesian inference. To study the relationships within each lineage, haplotype networks were calculated using NeighborNet and TCS algorithms. Moreover, data were combined with fossil information to construct time-calibrated trees for the Macaronesian Campanulaceae species. The phylogenetic analyses are largely congruent with current taxon circumscriptions, and all the endemic genera formed monophyletic clades, namely Azorina in Azores; Musschia in Madeira; and Campanula in Cape Verde. The Azorina clade and the Cape Verde endemic Campanula may share a common ancestor in North Africa, and the divergence was dated ca. 12.3 million years ago (Mya). The divergence of the Musschia clade began in the Pliocene ca. 3.4 Mya. Moreover, several examples of intraspecific variation were revealed among the native species with a clear geographic structured patterns, suggesting that cryptic diversity might exist within the native Macaronesian Campanulaceae when compared to the close mainland taxa (e.g., Campanula erinus, Trachelium caeruleum), but additional studies are needed to support the molecular data. This study highlights the power of combining data (e.g., phylogeny and divergence times, with species distribution data) for testing diversification hypotheses within the unique Macaronesian flora, providing useful information for future conservation efforts.