Phylogenomics and morphological evolution of the mega-diverse genus Artemisia (Asteraceae: Anthemideae): implications for its circumscription and infrageneric taxonomy.

BACKGROUND AND AIMS: Artemisia is a mega-diverse genus consisting of ~400 species. Despite its medicinal importance and ecological significance, a well-resolved phylogeny for global Artemisia, a natural generic delimitation and infrageneric taxonomy remain missing, owing to the obstructions from limited taxon sampling and insufficient information on DNA markers. Its morphological characters, such as capitulum, life form and leaf, show marked variations and are widely used in its infrageneric taxonomy. However, their evolution within Artemisia is poorly understood. Here, we aimed to reconstruct a well-resolved phylogeny for global Artemisia via a phylogenomic approach, to infer the evolutionary patterns of its key morphological characters and to update its circumscription and infrageneric taxonomy. METHODS: We sampled 228 species (258 samples) of Artemisia and its allies from both fresh and herbarium collections, covering all the subgenera and its main geographical areas, and conducted a phylogenomic analysis based on nuclear single nucleotide polymorphisms (SNPs) obtained from genome skimming data. Based on the phylogenetic framework, we inferred the possible evolutionary patterns of six key morphological characters widely used in its previous taxonomy. KEY RESULTS: The genus Kaschgaria was revealed to be nested in Artemisia with strong support. A well-resolved phylogeny of Artemisia consisting of eight highly supported clades was recovered, two of which were identified for the first time. Most of the previously recognized subgenera were not supported as monophyletic. Evolutionary inferences based on the six morphological characters showed that different states of these characters originated independently more than once. CONCLUSIONS: The circumscription of Artemisia is enlarged to include the genus Kaschgaria. The morphological characters traditionally used for the infrageneric taxonomy of Artemisia do not match the new phylogenetic tree. They experienced a more complex evolutionary history than previously thought. We propose a revised infrageneric taxonomy of the newly circumscribed Artemisia, with eight recognized subgenera to accommodate the new results.

Novel Insights into the Nature of Intraspecific Genome Size Diversity in Cannabis sativa L.

Cannabis sativa has been used for millennia in traditional medicine for ritual purposes and for the production of food and fibres, thus, providing important and versatile services to humans. The species, which currently has a worldwide distribution, strikes out for displaying a huge morphological and chemical diversity. Differences in Cannabis genome size have also been found, suggesting it could be a useful character to differentiate between accessions. We used flow cytometry to investigate the extent of genome size diversity across 483 individuals belonging to 84 accessions, with a wide range of wild/feral, landrace, and cultivated accessions. We also carried out sex determination using the MADC2 marker and investigated the potential of flow cytometry as a method for early sex determination. All individuals were diploid, with genome sizes ranging from 1.810 up to 2.152 pg/2C (1.189-fold variation), apart from a triploid, with 2.884 pg/2C. Our results suggest that the geographical expansion of Cannabis and its domestication had little impact on its overall genome size. We found significant differences between the genome size of male and female individuals. Unfortunately, differences were, however, too small to be discriminated using flow cytometry through the direct processing of combined male and female individuals.

The Use of Cannabis sativa L. for Pest Control: From the Ethnobotanical Knowledge to a Systematic Review of Experimental Studies.

Background: Despite the benefits that synthetic pesticides have provided in terms of pest and disease control, they cause serious long-term consequences for both the environment and living organisms. Interest in eco-friendly products has subsequently increased in recent years. Methods: This article briefly analyzes the available ethnobotanical evidence regarding the use of Cannabis sativa as a pesticide and offers a systematic review of experimental studies. Results: Our findings indicate that both ethnobotanical and experimental procedures support the use of C. sativa as a pesticide, as remarkable toxicity has been observed against pest organisms. The results included in the systematic review of experimental studies (n=30) show a high degree of heterogeneity, but certain conclusions can be extracted to guide further research. For instance, promising pesticide properties were reported for most of the groups of species tested, especially Arachnida and Insecta; the efficacy of C. sativa as a pesticide can be derived from a wide variety of compounds that it contains and possible synergistic effects; it is crucial to standardize the phytochemical profile of C. sativa plants used as well as to obtain easily reproducible results; appropriate extraction methods should be explored; and upper inflorescences of the plant may be preferred for the production of the essential oil, but further studies should explore better other parts of the plant. Conclusion: In the coming years, as new findings are produced, the promising potential of C. sativa as a pesticide will be elucidated, and reviews such as the present one constitute useful basic tools to make these processes easier.

Traditional uses of Cannabis: An analysis of the CANNUSE database.

ETHNOPHARMACOLOGICAL RELEVANCE: Cannabis is one of the most versatile genera in terms of plant use and has been exploited by humans for millennia. Nowadays, Cannabis is the centre of many scientific studies, most of them focusing on chemical composition and medicinal values. While new and varied applications are continuously being developed, the knowledge surrounding less common uses of the plant is slowly disappearing. AIM OF THE REVIEW: We have analysed diversity of global data of Cannabis traditional uses, to investigate if certain plant parts are significantly associated with particular Cannabis use. We wanted to uncover potential associations between the plant parts used for the treatment of different body systems and ailments. MATERIALS AND METHODS: We have analysed the extensive database of Cannabis traditional uses (CANNUSE). This database contains 2330 data entries of Cannabis ethnobotanical uses from over 40 countries across the world. The dataset was divided into five general groups based on the type of use: medicinal, alimentary, psychoactive, fibre and other uses. Given the abundance of human medicinal uses, detailed analysis was done on the subset of 1167 data entries. We analysed the relationship between 16 body system categories and ailments treated with Cannabis plant parts. We used a Pearson's chi-square and Fisher's exact test, to determine which Cannabis parts are characteristic of treatment for specific ailments. RESULTS: In this dataset, the majority of reports were represented by medicinal (75.41%), followed by psychoactive (8.35%) and alimentary (7.29%) use. The most commonly used plant parts were leaf (50.51%), seed (15.38%) and inflorescence (11.35%). We found that different Cannabis plant parts were significantly associated with different uses; the leaf was typically used for medicinal, seed for alimentary and inflorescence for psychoactive use. Regarding the human medicinal uses, most common were reports for treatments of the digestive system and nutritional disorders (17.66%), nervous system and mental disorders (16.24%), followed by pain and inflammations (12.21%). We found a significant relationship between the use of certain Cannabis parts and treatment of ailments and body systems categories; leaf was significantly associated with treatment of two categories: skin and subcutaneous tissue disorders and circulatory system and blood disorders; seed use was associated with musculoskeletal system disorders and traumas; while inflorescence use shows a statistical support for treatment of nervous system and mental disorders. CONCLUSION: Several pharmaceutical companies are intensely working on developing new drugs with isolated chemical compounds or crude extracts, almost exclusively from Cannabis inflorescences. However, our review revealed that use of leaf or seed in traditional medicine is often more important than use of inflorescence for the treatment of certain ailments. A review of traditional medicine provides a body of knowledge and an initial pathway to identify landraces and plant parts that could have an important role in future medicinal research. We are confident that traditional medicine still has a large potential for modern medicine. As more information on Cannabis diversity (genetics, biochemistry, and clinical studies) becomes available, ethnobotanical data are poised to be of much greater significance.

CANNUSE, a database of traditional Cannabis uses-an opportunity for new research.

Cannabis is one of the most versatile genera in terms of plant uses and has been exploited by humans for millennia due to its medicinal properties, strong fibres, nutritious seeds and psychoactive resin. Nowadays, Cannabis is the centre of many scientific studies, which mainly focus on its chemical composition and medicinal properties. Unfortunately, while new applications of this plant are continuously being developed, some of its traditional uses are becoming rare and even disappearing altogether. Information on traditional uses of Cannabis is vast, but it is scattered across many publication sources in different formats, so synthesis and standardization of these data are increasingly important. The CANNUSE database provides an organized information source for scientists and general public interested in different aspects of Cannabis use. It contains over 2300 entries from 649 publications related to medicinal, alimentary, fibre and other uses from different geographical areas and cultures around the world. We believe this database will serve as a starting point for new research and development strategies based on the traditional knowledge. Database URL: http://cannusedb.csic.es.

Phenolic Compounds Content and Genetic Diversity at Population Level across the Natural Distribution Range of Bearberry (Arctostaphylos uva-ursi, Ericaceae) in the Iberian Peninsula.

Bearberry (Arctostaphylos uva-ursi) is a medicinal plant traditionally employed for the treatment of urinary tract infections due to high contents of arbutin (hydroquinone ß-D-glucoside), which is now mainly used as a natural skin-whitening agent in cosmetics. Bearberry has also been proposed as a natural antioxidant additive due to the high contents of phenolic compounds in leaves. We studied the variation on phenolic compounds in 42 wild populations of bearberry, aiming to elucidate if intrinsic biological, climatic, and/or geographic factors affect phenolic contents across its natural distribution in the Iberian Peninsula. Bearberry leaves were collected during autumn over a three-year period (2014-2016) in populations across a latitude and altitude gradient. Methanolic extracts showed a wide range of variation in total phenols content, and different phenolic profiles regarding arbutin (levels of this major constituent varied from 87 to 232 mg/g dr wt), but also catechin and myricetin contents, which were affected by geographic and climatic factors. Moderate levels of variation on genome size-assessed by flow cytometry-and on two plastid DNA regions were also detected among populations. Genetic and cytogenetic differentiation of populations was weakly but significantly associated to phytochemical diversity. Elite bearberry genotypes with higher antioxidant capacity were subsequently identified.

A phylogenetic road map to antimalarial Artemisia species.

ETHNOPHARMACOLOGICAL RELEVANCE: The discovery of the antimalarial agent artemisinin is considered one of the most significant success stories of ethnopharmacological research in recent times. The isolation of artemisinin was inspired by the use of Artemisia annua in traditional Chinese medicine (TCM) and was awarded a Nobel Prize in 2015. Antimalarial activity has since been demonstrated for a range of other Artemisia species, suggesting that the genus could provide alternative sources of antimalarial treatments. Given the stunning diversity of the genus (c. 500 species), a prioritisation of taxa to be investigated for their likely antimalarial properties is required. MATERIALS AND METHODS: Here we use a phylogenetic approach to explore the potential for identifying species more likely to possess antimalarial properties. Ethnobotanical data from literature reports is recorded for 117 species. Subsequent phylogenetically informed analysis was used to identify lineages in which there is an overrepresentation of species used to treat malarial symptoms, and which could therefore be high priority for further investigation of antimalarial activity. RESULTS: We show that these lineages indeed include several species with documented antimalarial activity. To further inform our approach, we use LC-MS/MS analysis to explore artemisinin content in fifteen species from both highlighted and not highlighted lineages. We detected artemisinin in nine species, in eight of them for the first time, doubling the number of Artemisia taxa known to content this molecule. CONCLUSIONS: Our findings indicate that artemisinin may be widespread across the genus, providing an accessible local resource outside the distribution area of Artemisia annua.