Identification of the relationship between Chinese Adiantum reniforme var. sinense and Canary Adiantum reniforme.

BACKGROUND: There are different opinions about the relationship of two disjunctively distributed varieties Adiantum reniforme L. var. sinense Y.X.Lin and Adiantum reniforme L. Adiantum reniforme var. sinense is an endangered fern only distributed in a narrowed region of Chongqing city in China, while Adiantum reniforme var. reniforme just distributed in Canary Islands and Madeira off the north-western African coast. To verify the relationship of these two taxa, relative phylogenetic analyses, karyotype analyses, microscopic spore observations and morphological studies were performed in this study. Besides, divergence time between A. reniforme var. sinense and A. reniforme var. reniforme was estimated using GTR model according to a phylogeny tree constructed with the three cpDNA markers atpA, atpB, and rbcL. RESULTS: Phylogenetic results and divergence time analyses--all individuals of A. reniforme var. sinense from 4 different populations (representing all biogeographic distributions) were clustered into one clade and all individuals of A. reniforme var. reniforme from 7 different populations (all biogeographic distributions are included) were clustered into another clade. The divergence between A. reniforme var. reniforme and A. reniforme var. sinense was estimated to be 4.94 (2.26-8.66) Myr. Based on karyotype analyses, A. reniforme var. reniforme was deduced to be hexaploidy with 2n = 180, X = 30, while A. reniforme var. sinense was known as tetraploidy. Microscopic spore observations suggested that surface ornamentation of A. reniforme var. reniforme is psilate, but that of A. reniforme var. sinense is rugate. Leaf blades of A. reniforme var. sinense are membranous and reniform and with several obvious concentric rings, and leaves of A. reniforme var. reniforme are pachyphyllous and coriaceous and are much rounder and similar to palm. CONCLUSION: Adiantum reniforme var. sinense is an independent species rather than the variety of Adiantum reniforme var. reniforme. As a result, we approve Adiantum nelumboides X. C. Zhang, nom. & stat. nov. as a legal name instead of the former Adiantum reniforme var. sinense. China was determined to be the most probable evolution centre based on the results of phylogenetic analyses, divergence estimation, relative palaeogeography and palaeoclimate materials.

A new phylogenetic analysis sheds new light on the relationships in the Calanthe alliance (Orchidaceae) in China.

The taxonomy of the Calanthe alliance (Epidendroideae, Orchidaceae), consisting of Calanthe, Cephalantheropsis, and Phaius, has been difficult for orchidologists to understand because of the presence of common morphological features. In this study, in addition to morphological and geographical analyses, maximum parsimony and Bayesian inference analyses were performed based on nucleotide sequences of the nuclear internal transcribed spacer and cpDNA genes of 88 taxa representing the major clades of the Calanthe alliance in China. The results indicated that Cephalantheropsis is monophyletic, while both Phaius and Calanthe are polyphyletic. In Phaius, a total of three species, P. flavus, P. columnaris, and P. takeoi, were segregated to form a new genus, Paraphaius. In Calanthe, subgenus Preptanthe and sect. Styloglossum were both categorized as distinct genera from Calanthe. Our results also confirm that Calanthe delavayi and C. calanthoides are members of Calanthe. Previous studies assigned C. delavayi to Phaius and C. calanthoides to Ghiesbrechtia. Five sections, namely, Alpinocalanthe, Puberula, Ghiesbrechtia, Tricarinata, and Calanthe, three of which are new taxa, were recognized in Calanthe. Therefore, we propose that the Calanthe alliance is composed of six genera: Calanthe, Cephalantheropsis, Paraphaius, Phaius, Preptanthe and Styloglossum.

Microsatellite markers for Kleinia neriifolia, an endemic Asteraceae species on the Canary Islands.

PREMISE OF THE STUDY: Microsatellite makers were developed in Kleinia neriifolia, an endemic Asteraceae species on the Canary Islands, for investigation of the population genetic structure and colonization history of this insular species. • METHODS AND RESULTS: Seventeen polymorphic and seven monomorphic microsatellite loci were isolated and characterized from K. neriifolia using the protocol of Fast Isolation by AFLP of Sequences COntaining repeats (FIASCO). At the population level, the number of alleles detected per locus varied from two to 16, and the observed and expected heterozygosities were 0.105-1.000 and 0.100-0.923, respectively. • CONCLUSIONS: These highly informative loci are potentially useful to obtain a deeper understanding of the evolutionary process of K. neriifolia.

Development of microsatellite markers for the endangered medicinal plant Launaea arborescens (Asteraceae).

PREMISE OF THE STUDY: Microsatellite loci were developed in Launaea arborescens, an endangered and medicinal Asteraceae species in North Africa, for further investigation of its conservation genetics. • METHODS AND RESULTS: We isolated and characterized 10 polymorphic and nine monomorphic microsatellite loci from L. arborescens using the protocol of Fast Isolation by AFLP of Sequences COntaining repeats (FIASCO). For the 10 polymorphic loci, the number of alleles detected per locus varied from two to six, and the observed and expected heterozygosities ranged from 0.000 to 0.833 and 0.059 to 0.713, respectively. • CONCLUSIONS: The polymorphic markers provide a useful tool for conservation genetics studies of L. arborescens, including analysis of mating system, estimating gene flow, and identifying discrete genetic units within the species.

A revised classification of Chinese Davalliaceae based on new evidence from molecular phylogenetics and morphological characteristics.

Although the phylogenetic framework of Davalliaceae is known, the classification of Chinese Davalliaceae is still controversial. In this study, a molecular phylogenetic tree of 60 accessions, including 29 species produced in China, was constructed using five plastid DNA markers-atpB, atpB-rbcL, rbcL, rbcL-accD, and accD. New data on studied specimens, field investigations, and scanning electron microscopy analysis of leaf epidermis and spores were used to reclassify Chinese Davalliaceae. The taxonomic position of Davallia canariensis was confirmed based on new evidence and a new key to sections of Chinese Davalliaceae was proposed. The taxonomically controversial genus Paradavallodes was confirmed as a polyphyletic group, and it was assigned to Davallia sect. Trogostolon and Davallia sect. Davallodes. Further, species endemic to China were delimited, 21 species were admitted to six sections of Davallia, two new combinations were proposed, two new synonyms were defined and a new key to Chinese species of Davalliaceae was presented.

Ethnobotanical survey of herbal tea plants from the traditional markets in Chaoshan, China.

ETHNOPHARMACOLOGICAL RELEVANCE: Herbal tea, which refers to "cooling tea", "cool beverage", or "liáng chá" in China, includes a range of drinks with heat-clearing and detoxification qualities. Herbal tea plants are great contributive to the health and prosperity of Chaoshan people. The aim of the study was to document herbal tea plant species used and commercialized as "liáng chá" in Chaoshan area, to facilitate the use and development of herbal tea enterprises, and to promote the further development of national herbal tea. MATERIALS AND METHODS: Information and data were obtained from all 83 stall holders in 12 traditional markets, semi-structured informant interviews were carried out individually with the stall holders, 10 questions were asked. RESULTS: In this study, 186 species of herbal tea plants belonging to 65 families and 156 genera were indicated by 83 stall holders, with Asteraceae being the most prevalent family with 22 species. Herbs are main sources of herbal tea plants in Chaoshan area, with whole plants (97 species) being the most used parts. Herbal drinks are mostly consumed for heat-clearing and detoxification, and a large number of plant species were reported to treat coughs, colds, dysentery, dampness and sore throats. The most cited species were Hedyotis corymbosa (L.) Lam. (47 times mentioned), Hedyotis diffusa Willd. (46), Plantago asiatica L. (43), Houttuynia cordata Thunb (42), Centella asiatica (L.) Urban (36), Desmodium styracifolium (Osbeck) Merr. (35) and Morus alba L. (31), and 5 protected species were recorded in the list of the nationally protected species of China: Dendrobium officinale Kimura et Migo, Dendrobium nobile Lindl., Anoectochilus formosanus Hayata, Bulbophyllum odoratissimum (J. E. Smith) Lindl. and Pholidota chinensis Lindl. The selling price of most fresh herbal tea plants in the market varied fromï¿¥10-16/kg, with the profit margin of sales ranging from 12.5% to 20%. The consumption of herbal tea for one family costs about ï¿¥3-5/day. CONCLUSION: Chaoshan herbal teas, prepared by diverse plant species, are regarded as a long tradition inseparable part in daily life of local people. Although they can effectively prevent and treat different kinds of diseases, they are not suitable for everyone (e.g. the elderly, children, and pregnant or breastfeeding women), even in healthy people, improper use of herbal tea can lead to physical discomfort. Furthermore, in order to appropriately understand the function, efficacy, and safety of herbal tea plants, additional research of traditional practices and phytochemistry, nutrient, physiological and toxicity properties should be analyzed.

Revision of series Gravesiana (Adiantum L.) based on morphological characteristics, spores and phylogenetic analyses.

Since the adoption of some ambiguous and quantitative characters in Flora Republicae Popularis Sinicae 3(1), species identifications of the series Gravesiana have been in disarray, requiring clarification. Two hundred and fifty-nine individuals from 47 different populations were collected for the estimation of morphological characters and phylogenetic analyses. Spores of 26 populations were observed through scanning electron microscope. Our results were different from those of previous research: (1) six identifiable species, rather than five species observed previously, were confirmed in the series Gravesiana, they are A. gravesii, A. juxtapositum, A. mariesii, A. dentatum, A. longzhouensis and A. obovatum, of which the latter three are newly recognized species. (2) Thirteen characters were measured and estimated through the program Mesquite v. 2.71. The character whether the pinna stalks were 1/3-1/2 times longer than the pinna was used to distinguish A. gravesii and A. lianxianense previously and was found to be unreliable here, whereas such characters as the height of the plant (H), pinna aligned forms (FP), number of pinna (NP), pinna margin (M), number of veins flabellate at base (NV), sori number and shape per pinna (NSS), pinna texture (T), and powder-covered or not on the abaxial surface of the pinna (P) are estimated to be stable and reliable characters useful for identification. Descriptions of new species and their retrieve keys are also listed. (3) Surface ornamentations and spore sizes are helpful for us to distinguish species in series Gravesiana.

Ethnobotanical study on medicinal plants used by local Hoklos people on Hainan Island, China.

AIMS OF THE STUDY: The aim of this study is to collect information on the use of medicinal plants by the local Hoklos people on Hainan Island, and compare medicinal traditions in the study area with Li medicines (LM) and traditional Chinese medicines (TCM). MATERIALS AND METHODS: Ethnobotanical data were collected by 3 means: semi-structured interviews, personal conversation and guided field trips. There were 27 informants from 9 towns and 14 villages who were visited. Quantitative indices (Informant Consensus Factor - FIC, Use Value - UV, and Fidelity Level - FL) were calculated. RESULTS: In the present study, 264 species from 92 families and 233 genera were recorded, with Compositae (20 species), Leguminosae (19 species), Rubiaceae (12 species) and Gramineae (11 species) as predominate families. Leaves were the most frequently used parts in the preparation of local medicines. The most common preparation method was decoction (452 mentions). The plant with the highest values was Eclipta prostrata (0.46). The 6 plant species with the maximum FL (100%) were Atalantia buxifolia (Poir.) Oliv., Garcinia oblongifolia Champ. ex Benth., Hypericum japonicum Thunb. ex Murray, Imperata cylindrica (L.) Beauv., Microcos paniculata L., and Psidium guajava L. In addition, 120 investigated human ailments were grouped into 10 categories, within which symptoms and signs (184 mentions), diseases of the skin and subcutaneous tissue (139 mentions) and diseases of the digestive system (94 mentions) were the most mentioned in our investigation. The informant consensus about using medicinal plants ranged from 0.27 to 0.48, which showed a high level of agreement among the informants on symptoms and signs (0.48) and certain infectious and parasitic diseases (0.43). In comparison with TCM and LM, the results reflected a closer connection between local medicine and TCM. CONCLUSION: The information reported by Hoklos people is of great value to ethnic medicinal culture. However, this precious medicinal knowledge is at risk of being lost due to rapid degradation of the environment. It is essential that more people engage in in-depth studies on local medicinal plants and relevant organizations address this serious problem before the damage is irreversible.

Re-establishment of the genus Ania Lindl. (Orchidaceae).

Ania Lindl. is a small genus of the tribe Collabieae subtribe Collabiinae (Orchidaceae). For the last 150 years, it has generally been treated as a synonym of Tainia Blume. In this study, we critically re-examined morphological characters that have been used to distinguish Ania from Tainia, and assessed the phylogeny of Tainia using morphological and palynological characters. Sequences of the nuclear ribosomal ITS, chloroplast trnL intron and combined DNA data sets were analysed to clarify the delimitation and the phylogeny of these groups. The morphological and palynological survey revealed a number of useful diagnostic characters which permit a clear definition of Ania, after the exclusion of a single taxonomically questionable species. Results confirmed that Ania is distinct from Tainia. Phylogenetic reconstructions based on molecular data provided the greatest resolution and produced a morphologically well differentiated clade of Ania. In addition to morphological and suggested palynological characters, the phylogenies were also supported by karyological evidence. Our results support the independent generic status of Ania. The genus name Ania is revived and re-established.

A new orchid genus, Danxiaorchis, and phylogenetic analysis of the tribe Calypsoeae.

BACKGROUND: Orchids have numerous species, and their speciation rates are presumed to be exceptionally high, suggesting that orchids are continuously and actively evolving. The wide diversity of orchids has attracted the interest of evolutionary biologists. In this study, a new orchid was discovered on Danxia Mountain in Guangdong, China. However, the phylogenetic clarification of this new orchid requires further molecular, morphological, and phytogeographic analyses. METHODOLOGY/PRINCIPAL FINDINGS: A new orchid possesses a labellum with a large Y-shaped callus and two sacs at the base, and cylindrical, fleshy seeds, which make it distinct from all known orchid genera. Phylogenetic methods were applied to a matrix of morphological and molecular characters based on the fragments of the nuclear internal transcribed spacer, chloroplast matK, and rbcL genes of Orchidaceae (74 genera) and Calypsoeae (13 genera). The strict consensus Bayesian inference phylogram strongly supports the division of the Calypsoeae alliance (not including Dactylostalix and Ephippianthus) into seven clades with 11 genera. The sequence data of each species and the morphological characters of each genus were combined into a single dataset. The inferred Bayesian phylogram supports the division of the 13 genera of Calypsoeae into four clades with 13 subclades (genera). Based on the results of our phylogenetic analyses, Calypsoeae, under which the new orchid is classified, represents an independent lineage in the Epidendroideae subfamily. CONCLUSIONS: Analyses of the combined datasets using Bayesian methods revealed strong evidence that Calypsoeae is a monophyletic tribe consisting of eight well-supported clades with 13 subclades (genera), which are all in agreement with the phytogeography of Calypsoeae. The Danxia orchid represents an independent lineage under the tribe Calypsoeae of the subfamily Epidendroideae. This lineage should be treated as a new genus, which we have named Danxiaorchis, that is parallel to Yoania. Both genera are placed under the subtribe Yoaniinae.

Ethnobotanical study on medicinal plants around Mt.Yinggeling, Hainan Island, China.

AIM OF THE STUDY: The main objectives were to collect information on the use of medicinal plants and compare medicinal plant traditions between Run and Qi. MATERIALS AND METHODS: Information was obtained from semi-structured interviews, personal conversation and guided fieldtrips with herbalists. RESULTS: 385 species belonging to 290 genera in 104 families were used for the treatment of various diseases. Rubiaceae (20 species), Euphorbiaceae and Compositae (19 species respectively) were predominant families used by herbalists. The most species were used for injuries (20.1%), muscular-skeletal system disorders (18.3%) and infections/infestations (18.0%). The coefficient of similarity (36.6%) shown a high consensus of plant species used by Run and Qi. The 'informant agreement ratio' values for both Run and Qi are rather low (less than 0.3). CONCLUSIONS: Traditional medicinal plants still play an important role in medical practices of Li Ethnic Group around Mt.Yinggeling. There is a close relationship of medicinal plant traditions between Run and Qi. Further investigation is necessary to record this valuable knowledge before its disappearance.