Study of anti-diabetic, beta-carotene-bleaching inhibiting and iron chelating properties of Carissa opaca root extracts

Abstract Degenerative diseases diabetes and oxidative stress constitute a major health concern worldwide. Medicinal plants are expected to provide effective and affordable remedies. The present research explored antidiabetic and antioxidant potential of extracts of Carissa opaca roots. Methanolic extract (ME) was prepared through maceration. Its fractions were obtained, sequentially, in hexane, chloroform, ethyl acetate and n-butanol. An aqueous decoction (AD) of the finely ground roots was obtained by boiling in distilled water. The leftover biomass with methanol was boiled in water to obtain biomass aqueous decoction (BAD). The extracts and fractions showed considerable porcine pancreatic α-amylase inhibitory activity with IC50 in the range of 5.38-7.12 mg/mL while acarbose had 0.31 mg/mL. The iron chelating activity in terms of EC50 was 0.2939, 0.3429, 0.1876, and 0.1099 mg/mL for AD, BAD, ME, and EDTA, respectively. The EC50 of beta-carotene bleaching activity for AD, BAD, ME, and standard BHA were 4.10, 4.71, 3.48, and 2.79 mg/mL, respectively. The total phenolic content (TPC) and total flavonoid content (TFC) of AD and BAD were also considerable. In general, ethyl acetate fraction proved to be the most potent. Thus, the C. opaca roots had excellent antioxidant activity while having moderate α-amylase inhibitory potentia

Decalepis salicifolia (Bedd. ex Hook. f.) Venter: A steno-endemic and critically endangered medicinal and aromatic plant from Western Ghats, India.

Decalepis salicifolia (Bedd. ex Hook. f.) Venter is a potential medicinal and highly aromatic plant species confined to the southernmost part of the Western Ghats of India. The plant is well known for its traditional uses among the various tribal communities of south India. The tubers of the plant possess characteristic vanillin-like aroma due to the presence of the compound 2-hydroxy-4-methoxybenzaldehyde. The tubers are used to substitute Hemidesmus indicus in various herbal formulations. The plants in the wild are continuously uprooted for their roots, leading to the irreversible destruction of the whole plant. The resulting tremendous loss of populations in the wild led to the species being declared as critically endangered by IUCN. Our group is working on the various aspects of this species including population status, distribution mapping, prospection, and conservation management. In the present review, we have brought out the available information till date on D. salicifolia, including taxonomy, ethno-medicinal uses, phytochemistry, pharmacology, population status, and conservation efforts along with research gap and lacunae to provide direction for further research into this less explored medicinal and aromatic plant.

Chemical Diversity and Bioactivities of Monoterpene Indole Alkaloids (MIAs) from Six Apocynaceae Genera.

By the end of the twentieth century, the interest in natural compounds as probable sources of drugs has declined and was replaced by other strategies such as molecular target-based drug discovery. However, in the recent times, natural compounds regained their position as extremely important source drug leads. Indole-containing compounds are under clinical use which includes vinblastine and vincristine (anticancer), atevirdine (anti-HIV), yohimbine (erectile dysfunction), reserpine (antihypertension), ajmalicine (vascular disorders), ajmaline (anti-arrhythmic), vincamine (vasodilator), etc. Monoterpene Indole Alkaloids (MIAs) deserve the curiosity and attention of researchers due to their chemical diversity and biological activities. These compounds were considered as an impending source of drug-lead. In this review 444 compounds, were identified from six genera belonging to the family Apocynaceae, will be discussed. These genera (Alstonia, Rauvolfia, Kopsia, Ervatamia, and Tabernaemontana, and Rhazya) consist of 400 members and represent 20% of Apocynaceae species. Only 30 (7.5%) species were investigated, whereas the rest are promising to be investigated. Eleven bioactivities, including antibacterial, antifungal, anti-inflammatory and immunosuppressant activities, were reported. Whereas cytotoxic effect represents 47% of the reported activities. Convincingly, the genera selected in this review are a wealthy source for future anticancer drug lead.

Identification of medicinal plants within the Apocynaceae family using ITS2 and psbA-trnH barcodes.

To ensure the safety of medications, it is vital to accurately authenticate species of the Apocynaceae family, which is rich in poisonous medicinal plants. We identified Apocynaceae species by using nuclear internal transcribed spacer 2 (ITS2) and psbA-trnH based on experimental data. The identification ability of ITS2 and psbA-trnH was assessed using specific genetic divergence, BLAST1, and neighbor-joining trees. For DNA barcoding, ITS2 and psbA-trnH regions of 122 plant samples of 31 species from 19 genera in the Apocynaceae family were amplified. The PCR amplification for ITS2 and psbA-trnH sequences was 100%. The sequencing success rates for ITS2 and psbA-trnH sequences were 81% and 61%, respectively. Additional data involved 53 sequences of the ITS2 region and 38 sequences of the psbA-trnH region were downloaded from GenBank. Moreover, the analysis showed that the inter-specific divergence of Apocynaceae species was greater than its intra-specific variations. The results indicated that, using the BLAST1 method, ITS2 showed a high identification efficiency of 97% and 100% of the samples at the species and genus levels, respectively, via BLAST1, and psbA-trnH successfully identified 95% and 100% of the samples at the species and genus levels, respectively. The barcode combination of ITS2/psbA-trnH successfully identified 98% and 100% of samples at the species and genus levels, respectively. Subsequently, the neighbor joining tree method also showed that barcode ITS2 and psbA-trnH could distinguish among the species within the Apocynaceae family. ITS2 is a core barcode and psbA-trnH is a supplementary barcode for identifying species in the Apocynaceae family. These results will help to improve DNA barcoding reference databases for herbal drugs and other herbal raw materials.

Hancornia speciosa serum fraction latex stimulates the angiogenesis and extracellular matrix remodeling processes.

The Hancornia speciosa latex reveals angiogenic, osteogenic, and anti-inflammatory properties, which present its potential for developing of wound healing drugs; however, the latex compounds responsible for angiogenesis remain unknown. One strategy to screen these active compounds is evaluation of latex fractions. This study aimed to obtain different fractions of latex and evaluate its angiogenic activity separately using the chick chorioallantoic membrane (CAM) assay. The serum (SE) fraction was responsible for angiogenesis, which was subject to biochemical characterization and computational simulations in order to understand the contribution of H. speciosa latex in wound healing process. Our results revealed weak antioxidant potential and absence of antimicrobial activity in the SE fraction. Phytochemical analysis identified chlorogenic acids (CGA) as the main compound of SE fraction. CGA bioactivity predictions identify different molecules associated with extracellular matrix (ECM) remodeling, such as metalloproteinases, which also are overexpressed in our CAM assay experiment. Docking simulations revealed the interactions between CGA and matrix metalloproteinase 2. In conclusion, SE latex fraction stimulates angiogenesis and may influence ECM remodeling. These properties may contribute to the wound healing process, and also confirm the widespread use of this plant.

Microscopic investigations and pharmacognostic techniques for the standardization of Caralluma edulis (Edgew.) Benth. ex Hook.f.

Herbal medicines frequently suffer with quality controversies because of similar species or varieties. This often leads to sophistication or admixture of the crude drug as they share various look alike physical features. Commercially, stalks of Caralluma edulis are commonly adulterated with other species because of their analogous morphology. In the present research, the microscopic standardization of the drug was performed by structural, histological, and anatomical features. Pharmacognostical characterization was carried out by physicochemical (moisture content, swelling index, pH value, ash values, and extractive values) and fluorescence analysis. The chemical characterization by UV-Visible, Fourier transform infrared spectroscopy, and high-performance liquid chromatography was also performed that disclosed the chief phytoconstituents of the herbal drug. The above mentioned constraints, being reported for the first time in C. edulis, are substantial in establishing the microscopic and pharmacopoeial standards for future identification and authentication.

Ethnopharmacology, phytochemistry, and biotechnological advances of family Apocynaceae: A review.

The family Apocynaceae is one of the largest and important families in angiosperm. Several members of this family have medicinal properties and have been in the treatment of various ailments. Most of them are consumed as food by tribal people whereas a few plants are used as source of poison. Members of family Apocynaceae are rich in alkaloids, terpenoids, steroids, flavonoids, glycosides, simple phenols, lactones, and hydrocarbons. Other compounds such as sterols, lignans, sugars, lignans, and lactones have been isolated and systematically studied. Few studies have reported antioxidant, anti-inflammatory, antimicrobial, and cytotoxic activities of crude extracts as well as single compound(s) isolated from various members of the family Apocynaceae. Holarrhena antidysenterica, Rauvolfia serpentina, Carissa carandas, and Tabernaemontana divaricata are the extensively studied plants in this family. The present review provides a detailed outlook on ethnopharmacology, phytochemistry, and biological activities of selected members of this family. Moreover, it also covers the biotechnological advances used for large-scale production of bioactive compounds of therapeutic interest along with plant tissue culture-based approaches for conservation of this medicinally valuable family.

Leptadenia reticulata (Retz.) Wight & Arn. (Jivanti): Botanical, Agronomical, Phytochemical, Pharmacological, and Biotechnological Aspects.

Leptadenia reticulata (Retz.) Wight & Arn. (Apocynaceae), is a traditional medicinal plant species widely used to treat various ailments such as tuberculosis, hematopoiesis, emaciation, cough, dyspnea, fever, burning sensation, night blindness, cancer, and dysentery. In Ayurveda, it is known for its revitalizing, rejuvenating, and lactogenic properties. This plant is one of the major ingredients in many commercial herbal formulations, including Speman, Envirocare, Calshakti, Antisept, and Chyawanprash. The therapeutic potential of this herb is because of the presence of diverse bioactive compounds such as α-amyrin, ß-amyrin, ferulic acid, luteolin, diosmetin, rutin, ß-sitosterol, stigmasterol, hentricontanol, a triterpene alcohol simiarenol, apigenin, reticulin, deniculatin, and leptaculatin. However, most biological studies on L. reticulata are restricted to crude extracts, and many biologically active compounds are yet to be identified in order to base the traditional uses of L. reticulata on evidence-based data. At present, L. reticulata is a threatened endangered plant because of overexploitation, unscientific harvesting, and habitat loss. The increased demand from pharmaceutical, nutraceutical, and veterinary industries has prompted its large-scale propagation. However, its commercial cultivation is hampered because of the non-availability of genuine planting material and the lack of knowledge about its agronomical practices. In this regard, micropropagation techniques will be useful to obtain true-to-type L. reticulata planting materials from an elite germplasm to meet the current demand. Adopting other biotechnological approaches such as synthetic seed technology, cryopreservation, cell culture, and genetic transformation can help conservation as well as increased metabolite production from L. reticulata. The present review summarizes scientific information on the botanical, agronomical, phytochemical, pharmacological, and biotechnological aspects of L. reticulata. This comprehensive information will certainly allow better utilization of this industrially important herb towards the discovery of lead drug molecules.

Apocynaceae species with antiproliferative and/or antiplasmodial properties: a review of ten genera.

Apocynaceae is a large family of tropical trees, shrubs and vines with most species producing white latex. Major metabolites of species are triterpenoids, iridoids, alkaloids and cardenolides, which are known for a wide range of biological and pharmacological activities such as cardioprotective, hepatoprotective, neuroprotective, anti-inflammatory, anticancer and antimalarial properties. Prompted by their anticancer and antimalarial properties, the current knowledge on ten genera (Allamanda, Alstonia, Calotropis, Catharanthus, Cerbera, Dyera, Kopsia, Nerium, Plumeria and Vallaris) is updated. Major classes of metabolites are described using some species as examples. Species with antiproliferative (APF) and/or antiplasmodial (APM) properties have been identified. With the exception of the genus Dyera, nine genera of 22 species possess APF activity. Seven genera (Alstonia, Calotropis, Catharanthus, Dyera, Kopsia, Plumeria and Vallaris) of 13 species have APM properties. Among these species, Alstonia angustiloba, Alstonia macrophylla, Calotropis gigantea, Calotropis procera, Catharanthus roseus, Plumeria alba and Vallaris glabra displayed both APF and APM properties. The chemical constituents of these seven species are compiled for assessment and further research.

Antinociceptive and anti-inflammatory effects of a Geissospermum vellosii stem bark fraction.

Geissospermum vellosii (Pao pereira) is a Brazilian tree whose stem barks are rich in indole alkaloids that present intense anticholinesterase activity. The present study evaluated the effects of a stem bark fraction (PPAC fraction) and ethanolic extract (EE) of Pao pereira in classic murine models of inflammation and pain. The EE and PPAC fraction, both at a dose of 30 mg/kg, significantly reduced mice abdominal constriction induced by acetic acid by 34.8% and 47.5%, respectively. In the formalin test, EE (30 mg/kg) and PPAC fraction (30 and 60 mg/kg) inhibited only the second phase, by 82.8%, 84.9% and 100%, respectively. Compared with indomethacin, similar doses of EE or PPAC fraction were approximately twice as effective in causing antinociception. PPAC fraction was not effective in the hot plate test but reduced the inflammatory response at the second (50.6%) and third (57.8%) hours of rat paw edema induced by carrageenan. Antihyperalgesic activity was observed within 30 min with a peak at 2 h (60.1%). These results demonstrate that compounds in PPAC fraction have anti-inflammatory and antinociceptive activity by a mechanism apparently unrelated to the opioid system. Regardless of similar responses to indomethacin, the effects of PPAC fraction are mainly attributed to acetylcholine actions.

Uzara - A quality control perspective of Xysmalobium undulatum.

CONTEXT: Xysmalobium undulatum (L.) Aiton f var. (Asclepiadaceae), commonly known as uzara, is an ethnomedicinally important plant from southern Africa used to treat a variety of ailments. In addition to local use in African Traditional Medicine (ATM), formulations containing uzara have been successfully marketed by a number of pharmaceutical companies. Despite its commercialization, published adequate quality control (QC) protocols are lacking. OBJECTIVE: The study was conducted to develop QC protocols for uzara based on chromatographic and spectroscopic techniques. MATERIALS AND METHODS: High performance thin layer chromatography (HPTLC) and liquid chromatography coupled to mass spectrometry (LC-MS) were used to develop phytochemical fingerprints of ethanolic root extracts of 47 uzara samples collected from eight distinct localities in South Africa. Mid-infrared (MIR) spectroscopy was also explored as a suitable alternative technique for rapid and economic quantification of uzarin. RESULTS: Adequate chromatographic profiles were obtained using both HPTLC and LC-MS analyses. The chromatographic patterns showed qualitative similarities among plants collected from different locations. The levels of uzarin, the major constituent of uzara, were highly variable between locations, ranging from 17.8 to 139.9 mg/g (dry weight). A good coefficient of determination (R(2 )= 0.939) and low root mean square error of prediction (RMSEP = 7.9 mg/g) confirmed the accuracy of using MIR-PLS calibration models for the quantification of uzarin. DISCUSSION AND CONCLUSION: Both HPTLC and LC-MS can be used as tools in developing quality control procedures for uzara. MIR in combination with chemometrics provides a fast alternative method for the quantification of uzarin.

The complete chloroplast genome sequence of Cynanchum auriculatum Royle ex Wight (Apocynaceae).

Cynanchum auriculatum is a climbing vine belonging to the Apocynaceae family and shows very similar morphology to Cynanchum wilfordii, a medicinal plant. The complete chloroplast genome of C. auriculatum was generated by de novo assembly using the small amount of whole genome sequencing data. The chloroplast genome of C. auriculatum was 160 840 bp in length and consisted of four distinct regions, such as large single copy region (91 973 bp), small single copy region (19 667 bp), and a pair of inverted repeat regions (24 600 bp). The overall GC contents of the chloroplast genome were 37.8%. A total of 114 genes were predicted and included 80 protein-coding genes, 30 tRNA genes, and four rRNA genes. Phylogenetic analysis with the reported chloroplast genomes revealed that C. auriculatum is most closely related to Cynanchum wilfordii, a medicinal plant.

Estudo etnofarmacológico e etnobotânico de Himatanthus drasticus (Mart. ) Plumel (janaguba) / Ethnopharmacological and ethnobotanical study of Himatanthus drasticus (Mart. ) Plumel (janaguba).

RESUMO Himatanthus drasticus (Mart.) Plumel, janaguba, produz um látex em seu caule que é usado para o tratamento de alguns tipos de câncer, vermes intestinais, febre, menstruações irregulares, infertilidade feminina e úlceras gástricas. Estudos fitoquímicos revelaram a presença de acetato de lupeol e proteínas; e estudos farmacológicos, os efeitos antiúlcera, antitumoral, imunomodulador, analgésico e anti-inflamatório. O presente trabalho objetivou realizar levantamento etnobotânico e etnofarmacológico sobre H. drasticus como contribuição ao conhecimento científico da referida espécie. Foi elaborado um questionário, que foi aplicado em entrevista a 21 raizeiros nos mercados das cidades de Crato, Fortaleza e Pacajus do estado do Ceará (CE). Observou-se que 38,1% dos raizeiros têm mais de 10 anos de serviço e 47,6%, estão na faixa etária de 40 a 59 anos, o que representa um fator positivo quanto à experiência com plantas medicinais. Aproximadamente 85,7% dos raizeiros relataram o uso do látex da H. drasticus para tratamento de úlcera; 66,7% para inflamação; 66,7% para câncer. Verificou-se que as principais indicações terapêuticas relatadas pelos raizeiros sobre esta espécie estão de acordo com as informações científicas. Os levantamentos etnobotânicos e etnofarmacológicos exercem um papel primordial no resgate do conhecimento tradicional, nos meios rurais e urbanos, tanto por seu valor histórico-cultural, como também pela necessidade de confirmação das indicações de uso.

ABSTRACT The Himatanthus drasticus (Mart.) Plumel, janaguba, produces a latex whose stem is used to treat some types of cancer, intestinal worms, fever, irregular menses, female infertility and gastric ulcers. Phytochemical studies revealed the presence of lupeol acetate and proteins in the latex; and pharmacological studies indicated antiulcer, anti-tumor, immunomodulating, analgesic and anti-inflammatory effects. This study aimed to conduct surveys on ethnopharmacological and ethnobotanical species H. drasticus as a contribution to the scientific knowledge about the species. A questionnaire and interviews were made with 21 healers in the markets of the cities of Crato, Fortaleza and Pacajús, in state of Ceara (CE). It was observed that 38.1% of the healers have more than 10 years of service, and 47.6% are between 40-59 years old, which is a positive factor about the experience with medicinal plants. Approximately 85.7% of the healers reported to apply latex H. drasticus for the treatment of ulcers, 66.7% for inflammation, and 66.7% for cancer. It was found that the main therapeutic indications mentioned by the healers about this species are consistent with the scientific information available. The ethnobotanical and ethnopharmacological surveys play an essential role on keeping the traditional knowledge in rural and urban areas, both for its historical and cultural value, but also for the need of confirmation of the intended use.

Extratos vegetais no controle da antracnose e na conservação da qualidade em frutos de mamoeiro / Plant extracts in controlling anthracnose and quality conservation in papayas fruit

RESUMO O presente trabalho teve como objetivo avaliar a ação antifúngica dos extratos de Allamanda blanchetti A.DC. e Momordica charantia L. no controle de Colletotrichumgloesporioides e qualidade pós-colheita em frutos de mamoeiro da cultivar Sunrise solo. Os experimentos foram conduzidos nos Laboratório de Fitopatologia e Laboratório de Produtos de Origem Animal, CCA-UFPB, Campus.II, Areia-PB. Os frutos foram colhidos no estádio 1 de maturação, em seguida, sanitizados com hipoclorito de sódio a 1% por três minutos e pulverizados com os extratos etanólicos de A. blanchetti e M. charantia nas concentrações: 0, 10, 100, 500 e 1000 µg.mL-1. Os frutos foram distribuídos em bandejas de polipropileno expandido em temperatura ambiente (23±2 °C) e umidade relativa em torno de 70%. Após doze dias de armazenamento, os frutos foram avaliados quanto à incidência natural de C. gloesporioides, severidade, atividade da enzima peroxidase, sólidos solúveis, acidez titulável, relação SS/AT, perda de massa e aparência externa dos frutos. Foi realizada análise de regressão polinomial testando modelos até o terceiro grau para as concentrações dos extratos utilizando análise de variância pelo teste F enquanto as médias foram comparadas pelo teste de Tukey a 5% de probabilidade, através do programa SAS® 9.2. Não se observou diferença estatística quando avaliou-se a incidência natural de C. gloesporioides em frutos de mamoeiro utilizando os extratos de A. blanchetti e M. charantia. Já na avaliação da severidade da antracnose, verificou-se diferença para o tratamento com o extrato de A. blanchetti, que apresentou as menores escalas de notas em relação ao tratamento com a M. charantia. Observou-se atividade da peroxidase (POD) em frutos de mamoeiro nos extratos de A. blanchetti e M. charantia. Os frutos tratados com A. blanchetti apresentaram teor de acidez, com máximo de 0,12 g de ácido cítrico.100 g-1 e no de M. charantiaapresentou aumento para 0,15 g de ácido cítrico.100 g-1. A relação SS/AT diminuiu com o aumento das concentrações dos dois extratos. Os frutos tratados com A. blanchetti apresentaram maior perda de massa durante todo o armazenamento, sendo maior que 10% aos nove dias. Já a perda de massa dos frutos tratados com M. charantia apresentou comportamento linear, sendo superior a 12% aos 12 dias de armazenamento. A cor evoluiu de 1 (verde) para 6 (casca alaranjada) e a aparência dos frutos diminuiu com o decorrer do tempo de 5 (excelente) para 3 (regular) em todas concentrações dos extratos de A. blanchetti e M. charantia. Os extratos de A. blanchetti e M. charantia não reduziram a incidência natural, porém, só o extrato de A. blanchetti foi responsável por reduzir a severidade da antracnose. Ocorreu a atividade da enzima peroxidase em todos os tratamentos avaliados e conservaram a qualidade pós-colheita em frutos até os nove dias de armazenamento.

ABSTRACT This study aimed to evaluate the antifungal effect of the extracts Allamandablanchetti A.DC. and Momordica charantia L. in the control of Colletotrichum gloesporioides in papaya fruits of the Sunrise Solo cultivation, as well as their post-harvest quality. The trials were conducted in the Laboratory of Plant Pathology and Animal Products Laboratory, CCA-UFPB, Campus II, Areia-PB. The fruits were harvested at maturity stage 1, then sanitized with sodium hypochlorite 1% for three minutes and sprayed with ethanol extracts of A. blanchetti and M.charantia at concentrations of 0, 10, 100, 500 and 1000 μg.mL-1.The fruits were distributed in polypropylene trays expanded at room temperature (23±2 °C) and relative humidity around 70%. After twelve days of storage, the fruits were evaluated for natural incidence of C. gloesporioides, severity, peroxidase enzyme activity, soluble solids, titratable acidity, SS/TA ratio, weight loss and external appearance of the fruit. Polynomial regression testing models to the third degree were employed for the extracts of concentrations, using analysis of variance by F test. The averages were compared by Tukey test at 5% probability, through the SAS® 9.2 program. There was no statistical difference observed when we evaluated the natural incidence of C. gloesporioides in the papaya fruit, using extracts from A. blanchetti and M. charantia. In the assessment of the severity of the anthracnose, there was no statistical difference for the treatment with the extract of A. blanchetti, which had smaller scales of notes when compared with the treatment using the M. charantia. It was observed peroxidase activity (POD) in the papaya fruits treated with the extracts of A. blanchetti and M. charantia. The fruits treated with A. blanchetti presented a maximum dose of acidity of 0.12 g g-1 of citric acid.100 g-1. In the fruits treated with M. charantia this increase was of 0.15g of citric acid.100 g-1. The SS/TA ratio decreased with increasing concentrations of the two extracts. The fruits treated with A. blanchettipresented higher weight loss of approximately 10%, since the M. charantia was over 12%. The color evolved from 1 (green) to 6 (Orange Peel) and the appearance of the fruit got worse according to the time, from 5 (excellent) to 3 (regular) in all the concentrations of the extracts of A. blanchetti and M. charantia. The A. blanchetti and M. charantia extracts did not reduce the natural incidence, however, only the A. blanchetti extract was able to decrease the severity of the anthracnose. The peroxidase enzyme presented activity in all treatments and kept the postharvest quality of the fruits in the nine days of storage.

[Identification of herbal tea ingredient Plumeria rubra and its adulterants using DNA barcoding].

ITS2 sequence was used as a barcode to identify herbal tea ingredient Plumeria rubra and its adulterants. Genomic DNAs from forty eight samples were extracted, the ITS2 sequences were amplified and sequenced bi-direstionlly, and then assembled and obtained using CodonCode Aligner. The sequences were aligned using ClustalW, the genetic distances were computed by kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic trees were constructed using MEGA5.0. Results showed that the length of ITS2 sequence of P. rubra were 244 bp. The intra-specific genetic distances (0-0. 016 6) were much smaller than inter-specific ones between P. rubra and its adulterants(0.320 8-0.650 4). The NJ tree indicated that P. rubra and its adulterants could be distinguished clearly. Therefore, Using ITS2 barcode can accurately andeffectively distinguish herbal tea ingredient P. rubra from its adulterants, which providesa new molecular method to identify P. rubra and ensure its safety in use.

Phylogenetic signal detection from an ancient rapid radiation: Effects of noise reduction, long-branch attraction, and model selection in crown clade Apocynaceae.

Crown clade Apocynaceae comprise seven primary lineages of lianas, shrubs, and herbs with a diversity of pollen aggregation morphologies including monads, tetrads, and pollinia, making them an ideal group for investigating the evolution and function of pollen packaging. Traditional molecular systematic approaches utilizing small amounts of sequence data have failed to resolve relationships along the spine of the crown clade, a likely ancient rapid radiation. The previous best estimate of the phylogeny was a five-way polytomy, leaving ambiguous the homology of aggregated pollen in two major lineages, the Periplocoideae, which possess pollen tetrads, and the milkweeds (Secamonoideae plus Asclepiadoideae), which possess pollinia. To assess whether greatly increased character sampling would resolve these relationships, a plastome sequence data matrix was assembled for 13 taxa of Apocynaceae, including nine newly generated complete plastomes, one partial new plastome, and three previously reported plastomes, collectively representing all primary crown clade lineages and outgroups. The effects of phylogenetic noise, long-branch attraction, and model selection (linked versus unlinked branch lengths among data partitions) were evaluated in a hypothesis-testing framework based on Shimodaira-Hasegawa tests. Discrimination among alternative crown clade resolutions was affected by all three factors. Exclusion of the noisiest alignment positions and topologies influenced by long-branch attraction resulted in a trichotomy along the spine of the crown clade consisting of Rhabdadenia+the Asian clade, Baisseeae+milkweeds, and Periplocoideae+the New World clade. Parsimony reconstruction on all optimal topologies after noise exclusion unambiguously supports parallel evolution of aggregated pollen in Periplocoideae (tetrads) and milkweeds (pollinia). Our phylogenomic approach has greatly advanced the resolution of one of the most perplexing radiations in Apocynaceae, providing the basis for study of convergent floral morphologies and their adaptive value.

Complete sequences of organelle genomes from the medicinal plant Rhazya stricta (Apocynaceae) and contrasting patterns of mitochondrial genome evolution across asterids.

BACKGROUND: Rhazya stricta is native to arid regions in South Asia and the Middle East and is used extensively in folk medicine to treat a wide range of diseases. In addition to generating genomic resources for this medicinally important plant, analyses of the complete plastid and mitochondrial genomes and a nuclear transcriptome from Rhazya provide insights into inter-compartmental transfers between genomes and the patterns of evolution among eight asterid mitochondrial genomes. RESULTS: The 154,841 bp plastid genome is highly conserved with gene content and order identical to the ancestral organization of angiosperms. The 548,608 bp mitochondrial genome exhibits a number of phenomena including the presence of recombinogenic repeats that generate a multipartite organization, transferred DNA from the plastid and nuclear genomes, and bidirectional DNA transfers between the mitochondrion and the nucleus. The mitochondrial genes sdh3 and rps14 have been transferred to the nucleus and have acquired targeting presequences. In the case of rps14, two copies are present in the nucleus; only one has a mitochondrial targeting presequence and may be functional. Phylogenetic analyses of both nuclear and mitochondrial copies of rps14 across angiosperms suggests Rhazya has experienced a single transfer of this gene to the nucleus, followed by a duplication event. Furthermore, the phylogenetic distribution of gene losses and the high level of sequence divergence in targeting presequences suggest multiple, independent transfers of both sdh3 and rps14 across asterids. Comparative analyses of mitochondrial genomes of eight sequenced asterids indicates a complicated evolutionary history in this large angiosperm clade with considerable diversity in genome organization and size, repeat, gene and intron content, and amount of foreign DNA from the plastid and nuclear genomes. CONCLUSIONS: Organelle genomes of Rhazya stricta provide valuable information for improving the understanding of mitochondrial genome evolution among angiosperms. The genomic data have enabled a rigorous examination of the gene transfer events. Rhazya is unique among the eight sequenced asterids in the types of events that have shaped the evolution of its mitochondrial genome. Furthermore, the organelle genomes of R. stricta provide valuable genomic resources for utilizing this important medicinal plant in biotechnology applications.

Prospecção fitoquímica de Himatanthus drasticus Plumel (Apocynaceae), da mesorregião leste maranhense / Phytochemical screening of Himatanthus drasticus Plumel (Apocynaceae), from the eastern mesoregion of Maranhão, Brazil

O uso indiscriminado de Himatanthus drasticus (Janaúba) por pequenos caprinocultores no controle de verminose em pequenos ruminantes foi o que motivou a realização do estudo fitoquímico do mesmo. Cascas da janaúba foram coletadas na mesorregião Leste do Maranhão e conduzidas aos laboratórios de Nutrição da Universidade Estadual do Maranhão e de Produtos Naturais da Universidade Federal do Maranhão para a identificação botânica e fitoquímica pela metodologia da Prospecção Preliminar e CCD, realizando testes para as diversas classes de metabólitos secundários. A partir das cascas do vegetal moído e desidratado foi realizado o preparo do extrato bruto (EB). O material foi colocado em uma mistura hidroalcoólica de EtOH: H2O (7:3 v:v), e submetido a agitação mecânica esporádica. Os Subextratos foram obtidos a partir do EBHA pelo processo de partição líquido-líquido, ETOH: H2O (2:1, v:v). As misturas foram preparadas com os seguintes solventes orgânicos de polaridades crescentes: hexano, acetato de etila e butanol. As análises cromatográficas evidenciaram a presença de grupos de metabólitos secundários no extrato e nos subextratos. As classes de metabólitos secundários que apresentaram maior expressividade na análise de prospecção foram os alcaloides e taninos, enquanto que na analise por CCD foram os flavonoides e terpenos, indicando o potencial da ação farmacológica das cascas de H. drasticus.

The indiscriminate use of the plant Himatanthus drasticus (Janaúba) by small goat farmers to control nematode parasites in small ruminants was the aim of this phytochemical study of the plant. Barks were collected in the eastern mesoregion of the state of Maranhão, Brazil, and sent to the laboratory for botanical and phytochemical identification by the methodology of Preliminary Prospecting and TLC, testing for the various classes of secondary metabolites. From the ground, dehydrated bark, we conducted the preparation of the crude extract (CE). The material was placed in a water-alcohol mixture of EtOH:H2O (7:3 v:v) and subjected to sporadic mechanical agitation. The subextracts were obtained using the HACE process by liquid-liquid partition, ETOH:H2O (2:1, v:v). The mixtures were prepared with organic solvents of increasing polarity, such as hexane, ethyl acetate and butanol. The GC analysis showed the presence of groups of secondary metabolites in the extract and subextractes. The classes of secondary metabolites that showed greater expressiveness are the alkaloids and tannins, when analyzed by prospecting, and flavonoids and terpenes, when analyzed by TLC, and H. drasticus presents elements indicative of pharmacological action.

Estruturas secretoras em órgãos vegetativos e florais de Secondatia densiflora A. DC. (Apocynaceae - Apocynoideae - Odontadenieae) / Secretory structures in vegetative and floral organs of Secondatia densiflora A. DC. (Apocynaceae - Apocynoideae - Odontadenieae)

A família Apocynaceae é caracterizada por possuir grande diversidade de estruturas secretoras como idioblastos, coléteres, laticíferos e nectários florais. Este estudo teve por objetivo caracterizar anatomicamente as estruturas secretoras nos órgãos vegetativos e reprodutivos de Secondatia densiflora e identificar as principais classes de compostos químicos nos idioblastos e na secreção dos coléteres vegetativos. Os idioblastos estão distribuídos por todos os órgãos aéreos da planta possuindo conteúdo na maioria das vezes denso e fortemente corado pela safranina. Alguns idioblastos apresentam secreção de aspecto granulado. Os testes histoquímicos evidenciaram apenas compostos fenólicos. Os laticíferos são do tipo anastomosado podendo ser observados em todos os órgãos estudados e identificados pelo conteúdo diferenciado e, em alguns casos, pela presença de paredes celulares mais espessas que as paredes das células parenquimáticas. O látex tem cor branca e aspecto leitoso, sendo extravasado logo que a planta é lesionada. Os coléteres vegetativos são do tipo padrão, formados por uma porção alongada que se afina em direção à extremidade. A epiderme secretora em paliçada delimita uma região parenquimática e o curto pedúnculo é coberto por epiderme não secretora de formato retangular. Todo coléter é recoberto por cutícula fina. A mucilagem é constatada tanto no interior das células secretoras quanto no meio extracelular pelo vermelho de rutênio e pela reação PAS. Os nectários florais têm origem receptacular, possuem uma epiderme que reveste toda estrutura, parênquima nectarífero, e feixes vasculares; são fundidos na base e livres na região apical constituindo cinco unidades distintas.

The family Apocynaceae is characterized for showing a wide variety of secretory structures such as idioblasts, colleters, laticiferous glands and floral nectaries. The present study aimed to anatomically characterize the secretory structures in vegetative and reproductive organs of Secondatia densiflora and to identify the major classes of chemical compounds in idioblasts and in the secretion of vegetative colleters. Idioblasts are distributed all over the aerial organs of the plant and their content is usually dense and strongly stained with safranin. Some idioblasts have secretion of granulated aspect. Histochemical tests evidenced phenolic compounds only. Laticiferous glands are of the anastomosed type and can be seen in all the studied organs and identified by their distinct content and, in some cases, by the presence of cell walls that are thicker than those of parenchyma cells. Latex is white, milky and leaks as the plant is injured. Vegetative colleters are of the standard type and formed of an elongated portion that becomes thinner towards the end. The secretory palisade epidermis delimits a parenchymatic region, while the short peduncle is covered by rectangular non-secretory epidermis. A thin cuticle covers the entire colleter. Mucilage is detected both inside the secretory cells and in the extracellular medium by ruthenium red staining and PAS reaction. Floral nectaries have receptacular origin and epidermis covering their entire structure, nectariferous parenchyma, and vascular bundles; they are fused at their bases but have free apical region, forming five distinct units.

Chemical constituents of Cynanchum wilfordii and the chemotaxonomy of two species of the family Asclepiadacease, C. wilfordii and C. auriculatum.

Definitive identification of original plant species is important for standardizing herbal medicine. Although only the dried roots of Cynanchum wilfordii (Asclepiadaceae) are prescribed as Cynanchi Wilfordii Radix in Korean Pharmacopoeia, the roots of C. wilfordii and C. auriculatum are often misused in the Korean herbal market due to their morphological similarity and similar name. Therefore, it would be very useful to discover an effective chemical marker for the identification of the two species. To this end, we carried out a phytochemical study on the roots of C. wilfordii. As a result, twenty compounds were isolated from the roots of C. wilfordii and their chemical structures were identified as ß-sitosterol (1), wilfoside C1N (2), wilfoside C3N (3), wilfoside K1N (4), methyleugenol (5), wilfoside C1G (6), cynauriculoside A (7), daucosterol (8), 2,4-dihydroxyacetophenone (9), cynandione A (10), 2,5-dihydroxyacetophenone (11), acetovanillone (12), p-hydroxyacetophenone (13), sucrose (14), conduritol F (15), geniposide (16), succinic acid (17), 3-(ß-D-ribofuranosyl)-2,3-dihydro-6H-1,3-oxazine-2,6-dione (18), bungeiside A (19), cynanoneside B (20). Among them, compounds 15, 16, 18, 19, and 20 were isolated for the first time from this species. Furthermore, conduritol F (15) was demonstrated to be contained only in C. wilfordii. Therefore, it may be useful as a chemical marker to identify the two species C. wilfordii and C. auriculatum.