Authentication of Garcinia fruits and food supplements using DNA barcoding and NMR spectroscopy.

Garcinia L. (Clusiaceae) fruits are a rich source of (-)-hydroxycitric acid, and this has gained considerable attention as an anti-obesity agent and a popular weight loss food supplement. In this study, we assessed adulteration of morphologically similar samples of Garcinia using DNA barcoding, and used NMR to quantify the content of (-)-hydroxycitric acid and (-)-hydroxycitric acid lactone in raw herbal drugs and Garcinia food supplements. DNA barcoding revealed that mostly G. gummi-gutta (previously known as G. cambogia) and G. indica were traded in Indian herbal markets, and there was no adulteration. The content of (-)-hydroxycitric acid and (-)-hydroxycitric acid lactone in the two species varied from 1.7% to 16.3%, and 3.5% to 20.7% respectively. Analysis of ten Garcinia food supplements revealed a large variation in the content of (-)-hydroxycitric acid, from 29 mg (4.6%) to 289 mg (50.6%) content per capsule or tablet. Only one product contained quantifiable amounts of (-)-hydroxycitric acid lactone. Furthermore the study demonstrates that DNA barcoding and NMR could be effectively used as a regulatory tool to authenticate Garcinia fruit rinds and food supplements.

Isolation and characterization of microsatellite markers in Garcinia gummi-gutta by next-generation sequencing and cross-species amplification.

Garcinia gummi-gutta (L.) Roxb. (Clusiaceae) is an endemic, semidomesticated, fruit-yielding tree species distributed in the Western Ghats of India and Sri Lanka. Various bioactive phytochemicals, such as garcinol, benzophenones and xanthones are isolated from G. gummi-gutta and have shown antibacterial, antiviral and antioxidant activities. We sequenced the total genomic DNA using Illumina Hiseq 2000 platform and examined 241,141,804 bp high quality data, assembled into 773,889 contigs. In these contigs, 27,313 simple-sequence repeats (SSRs) were identified, among which mononucleotide repeats were predominant (44.98%) followed by dinucleotide and trinucleotide repeats. Primers were designed for 9964 microsatellites among which 32 randomly selected SSR primer pairs were standardized for amplification. Polymerase chain reaction (PCR) amplification of genomic DNA in 30 G. gummi-gutta genotypes revealed polymorphic information content (PIC) across all 32 loci ranging from 0.867 to 0.951, with a mean value of 0.917. The observed and expected heterozygosity ranged from 0.00 to 0.63 and 0.896 to 0.974, respectively. Alleles per locus ranged from 12 to 27. This is the first report on the development of genomic SSR markers in G. gummi-gutta using next-generation sequencing technology. The genomic SSR markers developed in this study will be useful in identification, mapping, diversity and breeding studies.

Garmultins A-G, Biogenetically Related Polycyclic Acylphloroglucinols from Garcinia multiflora.

Garmultins A and B (1 and 2), two polycyclic polyprenylated acylphloroglucinols characterized by the coupling of two novel cages, 2,11-dioxatricyclo[4.4.1.03,9]undecane and tricyclo[4.3.1.03,7]decane, along with five biogenetically related analogues (3-7), were isolated from Garcinia multiflora. Their structures and absolute configurations were determined by extensive NMR analysis, X-ray crystallography, and electronic circular dichroism calculations. Three compounds were capable of inhibiting oncogene expression and inducing apoptosis in human erythroleukemia cells.

Identification of molecular markers to study the Garcinia spp. diversity.

The genus Garcinia shows a considerable variation in its morphological characters such as leaf, flower and fruit with taxonomic ambiguity. It is a potential under-exploited multipurpose crop that gained considerable attention for the presence of (-) hydroxycitric acid, an anti-obesity compound, in its fruit rind and leaves. Here, we evaluated the genetic relationship through molecular markers among the selected 9 species commonly available in the Western Ghats and the Northeastern Himalayan foot hills of India. The nucleotide sequence data obtained from two prominent monomorphic bands generated in ISSR profiling of the species was utilized for the study. The selected bands were found to be of ITS region (700 bp) and partial region of KNOX-1 gene (600 bp). The evolutionary cluster was formed using MEGA5 software. The study indicated 2 major clusters, influenced by floral morphology of the species and availability of (-) hydroxycitric acid in their fruit rinds. In the subclusters, one species from the Western Ghats were paired with another from Northeastern Himalayas with relatively similar morphological traits.

A comparative study of karyomorphology among three populations of Garcinia indica (Clusiaceae) (Thomas-Dupetite) Choisy.

Garcinia indica is a tree species of the family Clusiaceae. This species is endemically distributed only in Ratnagiri and Sindhudurga district of Maharashtra. Plants collected in particularly Dive Agar and Sawantwadi show morphological variations. Mitotic chromosomes from three different population from the Konkan region were studied using propionic orcein stain. Chromosome numbers 2n = 54 is reported for all the populations. Three populations differed from each other in some aspects. Karyotype analysis showed that Deorukh population from Ratnagiri region showed one pair of satellite and one pair of secondary constrictions while Otawane population from Sawantwadi showed one satellite chromosome but differ in pattern of secondary constriction. Third, Diveagar population from Raigad district showed three pairs of satellite chromosomes and one pair of secondary constriction. So, it can be concluded that, plants from Bapat Garden of Dive Agar and Otawane region can be considered as cytotypes.

Estudo farmacobotânico das folhas de Garcinia brasiliensis Mart. (Clusiaceae) / Pharmacobotanical studies of Garcinia brasiliensis Mart. (Clusiaceae) leaves

A espécie Garcinia brasiliensis Mart. (Clusiaceae), nativa da região Amazônica e cultivada em todo o território brasileiro, vem sendo bastante estudada devido seu potencial farmacológico, porém são escassos estudos que tratam da caracterização farmacobotânica desta espécie. Considerando as propriedades terapêuticas para tornar-se um medicamento fitoterápico, o presente trabalho teve como objetivos estudar a anatomia e histoquímica da folha e do pecíolo e elaborar dados macroscópicos e microscópicos que forneçam características marcantes para sua identificação além de dar subsídios para a análise farmacognóstica no controle de qualidade da droga vegetal. O material vegetal foi fixado e submetido às técnicas usuais de microscopia de luz e a testes histoquímicos. As folhas de G. brasiliensis são opostas, simples, descolores, forma elíptica com nervação peninérvia. As células epidérmicas, em vista frontal, apresentam contorno sinuoso e estômatos paracíticos somente na face abaxial. O mesofilo é dorsiventral, a nervura central apresenta contorno biconvexo e feixe vascular em forma de semi-arco fechado envolto por bainha esclerenquimática. Inclusões inorgânicas de cristais na forma de drusas e orgânicas representadas por compostos fenólicos e grãos de amidos estão dispersos ao longo de toda lâmina foliar e pecíolo. Observa-se com frequência a presença de canais secretores preenchidos por um conteúdo lipídico dispersos pelo parênquima fundamental e próximos aos feixes vasculares. Estes dados fornecem subsídios para o controle de qualidade da matéria-prima utilizada para a produção de fitoterápicos.

The Garcinia brasiliensis Mart. (Clusiaceae) species, native of the Amazon region and cultivated throughout the Brazilian territory, has been widely studied due to its pharmacological potential, but there are few studies dealing with the pharmacobotanic characterization of this species. Considering the therapeutic properties in order to become an herbal medicine, the present paper had the purpose of studying the anatomical and histochemical characterization of the leaf and petiole, as well as producing macroscopic and microscopic data that provide important characteristics for its identification, in addition to providing subsidies for the pharmacognostical analysis in order to offer elements for the quality assurance of the drug. The botanical material was prepared through the usual optical and histochemical microtechniques. The leaves of G. brasiliensis are simple, opposed, colorless, and they show an elliptical shape. As seen from the front, the epidermal cells have a sinuous contour, and paracytic stomata occur on the low surface. The leaves are hipostomatic and dorsiventral with heterogeneous mesophile. The mesophile is dorsiventral, the central midrib shows a biconvex contour and vascular system in a semi-closed arch shape surrounded by a sclerenchymatic sheath. Inorganic inclusions of crystals in the shape of druses, and organic inclusions represented by phenolic compounds and starch grains are found throughout the leaf blade and petiole. It is common to find secretory canals filled with a lipid content dispersed throughout the parenchyma and near the vascular bundles. These data support the quality assurance of the elements used to produce herbal medicines.