Important Herbal Plant Research Articles

Transcriptome analysis of Panax vietnamensis var. fuscidicus discovers putative ocotillol-type ginsenosides biosynthesis genes and genetic markers.

BackgroundP. vietnamensis var. fuscidiscus, called “Yesanqi” in Chinese, is a new variety of P. vietnamensis, which was first found in Jinping County, the southern part of Yunnan Province, China. Compared with other Panax plants, this species contains higher content of ocotillol-type saponin, majonoside R2. Despite the pharmacological importance of ocotillol-type saponins, little is known about their biosynthesis in plants. Hence, P. vietnamensis var. fuscidiscus is a suitable medicinal herbal plant species to study biosynthesis of ocotillol-type saponins. In addition, the available genomic information of this important herbal plant is lacking.ResultsTo investigate the P. vietnamensis var. fuscidiscus transcriptome, Illumina HiSeq™ 2000 sequencing platform was employed. We produced 114,703,210 clean reads, assembled into 126,758 unigenes, with an average length of 1,304 bp and N50 of 2,108 bp. Among these 126,758 unigenes, 85,214 unigenes (67.23%) were annotated based on the information available from the public databases. The transcripts encoding the known enzymes involved in triterpenoid saponins biosynthesis were identified in our Illumina dataset. A full-length cDNA of three Squalene epoxidase (SE) genes were obtained using reverse transcription PCR (RT-PCR) and the expression patterns of ten unigenes were analyzed by reverse transcription quantitative real-time PCR (RT-qPCR). Furthermore, 15 candidate cytochrome P450 genes and 17 candidate UDP-glycosyltransferase genes most likely to involve in triterpenoid saponins biosynthesis pathway were discovered from transcriptome sequencing of P. vietnamensis var. fuscidiscus. We further analyzed the data and found 21,320 simple sequence repeats (SSRs), 30 primer pairs for SSRs were randomly selected for validation of the amplification and polymorphism in 13 P. vietnamensis var. fuscidiscus accessions. Meanwhile, five major triterpene saponins in roots of P. vietnamensis var. fuscidicus were determined using high performance liquid chromatography (HPLC) and evaporative light scattering detector (ELSD).ConclusionsThe genomic resources generated from P. vietnamensis var. fuscidiscus provide new insights into the identification of putative genes involved in triterpenoid saponins biosynthesis pathway. This will facilitate our understanding of the biosynthesis of triterpenoid saponins at molecular level. The SSR markers identified and developed in this study show genetic diversity for this important crop and will contribute to marker-assisted breeding for P. vietnamensis var. fuscidiscus.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1332-8) contains supplementary material, which is available to authorized users.

LC–MS Based Workflows for Qualitative and Quantitative Analysis for Homeopathic Preparation of Hydrastis canadensis

Hydrastis canadensis L. is one of the important herbal medicinal plants, which is used for homeopathic preparations. It contains berberine, hydrastine as major and canadine, hydrastinine, berberastine and canadaline as minor bioactive compounds. The aim was to develop (I) a sensitive mass spectrometric-based quantitative/qualitative method for berberine in homeopathic dilutions of H. canadensis, (II) with identification of berberine and its structurally related compound present in the homeopathic preparation of H. canadensis using predictive MRM workflow, (III) confirmatory analysis for the structurally related compound in homeopathic dilutions of H. canadensis using high-resolution mass spectrometer. Different mass spectrometric-based workflows (MRM to EPI; pMRM to EPI and IDA-DBS) were used to develop the method to analyze the homeopathic dilutions of H. canadensis. A high-throughput liquid chromatography-tandem mass spectrometry method using MRM to EPI workflow was developed to analyze berberine in homeopathic preparation of H. canadensis. Multiple reaction monitoring transitions of 336.0/292.08 and 336.0/277.9 were selected for developing the quantitation method of berberine. The method was linear from 5.0 to 2000.0 pg/mL. The validated method has been successfully applied for the analysis of different potencies of homeopathic preparation of H. canadensis. Predictive MRM to enhanced product ion workflows was used to identify and confirm the structurally related molecules of berberine. Non targeted information dependent acquisition-DBS method was developed to identify the compounds in H. canadensis mother tincture using high-resolution mass spectrometry. The established method using different mass spectrometric workflows facilitates the convenient and fast quality control analysis for the herbal preparation of H. canadensis.

Identification of Agronomic Traits of Centella asiatica (L.) Urban. Naturally Grown at Regions With Different Altitudes

The Centella asiatica is an important herbal plant in Indonesia. The research was done to describe severalagronomic traits that related to yield of the plants grown at regions with different altitudes. The research wascarried out at three different regions of highland, midland, and lowerland, each at >500m, 250-500m, and < 250mabove sea level. The research location was Pagar Alam, Musi Rawas, and Ogan Ilir, respectively. Research indicatedthat the agronomic traits of the plants grown at highland, midland, and lowerland, regions were about the same forgeneral appearance of leaf shape and flower color. The leaf size, number and length of stolone, number of rossets,length of rhizomes, and number of flower were varied depending on the region. The plants grown at highland andmidland region tend to have larger leaf size, shorter petiole and stolone, more number of rossets and stolone,longer rhizomes, and more number of flowers per plant, compared to the plants of lowerland region. This could beindication that Centella asiatica plants grown on midland, and highland were more productive than plants oflowerland region.

Effects of tissue-cultured mountain ginseng (Panax ginseng CA Meyer) extract on male patients with erectile dysfunction

Korean ginseng and mountain ginseng (Panax ginseng CA Meyer) are important traditional herbal plants whose ginsenosides are generally accepted as serving to improve sexual functions, such as penile erection. We investigated the effects of tissue-cultured mountain ginseng extract (TMGE) on male patients with erectile dysfunction (ED). A double-blind, placebo-controlled study was conducted with 143 patients experiencing ED. Over the course of 8 weeks, one group took 1 000 mg of TMGE twice a day, and the other group took 1,000 mg of placebo twice a day. The effects of the TMGE and the placebo were analyzed using the Korean version of the International Index of Erectile Function (IIEF) questionnaire. A total of 86 patients completed 8 weeks of treatment. The scores on the five domains of the IIEF after medication were significantly higher than the baseline scores in the group treated with TMGE (P<0.05), whereas no significant improvement was observed in the placebo group (P>0.05). Erectile function and overall satisfaction scores after medication were significantly higher in the TMGE group than in the placebo group (P<0.05). Erectile function of patients in the TMGE-treated group significantly improved, suggesting that TMGE could be utilized for improving erectile function in male patients.

Heavy trace metals and macronutrients status in herbal plants of Nigeria

Abstract The concentration levels (ppm) of selected toxic trace metals (Fe, Mn, Cu, Pb and Zn) and macronutrients (Na, K, Mg and Ca), along with P, were estimated in some of the important herbal plants of the southwest part of Nigeria. The atomic absorption spectrophotometer was employed for the estimation conducted on 10 plant species collected from different locations within Ogbomoso. The plants were Anacardium occidentale, Azadirachta indica, Butyrospermum paradoxum, Mangifera indica, Morinda lucida, Ocimum canum, Solanum erianthum, Solanum torvum, Zingiber officinale and Hyptis suaveolens . The metal contents in the samples were found at different levels. The highest mean levels (ppm) of Zn (35.1±0.01) and Cu (24.4±0.01) were found in Hyptis suaveolens while those of Mn (685±0.02) and Ca (51 340±21) were found in Morinda lucida . The result also showed that Ocimum canum had the highest amounts of K (36 600±350), P (3700±35) and Fe (241±0.05). Anacardium occidentale had the highest concentration of Na (613±0.60) while Azadirachta indica had the highest mean concentrations of Pb (0.49±0.03) and Mg (5630±12).

Micropropagation of Leptadenia reticulata—A medicinal plant

Leptadenia reticulata (Retz.) Wight. & Arn., an important herbal medicinal plant, belongs to the family Asclepiadaceae. This plant has been known for its medicinal uses since 4500 BC. Presently this is an endangered species. There is a need for applying non-conventional methods of propagation for conservation and sustainable utilization of biodiversity of Leptadenia reticulata. We developed a micropropagation method for mass multiplication of L. reticulata. Explants harvested from greenhouse-maintained and field-grown plants were used to establish cultures of L. reticulata. The nodal shoot segments were surface-sterilized and cultured on Murashige and Skoog (MS) medium along with additives (25 mg l−1 each of adenine sulfate, arginine, and citric acid, and 50 mg l−1 ascorbic acid) containing 0.6 μM indole-3-acetic acid (IAA) and 9 μM N6-benzyladenine (BA). Three to four shoots differentiated from each node within 25–30 d at 26±2°C and 36 μmol m−2 s−1 spectral flux photon (SFP) for 12 hd−1. Shoots were further multiplied by (1) repeated transfer of mother explant on fresh medium containing 0.6 μM IAA and 2.2 μM BA, and (2) subculture of in vitro-differentiated shoots on MS medium with 6.6 μM BA and 0.6 μM IAA. After three or four subcultures, the basal clump with shoot bases was divided into three or four subclumps and multiplied on the fresh medium. From each clump 15–20 shoots regenerated within 25 d. Ninety percent of the in vitro-produced shoots rooted ex vitro if these were pulse-treated with 123 μM each of indole-3-butyric acid and β-naphthoxyacefic acid. The plantlets were transferred to bottles containing sterile ‘soilrite’ (soilless compost and soil conditioner) moistened with half-strength MS macrosalts. Ninety-five percent of the plantlets were hardened in the bottles within 15 d. The hardened plants were then transferred to black polybags in the nursery. Field transferred plants are growing normally and have flowered. The protocol developed is reproducible. From a single nodal segment about 1700 hardened plants could be regenerated within 174 d.

Studies with plant‐cell cultures of the Chinese herbal plant, Tripterygium wilfordii. Isolation and characterization of diterpenes

AbstractA detailed analysis of the established cell‐culture line (coded as TRP4a) of the important Chinese herbal plant, Tripterygium wilfordii, in terms of diterpene production, is presented. In this study, 28 diterpenes have been isolated and characterized. Of these, 26 were isolated for the first time from the TRP4a cell line, 8 were novel compounds. From an analysis of the diterpenes obtained, a biosynthetic pathway of the pharmacologically interesting triepoxide, tripdiolide 29, is proposed.

Biotechnology and synthetic chemistry: Routes to clinically important natural products

Abstract