In Vitro Propagation of Cannabis sativa L. and Evaluation of Regenerated Plants for Genetic Fidelity and Cannabinoids Content for Quality Assurance
Cannabis sativa L. (Marijuana; Cannabaceae), one of the oldest medicinal plants in the world, has been used throughout history for fiber, food, as well as for its psychoactive properties. The dioecious and allogamous nature of C. sativa is the major constraint to maintain the consistency in chemical profile and overall efficacy if grown from seed. Therefore, the present optimized in vitro propagation protocol of the selected elite germplasm via direct organogenesis and quality assurance protocols using genetic and chemical profiling provide an ideal pathway for ensuring the efficacy of micropropagated Cannabis sativa germp...
Source: Springer protocols feed by Plant Sciences - April 8, 2016 Category: Biology Source Type: news

In Vitro Shoot Cultures and Analysis of Steroidal Lactones in Withania coagulans (Stocks) Dunal
Withania coagulans (Stocks) Dunal (Solanaceae), also known as ‘Panir Bandh’ is an important medicinal plant that is extensively used as a home remedy for several diseases in the Indian subcontinent. The plant possesses specific steroidal lactones known as withanolides which show high level of pharmaceutical activity against a broad spectrum of microorganisms. Natural propagation of the plant occurs through Seed but due to unisexual nature of the flowers; chances of Seed setting are very limited and the plant is on the verge of extinction because of overexploitation and reproductive failure. Plant tissue culture...
Source: Springer protocols feed by Plant Sciences - April 8, 2016 Category: Biology Source Type: news

Scale-Up of Agrobacterium rhizogenes-Mediated Hairy Root Cultures of Rauwolfia serpentina: A Persuasive Approach for Stable Reserpine Production
Roots of Rauwolfia serpentina, also known as “Sarpagandha” possess high pharmaceutical value due to the presence of reserpine and other medicinally important terpene indole alkaloids. Ever increasing commercial demand of R. serpentina roots is the major reason behind the unsystematic harvesting and fast decline of the species from its natural environment. Considering Agrobacterium rhizogenes-mediated hairy root cultures as an alternative source for the production of plant-based secondary metabolites, the present optimized protocol offers a commercially feasible method for the production of reserpine, the most p...
Source: Springer protocols feed by Plant Sciences - April 8, 2016 Category: Biology Source Type: news

Production of Gymnemic Acid from Cell Suspension Cultures of Gymnema sylvestre
Gymnema sylvestre R. Br. is a popular herbal medicine. It has been used in ayurvedic system of medicine for thousands of years. It is popularly called as “Gur-mar” for its distinctive property of temporarily destroying the taste of sweetness and is used in the treatment of diabetes. The leaves of gymnema possess antidiabetic, antimicrobial, anti-hypercholesterolemic, anti-sweetener, anti-inflammatory, and hepatoprotective properties and have traditional uses in the treatment of asthma, eye complaints, and snake bite. The leaves contain triterpene saponins such as gymnemic acid which is an active ingredient of G...
Source: Springer protocols feed by Plant Sciences - April 8, 2016 Category: Biology Source Type: news

In Vitro Propagation of Sambong (Blumea balsamifera Linn.)
Terminal shoot tips of sambong (Blumea balsamifera Linn.) are cultured to initiate and regenerate shoots on Murashige and Skoog (MS) medium containing 1.0 mg/L benzyl adenine (BA). After 1 month, shoots, usually 4.5 cm long are separated and subcultured for multiplication. Regenerated shoots, about 6 cm long are rooted on MS medium supplemented with 1.0 mg/L naphthalene acetic acid (NAA). Exposure of shoots to high humidity for the first 2 weeks and equal proportion (1:1:1) of sterile sand, compost, and coir dust as potting mix favors the development of whole sambong plants. Young shoots from in vitro-derived sambong plant...
Source: Springer protocols feed by Plant Sciences - April 8, 2016 Category: Biology Source Type: news

In Vitro Propagation of Withania somnifera (L.) Dunal
Withania somnifera (L.) Dunal known as Ashwagandha is commonly used in traditional Indian medicine system. It possesses immense therapeutic value against a large number of ailments such as mental diseases, asthma, inflammation, arthritis, rheumatism, tuberculosis, and a variety of other diseases including cancer. The therapeutic potential of W. somnifera is due to the presence of secondary metabolites mainly, tropane alkaloids and withanolides (steroidal lactones). The growing realization of commercial value of the plant has initiated a new demand for in vitro propagation of elite chemotypes of Withania. Micropropagation w...
Source: Springer protocols feed by Plant Sciences - April 8, 2016 Category: Biology Source Type: news

Micropropagation, Acclimatization, and Greenhouse Culture of Veratrum californicum
Micropropagation and production of Veratrum californicum is most successful when using a premixed Murishage and Skoog basal medium with vitamins and a 5-week subculture cycle at 16 °C for multiplication. These culture conditions provide the best percent survival after acclimatization in the greenhouse. However, clone response to temperature and light quality within culture conditions varies. Micropropagated plants have mass and morphology similar to 2- or 3-year-old seedlings. Acclimatized plantlets can then be grown in the greenhouse using sub-irrigation (ebb and flood) to maintain substrate volumetric water content&t...
Source: Springer protocols feed by Plant Sciences - April 8, 2016 Category: Biology Source Type: news

Establishment, Culture, and Scale-up of Brugmansia candida Hairy Roots for the Production of Tropane Alkaloids
Brugmansia candida (syn. Datura candida) is a South American native plant that produces tropane alkaloids. Hyoscyamine, 6β-hydroxyhyoscyamine (anisodamine), and scopolamine are the most important ones due to their anticholinergic activity. These bioactive compounds have been historically and widely applied in medicine and their demand is continuous. Their chemical synthesis is costly and complex, and thereby, these alkaloids are industrially produced from natural producer plants. The production of these secondary metabolites by plant in vitro cultures such as hairy roots presents certain advantages over the natural so...
Source: Springer protocols feed by Plant Sciences - April 8, 2016 Category: Biology Source Type: news

In Vitro Propagation and Conservation of Bacopa monnieri L.
Bacopa monnieri L. (common name brahmi) is a traditional and renowned Indian medicinal plant with high commercial value for its memory revitalizer potential. Demand for this herb has further escalated due to popularization of various brahmi-based drugs coupled with reported anticancer property. Insufficient seed availability and problems associated with seed propagation including short seed viability are the major constraints of seed conservation in the gene banks. In vitro clonal propagation, a prerequisite for in vitro conservation by enhanced axillary branching was standardized. We have developed a simple, single step p...
Source: Springer protocols feed by Plant Sciences - April 8, 2016 Category: Biology Source Type: news

Immobilization of Rubia tinctorum L. Suspension Cultures and Biomass Production
Plants are natural sources of valuable secondary metabolites used as pharmaceuticals, agrochemicals, flavors, fragrances, colors, biopesticides, and food additives. There is an increasing demand to obtain these metabolites through more productive plant tissue applications and cell culture methods due to the importance of secondary metabolites. (Source: Springer protocols feed by Plant Sciences)
Source: Springer protocols feed by Plant Sciences - April 8, 2016 Category: Biology Source Type: news

Isolation of Plant Nuclei at Defined Cell Cycle Stages Using EdU Labeling and Flow Cytometry
We present protocols for EdU pulse labeling, tissue fixation and harvesting, nuclei preparation, and flow sorting. Although developed for Arabidopsis suspension cells and maize root tips, these protocols should be modifiable to many other plant systems. (Source: Springer protocols feed by Plant Sciences)
Source: Springer protocols feed by Plant Sciences - December 14, 2015 Category: Biology Source Type: news

Imaging Nuclear Morphology and Organization in Cleared Plant Tissues Treated with Cell Cycle Inhibitors
Synchronization of root cells through chemical treatment can generate a large number of cells blocked in specific cell cycle phases. In plants, this approach can be employed for cell suspension cultures and plant seedlings. To identify plant cells in the course of the cell cycle, especially during mitosis in meristematic tissues, chemical inhibitors can be used to block cell cycle progression. Herein, we present a simplified and easy-to-apply protocol to visualize mitotic figures, nuclei morphology, and organization in whole Arabidopsis root apexes. The procedure is based on tissue clearing, and fluorescent staining of nuc...
Source: Springer protocols feed by Plant Sciences - December 14, 2015 Category: Biology Source Type: news

Cytrap Marker Systems for In Vivo Visualization of Cell Cycle Progression in Arabidopsis
This chapter focuses on visualization of cell cycle progression in plant tissues using the dual-color marker system “Cytrap.” The Cytrap line carries a part of Arabidopsis CDT1a fused to the red fluorescent protein (RFP) gene, which monitors the cell cycle phases from S to late G2 or early mitosis, and the G2/M-specific cyclin B1 marker fused to the green fluorescent protein (GFP) gene. We introduce growth conditions of Arabidopsis roots, the setup for microscopic observation, and analysis of obtained images. (Source: Springer protocols feed by Plant Sciences)
Source: Springer protocols feed by Plant Sciences - December 14, 2015 Category: Biology Source Type: news

Identifying Gene Regulatory Networks in Arabidopsis by In Silico Prediction, Yeast-1-Hybrid, and Inducible Gene Profiling Assays
A system-wide understanding of gene regulation will provide deep insights into plant development and physiology. In this chapter we describe a threefold approach to identify the gene regulatory networks in Arabidopsis thaliana that function in a specific tissue or biological process. Since no single method is sufficient to establish comprehensive and high-confidence gene regulatory networks, we focus on the integration of three approaches. First, we describe an in silico prediction method of transcription factor–DNA binding, then an in vivo assay of transcription factor–DNA binding by yeast-1-hybrid and lastly ...
Source: Springer protocols feed by Plant Sciences - December 14, 2015 Category: Biology Source Type: news

Plant Cell Division Analyzed by Transient Agrobacterium-Mediated Transformation of Tobacco BY-2 Cells
The continuing analysis of plant cell division will require additional protein localization studies. This is greatly aided by GFP-technology, but plant transformation and the maintenance of transgenic lines can present a significant technical bottleneck. In this chapter I describe a method for the Agrobacterium-mediated genetic transformation of tobacco BY-2 cells. The method allows for the microscopic analysis of fluorescence-tagged proteins in dividing cells in within 2 days after starting a coculture. This transient transformation procedure requires only standard laboratory equipment. It is hoped that this rapid method ...
Source: Springer protocols feed by Plant Sciences - December 14, 2015 Category: Biology Source Type: news