Cannabis sativa L. is an important herbaceous species originating from Central Asia, which has been used in folk medicine and as a source of textile fiber since the dawn of times. This fast-growing plant has recently seen a resurgence of interest because of its multi-purpose applications: it is indeed a treasure trove of phytochemicals and a rich source of both cellulosic and woody fibers. Equally highly interested in this plant are the pharmaceutical and construction sectors, since its metabolites show potent bioactivities on human health and its outer and inner stem tissues can be used to make bioplastics and concrete-like material, respectively. In this review, the rich spectrum of hemp phytochemicals is discussed by putting a special emphasis on molecules of industrial interest, including cannabinoids, terpenes and phenolic compounds, and their biosynthetic routes. Cannabinoids represent the most studied group of compounds, mainly due to their wide range of pharmaceutical effects in humans, including psychotropic activities. The therapeutic and commercial interests of some terpenes and phenolic compounds, and in particular stilbenoids and lignans, are also highlighted in view of the most recent literature data. Biotechnological avenues to enhance the production and bioactivity of hemp secondary metabolites are proposed by discussing the power of plant genetic engineering and tissue culture. In particular two systems are reviewed, i.e., cell suspension and hairy root cultures. Additionally, an entire section is devoted to hemp trichomes, in the light of their importance as phytochemical factories. Ultimately, prospects on the benefits linked to the use of the -omics technologies, such as metabolomics and transcriptomics to speed up the identification and the large-scale production of lead agents from bioengineered Cannabis cell culture, are presented.

/pdf/cannabis-sativa-the-plant-of-the-thousand-and-one-molecules-1ud0br1diz.pdf

Cannabis sativa: The Plant of the Thousand and One Molecules

Medicinal plants are globally valuable sources of herbal products, and they are disappearing at a high speed. This article reviews global trends, developments and prospects for the strategies and methodologies concerning the conservation and sustainable use of medicinal plant resources to provide a reliable reference for the conservation and sustainable use of medicinal plants. We emphasized that both conservation strategies (e.g. in situ and ex situ conservation and cultivation practices) and resource management (e.g. good agricultural practices and sustainable use solutions) should be adequately taken into account for the sustainable use of medicinal plant resources. We recommend that biotechnical approaches (e.g. tissue culture, micropropagation, synthetic seed technology, and molecular marker-based approaches) should be applied to improve yield and modify the potency of medicinal plants.

/pdf/conservation-and-sustainable-use-of-medicinal-plants-3cr53p4qye.pdf

Conservation and sustainable use of medicinal plants: problems, progress, and prospects.

Plant growth promoting endophytic bacteria have been identified as potential growth regulators of crops. Endophytic bacterium, Sphingomonas sp. LK11, was isolated from the leaves of Tephrosia apollinea. The pure culture of Sphingomonas sp. LK11 was subjected to advance chromatographic and spectroscopic techniques to extract and isolate gibberellins (GAs). Deuterated standards of [17, 17-(2)H2]-GA4, [17, 17-(2)H2]-GA9 and [17, 17-(2)H2]-GA20 were used to quantify the bacterial GAs. The analysis of the culture broth of Sphingomonas sp. LK11 revealed the existence of physiologically active gibberellins (GA4: 2.97 ± 0.11 ng/ml) and inactive GA9 (0.98 ± 0.15 ng/ml) and GA20 (2.41 ± 0.23). The endophyte also produced indole acetic acid (11.23 ± 0.93 μM/ml). Tomato plants inoculated with endophytic Sphingomonas sp. LK11 showed significantly increased growth attributes (shoot length, chlorophyll contents, shoot, and root dry weights) compared to the control. This indicated that such phyto-hormones-producing strains could help in increasing crop growth.

Bacterial endophyte Sphingomonas sp. LK11 produces gibberellins and IAA and promotes tomato plant growth

Cannabis (Cannabis sativa, or hemp) and its constituents—in particular the cannabinoids—have been the focus of extensive chemical and biological research for almost half a century since the discovery of the chemical structure of its major active constituent, Δ9-tetrahydrocannabinol (Δ9-THC). The plant’s behavioral and psychotropic effects are attributed to its content of this class of compounds, the cannabinoids, primarily Δ9-THC, which is produced mainly in the leaves and flower buds of the plant. Besides Δ9-THC, there are also non-psychoactive cannabinoids with several medicinal functions, such as cannabidiol (CBD), cannabichromene (CBC), and cannabigerol (CBG), along with other non-cannabinoid constituents belonging to diverse classes of natural products. Today, more than 560 constituents have been identified in cannabis. The recent discoveries of the medicinal properties of cannabis and the cannabinoids in addition to their potential applications in the treatment of a number of serious illnesses, such as glaucoma, depression, neuralgia, multiple sclerosis, Alzheimer’s, and alleviation of symptoms of HIV/AIDS and cancer, have given momentum to the quest for further understanding the chemistry, biology, and medicinal properties of this plant.

Phytochemistry of Cannabis sativa L.

Podophyllotoxin is the natural product precursor of the chemotherapeutic etoposide, yet only part of its biosynthetic pathway is known. We used transcriptome mining in Podophyllum hexandrum (mayapple) to identify biosynthetic genes in the podophyllotoxin pathway. We selected 29 candidate genes to combinatorially express in Nicotiana benthamiana (tobacco) and identified six pathway enzymes, including an oxoglutarate-dependent dioxygenase that closes the core cyclohexane ring of the aryltetralin scaffold. By coexpressing 10 genes in tobacco-these 6 plus 4 previously discovered-we reconstitute the pathway to (-)-4'-desmethylepipodophyllotoxin (the etoposide aglycone), a naturally occurring lignan that is the immediate precursor of etoposide and, unlike podophyllotoxin, a potent topoisomerase inhibitor. Our results enable production of the etoposide aglycone in tobacco and circumvent the need for cultivation of mayapple and semisynthetic epimerization and demethylation of podophyllotoxin.

http://science.sciencemag.org/content/sci/349/6253/1224.full.pdf

Six enzymes from mayapple that complete the biosynthetic pathway to the etoposide aglycone

A comparison of the product yield, total phenolics, total flavonoids, and antioxidant properties was done in different leafy vegetables/herbs (basil, chard, parsley, and red kale) and fruit crops (bell pepper, cherry tomatoes, cucumber, and squash) grown in aeroponic growing systems (AG) and in the field (FG). An average increase of about 19%, 8%, 65%, 21%, 53%, 35%, 7%, and 50% in the yield was recorded for basil, chard, red kale, parsley, bell pepper, cherry tomatoes, cucumber, and squash, respectively, when grown in aeroponic systems, compared to that grown in the soil. Antioxidant properties of AG and FG crops were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DDPH) and cellular antioxidant (CAA) assays. In general, the study shows that the plants grown in the aeroponic system had a higher yield and comparable phenolics, flavonoids, and antioxidant properties as compared to those grown in the soil.

/pdf/assessment-of-total-phenolic-and-flavonoid-content-1dujuy2ak3.pdf

Assessment of Total Phenolic and Flavonoid Content, Antioxidant Properties, and Yield of Aeroponically and Conventionally Grown Leafy Vegetables and Fruit Crops: A Comparative Study

Cannabis sativa L (Cannabaceae) is an important medicinal plant well known for its pharmacologic and therapeutic potency Because of allogamous nature of this species, it is difficult to maintain its potency and efficacy if grown from the seeds Therefore, chemical profile-based screening, selection of high yielding elite clones and their propagation using biotechnological tools is the most suitable way to maintain their genetic lines In this regard, we report a simple and efficient method for the in vitro propagation of a screened and selected high yielding drug type variety of Cannabis sativa, MX-1 using synthetic seed technology Axillary buds of Cannabis sativa isolated from aseptic multiple shoot cultures were successfully encapsulated in calcium alginate beads The best gel complexation was achieved using 5 % sodium alginate with 50 mM CaCl22H2O Regrowth and conversion after encapsulation was evaluated both under in vitro and in vivo conditions on different planting substrates The addition of antimicrobial substance — Plant Preservative Mixture (PPM) had a positive effect on overall plantlet development Encapsulated explants exhibited the best regrowth and conversion frequency on Murashige and Skoog medium supplemented with thidiazuron (TDZ 05 μM) and PPM (0075 %) under in vitro conditions Under in vivo conditions, 100 % conversion of encapsulated explants was obtained on 1:1 potting mix- fertilome with coco natural growth medium, moistened with full strength MS medium without TDZ, supplemented with 3 % sucrose and 05 % PPM Plantlets regenerated from the encapsulated explants were hardened off and successfully transferred to the soil These plants are selected to be used in mass cultivation for the production of biomass as a starting material for the isolation of THC as a bulk active pharmaceutical

/pdf/propagation-through-alginate-encapsulation-of-axillary-buds-1sr3nn5fh5.pdf

Propagation through alginate encapsulation of axillary buds of Cannabis sativa L. — an important medicinal plant

Classification of Cannabis sativa L. in relation to agricultural, biotechnological, medical and recreational utilization -- History of Cannabis as Medicine: Nineteenth Century Irish Physicians and Correlations of their Observations to Modern Research -- Cannabis Sativa L.: Botany and Horticulture -- Cannabis sativa and Cannabis indica versus “Sativa” and “Indica” -- Morpho-anatomy of Marijuana (Cannabis sativa L.).-Chemical and Morphological Phenotypes in Breeding of Cannabis sativa L -- Natural Cannabinoids of Cannabis and Methods of Analysis -- Cannabinoids: Biosynthesis and Biotechnological Applications -- The Pharmacology and Therapeutic Potential of Plant Cannabinoids.-Cannabinoid CB2 Receptor Mechanism of  Cannabis sativa L. -- Cannabidiol as a Treatment for Seizures, Convulsions and Epilepsy -- Allergenicity to Cannabis sativa L. and Methods to Assess Personal Exposure -- Micropropagation of Cannabis sativa L. – An Update -- Hairy Root Culture as a Biotechnological Tool in C. sativa -- Genomics and Molecular Markers in Cannabis sativa L -- The Role of Agrobacterium-Mediated and Other Gene-Transfer Technologies in Cannabis Research and Product Development -- Induction of Polyploidy and Its Effect on Cannabis sativa L.- Classical and Molecular Cytogenetics of Cannabis sativa L -- Assessing Genetic Diversity in Cannabis sativa Using Molecular Approaches -- Cannabis Endophytes and Their Application in Breeding and Physiological Fitness -- Chemical and physical elicitation for enhanced cannabinoid production in Cannabis -- Contaminants of Concern in Cannabis: Microbes, Heavy Metals and Pesticides.

Cannabis sativa L. - Botany and Biotechnology

In vitro mass propagation of Cannabis sativa L.: A protocol refinement using novel aromatic cytokinin meta-topolin and the assessment of eco-physiological, biochemical and genetic fidelity of micropropagated plants

Induction of high-frequency shoot regeneration using nodal segments containing axillary buds from a 1-yr-old mother plants of Cannabis sativa was achieved on Murashige and Skoog (MS) medium containing 0.05–5.0 μM thidiazuron. The quality and quantity of regenerants were better with thidiazuron (0.5 μM thidiazuron) than with benzyladenine or kinetin. Adding 7.0 μM of gibberellic acid into a medium containing 0.5 μM thidiazuron slightly increased shoot growth. Elongated shoots when transferred to half-strength MS medium supplemented with 500 mg l−1 activated charcoal and 2.5 μM indole-3-butyric acid resulted in 95% rooting. The rooted plants were successfully acclimatized in soil. Following acclimatization, growth performance of 4-mo-old in vitro propagated plants was compared with ex vitro vegetatively grown plants of the same age. The photosynthesis and transpiration characteristics were studied under different light levels (0, 500, 1,000, 1,500, or 2,000 μmol m−2 s−1). An increase in photosynthesis was observed with increase in the light intensity up to 1,500 μmol m−2 s−1 and then decreased subsequently at higher light levels in both types of plants. However, the increase was more pronounced at lower light intensities below 500 μmol m−2 s−1. Stomatal conductance and transpiration increased with light intensity up to highest level (2000 μmol m−2 s−1) tested. Intercellular CO2 concentration (C
 i) and the ratio of intercellular CO2 concentration to ambient CO2 (C
 i/C
 a) decreased with the increase in light intensity in both in vitro as well as ex vitro raised plants. The results show that in vitro propagated and hardened plants were functionally comparable to ex vitro plants of same age in terms of gas and water vapor exchange characteristics, within the limits of this study.

/pdf/thidiazuron-induced-high-frequency-direct-shoot-4kyltp92nj.pdf

Hemant Lata

Papers

Assessment of Total Phenolic and Flavonoid Content, Antioxidant Properties, and Yield of Aeroponically and Conventionally Grown Leafy Vegetables and Fruit Crops: A Comparative Study

Propagation through alginate encapsulation of axillary buds of Cannabis sativa L. — an important medicinal plant

Cannabis sativa L. - Botany and Biotechnology

In vitro mass propagation of Cannabis sativa L.: A protocol refinement using novel aromatic cytokinin meta-topolin and the assessment of eco-physiological, biochemical and genetic fidelity of micropropagated plants

Thidiazuron-induced high-frequency direct shoot organogenesis of Cannabis sativa L.