In Vitro Propagation and Secondary Metabolite Production from Medicinal Plants: Current Trends (Part 2) E-Book
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- Herausgeber: Bentham Science Publishers
- Kategorie: Wissenschaft und neue Technologien
- Sprache: Englisch
This book is a comprehensive review of secondary metabolite production from plant tissue culture. The editors have compiled 12 meticulously organized chapters that provide the relevant theoretical and practical frameworks in this subject using empirical research findings. The goal of the book is to explain the rationale behind in vitro production of secondary metabolites from some important medicinal plants. Biotechnological strategies like metabolic engineering and the biosynthesis, transport and modulation of important secondary metabolites are explained along with research studies on specific plants. In addition to the benefits of secondary metabolites, the book also aims to highlight the commercial value of medicinal plants for pharmaceutical and healthcare ventures.
Topics covered in this part include:
1. In vitro propagation and tissue culture for several plants including Withania somnifera (L.) Dunal, Aloe vera, Oroxylum indicum (L) Kurz, Ocimum basilicum L, Rhubarb, Tea, and many others (including plants in Northern India).
2. Genetic Improvement of Pelargonium
3. Bioactive Components in Senna alata L. Roxb
4. Plant tissue culture techniques
The book caters to a wide readership. It primarily prepares graduate students, researchers, biotechnologists, giving them a grasp of the key methodologies in the secondary metabolite production. It is a secondary reference for support executives, industry professionals, and policymakers at corporate and government levels to understand the importance of plant tissue culture and maximizing its impact in the herbal industry.
Readership
Graduate students and researchers in plant biotechnology courses; industry professionals and policymakers in the herbal industry.
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Seitenzahl: 644
Veröffentlichungsjahr: 2024
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PREFACE
The extinction of plant species is progressively taking place due to their being trapped in the vicious circle of ever-increasing industrialization, deforestation, global warming, climate change and also unscrupulous human activities. This has led to many species being listed in the Red Data Book or /in the various threat categories of IUCN. Of the total 3000 medicinal plants reported from India, over 1700 species of medicinal value are found in the Indian Himalayan region; nearly 47% are endemic to this region and about 62 species fall under different categories of threat. The situation warrants the acceleration of efforts to develop methods for germplasm preservation. The importance and applications of plant cell and tissue culture in plant science are vast and varied. The last few years of our research investigations have led to the emergence of this technique. Utilizing the biotechnological tools, many tissue culture protocols have been developed for rapid and mass multiplication of valuable medicinal plants to increase planting stock so as to meet the market demand. The rapid increase in knowledge of nutrition, medicine, agriculture, and plant biotechnologies has effectively changed the concept of food and health causing an overwhelming revolution.
India is known for its diverse climatic zones which are habituated of diverse flora having medicinal value, thus there is a wide scope for India to lead global herbal market. The National Medicinal Plant Board of India has recognized more than 7000 medicinal plants, which are currently used in different systems of medicines. The Ayurveda market in India has been valued at INR 300 billion in 2018 and is expected to reach INR 710 billion by 2024. Plants are active biochemical factories of a vast group of secondary metabolites which are indeed the basic source of various commercial pharmaceutical drugs. There are possibilities for year round production of biomass with reduced cost and time. Elicitation and precursor feeding are two important strategies of the in-vitro techniques to enhance metabolite production to meet the demands of mankind. Utilization of the existing genetics resources and understanding the biosynthesis, transport, accumulation and modulation of important secondary metabolites are critical issues linked to its improvement.
Overall, the rapid propagation of elite plants will provide high dividends to farmers and the associated herbal industry.
This book provides comprehensive coverage of the fundamental principles, current practices and trends in the field of pharmaceutical industry and provides baseline data for further research in the field. We are grateful to all the contributors and hope the book will be beneficial to students, researchers, scientists and other concerned stake holders who are working in the respective fields. MA acknowledges the much needed moral support of his wife, Humera Anis. We would also like to place on record our sincere thanks to Mr. Mohammad Zohaib Siddiqui for preparing the layout of the contents.
The cooperation and help received from Bentham Science Publishers is duly appreciated.
List of Contributors
Bioactive Components in Senna Alata L. Roxb
Archana Pamulaparthi1,Vamshi Ramana Prathap2,Ramaswamy Nanna1,*Abstract
Senna alata is an ethnomedicinal plant. The crude extracts of the plants are said to have a large number of medicinal properties due to their phytochemicals. In the present study, we made an attempt to isolate and screen the phytochemical constituents present in the species. In order to determine the bioactive constituents present in S. alata, and the effect of drying on the loss of bioactive constituents, studies on a set of pharmacognostical parameters were conducted on seeds, shade and sun-dried leaves of S. alata as per US pharmacopeia and WHO guidelines. The results of the present studies showed the presence of various important bioactive molecules that are responsible for the medicinal properties of the species. The phytochemical analysis of seed extracts revealed the presence of alkaloids, flavonoids, tannins, saponins, anthraquinones, resins and glycosides in all the extracts, while coumarins, phenols, terpenoids, phlobatannins and quinines are completely absent in all the seed extracts. Preliminary phytochemical investigations from shade and sun-dried leaf extracts showed alkaloids, flavonoids, anthraquinones, saponins, glycosides and tannins in high amounts in all the extracts, resins and phenols are present in moderate amounts. Terpenoids and phlobatannins are present only in fresh leaf extracts. Studies were also conducted on the physicochemical and organoleptic properties of leaves of S. alata that help in the identification and standardization of the leaf extracts for manufacturing of plant-based drugs of S. alata.
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Madhukar Garg1,Soumi Datta2,Sayeed Ahmad3,*Abstract
Plants are an immense source of phytochemicals with therapeutic effects and are widely used as life-saving drugs, and other products of varied applications. Plant tissue culture is a unique technique employed under aseptic conditions from different plant parts called explants (leaves, stems, roots, meristems, etc.) for in vitro regeneration and multiplication of plants and synthesis of secondary metabolites (SMs). Selection of elite germplasm, high-producing cell lines, strain enhancements, and optimization of media and plant growth regulators may lead to increased in vitro biosynthesis of SMs. Interventions in plant biotechnology, like the synthesis of natural and recombinant bioactive molecules of commercial importance, have attracted attention over the past few decades; and the rate of SMs biosynthesis has increased manifold than the supply of intact plants, leading to a quick acceleration in its production through novel plant cultures. Over the years, the production of SMs in vitro has been enhanced by standardising cultural conditions, selection of high-yielding varieties, application of transformation methods, precursor feeding, and various immobilization techniques; however, most often, SM production is the result of abiotic or biotic stresses, triggered by elicitor molecules like natural polysaccharides (pectin and chitosan) that are used to immobilize and cause permeabilization of plant cells. In vitro synthesis of SMs is especially promising in plant species with poor root systems, difficulty in harvesting, unavailability of elite quality planting material, poor seed set and germination, and difficult to propagate species. Thus, the present article reviews various biotechnological interventions to enhance commercially precious SMs production in vitro.
Introduction
Plants are renewable sources and form an important part of our daily diet, and provide essential primary metabolites (e.g., carbohydrates, lipids and amino acids) [1] and phytochemicals (low molecular weight compounds-SMs) for different industrial applications like pharmaceuticals, nutraceutical, textile, construction and cosmetic sectors [2]. The majority of the world population’s health and wellness relies on plant-derived components. Therefore, plants with medicinal properties are considered important to support the transition to a bio-economy that is less dependent on fossil resources. The SMs not only play a pivotal role in plants’ adaptation to their environment but also represent an important source of active pharmaceuticals [3] and are synthesised by plants to defend themselves against exogenous stresses, both biotic and abiotic. A study [4] proposed the concept of SMs that were known as opposed to primary ones and an entire volume of “plant biochemistry” series named as “endproduct” [5]. It is known that higher plants are a rich source of phyto-pharmaceuticals and are used in the pharmaceutical industry. Some of the plant-derived products include drugs like morphine, codeine, cocaine, pilocarpine, belladonna alkaloids, colchines, phytostigminine, L-DOPA, berberine, reserpine, capsaicin, podophyllotoxin, shikonin derivatives, ajmalicine, vincristine and vinblastine [6] and steroids like ginsenosides, anti-cancer (taxol), diosgenin, digoxin and digitoxin. Significant synthetic substitutes of these drugs with the same efficacy and pharmacological specificity are yet to be found [7].
Previously, chemical synthesis for the production of SMs was achieved through field cultivation; however, the plants originating from particular biotypes were difficult to grow outside their ecosystems and thus led scientists and biotechnologists to consider plant cell, tissue and organ cultures as an alternative to produce secondary metabolites. The major advantages of in vitro synthesis of bioactive secondary metabolites within controlled conditions include: these are climatic and soil stipulations independent, minimal inferences of negative biological parameters affecting the SM production, possible choice of elite germplasm with respect to the presence of SMs, computerization of cell growth control, metabolic processes regulation, and cost price, which can be decreased with increased production. Plants produce alkaloids, flavonoids, lactones, glycosides, quinines, phenylpropanoids, resins, tannins, terpenoids, saponins, sesquiterpene, and steroids [8]. The first large-scale production of commercial plant cells application was carried out in stirred tank reactors to synthesis shikonin by cell cultures of Lithospermum erythrorhizon [9, 10].
Secondary metabolites
Plants are capable of producing different organic molecules called secondary metabolites, having unique carbon skeletons with basic properties. SMs are not necessarily for a cell (organism) to live but also for interaction with its environment. These are organ, tissue and cell-specific with low molecular weights and often differ amid individuals from the same population with respect to their type. SMs protect plants against stress; and are used as drugs, flavors, fragrances, insecticides and dyes and hence are of great economic value. SMs have evolved as molecules imperative for organisms producing them, the majority of these interfere with the pharmacological targets, and thus make them significant for several biotechnological applications.
Primary vs Secondary Metabolites
Primary metabolites (PMs) are compounds that are universally present in all plants, but are not species-specific and, thus might be identical in some organisms. These are directly involved in metabolic activities like growth, development, nutrition and reproduction of a plant whereas secondary metabolites are produced in other metabolic pathways that, although important, but are not essential to the functioning of the plant. Whereas, SMs are species specific and, therefore, unique for each species. The major differences between PMs and SMs are listed in Table 1.
The Biosynthesis of Secondary Metabolites
SMs are synthesized by diverting energy-generating directions in metabolic pathways like photosynthesis, glycolysis, and Krebs cycle to biosynthetic intermediates; and are classified in separate categories depending upon their biosynthesis, structures and functions. SMs are mostly biosynthesized from acetyl coenzyme A, mevalonic acid, shikimic acid, deoxyxylulose 5-phosphate or various combined pathways [11]. Accordingly, they are classified into terpenoids, steroids, alkaloids, saponin, terpenes, lipids and enzyme cofactors [12, 13]. There are three major pathways to produce SMs-Shikimate, isoprenoid and polyketide. Formation of the fundamental skeleton is followed by further modifications, resulting in synthesis of plant specific compounds. The shikimate pathway is mostly found in microorganisms and plants, but not in mammals, and is a major source of aromatic compounds; thus making it an important target for insecticides and antibiotics, which are harmless on mammalian systems. In the glyphosate pathway, enzymes chorismate mutase and anthranilate synthase channelise chorismate into aromatic amino acids. However, the most popular pathway for the synthesis of SMs is the phenylpropanoid pathway as it leads to the synthesis of major SMs like lignin, lignans, flavonoids, and anthocyanins; it is found in all plants, while others may have several co-enzymes. Phenylalanine Ammonia Lyase (PAL) is the key enzyme that converts phenylalanine into trans-cinnamic acid by non-oxidative deamination. Few others like isoprenoid pathways are involved in the synthesis of terpenoids. The C5 building block when incorporated into other skeletons forms an array of SMs like anthraquinones, cannabinoids, furanocoumarines, indole alkaloids, and napthaquinones; while incorporation into
basic skeletons results in the synthesis of hop bitter acids, flavonoids and isoflavonoids [14].
The Physiological Function of Secondary Metabolites
Secondary plant metabolites are abundant chemical compounds produced by the plant cell through different metabolic pathways derived from the primary metabolic pathways. Some of the major roles they play in plants are:
Act as signalling functions that influence the activities of other cells, control their metabolic activities and coordinate the development of the complete plant.SMs like terpenoids, alkaloids and flavonoids have therapeutic applications in the pharmaceutical industry as drugs and dietary supplements.Protection against harmful environmental conditions.Protection against pathogens and herbivores in the form of volatile monoterpenes or essential oils.The volatile terpenoids also play a major function in plant-plant interactions and serve as attractants for pollinators [15].SMs are often used as flower colors which serve to communicate with pollinators or protect plants from feeding deterrence by producing specific phytoalexines after fungi infections that inhibit the spreading of the fungi mycelia in plants.Plants use SMs in the form of volatile essential oils colored flavonoids or tetraperpenes to attract insects for pollination and seed disposal.They constitute important UV absorbing compounds, thus preventing serious leaf damage from the light.Classification of Secondary Metabolites
The different classes of SMs synthesized in vitro (callus and cell suspension) by various culture conditions are:
Alkaloids
Acridines, Betalaines, Furoquinolines, Galanthamine, Harringtonines, Isoquinolines, Lobeline, Quinolizidines, Indole alkaloids, Isoquinoline alkaloids, Piperidine, Thebaine, Trigonelline, and Tropane alkaloids.
Terpenoids
Artemisinin, Cucurbitacins, Diterpenes, Ginsenoside, Meroterpenes, Monoterpenes. Paclitaxel, Sesquiterpenes, Thapsigargin, Triterpenes, Ursane, and Withanolides.
Steroids
Ajugalactone, Asiaticosid, Asiatic acid, β-sitosterol, Brassinolid, Bufadienolides, Catasterone, Conesine, Cyasterone, Cardenolides, Digoxin, Digitoxin, Digitoxigenin, Diosgenin, Ecdysteroids, Ecdysone, Feruginol, Gagaminine, Gentipicroside, Guggulusterones, Harpagoside, Helleborin, Paclitaxel, Physodine, Polypodine B, Phytosterols, Pterosterone, Ponasterone, Madecassic Acid, Madecassoside, Ouabain, Saponins, Saikosaponins, Scillaridine, Sengoterone, Solasodine, Solarmargine, Spirostanol Saponin, Steroidal glycosides, Steroidal lactones, Swertiamarin, Tanshinone, Taxol, Taxoids, Turkesteron, Typhasterol, 29-norcyasterone, and 20 E Cryptotanshinone.
Quinones
Aloe-emodin, Anthraquinones, Benzoquinones, Chrysophanol, Emodin, β-Lapacho, Naphthoquinones, Phenanthrenequinone, Plumbagin, Rhein, Shikonin, Thymoquinone, and Uglone.
Phenylpropanoid
Anthocyanins, Caffeic acid, Coumarins, Eugenol, Ferulic acid, Flavonoids, Hydroxycinnamoyl derivatives, Isoflavonoids, Lignans, Phenalinones, Proanthocyanidins, Stilbenes, and Tannins.
Secondary Metabolites and Plant Tissue Culture
The primary and age-old practice is to grow selected plants in greenhouses or protected areas and to extract the biomolecule from them. In this context, plant tissue culture (especially cell and organ culture) plays an immense role in in vitro propagation of desired germplasm with selected traits. For example, genetic engineering has directly enhanced the production of scopolamine in Atropa belladonna by transforming a gene that encodes the enzyme converting L-hyoscyamine into L-scopolamine; similarly, the metabolism of other SMs has been genetically altered. Therefore, there is a gamut to isolate the genes of biosynthetic pathways and to express them either in transgenic plants or in microorganisms; thus combinatorial biosynthesis is the cutting-edge technology, where successful recombinant bacteria or yeasts are used to produce desired SMs. Undifferentiated cell cultures have often failed to produce desirable SMs, whereas differentiated organ cultures (transformed root cultures) have been successful, as cell and tissue-specific gene expression appears to control such mechanisms. The key merits of an in vitro synthesis over the traditional cultivation of whole plants are as follows:
Under traditional conditions, desired plant metabolites may take many years to reach the point where they produce SMs commercially. Alternatively, PTC techniques may be used to surpass such situations and effectively produce SM commercially.Desired biomolecules are produced under controlled aseptic conditions independent of environmental parameters (climate and soil).Obtained culture cells are microbes/ virus-free.The desired trait can be used via selective cell culture and multiplied to produce their specific metabolites.Automation of cell growth and rational regulation of SMs processes would reduce labor costs and improve productivity.Desired SMs are easily extractable from callus cultures.
