Stress-responsive Factors and Molecular Farming in Medicinal Plants 9819944791, 9789819944798

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Table of contents :
Preface
Contents
Editors and Contributors
1: An Overview of Medicinal Plants: Drugs of Tomorrow
1.1 Introduction
1.2 Indian Traditional Medicine System
1.2.1 Herbal Remedies
1.2.2 Traditional Chinese Medicine
1.2.3 European Traditional Medicine System
1.2.4 Traditional Medicine of Arabic and North African
Documents of Shanidar IV
1.3 Ethnobotany and Ethnomedicine
1.3.1 Ethnobotany
1.3.2 Ethnobotany and the Search for New Drugs
1.4 Medicinal Plants of Future Prospects
1.5 Conclusion
References
2: Medicinal Properties of the Plant Under Adverse Environmental Conditions
2.1 Introduction
2.2 Current Status and Trends of Medicinal Plants
2.3 Effect of Adverse Environmental Conditions on Medicinal Plant
2.3.1 Temperature Extremities
2.3.2 Light
2.3.3 Salinity
2.3.4 Drought
2.4 Summary
References
3: Response of Secondary Metabolites of Ocimum gratissimum L. Under Copper Stress Condition
3.1 Introduction
3.2 Materials and Methods
3.2.1 Sapling Collection, Identification, Polyhouse Experimental Setup and Extraction of Oil
3.2.2 Essential Oil Yield
3.2.3 Essential Oil Analysis and Identification of the Volatile Constituents
3.2.4 Statistical Analysis
3.3 Result and Discussion
3.3.1 Essential Oil Yield
3.3.2 Impact of Copper Stress on the Secondary Metabolites of O. gratissimum L.
3.4 Conclusion
References
4: Resilience Mechanism of Medicinal Plants Under Harsh Environment
4.1 Introduction
4.2 Abiotic Stresses in Medicinal Plants: Effects and Consequences
4.3 Effects of Drought and Heat Stress on Medicinal Plants
4.4 Salt Stress in Medicinal Plants
4.5 Heavy Metal Stress in Medicinal Plants
4.6 Ultraviolet Radiation in Medicinal Plants
4.7 Genomics Approaches to Understand Abiotic Stress Responses in Medicinal Plants
4.8 Omics as Biotechnological Tools in Medicinal Plants
4.9 Genomics in Medicinal Plants
4.10 Transcriptomics in Medicinal Plants
4.11 Proteomics in Medicinal Plants
4.12 Metabolomics in Medicinal Plants
4.13 Use of New Biotechnological Tools in the Promotion of Medicinal and Aromatic Plants´ Tolerance to Stresses: The Example o...
4.14 Final Remarks
References
5: Nature Interpretation Sites: A New Hope of Ex-situ Garden for Conservation and Cultivation of Economically Important RET MA...
5.1 Introduction
5.2 Techniques for Conservation
5.3 Ex-situ Implementation
5.4 Ex-situ Institutions and Their Contribution to In-situ Conservation
5.5 List of MAP´s Ex-situ Gardens in Uttarakhand
5.6 Conclusion
5.7 About the Institute at a Glance
References
6: Gene Expression in Medicinal Plants in Stress Conditions
6.1 Introduction
6.2 Stress Factors and Their Impacts on Medicinal Plants
6.3 Gene Expression Study Methods and Their Uses
6.4 Studies on Gene Expressions in Medicinal Plants Under Stress Conditions
6.5 Conclusion
References
7: Revealing the Epigenetic Mechanisms Underlying the Stress Response in Medicinal Plants
7.1 Introduction
7.2 Abiotic Stresses
7.2.1 Heat Stress
7.2.2 Cold Stress
7.2.3 Salt Stress
7.2.4 Water Deficit Stress
7.3 Secondary Metabolites Produced in Plants in Response to Abiotic Stress
7.4 Pharmacological Properties of Secondary Metabolites
7.4.1 Antioxidant Activity
7.4.2 Anti-Inflammatory Activity
7.4.3 Antimicrobial Activity
7.4.4 Cytotoxic Activity
7.4.5 Neuroprotective Activity
7.4.6 Cardioprotective Activity
7.5 Epigenetic Responses against Stresses
7.5.1 Epigenetic Regulation by DNA Methylation
7.5.2 Mechanism of DNA Methylation
7.5.3 RNA-Mediated DNA Methylation
7.5.4 Epigenetic Regulation by Histone Modification
7.6 Conclusions
References
8: Transcriptional Regulation in Biosynthesis of Phytochemicals in Medicinal Plants Under Stress Conditions
8.1 Introduction
8.2 Biosynthetic Pathways of SMs
8.3 Transcriptional Regulation of SM Biosynthesis Via TFs
8.3.1 WRKY TFs
8.3.2 MYB TFs
8.3.3 HLH TFs
8.3.4 bZIP TFs
8.3.5 AP2/ERF TFs
8.3.6 NAC TFs
8.4 Techniques Intricated in Delineating the Regulatory Pathways
8.5 Conclusion
8.6 Future Directions
References
9: Role of miRNA in Medicinal Plants Under Stress Condition
9.1 Introduction
9.2 miRNA Biogenesis in Plants
9.3 Role of miRNAs in Plant under Stress Condition
9.4 Abiotic Stress
9.4.1 Role of miRNAs in ABA-Mediated Stress
9.4.2 miRNAs Activity under Drought and Salt Stress
9.4.3 Action of miRNAs in Cold and Heat Stress
9.4.4 miRNAs in Response to UV-B Radiation
9.5 Biotic Stress
9.6 Conclusions and Future Perspectives
References
10: Secondary Metabolites Biosynthesis and Related Gene Expression Under Ultraviolet-B Radiation
10.1 Introduction
10.2 Regulation of Secondary Metabolites Biosynthesis Through UV-B Mediated Signaling
10.3 Effects of UV-B on Gene Expression in Relation to Phenolics
10.4 Effects of UV-B on Gene Expression in Relation to Terpenes
10.5 Effects of UV-B on Gene Expression in Relation to Alkaloids
10.6 Conclusions
References
11: Signaling Molecules in Medicinal Plants Response to Cold Stress
11.1 Introduction
11.2 Cold Acclimation Mechanisms Acquired by Medicinal Plants
11.2.1 Biotic and Abiotic Elicitors: Rudiments Guiding a Catena of Events during the Cold Stress
11.2.1.1 Osmolytes/Osmoprotectants: Combatants of Cell Wall Damage
Sugars
Proline
Glycine Betaine
Polyamines
11.2.1.2 Stress Incited Generation of ROS and Activation of Anti-Oxidant Systems: Consorted Damage and Repair Mechanisms
11.2.1.3 Intracellular Calcium Responses: Key Mediators of Cross-Tolerance
Late Embryogenesis Abundant Proteins
Antifreeze Proteins
Cold Shock Domain Proteins
11.2.1.4 Dynamic Transcriptional Cascades and Alterations in Gene Expressions Are a Cue for Cold Acclimation
11.2.1.5 Phytohormones: Chemical Messengers Transmitting Cold Signals from Source to Sink
Abscisic Acid
Gibberellin
Methyl Jasmonates
Salicylic Acid
Ethylene
11.2.1.6 Secondary Metabolites: Prerequisites for the Sanative Actions of Medicinal Plants
11.3 Conclusion
References
12: Aquaporins Gene Expression in Plants Under Stress Condition
12.1 Introduction
12.2 Discovery of Aquaporin
12.3 Stress Response of Aquaporins
12.4 Expression of Aquaporin Gene
12.5 Aquaporin Expression in Response to Stress
12.6 Integrated Functions of Aquaporins in Plant Stresses
12.7 Conclusions
References
13: Genomic Instability in Medicinal Plants in Response to Heavy Metal Stress
13.1 Introduction
13.2 Heavy Metals
13.3 Effect of Heavy Metals on Plants
13.4 Effects of Heavy Metals on Plant Enzymes
13.5 Signaling Cascade in Plants in Response to Heavy Metals
13.6 Signaling Cascades and Response to Heavy Metals
13.7 Heavy Metal Transportation, Translocation, and Sequestration
13.8 Strategies to Combat Heavy Metal Stress
13.9 Chemical Toxicity from Heavy Metals Causes Oxidative and Genotoxic Stress
13.10 Transcriptional Response of Heavy Metal Toxicity in Plants
13.11 Genetic Mechanisms Upholding Heavy Metal Tolerance
13.12 Conclusion
References
14: Proteomics Response of Medicinal Plants to Salt Stress
14.1 Introduction
14.2 Medicinal Plants Under Salt Stress
14.3 Proteomic Studies of Medicinal Plants
14.4 Proteomics Approaches in Response to Salt Stress
14.5 Molecular Mechanism of Medicinal Plants Under Salt Stress
14.6 Limitations of Proteomics Studies
14.7 Conclusion
References
15: Regulation of PGPR-Related Genes in Medicinal Plants in Adverse Conditions
15.1 Introduction
15.2 PGPR and Plants: Mechanism of Actions
15.2.1 Direct Mechanisms
15.2.1.1 Auxins
15.2.1.2 Gibberellins
15.2.1.3 Cytokinin
15.2.1.4 ACC Deaminase
15.2.1.5 Nitrogen Fixation
15.2.1.6 Siderophores
15.2.2 Indirect Mechanisms
15.3 Status of Medicinal Plants in the World
15.4 Plant Growth-Promoting Rhizobacteria: Correlation with Medicinal Plants
15.5 PGPR-Induced Phytobeneficial Genes
15.6 PGPR in Mitigating Abiotic Stress
15.7 PGPR Genomics and Status of Research
15.8 Transcriptional Regulation of Rhizobacterial Genes
15.9 Modern Approaches Followed to Understand the Regulation of PGPR Genes
15.10 Biological Control of PGPR on Medicinal Plants
15.11 Growth and Yield of Medicinal Plants Induced by PGPR
15.12 Impact of PGPR Within a Diverse Range of Medicinal Plants
15.13 Therapeutic Benefits of Medicinal Plants in Association with Rhizobacteria
15.14 Challenges Faced by Plants to Withstand the Harsh Conditions: PGPR Perspective
15.15 Conclusions and Future Prospects
References
16: Role of Phytomelatonin in Plant Tolerance Under Environmental Stress
16.1 Introduction
16.2 Phytomelatonin Synthesis and Metabolism
16.3 Phytomelatonin Signaling Pathway and Its Mechanism of Action
16.4 The Role of Phytomelatonin in Response to Abiotic Stress
16.4.1 Light as a Stress Factor
16.4.2 Cold and Heat Stress
16.4.3 Drought/Water Stress Response
16.4.4 Salt Stress Response
16.4.5 Heavy Metal Stress Response
16.5 The Role of Phytomelatonin in Response to Biotic Stress
16.5.1 Bacteria-Mediated Response
16.5.2 Fungi-Mediated Response
16.5.3 Virus-Mediated Response
16.6 Conclusion and Future Prospects
References
17: Omics´ Approaches to Analysis of Stress Response Genes in Medicinal Plants
17.1 Introduction
17.2 Plant Abiotic Stress: Omics Perspectives
17.2.1 Plant Genomic Response to Abiotic Stress
17.2.1.1 The Impact of Drought Stress on Gene Expression
17.2.1.2 Expression of Genes in Response to Salt Stress
17.2.1.3 Nutritional Stress-Induced Gene Expression
17.2.1.4 Expression of Genes in Response to Heavy Metal Stress
Structural Genomics
Functional Genomics
17.3 Transcriptomics
17.4 Proteomics
17.5 Metabolomics
17.6 Bioinformatics
17.7 Data Integration and Mining
17.8 Conclusion and Future Perspective
References
18: Next-Generation Sequencing (NGS) for Metabolomics Study in Medicinal Plants Under Stress Condition
18.1 Introduction
18.1.1 Phytochemical Profile of Medicinal Plants
18.1.2 Metabolomics Study of Phytoconstituents Synthesized Under Stress Condition (POIUS)
18.1.3 Generation of PDB Files of POIUS
18.1.4 Next-Gen Sequencing Using Whole-Genome Approach
18.1.4.1 Genome Sequencing Methods
18.1.4.2 Genome Sequencing Strategies Using NGS
18.1.5 Whole-Genome Sequencing Using NGS for POIUS of Medicinal Plants
18.1.5.1 Authentication of Herbal Supplement
18.1.5.2 Genomics, Transcriptomics, and Metabolomics of POIUS
18.1.6 WGS of Genomes of Representative Medicinal and Aromatic Plant Species with Defined Antimicrobial Activity
18.2 Conclusion
References
19: Targeted Improvement of Medicinal Plant Under Stress Condition Through CRISPR-Cas-Mediated Genome Engineering
19.1 Introduction
19.2 Types of Abiotic Stress Affecting Plant Growth
19.3 History of CRISPR
19.4 Structure and Mechanism of CRISPR-Cas
19.5 Application of CRISPR-Cas on Medicinal Plants
19.5.1 Camelina sativa (L.) Crantz
19.5.2 Salvia miltiorrhiza
19.5.3 Dendrobium officinale Kimura and Migo
19.5.4 Cannabis sativa
19.5.5 Nicotiana tabacum
19.5.6 Papaver somniferum
19.5.7 Comfrey (Symphytum officinale)
19.5.8 Chrysanthemum morifolium
19.5.9 Grewia asiatica
19.6 Advantage and Disadvantage of CRISPR-Cas in Abiotic Stress
19.7 Conclusion
19.8 Future Prospective
References
20: Molecular Farming of Medicinal Plants in Face of Environmental Challenges
20.1 Introduction
20.2 Medicinal Plants
20.3 Medicinal Plant and Biotechnology
20.4 Threats for Medicinal Plants: Destruction of Habitat, Bioprospecting, and Overharvesting
20.5 Habitat Destruction
20.6 Bioprospecting and ``Biopiracy´´
20.7 Medicinal Plants in Industrialized Societies
20.8 Effect of Climate on Medicinal Plant
20.9 Examples of Endangered Species
20.10 Medicinal Plant Information Databases
20.11 Molecular Farming
20.12 History of Molecular Farming (Fig. 20.3)
20.13 Metabolomics and Metabolite Engineering
20.14 Technology for Hairy Root Culture
20.15 Production of Pharmaceutical Compounds and Their Prospective Applications
20.16 Production of Plant-Derived Antibodies
20.17 Plant-Derived Human and Animal Vaccines for Immunotherapy
20.18 Pharmaceutical and Nutraceutical Protein Production
20.19 Industrial Enzymes and Other Molecules for Industrial Use
20.20 Human Proteins and Biopharmaceuticals Generated from Plants
20.21 Conclusion
References
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Divya Singh Amit Kumar Mishra Akhileshwar Kumar Srivastava   Editors

Stress-responsive Factors and Molecular Farming in Medicinal Plants

Stress-responsive Factors and Molecular Farming in Medicinal Plants

Divya Singh • Amit Kumar Mishra • Akhileshwar Kumar Srivastava Editors

Stress-responsive Factors and Molecular Farming in Medicinal Plants

Editors Divya Singh Department of Mulberry Physiology Central Sericultural Research and Training Institute Mysore, India

Amit Kumar Mishra Department of Botany Mizoram University Aizawl, India

Akhileshwar Kumar Srivastava Department of Plant Cell Biotechnology Central Food Technological Research Institute Mysore, India

ISBN 978-981-99-4480-4 ISBN 978-981-99-4479-8 https://doi.org/10.1007/978-981-99-4480-4

(eBook)

# The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 This work is subject to copyright. All rights are solely and exclusively licensed by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors, and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, expressed or implied, with respect to the material contained herein or for any errors or omissions that may have been made. The publisher remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. This Springer imprint is published by the registered company Springer Nature Singapore Pte Ltd. The registered company address is: 152 Beach Road, #21-01/04 Gateway East, Singapore 189721, Singapore

Preface

Plants have provided all major requirements for human life such as shelter, clothing, food, flavors, and fragrances including medicines. Plants have made the basis of traditional medicine systems like Ayurvedic, Unani, Chinese, and others. Such traditional systems of medicine have provided to some crucial drugs that are still used today for the treatment of several ailments. Medicinal plants, a source of various phytochemical components, are currently faced with different environmental stresses during their growth and development. Several ecologically limiting factors such as temperature, carbon dioxide, lighting, ozone, soil water, soil salinity, and soil fertility have consequential effect on secondary metabolic processes of medicinal plants whereas the secondary metabolites are used as vital naturally derived drugs such as immune suppressants, antibiotics, antidiabetics, and anticancer agents. Plants are capable of synthesizing different kinds of secondary metabolites to combat the adverse impacts of stress. At present, the harsh environmental conditions negatively impact the sustainable potentiality of the plant system. For instance, the continuous increase in global temperature every year exposes plants to high heat waves during their entire life cycle. In the near future, it is expected that the integration of drought, salt stress, and heat stresses at the global level might severely impact the health of plants and their gross yield. Several metabolic pathways of plants and their products are impacted by climate changes with subsequent changes in quality. Overall, under adverse conditions, plants are obligated to adapt to environmental stresses and withstand the environmental changes, which in turn alter their physiological, metabolic, and molecular functions. They also protect and improve the physical development of plants even under conditions of water scarcity, soil salinity, temperature variation, and metal toxicity. Therefore, understanding the response of environmental stress signals and the balance between defensive and developing mechanism in plants establishes the basis for improving plant resilience. The book explains the molecular resilience of medicinal plants in adverse environmental conditions by explaining the medicinal properties under harsh conditions (such as UV-B, water deficit, cold stress, heavy metals, and salt stress), importance of gene expressions, epigenetic regulation, the role of melatonin, gene regulation for PGPR status of aquaporins-related genes, genomic instability under salt stress, cell signaling in cold stress, and the role of miRNA in plant adaptation. In addition, some v

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of the chapters have been designed to describe the next-generation sequencing of genes and the CRISPR/Cas editing tool associated with metabolic pathways for molecular farming of medicinal plants to overcome future challenges from environmental stress. The current perceptive on the relevant information explained in the book will be able to fulfill the requirements of students, researchers, and scientists from diverse fields, such as environmentalists, agricultural scientists, and pharmacologists working in the areas of environmental stress and medicinal plants. Mysore, India Aizawl, India Mysore, India

Divya Singh Amit Kumar Mishra Akhileshwar Kumar Srivastava

Contents

1

An Overview of Medicinal Plants: Drugs of Tomorrow . . . . . . . . . . Ramesh Kumar Ahirwar

2

Medicinal Properties of the Plant Under Adverse Environmental Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Abhijeet Mahana

17

Response of Secondary Metabolites of Ocimum gratissimum L. Under Copper Stress Condition . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mamta Bisht, Geeta Tewari, Chitra Pande, and Ankit Joshi

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Resilience Mechanism of Medicinal Plants Under Harsh Environment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Tiago Benedito dos Santos, Silvia Graciele Hülse de Souza, Hélida Mara Magalhães, Ilara Gabriela Frasson Budzinski, and Ana Cláudia Pacheco Santos Nature Interpretation Sites: A New Hope of Ex-situ Garden for Conservation and Cultivation of Economically Important RET MAPs in Higher Himalayan Regions . . . . . . . . . . . . . . . . . . . . . . . . Jaidev Chauhan, Vijay Kant Purohit, Pradeep Dobhal, Rajeev Ranjan Kumar, Ajay Hemdan, P. Prasad, and M. C. Nautiyal

1

43

69

6

Gene Expression in Medicinal Plants in Stress Conditions . . . . . . . Sadashivaiah, L. Sunil, and R. Chandrakanth

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Revealing the Epigenetic Mechanisms Underlying the Stress Response in Medicinal Plants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107 Chandrashekhar Singh, Rajesh Saini, Richa Upadhyay, and Kavindra Nath Tiwari

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Transcriptional Regulation in Biosynthesis of Phytochemicals in Medicinal Plants Under Stress Conditions . . . . . . . . . . . . . . . . . . . . 123 Akruti Gupta, Kishore Kumar Gupta, Sanjay Kumar Gupta, and Prashant Kumar Mishra

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Contents

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Role of miRNA in Medicinal Plants Under Stress Condition . . . . . . 141 Akhileshwar Kumar Srivastava, Ishita Chatterjee, Shreshtha Mishra, Vaishnavi Tripathi, Wafia Zehra, Khushboo Chakrwal, and Vibha Agrawal

10

Secondary Metabolites Biosynthesis and Related Gene Expression Under Ultraviolet-B Radiation . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155 Avantika Pandey, Deepanshi Jaiswal, Madhoolika Agrawal, and Shashi Bhushan Agrawal

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Signaling Molecules in Medicinal Plants Response to Cold Stress . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169 Guru Kumar Dugganaboyana, Sahana Shivaramakrishna, Jajur Ramanna Kumar, Gopalakrishnan Velliyur Kanniappan, Chethan Kumar Mukunda, and Rathi Muthaiyan Ahalliya

12

Aquaporins Gene Expression in Plants Under Stress Condition . . . 193 Aradhana Mishra, Preksha Jaiswal, Akhileshwar Kumar Srivastava, and Divya Singh

13

Genomic Instability in Medicinal Plants in Response to Heavy Metal Stress . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207 W. Jabez Osborne and Shivangi Sharma

14

Proteomics Response of Medicinal Plants to Salt Stress . . . . . . . . . . 227 L. Sunil, Sadashivaiah, R. Chandrakanth, and Darshan Dorairaj

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Regulation of PGPR-Related Genes in Medicinal Plants in Adverse Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 243 Kanti Kiran and Gunjan Sharma

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Role of Phytomelatonin in Plant Tolerance Under Environmental Stress . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 275 Sachin Kumar, Akanksha Pandey, Monika Singh, Sudhanshu Mishra, Sandeep Kumar, Navneet Bithel, and Minakshi Rajput

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Omics’ Approaches to Analysis of Stress Response Genes in Medicinal Plants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303 Diksha Sharma

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Next-Generation Sequencing (NGS) for Metabolomics Study in Medicinal Plants Under Stress Condition . . . . . . . . . . . . . . . . . . . . 323 Smaranika Pattnaik

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Targeted Improvement of Medicinal Plant Under Stress Condition Through CRISPR-Cas-Mediated Genome Engineering . . . . . . . . . . 345 Priyanka Shah, Priya Patel, Manisha Hariwal, Shweta Verma, Rahul Yadav, and Sanjay Kumar

Contents

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Molecular Farming of Medicinal Plants in Face of Environmental Challenges . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359 Priya Patel, Priyanka Shah, Manisha Hariwal, Shweta Verma, Rahul Yadav, and Sanjay Kumar

Editors and Contributors

About the Editors Divya Singh is presently working as Scientist-B in the Department of Mulberry Physiology, CSRTI, Mysore, India. Her research specialization is primarily in genetics, proteomics, metabolomics, toxicology, bioinformatics, and molecular biological evaluation in various plant models to elucidate the physiological changes under stress condition. In her 11 years of research experiences, she has published nine peer-reviewed publications, two books (one edited and one authored) including six book chapters, and several presentations at local and international scientific meetings. Amit Kumar Mishra is currently working as assistant professor in the Department of Botany at Mizoram University, Aizawl, India. He has 12 years of research experience in physiological and molecular mechanisms of plant responses to high carbon dioxide levels, tropospheric ozone, drought, heat, nitrogen limitation, ultraviolet-B radiation, and other abiotic stresses. He has published 13 research articles in reputed journals and one edited book including three chapters during his research work. He has also visited Ben-Gurion University of the Negev, Israel, and Texas A&M University, USA. Akhileshwar Kumar Srivastava works as a Research Associate (ICMR) in CSIRCentral Food Technological Research Institute, Mysore, India. In his 11 years of research, he has published more than 17 research articles in international journals of repute and two books (Elsevier publication) including nine book chapters. His research specialization is primarily in the area of pharmacognosy with genetics, metabolomics, bioinformatics, and molecular biology-associated targeting virulent factors of diseases. Also, he has studied at Augusta University (formerly, Georgia Regents University) in Augusta, GA, USA, on a J-1 Exchange Scholar Visa and at Ben-Gurion University, Israel. He is also a life member fellow in Indian Science Congress and Agriculture, Nutrition and Health Academy, UK.

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Editors and Contributors

Contributors Madhoolika Agrawal Laboratory of Air Pollution and Global Climate Change, Department of Botany, Institute of Science, Banaras Hindu University, Varanasi, India Shashi Bhushan Agrawal Laboratory of Air Pollution and Global Climate Change, Department of Botany, Institute of Science, Banaras Hindu University, Varanasi, India Vibha Agrawal Department of Biotechnology, Shri Agrasen Kanya P. G. College, Varanasi, India Rathi Muthaiyan Ahalliya Department of Biochemistry, Karpagam Academy of Higher Education, Coimbatore, Tamil Nadu, India Ramesh Kumar Ahirwar Department of Botany, Guru Vishwavidyalaya (A Central University), Bilaspur, Chhattisgarh, India

Ghasidas

Mamta Bisht Department of Chemistry, School of Applied & Life Sciences, Uttaranchal University, Dehradun, India Department of Chemistry, D. S. B. Campus, Kumaun University, Nainital, India Navneet Bithel Department of Botany and Microbiology, Gurukula Kangri (Deemed to be University), Haridwar, Uttarakhand, India Ilara Gabriela Frasson Budzinski Department of Genetics, Max Feffer Laboratory of Plant Genetics, “Luiz de Queiroz” College of Agriculture, University of São Paulo, Piracicaba, SP, Brazil Khushboo Chakrwal Department of Biotechnology, Shri Agrasen Kanya P. G. College, Varanasi, India R. Chandrakanth Department of Molecular Biology, Yuvaraja’s College, University of Mysore, Mysuru, Karnataka, India Ishita Chatterjee Department of Biotechnology, Shri Agrasen Kanya P. G. College, Varanasi, India Jaidev Chauhan High Altitude Plant Physiology Research Centre, HNB Garhwal University, Srinagar Garhwal, Uttarakhand, India Pradeep Dobhal High Altitude Plant Physiology Research Centre, HNB Garhwal University, Srinagar Garhwal, Uttarakhand, India Darshan Dorairaj Department of Plant Cell Biotechnology, CSIR-Central Food Technological Research Institute, Mysuru, India Akruti Gupta Vinoba Bhave University, Hazaribag, Jharkhand, India ICAR-Indian Institute of Agricultural Biotechnology, Ranchi, India

Editors and Contributors

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Kishore Kumar Gupta Vinoba Bhave University, Hazaribag, Jharkhand, India Sanjay Kumar Gupta ICAR-Indian Institute of Agricultural Biotechnology, Ranchi, India Manisha Hariwal Department of Botany, Banaras Hindu University, Varanasi, India Ajay Hemdan High Altitude Plant Physiology Research Centre, HNB Garhwal University, Srinagar Garhwal, Uttarakhand, India Deepanshi Jaiswal Department of Botany, Mahatma Gandhi Balika Vidyalaya (P.G.) College, Firozabad, India Preksha Jaiswal Babasaheb Bhimrao Ambedkar Central University, Lucknow, India Ankit Joshi Department of Chemistry, School of Applied & Life Sciences, Uttaranchal University, Dehradun, India Gopalakrishnan Velliyur Kanniappan School of Medicine, Bule Hora University Institute of Health, Bule Hora University, Bule Hora, Ethiopia Kanti Kiran Department of Plant Biotechnology, Gujarat Biotechnology University, Gandhinagar, Gujarat, India Dugganaboyana Guru Kumar Division of Biochemistry, School of Life Sciences, Mysuru, JSS Academy of Higher Education and Research (Deemed to be University), Mysuru, Karnataka, India Jajur Ramanna Kumar Division of Biochemistry, School of Life Sciences, Mysuru, JSS Academy of Higher Education and Research (Deemed to be University), Mysuru, Karnataka, India Rajeev Ranjan Kumar High Altitude Plant Physiology Research Centre, HNB Garhwal University, Srinagar Garhwal, Uttarakhand, India Sachin Kumar Department of Botany and Microbiology, Gurukula Kangri (Deemed to be University), Haridwar, Uttarakhand, India Sandeep Kumar Department of Botany and Microbiology, Gurukula Kangri (Deemed to be University), Haridwar, Uttarakhand, India Sanjay Kumar Department of Botany, Banaras Hindu University, Varanasi, India Hélida Mara Magalhães Postgraduate Program in Biotechnology Applied to Agriculture, Universidade Paranaense (UNIPAR), Umuarama, PR, Brazil Abhijeet Mahana French Associates Institutes for Agriculture and Biotechnology of Drylands, Jacob Blaustein Institute for Desert Research (BIDR), Ben-Gurion University of the Negev, Negev, Israel Aradhana Mishra Veer Bahadur Singh Purvanchal University, Jaunpur, India

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Editors and Contributors

Prashant Kumar Mishra Vinoba Bhave University, Hazaribag, Jharkhand, India Shreshtha Mishra Department of Biotechnology, Shri Agrasen Kanya P. G. College, Varanasi, India Sudhanshu Mishra Department of Biotechnology, School of Applied and Life Sciences, Uttaranchal University, Dehradun, Uttarakhand, India Chethan Kumar Mukunda Department of Biochemistry, JSS College of Arts Commerce and Science, Mysore, Karnataka, India M. C. Nautiyal High Altitude Plant Physiology Research Centre, HNB Garhwal University, Srinagar Garhwal, Uttarakhand, India W. Jabez Osborne Department of Biosciences, School of Bio Sciences and Technology, Vellore Institute of Technology, Vellore, Tamil Nadu, India Chitra Pande Department of Chemistry, D. S. B. Campus, Kumaun University, Nainital, India Akanksha Pandey Department of Botany and Microbiology, Gurukula Kangri (Deemed to be University), Haridwar, Uttarakhand, India Avantika Pandey Laboratory of Air Pollution and Global Climate Change, Department of Botany, Institute of Science, Banaras Hindu University, Varanasi, India Priya Patel Department of Botany, Banaras Hindu University, Varanasi, India Smaranika Pattnaik Laboratory of Medical Microbiology, School of Life Sciences, Sambalpur University, Sambalpur, India P. Prasad High Altitude Plant Physiology Research Centre, HNB Garhwal University, Srinagar Garhwal, Uttarakhand, India Vijay Kant Purohit High Altitude Plant Physiology Research Centre, HNB Garhwal University, Srinagar Garhwal, Uttarakhand, India Minakshi Rajput Department of Biotechnology, School of Applied and Life Sciences, Uttaranchal University, Dehradun, Uttarakhand, India Sadashivaiah Department of Molecular Biology, Yuvaraja’s College, University of Mysore, Mysuru, Karnataka, India Rajesh Saini Department of Botany, MMV, Banaras Hindu University, Varanasi, India Ana Cláudia Pacheco Santos Postgraduate Program in Agronomy, Universidade do Oeste Paulista (Unoeste), Presidente Prudente, SP, Brazil Tiago Benedito dos Santos Postgraduate Program in Agronomy, Universidade do Oeste Paulista (Unoeste), Presidente Prudente, SP, Brazil Priyanka Shah Department of Botany, Banaras Hindu University, Varanasi, India

Editors and Contributors

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Diksha Sharma School of Pharmacy, IEC University, Solan, Himachal Pradesh, India Gunjan Sharma Department of Plant Biotechnology, Gujarat Biotechnology University, Gandhinagar, Gujarat, India Shivangi Sharma Department of Biosciences, School of Bio Sciences and Technology, Vellore Institute of Technology, Vellore, Tamil Nadu, India Sahana Shivaramakrishna Division of Biochemistry, School of Life Sciences, Mysuru, JSS Academy of Higher Education and Research (Deemed to be University), Mysuru, Karnataka, India Chandrashekhar Singh Department of Pharmaceutical Engineering and Technology, Indian Institute of Technology, BHU, Varanasi, India Divya Singh Department of Mulberry Physiology, Central Sericultural Research and Training Institute, Mysore, India Monika Singh Department of Biotechnology, School of Applied and Life Sciences, Uttaranchal University, Dehradun, Uttarakhand, India Silvia Graciele Hülse de Souza Postgraduate Program in Biotechnology Applied to Agriculture, Universidade Paranaense (UNIPAR), Umuarama, PR, Brazil Akhileshwar Kumar Srivastava Department of Plant Cell Biotechnology, Central Food Technological Research Institute, Mysore, India L. Sunil Department of Plant Cell Biotechnology, CSIR-Central Food Technological Research Institute, Mysuru, India Geeta Tewari Department of Chemistry, D. S. B. Campus, Kumaun University, Nainital, India Kavindra Nath Tiwari Department of Botany, MMV, Banaras Hindu University, Varanasi, India Vaishnavi Tripathi Department of Biotechnology, Shri Agrasen Kanya P. G. College, Varanasi, India Richa Upadhyay Department of Botany, Mihir Bhoj Postgraduate College, G.B. Nagar, India Shweta Verma Department of Botany, Banaras Hindu University, Varanasi, India Rahul Yadav Department of Botany, Banaras Hindu University, Varanasi, India Wafia Zehra Department of Biotechnology, Shri Agrasen Kanya P. G. College, Varanasi, India

1

An Overview of Medicinal Plants: Drugs of Tomorrow Ramesh Kumar Ahirwar

Abstract

Instead of providing medication, plants have always provided humans with food, clothing, shelter and other basic requirements. Complex traditional medical systems, such as Ayurvedic, Unani, Chinese and others, have their roots in plants. Some of the most significant medications that are still used today were developed by these medical systems. African, Australian, Central and South American medical systems are among the lesser-known medical systems. Today, the search for novel molecules has gone a somewhat different route, with chemists employing the study of ethnography and ethnopharmacology as a guide to find new sources and classes of chemicals. Due to its richness, the flora of the tropics plays a significant role in offering fresh hints in this context. However, the Convention on Biological Diversity must also address the issues of sovereignty and property rights. This page emphasises this, offers an overview of plant molecule types and gives examples of molecules and secondary metabolites that have contributed to the creation of these pharmacologically potent extracts. The publication also discusses the necessity for plant extract validation and some information on the use of herbal products in the creation of functional foods, while consistently emphasizing the safety, effectiveness and quality of phytopharmaceutical products. Keywords

Traditional medicine · Ayurveda · Herbal medicine · Phytochemicals · Drug development · Medicinal plants R. K. Ahirwar (✉) Department of Botany, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur, Chhattisgarh, India e-mail: [email protected] # The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 D. Singh et al. (eds.), Stress-responsive Factors and Molecular Farming in Medicinal Plants, https://doi.org/10.1007/978-981-99-4480-4_1

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R. K. Ahirwar

Introduction

Humans have historically depended on nature to provide for their fundamental necessities, including the creation of food, housing, clothes, transportation, fertilizers, flea pores, fragrances and at least medicine. For thousands of years, comprehensive traditional medicinal systems have been built on the foundation of plants, and these systems still provide the human race cutting-edge therapies today. Although some of the therapeutic properties attributed to plants have been proven wrong, medicinal plant therapy is based on hundreds and thousands of years of empirical discoveries (Gurib-Fakim 2006). The oils of the Cedrus species (Cedar), Glycieriza glabra (Mulethi) and Papavar somniferum (Poppy juice) are still used today to treat conditions ranging from coughs and colds to cross-acetic infections and inflammation. The earliest written records, which are on cuneiform clay tablets, are from Mesopotamia and date back to about 2600 BC. According to Egyptian medical literature, bishop’s herb (Ammi majus) can be used to cure vitiligo, a skin disorder that is characterised by the loss of pigment. This plant has recently given rise to a medication (β-methoxyporalen) that is used to treat T-cell lymphoma, psoriasis and other skin conditions. The search for possible chemotherapeutic drugs in nature is still a topic of interest. More than 50% of all medications used in clinical settings across the world are made from natural materials or their derivatives. No less than 25% of the total is provided by the tallest plants. At least a dozen powerful medications have been made from flowering plants over the past 40 years, including the diosgenins made by Dioscorea spp., which are the source of all anovulatory tracers; Reserpine and other anti-hypertensive alkaloids and quilted by Rouwolphia spp.; Pilocarpin spp., which is used to treat glaucoma and “dry mouth,” made by a group of South Tropical woods are home to about half (125,000) of all kinds of flowering plants. A substantial pool of potential medicinal species continues to be supported by tropical rainforests. They continue to give natural product chemists essential compounds that can be used as building blocks for the creation of new drugs. Only 1% of tropical species have had their pharmacological potential studied thus far, which greatly increases the potential to discover new compounds. For species that are only found in tropical rainforests, this ratio is even lower. About 50 drugs have been derived from tropical plants so far. High annual extinction rates are a concern because one of the most significant justifications for protecting tropical forests has been associated with the existence of drugs unrecognised by modern medicine. Three primary sources of anticancer medications on the market have been derived from North American plants that have undergone clinical testing and have been utilised medicinally by Native Americans, despite their discovery through a sudden laboratory observation. These sources are Papav (Asimina spp.) and Western Yow (Taxus brevifolia), useful in the treatment of leukaemia and ovarian cancer, testicular cancer and pulmonary lymphoma.

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An Overview of Medicinal Plants: Drugs of Tomorrow

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Indian Traditional Medicine System

Perhaps the oldest medical system is Ayurveda. It is regarded as systemic medicine’s inception. In actuality, it is a realistic and universal manual for preserving harmony and balance in the system. Many of the theories attributed to Dioscorides, who inspired Hippocrates, are said to have originated in India. While Greek and Middle Eastern books discuss concepts and medications of an Indian provenance, ancient Hindu works on medicine do not mention foreign medicine. The Indian terms Ayar (life) and Veda are the origin of the name Ayurveda (knowledge or science). “Knowledge of life” is what it implies from here (Dharma). In India, songs and poetry that academics and doctors had to memorise and recite verbatim were used to transmit knowledge and wisdom. The Sama Veda, Yajur Veda and Atharva Veda are among the oldest, dating back to the year 2000 BC. In the first century BC, the first school that taught Ayurvedic medicine was established. The Great Samhita was authored about 500 AD at the University of Banara (or the Encyclopedia of Medicine). Ayurveda is comparable with galenic medicine as it is based on bodily humour (dosas) and inner life energy (prana), which are thought to support digestion and mental function. Earth (Prithvi), water (Jada), air (Vaju) and space (Akase) make up both inhabited and unoccupied surroundings, including man. Understanding the concepts of impurity and cleansing is crucial to understanding these traditions. Between various factors, disease is unequal. Popular Neem (Azadirachta indica), Centella asiatica (Gotu Kola), Cinnamomum camphora (Camphor), Elettaria cardamom, Rauwolfia serpentina (Indian snakeroot), Santalum album (Sandalwood), Terminalia kinds (Myrobolan) and Withania somnifera (Aswargandha).

1.2.1

Herbal Remedies

The use of plants for medical and therapeutic purposes to treat illness and enhance human health is known as herbal medicine or phytomedicine. Phytochemicals are secondary metabolites found in plants. Phyto is a Greek word that means “plant.” These substances shield plants against microbial diseases or insect infestations. Active components, known as phytochemicals, are substances that are used as drugs or medicines because they have therapeutic characteristics. Phytochemicals can be categorised according to their chemical makeup (Table 1.1, Fig. 1.1). Plants that are used as food and in traditional medicine are more likely to produce chemicals that have pharmacological activity. The standardization of raw materials comes as a significant problem for the herbal sector when the quality of an herbal product is questioned. During growth, processing and harvest, herbal plants are readily polluted. The two main issues with herbal medications that have been observed are adulteration and heavy metal poisoning. Therefore, in order to produce novel herbal drugs and stay up with other efforts in drug development, it is vital to increase the

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Table 1.1 Phytochemicals classification

S. No. 1.

Phytochemicals name Alkaloids

Name of chemicals Caffeine

2.

Glycosides

Andrographolide

3.

Polyphenols (phenolics, tannins and flavonoids)

Resveratrol

4. 5.

Saponins Terpenes (steroids and carotenoids) Anthraquinones

Diosgenin Artemisinin

6.

Rhein

Examples Morphine, caffeine, berberine and codeine Amygdalin, gentiopicrin, morpholine, polydalin and cinnamyl acetate Caffeic acid, flavones, rutin, naringin, hesperidin, chlorogenic, tannic, gallic, ellagic acids, as well as quercetin, resveratrol, kaempferol and quercitrin Hecogenin and diosgenin Lycopene, lutein, zeaxanthin, β-carotene, -carotene and artemisinin Rhein, salinos poramide and luteolin

Structure of chemicals Figure 1.1a Figure 1.1b

Figure 1.1c

Figure 1.1d Figure 1.1e

Figure 1.1f

Fig. 1.1 Chemical structure. (a) Caffeine; (b) andrographolide; (c) resveratrol; (d) diosgenin; (e) artemisinin; (f) rhein

quality and quantity of bioactive components. There are many therapeutic plants available today, and researchers have examined the bioactive components of these plants on a scientific and clinical level. Table 1.2 provides a list of plant species and the active ingredients that are used to treat various human disorders (Table 1.2) (Shakya 2016).

Aloe vera (L.) Burm. f.

Artemisia absinthium L. Azadirachta indica A. Juss.

Berberis vulgaris L. Bergenia ciliata (Haw.) Sternb.

2.

3.

5.

Catharanthus roseus (L.) G. Don Cinchona rugosa Pav. Curcuma longa L.

8.

11.

10.

9.

Camptotheca acuminata Decne.

7.

6.

4.

Scientific name Allium sativum L.

S. No. 1.

Haldi

Cinchona

Sadabahar

Frilly Bergenia/ hairy bergenia Happy tree

Barberry

Absinthe or wormwood Neem

Common name Lahsun/ garlic Gwarpatha/ Ghritkumari

Plantaginaceae

Zingiberaceae

Rubiaceae

Apocynaceae

Cornaceae

Saxifragaceae

Berberidaceae

Meliaceae

Compositae

Xanthorrhoeaceae

Family Amaryllidaceae

Table 1.2 Bioactive compounds of medicinal plants top 25

Digoxin

Flavonoid (Curcumin)

Alkaloid (Vinblastine and Vincristine) quinine

Irinotecan and Topotecan

Bergenin

Berberine

Di- and tri-terpenoids, limonoids (nimbidinin)

Artemisinin

Campesterol, aloin and emodin and β-Sisosterol

Bioactive compound Allicin

Anticancer, anti-inflammatory, hepatoprotective Used in heart diseases.

Antimalerial, antiparasitic effect

Anticancer agents (for the treatment of small cell lung and ovarian cancers) Anticancer

Chemo preventive, anti-colorectal cancer and carcinoma inhibitors purifier of blood Antidiabetic, hepatoprotective, antimicrobial. Anti-arthritis

Pharmacological action Cardioprotective, antiinflammatory Healing capabilities, anti-viral and anti-tumour properties, Antidiabetic, Hepatoprotective, effect of antiseptic Antimalarial drug

(continued)

Gajalakshmi et al. (2013) Titanji et al. (2008) Akram et al. (2010)

Li and Zhang (2014)

Rahimi-Madiseh et al. (2017) Chauhan et al. (2012)

Krishna et al. (2004) Biswas et al. (2002)

References Singh and Singh (2008) Sahu et al. (2013)

1 An Overview of Medicinal Plants: Drugs of Tomorrow 5

Ocimum tenuiflorum L.

Phyllanthus emblica L.

Piper nigrum L.

Podophyllum peltatum L. Ricinus communis L.

13.

14.

15.

16.

19.

18.

Silybum marianum (L.) Gaertn. Swertia chirayita (Roxb.) Buch.-

Nigella sativa L.

12.

17.

Scientific name Digitalis lanata Ehrh.

S. No.

Table 1.2 (continued)

Chiretta

Castor bean/ castor oil plant Milk thistle

Black pepper/ Kalimirch Mayapples

Amla

Tulsi

Common name Grecian foxglove/ woolly Foxglove Fennel flower/black cumin

Gentianaceae

Compositae

Euphorbiaceae

Berberidaceae

Piperaceae

Phyllanthaceae

Lamiaceae

Ranunculaceae

Family

Mangeferin, amarogenitine, ophelic acid and sawertiamarine

Silibinin (Flavonoid silymarin)

Alkaloid (Etoposide and Teniposide) Ricinine, an alkaloid and lectin (ricin)

Punigluconin, pedunculagin, emblicanin A and emblicanin B Piperidine, dehydropipernonaline

Taxol, Ursolic acid Apigenin and Citral

Thymoquinone

Bioactive compound

Das et al. (2008)

Anti-inflammatory, anti-cancer and liver tonic for hepatic diseases Antiviral, anti-diabetic and hepato-renal protective

Joshi and Dhawan (2005)

Jena and Gupta (2012)

Sharma (2013)

Ahmad et al. (2012)

Khan (2009)

Pradhan et al. (2022)

Paarakh (2010a)

References Negi et al. (2012)

Anti-tumour, hypoglycemic, antioxidant and hepatoprotective

Anticancer agents

Antidiabetic, anticancer, antimicrobial, Hepatorenalprotective and gastroprotective Anti-bacterial, anti-fungal, antibacterial, anti-pyretic and anticancer properties Hepatoprotective, anti-diabetic, anti-cancer, anti-microbial, antiviral and anti-cancer Anti-carcinogenic, antihyperlipidaemic, Epilepsy

Pharmacological action

6 R. K. Ahirwar

Tinospora sinensis (Lour.) Merr.

Withania somnifera (L.) Dunal

Zingiber officinale Roscoe

23.

24.

25.

22.

21.

20.

Ham. ex C. B. Clarke Taxus brevifolia Nutt. Terminalia arjuna (Roxb. ex DC.) Wight and Arn. Terminalia chebula Retz.

Zingiberaceae

Solanaceae

Ashagwanda

Ginger

Menispermaceae

Giloy

Combretaceae

Combretaceae

Arjuna or arjun tree

Harra

Taxaceae

Pacific yew

Zingerone, gingerols and mono but also sesquiterpenoids

Tannins, gallic acid, arjunic acid, tannic acid, tannins, saponins and phytosterols Triterpenoids, ellagic acid and derivatives of the shikimic acid Diterpenoid furanolactones (tinosporin), isoquinoline alkaloids Withanolides, particularly withaferin A and steroidal lactones

Taxol

Immunomodulator, chemopreventive, Cardioprotective, Antidiabetic Parkinson’s and Alzheimer’s disease are treated with immunomodulatory, cancerpreventive, memory-enhancing drugs and chemotherapy Hypercholesterolemic, antioxidant, hepatoprotective, anti-atherosclerotic, anticancerous

Gupta and Sharma (2014)

Umadevi et al. (2012)

Rathinamoorthy and Thilagavathi (2014) Mittal et al. (2014)

Paarakh (2010b)

Hepatoprotective, anticancer and cardioprotective Hepatoprotective, renoprotective, anti-diabetic, antioxidant

Rates (2001)

Anti-tumour

1 An Overview of Medicinal Plants: Drugs of Tomorrow 7

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R. K. Ahirwar

Traditional Chinese Medicine

When European cultures evolved with only some camouflage, Chinese and Indian civilizations grew and prospered. We anticipate seeing a lot of studies on both the beauty of greenery and therapeutic plants. This more than 5000-year-old medical system is founded on two distinct beliefs concerning the natural principles of health and longevity, namely, yin and yang and the five elements (wu xing). Herbs were mentioned by the legendary Emperor Shennong in his writings, which may have been penned about 2500 BP rather than the conventional 3500 BP, when Chinese medicine was systematised and recorded in 100 and 200 BP, nearly 6000 medications are listed; 4800 of which have a botanical origin. The opposites of yin and yang are complementary to one another. The four humours and elements of the Greeks as well as the three humours of Ayurveda are analogous to the five-element theory. Earth, metal, water, wood and fire are the five elements. They are related to the body’s five major organ systems (spleen, lungs, kidneys, heart and liver), emotions (reflection, sadness, fear, anger and joy), climate (wet, dry, cold, wind and cold), seasons (late summer, autumn, spring and summer) and taste (sweet, spicy, salty, sour and bitter), among other things. There are yin and yang aspects to the harmony between these components and the supply of life force (qi). Therefore, in addition to symptoms, therapy is based on an imbalanced image that is typically seen while taking the pulse or looking at the patient’s tongue. Herbs that are warm or hot, like ginger and cinnamon, are used to alleviate ailments like chilly hands, dyspepsia and stomach pain that are connected to cold symptoms. Chinese herbal medicine is often sold in set combinations or recipes of up to 20 herbs, just as Western and African traditional remedies. These recipes are painstakingly produced in accordance with age-old compendiums of traditional Chinese cooking. Such prescriptions are widely utilised in Western medicine and number in the hundreds. Similar to other medical cultures, traditional formulae are often employed to treat chronic illnesses, and Western medicine is utilised to treat acute or life-threatening illnesses. Unquestionably, the popularity of herbal medicines today is a result of Chinese medicine’s dissemination across the majority of the world’s continents. The polymorphic form of angelica is a well-known example of a Chinese medicinal plant: Panax ginseng (renshen), Ephedra sinica (ma hang), Artemisia annua (quing hao) and Rheum palmatum (de huang).

1.2.3

European Traditional Medicine System

The rational advancement of the use of herbal remedies in the ancient West was greatly influenced by the Greeks. However, Hippocrates (460–377 BC) and Aristotle (384–322 BC) are credited with developing the medical system in Europe, and both men’s theories drew inspiration from prehistoric Indian and Egyptian beliefs. In his History of Plants, the philosopher and naturalist Theophratus (300 BC) studied therapeutic aspects of plants and emphasised how cultivation may change their

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characteristics. The collection, storage and use of herbs were documented by the Greek physician Dioscorides (100 AD) while he travelled with the Roman army. Galen (130–200 AD), a pharmacy and medicine practitioner and teacher in Rome, published no less than 30 books on these topics and is renowned for the intricate formulae and recipes he used in complex medicines, which occasionally includes hundreds of substances (“Galenicals”). Based on the idea that the world is composed of four elements—earth, wind, fire and water—Greek and Roman medicines were developed. There are four significant bodily fluids that each of them corresponds to, and each has a unique chewing gum to go with it. The four bodily fluids—blood, phlegm, black bile and yellow bile—have an impact on mood and physical health (bloody, phlegm, melancholy and bile, respectively). Bloodletting and cleaning, which reduce excess blood and black bile, are extreme methods used to re-establish equilibrium. There is a matching cold, hot, wet or dry herb for each of these four body fluids that is supposed to correct imbalances. These four biological fluids are likewise related to the four elements of cold, hot, moist and dry. On regional folk customs and traditions, European traditions have also had a significant regional influence. The famous “De Materia Medica” (Medical Materia Medica) book by the Greek physician, Dioscorides, from the first century AD, had the most impact. It is widely considered as the first herbalist in Europe and has served as the norm there for more than a millennium, serving as the inspiration for the majority of succeeding herbs. Monasteries in Central Europe cultivated medicinal plants using a regular arrangement at the beginning of the year 800 AD. Hildegard of Bingen was one of the renowned healers of her time (1098–1179). Following this, the Swiss alchemist Paracelsus (1493–1541) highlighted the need of appropriate dose. Many European nations still use herbal medicine as a common practise, and it is still well-liked as a sophisticated and logical kind of disease therapy that is frequently seen as supportive rather than curative. Herbal tea is still a fairly common beverage in many European nations today. Additionally, sophisticated plant preparations (standardised and designed plant extracts, frequently bore-tested on people) and “natural goods” ingested in their original (unprocessed) form in tea or decoction continue to be well-liked alternatives to solely synthetic chemical-derived medicines. Due to commercialization, many traditional European herbs were widely utilised. Additionally, several active chemicals were extracted from medical plants and are still in use today as standalone chemical substances.

1.2.4

Traditional Medicine of Arabic and North African

The oldest written information in the Arabic tradition comes from the Sumerians and Akkadians of Mesopotamia and therefore comes from the same region as the archaeological records of Shanidar IV (Heinrich et al. 2017).

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Documents of Shanidar IV The earliest documented record, which presumably relates to medicinal plants, dates from 60,000 BCE in the grave of the Neanderthal man from Shanidar IV, an archeological site in Iraq. Pollen of several species of plants, presumably used as medicines, was discovered among which are: Centaurea solstitialis (Asteraceae), Ephedra altissima (Ephedraceae), Althea sp. (Malvaceae) amongst others. Although this may not be a finding with no direct bearing on the culture of Shanidar, these species or closely related ones from the same genus, are still important today in the phytotherapy of Iraq and also known from other cultural traditions. These species may well be typical for the Neanderthal people and may also be part of a tradition for which Shaidar IV represents the first available record. Several of the plants that are grown now were tamed in the middle East, which is regarded as the birthplace of civilization. Cuneiform was used by the Babylonians, Assyrians and Sumerians to engrave plants on a large number of clay tablets. The Code of Hammurabi, a complete collection of civil rules engraved in stone and ordered by the Babylonian monarchs, dates to around 1700 BC and is of particular relevance. It includes a number of therapeutic plants. Egypt has kept copies of such documents for thousands of years. For tomb frescoes and papyrus from the ancient kingdom, the latter created by Cyperus aquaticus, the Egyptians employed their expertise, including medicine and medicine. The most significant of these documents is the Ebers Papyrus, which is thought to have been created about 1500 BC and includes 3000 BC-era medical knowledge. The famous 20-m-high papyrus scroll with Egyptian hieroglyphics inscribed on it is said to have been found in the tomb and named after Professor Ebers George in Terrez in 1872. When it was sent to the University of Leipzig in 1873 for storage, G. Ebers published a facsimile of it 2 years later. The Ebers Papyrus is a book utilised as a medical reference that discusses all illnesses, their symptoms and symbolic methods of treatment. Impressive diagnostic precision is described in this paper. The Arabs were in charge of preserving the majority of Greco-Roman experience and expanding it to include the use of their own resources, as well as Chinese and Ayurvedic herbs that were previously unknown to the Greco-Roman world. If monasteries in nations like England, Ireland and Germany preserved artefacts of Western knowledge in the dark and mediaeval (fifth–twelfth centuries AD), the Arabs were responsible for doing the same for Western knowledge. In the eighth century, Arabs set the precedent for the construction of independent drug stores, and Avicenna, a Persian apothecary, physician, philosopher and poet, made significant contributions to the science of pharmacy and medicine in all of his writings, including canon medicine, which is regarded as “the final codification of all Greco-Roman medicine.” The independent Islamic medical system known today as Unani-Tibb was built on the foundation of Canon medicinae, which integrated components from different healing systems. Famous medicinal

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plants of the Middle East and Egypt include: Allium cepa (Onion), Astracantha gummifera (Tragacanth), Carthamus tinctorius (Safflower), Carum carvi (Carway), Ferula assafoetida (Asofoetida), Lawsonia inrmis (Henna), Papaver somniferum (Opium poppy), Peganum harmala (Syrian rue), Prunus dulcis (Almond), Punica gratum (Pomegranate), Rosa × damascene (Damascus rose), Ricinus communies (castor oil plant), Salvadora persica (Toothbrush tree), Senna Alexandria (Senna), Sesamum indicum (Sasame), Trachyspermum ammi (Ajawaan), Trigonella foenumgraecum (Fenugreek) and Vitis vinifera (Grape).

1.3

Ethnobotany and Ethnomedicine

1.3.1

Ethnobotany

Hasberg coined the term “ethnobotany” for the first time in 1896. He described it as the research of plants utilised by indigenous and prehistoric humans. When Robbins, Harrington and Freire-Mareco proposed expanding the definition of ethnobotanical science in 1916, they suggested including the analysis of all life stages in prehistoric societies as well as the impact of the plant environment on current tribal peoples’ living traditions, beliefs and histories. Twenty-five years later (Jones 1941), came up with a more succinct definition: the study of interrelationship between plants and primitive man. Schultes expanded it in 1967 to include “the relationship between man and the surrounding vegetation.”

1.3.2

Ethnobotany and the Search for New Drugs

As mentioned above, plants form the basis of traditional medicine systems and have been used for thousands or thousands of years in countries, such as China and India (Chang and But 1986; Kapoor 1990). It is well known that numerous civilizations throughout the world employ plants as part of their traditional medical practises. These plant-based systems continue to play an important role in healthcare, with the World Health Organization estimating that 80% of the world’s population continues to rely primarily on traditional medicine systems for healthcare (World Health Organization 2008). The remaining 20% of people, the majority of whom reside in industrialised nations, likewise depend heavily on plant-based meals for their health systems. An analysis of US community pharmacy prescription data from 1959 to 1980 showed that 25% contained plant extracts or active substances from higher plants, and at least 119 chemicals from 90 plants could be considered important drugs currently in use in one or more countries (Farnsworth et al. 1985); 74% of these 119 medications were found through chemical research focused at extracting active components from plants utilised in conventional treatment. Additionally, in recent decades, there has been an upsurge in the usage of purported supplementary or alternative botanical products. French pharmacists Caventoux and Pelletier published a paper in 1820 on the isolation of the antimalarial medication quinine

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from the bark of Cinchona species, such as Cincona officinalis. Indigenous populations in the Amazon have long utilised the bark to cure fever, and it was originally brought to Europe in the early 1600s to treat malaria. Modern medicine can assess the four primary ways that indigenous peoples use plants: 1. The alkaloid D-tubocurarine, which is derived from the South American jungle liana, is one example of a plant from the tropics that is occasionally employed as a source of direct medicinal compounds. In surgery, Condodendron tomentosum is frequently used as a muscle relaxant. Chemists still rely on wildlife collecting since they are unable to synthesis and create the medication in a way that has all the qualities of a natural product. Unexpectedly, gathering therapeutic herbs is frequently less expensive than purchasing manufactured, synthetic medications. Reserpine, a significant hypertensive drug derived from Rauwolfia, is another excellent illustration of this trait. Such molecules will require three times as much synthesis energy as they do collect energy. 2. Semi-synthetic chemicals are also developed from tropical plants as a starting point. An illustration of this is the chemical alteration of saponin extracts to manufacture the saponins required to make steroid medicines. Up until recently, extracts of the genus Diocarea’s neotropical yams accounted for 95% of all steroids. 3. Tropic plants can provide a supply of raw materials that can be exploited to create novel synthetic chemicals. For the synthesis of several local anaesthetics, including procaine, cocaine from the coca plant, Erythroxylum coca, serves as a model. Plants’ novel and uncommon molecules will continue to be used as “blueprints” for novel synthetic compounds and will become more and more significant in the future. 4. Additionally, plants can be utilised as taxonomic markers to find novel chemicals. The plant kingdom is randomly ordered from the perspective of phytochemistry; certain families have received a fair amount of research, while others have received absolutely little attention at all. For instance, the Liliaceae family has a long history of medicinal usage and is well-known for its abundance of alkaloids. Orchidaceae, on the other hand, is a little understood family. The close ties between several of these plants and the Liliaceae family have been investigated. Studies have revealed that in addition to being abundant in alkaloids, several of these alkaloids are also distinct and may one day be beneficial.

1.4

Medicinal Plants of Future Prospects

Medicinal plants have a promising future as there are about half a million plants worldwide and most of them have not yet been studied for their medical activities and their hidden potential for medical activities may be important in current and future study treatments (Singh 2015). Medical plants may have had a significant part in the evolution of human civilization, including in religion and other rites. Many of the many contemporary medications, like aspirin, are made in part from medicinal

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An Overview of Medicinal Plants: Drugs of Tomorrow

13

plants. Many food crops, like garlic, have therapeutic properties. Understanding plant toxicity and safeguarding people and animals from natural poisons are made possible, thanks to the study of therapeutic plants. The secondary metabolites that plants produce are what give them their medicinal properties. In light of this, interest in studying the chemistry of natural products is rising (Dar et al. 2017). The therapeutic need, the remarkable diversity of chemical structure and biological activities of naturally occurring secondary metabolites, the usefulness of new natural bioactive compounds like biochemical screening, and the availability of novel and sensitive detection methods are some of the possible causes of this interest. Development is a part of it. The need for very pure and structurally complex natural goods is being met by improvements in biologically active natural products and better methods to isolate these active substances (Clark 1996). The World Health Organization has established protocols, rules and standards for botanical medicine in recognition of the value of conventional medicine. When growing, manufacturing and processing medicinal plants, agro-industrial processes must be used (World Health Organization 1993). Many current pharmaceuticals are created in part by plants, and medicinal plants provide a source for developing novel medications. The development of new medications uses medicinal plants as natural resources (Fig. 1.2).

Fig. 1.2 Methods for obtaining active substances from plants

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R. K. Ahirwar

Conclusion

An overview of the significance of medicinal plants throughout history is provided in this work. Pharmaceuticals have undoubtedly had a rich history, but they have developed over time to become a staple in a number of industries, including pharmaceuticals, pharmaceuticals and natural products chemistry. The value of plants as a source of medicines is now acknowledged by all these scientific fields, and they have all started active research projects to find fresh leads or create appropriate chemicals. With between 10 and 100 million species thought to exist on Earth, higher plants make up a group of about 250,000 species, only 6% of which have been studied for their biological activities and 15% for their medicinal properties. It appears that we have only begun to scratch the surface of this incredible global resource. While the pharmaceutical industry in affluent nations will continue to look into potential leads in natural goods in an effort to produce new medicines, the creation of new medicines in underdeveloped nations may have completely different objectives. In some regions of the world, if a plant is easily accessible and capable of offering a cheap therapy for a disease, something may very well be done. It is anticipated that close cooperation between medical professionals and scientists would result in goods that are reliable, high-calibre and efficient. Pharmacognosy, a discipline or science that was formerly seen as being inactive, has a highly promising future since it will continue to produce novel molecular leads for the most important diseases we are currently facing. There is a lot of strong evidence in the literature to support this, and research can be expected to shed more light on these issues as long as key researchers are involved and adequate funding is provided. However, issues, such as conservation of plant and biodiversity data, should not be overlooked, as this would pose significant challenges to the search for new markers. There are more chances than ever to continue making a significant difference for health reasons, given the growing interest in natural goods among the general public, academic researchers and multinational corporations worldwide.

References Ahmad N, Fazal H, Abbasi BH, Farooq S, Ali M, Khan MA (2012) Biological role of Piper nigrum L.(Black pepper): a review. Asian Pac J Trop Biomed 2(3):S1945–S1953 Akram M, Shahab-Uddin AA, Usmanghani KH, Hannan AB, Mohiuddin E, Asif M (2010) Curcuma longa and curcumin: a review article. Rom J Biol Plant Biol 55(2):65–70 Biswas K, Chattopadhyay I, Banerjee RK, Bandyopadhyay U (2002) Biological activities and medicinal properties of neem (Azadirachta indica). Curr Sci 82:1336–1345 Chang HM, But PPH (1986) Pharmacology and applications of Chinese Materia Medica. World Scientific Publishing, Singapore Chauhan R, Ruby K, Dwivedi J (2012) Bergenia ciliata mine of medicinal properties: a review. Int J Pharm Sci Rev Res 15(2):20–23 Clark AM (1996) Natural products as a resource for new drugs. Pharm Res 13:1133–1141. https:// doi.org/10.1023/A:1016091631721 Dar RA, Shahnawaz M, Qazi PH (2017) Natural product medicines: a literature update. J Phytopharmacol 6(6):349–351

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Das SK, Mukherjee S, Vasudevan DM (2008) Medicinal properties of milk thistle with special reference to silymarin–an overview. Indian J Nat Prod Resour 7(2):182–192 Farnsworth NR, Akerele O, Bingel AS, Soejarto DD, Guo Z (1985) Medicinal plants in therapy. Bull World Health Organ 63(6):965–981 Gajalakshmi S, Vijayalakshmi S, Devi RV (2013) Pharmacological activities of Catharanthus roseus: a perspective review. Int J Pharm Bio Sci 4(2):431–439 Gupta SK, Sharma A (2014) Medicinal properties of Zingiber officinale Roscoe-a review. J Pharm Biol Sci 9:124–129 Gurib-Fakim A (2006) Medicinal plants: traditions of yesterday and drugs of tomorrow. Mol Asp Med 27(1):1–93. https://doi.org/10.1016/j.mam.2005.07.008 Heinrich M, Barnes J, Prieto-Garcia J, Gibbons S, Williamson EM (2017) Fundamentals of pharmacognosy and phytotherapy. Elsevier Health Sciences, Philadelphia, PA Jena J, Gupta AK (2012) Ricinus communis Linn: a phytopharmacological review. Int J Pharm Pharm Sci 4(4):25–29 Jones VH (1941) The nature and status of ethnobotany. Chronica Botanica: new series of plant science books. Chronica Botanica Co, Leyden Joshi P, Dhawan V (2005) Swertia chirayita–an overview. Curr Sci 89:635–640 Kapoor LD (1990) CRC Handbook of ayurvedic medicinal plants, 1st edn. CRC Press, Boca Raton, FL. https://doi.org/10.1201/9781351070997 Khan KH (2009) Roles of Emblica officinalis in medicine-a review. Bot Res Int 2(4):218–228 Krishna S, Uhlemann AC, Haynes RK (2004) Artemisinins: mechanisms of action and potential for resistance. Drug Resist Updat 7(4–5):233–244. https://doi.org/10.1016/j.drup.2004.07.001 Li S, Zhang W (2014) Ethnobotany of Camptotheca decaisne: new discoveries of old medicinal uses. Pharm Crop 5(1):140. https://doi.org/10.2174/2210290601405010140 Mittal J, Sharma MM, Batra A (2014) Tinospora cordifolia: a multipurpose medicinal plant. J Med Plants 2(2):1 Negi JS, Bisht VK, Bhandari AK, Sundriyal RC (2012) Determination of mineral contents of Digitalis purpurea L. and Digitalis lanata Ehrh. J Soil Sci Plant Nutr 12(3):463–470. https://doi. org/10.4067/S0718-95162012005000008 Paarakh PM (2010a) Nigella sativa Linn.–a comprehensive review. Indian J Nat Prod Resour 1(4): 409–429 Paarakh PM (2010b) Terminalia arjuna (Roxb.) Wt. and Arn.: a review. Int J Pharmacol 6(5): 515–534. https://doi.org/10.3923/ijp.2010.515.534 Pradhan D, Biswasroy P, Haldar J, Cheruvanachari P, Dubey D, Rai VK, Kar B, Kar DM, Rath G, Ghosh G (2022) A comprehensive review on phytochemistry, molecular pharmacology, clinical and translational outfit of Ocimum sanctum L. S Afr J Bot 150:342–360. https://doi.org/10. 1016/j.sajb.2022.07.037 Rahimi-Madiseh M, Lorigoini Z, Zamani-Gharaghoshi H, Rafieian-Kopaei M (2017) Berberis vulgaris: specifications and traditional uses. Iran J Basic Med Sci 20(5):569–587. https://doi. org/10.22038/ijbms.2017.8690 Rates SM (2001) Plants as source of drugs. Toxicon 39(5):603–613. https://doi.org/10.1016/s00410101(00)00154-9 Rathinamoorthy R, Thilagavathi G (2014) Terminalia chebula-review on pharmacological and biochemical studies. Int J Pharm Tech Res 6:97–116 Sahu PK, Giri DD, Singh R, Pandey P, Gupta S, Shrivastava AK, Kumar A, Pandey KD (2013) Therapeutic and medicinal uses of Aloe vera: a review. Pharmacol Pharm 4(8):599. https://doi. org/10.4236/pp.2013.48086 Shakya AK (2016) Medicinal plants: future source of new drugs. Int J Herb Med 4(4):59–64 Sharma V (2013) Part based HPLC-PDA quantification of podophyllotoxin in populations of Podophyllum hexandrum Royle “Indian Mayapple” from higher altitude Himalayas. J Med Plants Stud 1(3):176–183 Singh R (2015) Medicinal plants: a review. J Plant Sci 3(1-1):50–55

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Singh VK, Singh DK (2008) Pharmacological effects of garlic (Allium sativum L.). Annu Rev Biomed Sci 1:10 Titanji VP, Zofou D, Ngemenya MN (2008) The antimalarial potential of medicinal plants used for the treatment of malaria in Cameroonian folk medicine. Afr J Tradit Compl Altern Med 5(3): 302–321 Umadevi M, Rajeswari R, Rahale CS, Selvavenkadesh S, Pushpa R, Kumar KS, Bhowmik D (2012) Traditional and medicinal uses of Withania somnifera. Pharma Innov 1(9, A):102 World Health Organization (1993) Research guidelines for evaluating the safety and efficacy of herbal medicines. WHO Regional Office for the Western Pacific, Manila World Health Organization (2008) Media centre: traditional medicine. Fact sheet. WHO, Geneva, p 134

2

Medicinal Properties of the Plant Under Adverse Environmental Conditions Abhijeet Mahana

Abstract

Medicinal plants are used globally as a valuable source of medicine for various diseases. They have become an essential asset for life’s existence on earth since ancient times and play a crucial role to prepare medicines in the pharmaceutical industry of the modern era. However, poor environmental conditions have a negative impact on the growth, development, total yield, and production of bioactive compounds in medicinal plants. Therefore, the influence of various adverse environmental conditions on the medicinal properties of medicinal plants has been discussed in this chapter. Keywords

Medicinal plants · Temperature · Drought · Salinity · Light

2.1

Introduction

A medicinal plant is a type of plant in which one or more organs contain substances that can be used for therapeutic purposes or as precursors for the synthesis of valuable drugs. They exist in the environment as an ancient form of healing and act as a storekeeper for various bioactive compounds like vitamins, aldehydes, anthocyanins, flavonoids, tannins, terpenoids, alkaloids, and phenols (Ivanišová et al. 2021; Singh et al. 2022). These compounds increase therapeutic properties like antiviral, anticancer, antifungal, anti-inflammatory, antimalarial, and so forth A. Mahana (✉) French Associates Institutes for Agriculture and Biotechnology of Drylands, Jacob Blaustein Institute for Desert Research (BIDR), Ben-Gurion University of the Negev, Sede Boqer Campus, Israel # The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 D. Singh et al. (eds.), Stress-responsive Factors and Molecular Farming in Medicinal Plants, https://doi.org/10.1007/978-981-99-4480-4_2

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(El-Shahir et al. 2022; Loi et al. 2020; Rahman et al. 2022; Tran et al. 2020). A study by Lee and Goto (2022) suggested that exposure to ozone (200 ppb) up to 48 h, increased the concentration of total phenolics (2.1-fold), antioxidant capacity (1.07fold), flavonoids (1.83-fold), and anthocyanins (5.23-fold) in Lactuca sativa L., but after 72 h, it slightly decreased. Another study by Brima (2017) suggested that environmental pollution causes increased high levels of elements like Na, Mg, Cu, Zn, Fe, Pb, and Mn in various medicinal plant species. As a result, these medicinal plants can be considered another source to treat diabetes (Ngugi et al. 2015). However, in another side, the two major environmental pollutants like textile dyes and antibiotics negatively influence plant growth and development and affect the level of secondary metabolites (Copaciu et al. 2016). Copaciu et al. (2016) reported in their study that exposure to high concentrations of textile dyes (1.5 mg L-1) for 1-week results in a reduction in flavonoid content in Triticum aestivum L. Therefore, it can be concluded that the long-term exposure of plants in adverse condition decreases the presence of bioactive compounds, as a result, medicinal properties also affected. The application of medicinal plants has increased day by day due to their various medicinal properties, easy availability, affordability, accessibility, low cost, and adverse effect of standard synthetic drug agents. In the global scenario, about 80% of the world’s population totally depends upon traditional or herbal medicine for the treatment of diseases (WHO n.d.). Currently, medicinal plants are used to treat diabetes, hepatitis, ulcers, antipyretics, infertility, antioxidants, analgesics, inflammation, swelling, emetic, piles, smallpox, COVID-19, dermatological, tonic, antiburn, laxative, cancer, and earthworm diseases (Li et al. 2022; Abubakar et al. 2022; Bouyahya et al. 2021; Tonisi et al. 2020). Table 2.1 represents a list of some common medicinal plants and their therapeutic uses to cure various diseases. It is a widespread belief that medicines prepared from plants are natural and are therefore intrinsically harmless. However, among various herbal medicines extracted from medicinal plants, some are toxic to both humans and animals with potential damage to certain organs in the body whereas others are non-toxic in nature. Based on the toxic nature, the mode of uptake of plants as a medicine depends. The toxic effects of herbal medicine have been attributed to several factors, including the presence of toxic herbal ingredients, drug-herb interactions, or the contamination of herbal preparations by heavy metals, microorganisms, mycotoxins, and pesticide residues (Bateman et al. 1998). Basically, it can be said that adverse environmental conditions play a major role in the reduction of medicinal properties of plants that needs to be considered. Increasing the temperature of the earth from 2 to 4.9 °C by year 2100 leads to create abiotic stress (drought, salt stress, and extreme temperature) that causes a reduction in plant fitness and their overall productivity (Aref et al. 2016; Raftery et al. 2017). In association with abiotic stress, the change in weather, climate, and production risk causes physical damage to crop plants, and trees in the direct or indirect pathways (Das et al. 2003). Thus, understanding the adverse environmental condition of medicinal plants lays the foundation for the understanding of plant resilience. To mediate adverse environmental conditions, medicinal plants have also evolved

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Table 2.1 List of some common medicinal plants and their therapeutic uses to cure various diseases S. No. 1

Scientific name Abrus precatorius L. Acanthus pubescens (Oliv.) Engl. Achyranthes aspera L. Acmella paniculata (Wall. ex DC.) R. K. Jansen Acorus calamus L.

Family Fabaceae

Therapeutic use Painful bleeding

Acanthaceae

Chickenpox

Amaranthaceae

Toothache, kidney stone

Asteraceae

Toothache

Acoraceae

Headache and insomnia

Andrographis lineata Wall. ex Nees Andrographis paniculata (Burm. f.) Nees

Acanthaceae

Diabetics

Acanthaceae

8

Azadirachta indica A. Juss.

Meliaceae

9

Bambusa vulgaris Schrad. Barleria cristata L.

Poaceae

Malaria, chronic fever, leprosy, gonorrhea, scabies, boils, skin eruptions Vermifuge, antiseptic, anti-diabetics, astringent, antiperiodic Malaria

Acanthaceae

Cuts and wounds

Betulaceae

Cut and wounds

Crassulaceae

Kidney stones

Apocynaceae

Antiseptic, skin diseases

Lal et al. (2023)

Cannabaceae

Kumar et al. (2021) Tangjang et al. (2011) Kumar et al. (2021) Tangjang et al. (2011) Lal et al. (2023)

2

3 4

5 6

7

10 11 12

13

14 15 16 17 18

Betula utilis D. Don Bryophyllum pinnatum (Lam.) Oken Calotropis gigantea (L.) Dryand Cannabis sativa L. Capsicum frutescens L. Carissa spinarum L. Cassia alata L.

Solanaceae

Fabaceae

Body pain, cuts, burn, diabetes and dysentery Wounds healing, blood coagulant Fever, diarrhea and toothache Ringworm

Cassine glauca (Rottb.) Kuntze

Celastraceae

Snake and dog bite

Apocynaceae

Reference Balamurugan et al. (2018) Gumisiriza et al. (2021) Lal et al. (2023) Lal et al. (2023) Lal et al. (2023) Kadhirvel et al. (2010) Lal et al. (2023)

Kadhirvel et al. (2010) Kumar et al. (2021) Singh et al. (2017) Singh et al. (2017) Lal et al. (2023)

(continued)

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Table 2.1 (continued) S. No. 19 20 21 22

23 24 25

26

27

28

29 30 31

32

33 34

35

Scientific name Centella asiatica (L.) Urb. Centella asiatica (L.) Urb. Citrus limon (Linn.) Burm. f. Clitoria ternatea L.

Family Apiaceae

Therapeutic use Memory enhancer

Apiaceae

Diarrhea and dysentery

Rutaceae

Nail infection, body fitness Stomachache or related complications

Fabaceae

Coccinia indica W. and A. Coccinia indica. L.

Cucurbitaceae

Diabetics

Cucurbitaceae

Urinary obstruction

Colebrookea oppositifolia (Smith.) Cordia macleodii Hook. f. and Thomson. Drymaria cordata (L.) Willd. ex Schult. Dysphania ambrosioides (L.) Mosyakin and Clemants Emblica officinalis Gaertn. Hibiscus rosasinensisa L. Justicia wynaadensis B. Hyene Kalanchoe prolifera (Bowie ex Hook.) Raym.Hamet Litchi chinensis Sonn. Madhuca latifolia (Roxb.) J. F. Macbr. Megacarpaea polyandra Benth. ex Madden

Lamiaceae

cough, wounds and eye infection

Boraginaceae

Wound healing, cardiovascular disorders and high blood pressure Herpes, fever and headache

Caryophyllaceae

Reference Kumar et al. (2021) Lal et al. (2023) Kadhirvel et al. (2010) Bojjangada and Devi Prasad (2021) Kadhirvel et al. (2010) Balamurugan et al. (2018) Kumar et al. (2021) Lal et al. (2023) Singh et al. (2017)

Amaranthaceae

Vomiting

Gumisiriza et al. (2021)

Euphorbiaceae

Stomachache

Malvaceae

Boils

Acanthaceae

Ulcers

Tangjang et al. (2011) Tangjang et al. (2011) Gumisiriza et al. (2021)

Crassulaceae

Cough, malaria

Rakotoarivelo et al. (2015)

Sapindaceae

Diarrhea, dysentery, stomachache Itching, cysts, diabetes and ulcers

Rakotoarivelo et al. (2015) Srivastava et al. (2023)

Fever, abdominal problems, snake bite and scorpion sting

Singh et al. (2017)

Sapotaceae

Brassicaceae

(continued)

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Table 2.1 (continued) S. No. 36

Family Molluginaceae

Therapeutic use Malaria, stomachache

Moringaceae

Asthma, night blindness

Musaceae

Gallbladder stone

Nyctanthes arbortristis L. Ocimum sanctum L. Petchia erythrocarpa (Vatke) Leeuwenb. Phyllanthus amarus Schumach. and Th onn. Polyalthia longifolia (Sonn.) Thwaites Psidium guajava L.

Oleaceae

Apocynaceae

Arthritis, rheumatism, dyspepsia, flatulence Stomachache, loose motion Malaria

Phyllanthaceae

Jaundice

Annonaceae

Pyorrhoea, Hypotension, ulcers, diabetes

Myrtaceae

Diarrhea, dysentery

45

Saraca asoca (Roxb.) Willd

Fabaceae

46

Schefflera vinosa (Chan. and Schltdl.) Frodin and Fiaschi Smithia sensitive Aiton

Araliaceae

Menstrual disorders, Diabetes, haemorrhoids and fever Malaria

Fabaceae

Stomach ailments

Terminalia bellirica (Gaertn.) Roxb. Terminalia chebula Retz. Thottea siliquosa (Lam.) Ding Hou

Combretaceae

Cough and respiratory diseases

Combretaceae

Cough, skin diseases

Aristolochiaceae

Vomiting and diarrhea

37 38

39 40 41

42

43

44

47

48

49 50

Scientific name Mollugo nudicaulis Lam. Moringa oleifera Lam. Musa paradisiaca L.

Lamiaceae

Reference Rakotoarivelo et al. (2015) Srivastava et al. (2023) Bojjangada and Devi Prasad (2021) Srivastava et al. (2023) Tangjang et al. (2011) Rakotoarivelo et al. (2015) Bojjangada and Devi Prasad (2021) Srivastava et al. (2023) Rakotoarivelo et al. (2015) Srivastava et al. (2023) Lal et al. (2023)

Bojjangada and Devi Prasad (2021) Singh et al. (2017) Singh et al. (2017) Bojjangada and Devi Prasad (2021)

secondary metabolites as a defense to protect them from stress conditions (Berini et al. 2018; Isah 2019; Zaynab et al. 2018). Therefore, in this chapter, we are going to highlight the toxic effect of adverse environmental conditions on medicinal plants.

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Current Status and Trends of Medicinal Plants

In the global scenario, it is seen that more than 5000 research articles are published every year that are related to medicinal plants (Salmerón-Manzano et al. 2020). Out of the total available medicinal plants, 3000 wild species are already identified for their medicinal properties. India occupied the second position after China in the world for the supply of fine, pure, and best-quality herbal and natural medicines (Salmerón-Manzano et al. 2020; Agrawal and Paridhavi 2012). Figure 2.1 represents the temporal evolution of medicinal plant publications in top countries. Figure 2.1 indicated that the Indian traditional system of application of medicinal plants is one of the oldest systems of medicinal practice in the world and more than 80% of India’s population still use plants as medicine to provide an essential healthcare service to human and animal life (WHO n.d.; Ahmad et al. 2021).

2.3

Effect of Adverse Environmental Conditions on Medicinal Plant

Environment plays an important role in plant growth and development. The basic factors that affect plant growth include light, temperature, water, humidity, and nutrition. However, adverse environmental condition creates stress on plants and as a result, weakens the plants and makes them more susceptible to disease or insect attack. Due to various adverse environmental conditions, the physiological and biological parameters are altered. As a result, the medicinal properties of plants are reduced. A basic schematic illustration of the physiological and biochemical change in plant cells due to adverse conditions is represented in Fig. 2.2. Therefore, it is important to understand the basic factors and their possible manipulation to meet their needs. The most important factors that influence medicinal plants are described below.

2.3.1

Temperature Extremities

Temperature is one of the essential factors that influence various physiological processes of most plants, that is, photosynthesis, respiration, transpiration, germination, and flowering. In general, the growth of plants is best when the temperature is between 10 and 20 °C. However, several studies indicated that, at extreme temperatures, the plant gives a negative impact on growth and development, physiological processes, and lipid or hormonal signaling (Bita and Gerats 2013). Belmehdi et al. (2018), reported in their study that the highest percentage of seed germination of the medicinal plant Origanum elongatum showed at 20 °C (54%), whereas at temperatures of 10 °C, 15 °C, and 25 °C, the maximal germination was 12%, 44%, and 10%, respectively. Another study by Hatfield and Prueger (2015) also reported that the process of pollination in plants is the most sensitive phenological stage to extreme temperatures and due to extreme temperatures, the

Fig. 2.1 Figure depicts the evolution of research on medicinal plants in top countries in the recent eras. (Derived from Salmerón-Manzano et al. 2020)

2 Medicinal Properties of the Plant Under Adverse Environmental Conditions 23

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Fig. 2.2 Illustration of physiological and biochemical changes that occur in medicinal plant cells in adverse environmental conditions. ROS = Reactive oxygen species; ATP = Adenosine triphosphate; PSII = Photosystem II; PSI = Photosystem I

developmental stage world greatly affects production. Zhang et al. (2022) reported that the photosynthetically active radiation and air high temperature give a negative correlation with chlorophyll fluorescence parameter and at over 35 °C reduces the growth and physiology of tomato plants. Vicente et al. (2020) reported that the seeds of Hypericum ericoides exposed to intermediate temperatures of 10, 15, and 20 °C cause a high percentage of germination, while higher temperatures of 25, 30, and 21/30 °C completely inhibit the germination. Therefore, it can be suggested that the high temperature causes water deficit in soil, and as a result, future agricultural production, food security, and medicines that are extracted from plants are encountered.

2.3.2

Light

Light is another factor that refers to the intensity or concentration of sunlight that varies with season and plays an important role in plant growth through its quantity, quality, and duration. Vicente et al. (2020) resulted that light could not affect germination in plants alone, but at a maximal level, in combination with temperature, it gives a negative impact. In summer, excessive light stress in combination with temperature leads to the overproduction of chloroplastic reactive oxygen species

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(ROS) in plants as a result implicated to both signaling and oxidative damage (Exposito-Rodriguez et al. 2017).

2.3.3

Salinity

Salinity is one of the most detrimental environmental stresses that create a severe issue for plant growth, and reduction in crop yield worldwide. The salt stress on plants causes ionic toxicity, osmotic stress, and oxidative stress simultaneously and as a result causes various physiological and metabolic changes such as nutritional imbalance, inhibition of water uptake, seed germination, photosynthesis, DNA damage, production of ROS, induction of lipid peroxidation, and decrease in growth (Gohari et al. 2023; Haider et al. 2023; Rao et al. 2019; Sheldon et al. 2017; Tanveer et al. 2018). A study by Belmehdi et al. (2018) indicated that the low concentration of NaCl (1 g L-1) helps in the seed germination of Origanum elongatum plant by 88% compared with control (3.4%), but at high concentration causes a significant reduction of seed germination. High salinity (10 g L-1 of NaCl) causes a complete inhibition in seed germination (Belmehdi et al. 2018). Various other studies also give the agreement with the negative impact of salinity stress on plants (Kumar et al. 2021; Groome et al. 1991). Kumar et al. (2021) reported in their study that the increasing concentration of NaCl up to 200 mM significantly increases ROS (twofold) compared with control in the root and leaf of the plant Oenanthe javanica.

2.3.4

Drought

Drought is another condition that reduces leaf water potential, turgor pressure, and stomatal closure and decreases cell growth and enlargement. Due to drought stress cellular dehydration of plant cells occurs, water releases from cytosol and vacuole to apoplast. It influences various physiological and biological functions, such as photosynthesis, chlorophyll synthesis, nutrient metabolism, ion uptake and translocation, respiration, and carbohydrate metabolism in plants (Hussain et al. 2018; Li et al. 2011). Several studies indicated that drought stress induces a decrease in yield and yield component of maize, wheat, sugarcane, sunflower, peanut, and cotton (Barnabás et al. 2008; Furlan et al. 2012; Kamara et al. 2003; Vasantha et al. 2005). Vasantha et al. (2005) reported in their study that the sugarcane yield was reduced by 37% in drought treatment (67,770 ha-1).

2.4

Summary

The adverse environmental conditions on medicinal plants have been described with a focus on the physiological and biological functions of medicinal plants. Global climate change poses a threat to medicinal plants and therefore to traditional as well as modern health care systems. In general, medicinal plants have the potential to

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adjust stress conditions at a certain level due to the presence of their antioxidant system. However, according to available literature studies, it is seen that the adverse environmental conditions at a maximal level induce ROS in the cell and as a result damage the chloroplastic photosystem, mitochondrial respiration, induce lipid peroxidation, reduce water uptake, and so forth. As a result, plant growth and development are altered. However, rising interest and application of medicinal plants due to their non-toxic, cost-effective, and eco-friendly nature, it gives the challenge to protect and think about medicinal plants.

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Response of Secondary Metabolites of Ocimum gratissimum L. Under Copper Stress Condition Mamta Bisht

, Geeta Tewari, Chitra Pande, and Ankit Joshi

Abstract

Secondary metabolites obtained by the plants are important for evaluating the nature of the medicinal components, also influenced by different environmental stress factors. Metal toxicity is one of them. Copper being important cofactor for many enzymes, which are involved in lignin and photosynthesis, affects plants at higher level. At toxic level, it can accumulate in plant tissues resulting in an oxidative damage of plant cell. Under copper stress condition, secondary metabolites can precipitate metal by chelation or complex formation and defense themselves by antioxidative damage. Thus, antioxidative and metal chelating properties of secondary metabolites play a vital role in protecting against metal induced stress. This chapter describes the impact of copper stress on the secondary metabolites of Ocimum gratissimum L. O. gratissimum L. is one of precious aromatic plants for its essential oils and is widely used in the food, fragrance and pharmaceutical industries. A polyhouse study was conducted to measure the effects of different copper stress levels (270, 500, 700 and 900 mg kg-1) in triplicate along with an unmodified control on O. gratissimum volatiles. After 90 days, above-ground plant parts were harvested, hydrodistilled and analysed by GC and GC/MS. Analysis revealed the presence of 19–23 identifiable compounds M. Bisht (✉) Department of Chemistry, School of Applied & Life Sciences, Uttaranchal University, Dehradun, India Department of Chemistry, D. S. B. Campus, Kumaun University, Nainital, India G. Tewari · C. Pande Department of Chemistry, D. S. B. Campus, Kumaun University, Nainital, India A. Joshi Department of Chemistry, School of Applied & Life Sciences, Uttaranchal University, Dehradun, India # The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 D. Singh et al. (eds.), Stress-responsive Factors and Molecular Farming in Medicinal Plants, https://doi.org/10.1007/978-981-99-4480-4_3

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representing 85.61–96.20% of the total oil. Eugenol was the dominant compound in control and all amendments. After copper stress, eugenol content was found higher for the plants grown in higher level of copper stress along with essential oil yield when compared with the control. Thus, metal stress is one factor, which affects the metabolic pathway of secondary metabolites. Keywords

Ocimum gratissimum L. · Copper stress · Eugenol · Essential oil yield · GC · GCMS

3.1

Introduction

Environmental stress is a major problem growing rapidly nowadays due to industrialization and human activities, such as mining, smelting and electroplating. Copper is generally used in the manufacture of electrical wires, rings, machineries and containers, so raised concentrations of copper in the soil have been found near these regions. Further, use of copper bearing fertilizers to inhibit the attack of pathogens also contribute toward copper toxicity in soil (Shahid et al. 2014). Copper within a permissible limit is essential for plant growth and development. It activates some enzymes in plants, which are involved in lignin synthesis, and also participate in photosynthesis, electron transfer chain, nitrogen fixation, oxidative system and so forth. Copper also plays a significant role in transcriptional signalling, transmission and acceptance of ethylene signalling, oxidative phosphorylation and iron mobilization (Gonzalez and Giege 2014). Excessive amounts of copper can supress photosynthesis, inhibit the enzymatic activity, induce deficiencies of other element, like Fe, Ca, Mg, K and P, and affect the intracellular transport, distribution of substances between plants organs and perturbs the process (Penarrubia et al. 2015). Moreover, due to metal toxicity, health of human and domestic animals is disturbed. To reduce food chain contamination, non-food crops, like medicinal and aromatic plants, can be used as an alternate cash crop in metal elevated sites. Ocimum gratissimum is an important erect aromatic shrub, which belongs to the family Labiatae (Bisht et al. 2019). Leaves of this plant are used as folk medicine, and whole plant is used for medicinal purpose. The essential oil of the plant is well studied for its antimicrobial, antioxidant, antifungal, antimutagenic and antidiabetic activities. Further, O. gratissimum has been investigated for its volatile constituents; however, in the current scenario of adverse environmental conditions, the molecular studies of the secondary metabolites of the plant against metal stress still need to explore. Previous studies suggested that some plants like Celosia argentea, Mentha arvensis L. have ability to grow under copper stress condition (Bisht et al. 2021; Wang et al. 2021). Keeping above points in mind, this study was an attempt to explore the chemical profile of the plant under copper stress condition.

3

Response of Secondary Metabolites of Ocimum gratissimum L.. . .

3.2

Materials and Methods

3.2.1

Sapling Collection, Identification, Polyhouse Experimental Setup and Extraction of Oil

31

The saplings of O. gratissimum L. (Accession number 116136), purchased from Medicinal Research Development Centre (MRDC), Pantnagar. The saplings were transferred and cultivated in the earthen pots containing 5 kg of air-dried Cu amended soils having four amendments (270 mg kg-1: Cu270; 500 mg kg-1: Cu500; 700 mg kg-1: Cu700; 900 mg kg-1: Cu900) along with an unamended control (Cu0) used for the experiment. Nitrogen (N), phosphorous (P) and potassium (K) were also added to all the pots in appropriate amount as to meet the requirement of macronutrients (Bisht et al. 2021). Prior to the experiment, the modified soils were incubated for 1 month; 3 months after planting, the above-ground parts of O. gratissimum were harvested, and 100 g of each experimental material was cut into small pieces and then hydrodistilled in a Clevenger for 3 h, repeated three times for each supplement along with the control. The extracted oils were dried over anhydrous sodium sulphate and kept at 4 °C until analysis.

3.2.2

Essential Oil Yield

The percentage oil yield (% w/w) was computed on the fresh weight basis. Essential oil yield =

3.2.3

Weight of hydrodistilled oil ðgÞ × 100 Weight of the plant material taken for hydrodistillation ðgÞ

Essential Oil Analysis and Identification of the Volatile Constituents

The analysis of extracted oils was performed in a GC and GC/MS apparatus 2010 (Shimadzu) and under similar conditions applied by Bisht et al. (2019). The components of the essential oil were identified on the basis of retention index, NIST and Wiley library search and by comparing with the MS literature data (Adams 2007; Bisht et al. 2015).

3.2.4

Statistical Analysis

MS Excel was used to calculate the mean values and standard deviation, whereas SPSS 16.0 software was used for statistical analysis of the data (Bisht et al. 2022).

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3.3

Result and Discussion

3.3.1

Essential Oil Yield

A variety of environmental stresses faced by medicinal plants during growth and developmental stages. Various ecological limiting factors, such as soil fertility, soil water, carbon dioxide, temperature, light and salinity of soil greatly affect the physiological and biochemical responses of medicinal plants. Copper stress significantly affected the oil yield of the plant, leading to increased yield with elevated copper concentration with respect to control (Table 3.1). Our results were similar as reported by Alizadeh et al., where an increased application of copper increased the oil yield of summer savoury plant (Alizadeh et al. 2010). On the other hand, many authors reported that the foliar application of micronutrients increased the essential oil yield of some aromatic plants like fennel, Ocimum, chamomile and coriander (Al-Humaid 2004; Youssef et al. 2004; Nasiri et al. 2010; Sinta et al. 2015). In contrast, Ghorbanpour et al. and Zheljazkov and Nielsen recorded the essential oil yield of O. basilicum reduced with elevated concentration of copper and zinc (Ghorbanpour et al. 2016; Zheljazkov and Nielsen 1996a, b).

3.3.2

Impact of Copper Stress on the Secondary Metabolites of O. gratissimum L.

Metals play important roles in plant metabolic functions, but at high amount, they are toxic and disrupt physiological and biochemical functions (El-Sheekh et al. 2003; Parmar and Chanda 2005; Jayakumar and Vijayarengan 2006). During metal stress condition, oxidative stress defence mechanism was activated in the plants and affect the pathway of secondary metabolites (Wang et al. 2021). GC and GC/MS analysis showed the presence of 19–23 identifiable compounds representing 85.61–96.20% of the total oil (Table 3.2). In the current research, eugenol (64.62%) was the dominant compound in control followed by germacrene D (8.45%), (E)-caryophyllene (2.89%), α-copaene (2.71%), δ-cadinene (1.05%) and caryophyllene oxide (0.66%). Eugenol (68.80%) was found as a predominant

Table 3.1 Impact of copper on weight (g) and yield % (v/w) of O. gratissimum L.

Copper stress level mg kg-1 Cu0 Cu270 Cu500 Cu700 Cu900

Fresh weight (g) Mean ± SD 105d ± 2.0 55c ± 1.0 50b ± 1.0 40a ± 1.0 140e ± 1.0

Oil yield % (v/w) Mean ± SD 0.2a ± 0.0 0.4b ± 0.1 0.2a ± 0.1 0.5b ± 0.1 0.4b ± 0.1

Different alphabets in the columns are significantly different and alphabets (a–e) in each column are not significantly different ( p < 0.05) Duncan multiple range test

S. No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23

Compound name α-Thujene 1-Octen-3-ol Myrecene (Z )-β-Ocimene (E)-β-Ocimene γ-Terpinene cis-Sabinene hydrate Linalool Nonanol α-Cubenene Eugenol α-Copaene β-Cubebene Bicycloesequiphellandrene (E)-Caryophyllene β-Copaene γ-Cadiene Germacrene D δ-Cadinene Caryophyllene oxide Salviol4 (14)-ene-1-one Humulene epoxide Valencene hydroxy

RI (Adams 2007) 924 974 988 1032 1044 1056 1065 1095 1100 1348 1359 1376 1387 1417 1430 1513 1487 1522 1587 1596 1612 1767

Control (% of oil ± SD) ND 0.15 ND 0.01a ± 0.00 0.02 ND 1.70 ND 0.02 0.12 64.62d ± 0.03 2.71d ± 001 0.29 ND 2.89c ± 0.16 0.13 0.22 8.45a ± 0.05 1.05 0.66b ± 0.02 0.17 0.16 1.05

Cu270 (mg kg-1) (% of oil ± SD) 0.02 0.10 0.20 18.97e ± 0.08 0.72 0.10 0.68 0.11 ND 0.04 41.83b ± 0.02 1.07c ± 0.01 0.27 1.60 3.52d ± 0.04 0.33 0.32 24.20d ± 0.52 1.04 0.26a ± 0.08 0.25 0.41 0.03

Table 3.2 Effect of copper stress on the secondary metabolites of O. gratissimum L. Cu500 (mg kg1 ) (% of oil ± SD) ND ND 0.05 0.44b ± 0.02 0.03 ND 0.72 1.23 ND 0.08 62.74c ± 0.052 0.60a ± 002 ND ND 1.91b ± 0.03 0.16 0.17 12.28c ± 0.08 0.81 4.43d ± 1.79 0.39 0.36 0.47 Cu700 (mg kg-1) (% of oil ± SD) ND 0.05 0.05 7.99c ± 0.04 0.28 0.05 0.52 0.14 0.07 0.12 27.28a ± 0.04 ND ND 3.84 5.34e ± 1.66 0.39 0.34 41.41e ± 0.08 1.41 1.61c ± 0.02 0.21 0.23 0.37

Cu900 (mg kg-1) (% of oil ± SD) 0.19 0.09 0.23 13.97d ± 0.38 0.50 0.10 1.05 0.13 ND 0.03 68.30e ± 0.11 ND ND ND 1.44a ± 0.22 0.13 0.09 9.11b ± 0.15 0.39 0.10a ± 0.00 0.05 ND 0.01

Response of Secondary Metabolites of Ocimum gratissimum L.. . . (continued)

Mode of identification A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B A, B

3 33

Compound name Caryophyllane 4, 12, 8, 13 diene-5α ol Aromadrene (epoxide ) Muskatone Total compounds identified Total identified (%)

1676

1639

RI (Adams 2007) 1640

0.13 23 96.20

85.61

ND

ND 20

0.64

Control (% of oil ± SD) 0.55

Cu270 (mg kg-1) (% of oil ± SD) ND

87.21

ND 19

0.15

Cu500 (mg kg1 ) (% of oil ± SD) 0.19

92.24

0.11 23

0.25

Cu700 (mg kg-1) (% of oil ± SD) 0.18

95.93

0.02 19

ND

Cu900 (mg kg-1) (% of oil ± SD) ND

A, B

A, B

Mode of identification A, B

A = Retention index (RI) relative to homologous series of (C8-C33) on Rtx-5 non polar fused silica capillary column; B = Gas chromatography–mass spectrometry (GC-MS); ND = Not detected (