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Biotechnological Approaches
to Enhance Plant Secondary
Metabolites
Biotechnological Approaches
to Enhance Plant Secondary
Metabolites

Recent Trends and Future Prospects

Edited by

Mohd. Shahnawaz
First edition published 2022
by CRC Press
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and by CRC Press
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Names: Shahnawaz, Mohd., editor.

Title: Biotechnological approaches to enhance plant secondary metabolites :
recent trends and future prospects / edited by Mohd. Shahnawaz.
Description: First edition. | Boca Raton : CRC Press, 2022. | Includes bibliographical references and index.
Identifiers: LCCN 2021026997 | ISBN 9780367473365 (hardback) | ISBN 9781032122021 (paperback) |
ISBN 9781003034957 (ebook)
Subjects: LCSH: Phytochemicals–Biotechnology. | Plant metabolites–Biotechnology. |
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DOI: 10.1201/9781003034957

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 Dedication

To my parents (Mr. Shamas-ud-Din & Mrs. Muneera Begum)

and
my better half (Mrs. Mubeena Kousar)
Contents
Preface����������������������������������������������������������������������������������������������������������������������������������������������ix
Acknowledgements���������������������������������������������������������������������������������������������������������������������������xi
Editor��������������������������������������������������������������������������������������������������������������������������������������������� xiii
Contributors������������������������������������������������������������������������������������������������������������������������������������� xv

Chapter 1 Plant Secondary Metabolites—The Key Drivers of Plant’s Defence

Mechanisms: A General Introduction��������������������������������������������������������������������������1
D. Sruthi and C. Jayabaskaran

Chapter 2 Generation of Plant Mutant Lines Using Gamma Radiation with Enhanced
Secondary Metabolite Contents��������������������������������������������������������������������������������� 27
Lata I. Shukla, P. Vivek Vardhan, T. K. Devika, Sayan Roy
and Sourav Bhatacharya

Chapter 3 Salinity Stress and Plant Secondary Metabolite Enhancement:

An Overview�������������������������������������������������������������������������������������������������������������� 49
Bedabrata Saha, Bhaben Chowardhara, Jay Prakash Awasthi,
Sanjib Kumar Panda and Kishore C.S. Panigrahi

Chapter 4 Enhancement of Plant Secondary Metabolites Using Fungal Endophytes���������������� 61

Touseef Hussain, Mulla Javed, Samrin Shaikh, Bilquees Tabasum,
Kashif Hussain, Moh Sajid Ansari, Amir Khan and Abrar Ahmad Khan

Chapter 5 Hydroponic Cultivation Approaches to Enhance the Contents

of the Secondary Metabolites in Plants���������������������������������������������������������������������� 71
Yogesh Chandrakant Suryawanshi

Chapter 6 Tissue Culture Approaches to Enhance Plant Secondary

Metabolites Production���������������������������������������������������������������������������������������������� 89
Vishal N. Patil and Mohd. Shahnawaz

Chapter 7 Hairy Roots and Plant Secondary Metabolites Production: An Update��������������������99

Sharada L. Deore, Bhushan A. Baviskar and Anjali A. Kide

Chapter 8 Brassinosteroids: The Phytohormones with Potential to Enhance the

Secondary Metabolite Production in Plants������������������������������������������������������������� 125
Barket Ali, Zahoor A. Wani and Mudasir Irfan Dar

vii
viii Contents

Chapter 9 CRISPR-Cas9 Approaches to Enhance Contents of Plant

Secondary Metabolites��������������������������������������������������������������������������������������������� 133
Riddhi Rajyaguru, Nataraja Maheshala, Chandrashekar Mootapally, Neelam
Nathani, Rukamsingh Tomar, Hiren Bhalani and Priyanka Sharma

Chapter 10 RNA Interference for Improvement of Secondary Metabolite

Production in Plants������������������������������������������������������������������������������������������������� 153
Ashutosh Kumar Rai and Pramod Wasudeo Ramteke

Index���������������������������������������������������������������������������������������������������������������������������������������������� 165
Preface
We have been dependent on plants for food, furniture, construction, and fuel since prehistoric times.
Plants in all forms have tendered their services to mankind. We have a long history of plants cultiva-
tion and domestication to meet our basic needs. People living in different continents of the world
used plants to treat various diseases. Accordingly, various systems of medicines were established
viz. Ayurvedic, Unani, Siddha, Homeopathy, Tibetan, folklore and Chinese system of medicines,
etc. Due to the advancement of science and technologies, various other non-conventional methods
to treat the diseases were also identified. Each system of medicine is having advantages and dis-
advantages. The system which utilizes plant-based natural products for the discoveries of drugs
have attracted the attention globally, due to its least or no side effects. Plant synthesized various
secondary metabolites besides its primary metabolites to resist the changing environmental stress
and to cross-talk with other biological entities. These secondary metabolites have also been found to
have various activities against numerous human diseases. So, to harvest these medicinal principles
from the medicinal plants, people have exploited the plant germplasm from the natural habitat at
an alarming rate. Most of the medicinal plants are now considered endangered. As per the reports,
the contents of these naturally occurring medicinal components were found low in most of the
plants. So, it was needed to enhance the contents of such key principal components of the plants.
Various people around the globe tried to enhance the contents of the secondary metabolites in the
plants using various methods and were reported significant enhancement of the targeted second-
ary metabolites in the tested plants. The most common methods employed to enhance the contents
of the plant secondary metabolites, viz. abiotic and biotic elicitation at in vitro and in situ levels,
mutation breeding, plant tissue culture techniques, genetic engineering, RNAi, CRISPR-CAS9
technology etc.
Various literature on plant secondary metabolites is available; however, there is a lack of books
that provide an overview of all the methods involved in the enhancement of the plant secondary
metabolite.
This edited volume offers 10 excellent chapters, contributed by various experienced research-
ers, to fill the lacunae in the literature. Chapter 1 introduces plant secondary metabolites as the key
drivers of plant’s defence mechanisms. Chapter 2 describes the gamma radiations usage to enhance
the contents of plant secondary metabolites. Chapter 3 and 4 provides an overview of the impact of
salinity stress and the role of fungal endophytes to enhance secondary metabolite contents in plants.
Chapter 5 reports on the hydroponic cultivation approaches to enhance the contents of the second-
ary metabolites in plants. Chapters 6 and 7 highlight the tissue culture techniques and hairy root
cultures the enhancement of plant secondary metabolites in plants. Chapter 8 documents the impact
of brassinosteroids to enhance the secondary metabolite production in plants. Chapters 9 and 10
discuss the application of RNAi technology and CRISPR-CAS9 approaches to enhance contents of
plant secondary metabolites.

ix
Acknowledgements
I bow my head before Almighty Allah the most beneficent and most merciful for his endless bless-
ings on me to complete this book. The financial support provided by DST-SERB in the form of the
National Post-doctoral Fellowship (DST-SERB (PDF/2017/000178) award is dully acknowledged.
I am highly thankful to all the anonymous reviewers, who reviewed the chapters thoroughly and
allowed us to make wise decisions about the selection of the best chapters in this current volume.
It is of immense pleasure to acknowledge the help and support tendered by the editorial and
production team (especially Renu Upadhyay and Jyotsna Jangra) of CRC Press, Taylor & Francis
Group.
Last but not least, I am thankful to my parents (Mr. Shamas-ud-Din & Mrs. Muneera Begum)
and my better half (Mrs. Mubeena Kousar) for their kind love and support throughout this work.

Mohd. Shahnawaz ‘Khakii’

xi
Editor
Dr. Mohd. Shahnawaz is currently working as a Lecturer at the Department of Botany, Govt.
Degree College Paloura, Mishriwala, Jammu and Kashmir, India, on an academic arrangement
basis (2020–2021). In 2020, he was selected as a post-doctoral fellow at Yeungnam University,
Gyeongsan, North Gyeongsang, South Korea; however, due to COVID-19 scenario, he was not
able to join. He has also completed post-doctoral research (funded by DST-SERB, Govt. of India)
from the Plant Biotechnology Division, CSIR-Indian Institute of Integrative Medicine, Jammu
& Kashmir, India. He worked to enhance the contents of secondary metabolites in Coleus for-
skohli using in vitro salinity stress. Previously, he has worked as a Lecturer in Botany at the
Department of Botany, Govt. Degree College Kishtwar, Jammu and Kashmir, India (2016–2017)
on an academic arrangement basis. He has earned his M. Phil. and Ph.D. in Botany from the
Department of Botany, Savitribai Phule Pune University, Maharashtra, India under the guid-
ance of Prof. A. B. Nadaf and Prof. A. B. Ade in 2010 and 2016. He is the recipient of various
fellowships awarded by the Savitribai Phule Pune University, University Grants Commission
(UGC), and Department of Science and Technology (DST)-Science Engineering and Research
Board (SERB), India. His research interests are focused on ecology, microbiology, bioremedia-
tion of plastic and plant biotechnology. He has served as a referee and editorial board member of
various journals of International repute. In 2019, he was awarded the Young Scientist Award by
VDGOOD Professional Associations, Vishakhapatnam, India. He has published more than 20
research articles in peer-reviewed international journals and authored 7 books. Previously, he had
served as an Academic Editor of the Asian Journal of Biology (Science Domain International).

xiii
Contributors
Amir Khan Sharada L. Deore
Department of Botany Department of Pharmacognosy and
Aligarh Muslim University Phytochemistry
Aligarh, India Government College of Pharmacy
Amravati, India
Barket Ali
Department of Botany T. K. Devika
Government Degree College Kilhotran Department of Biotechnology
Kishtwar, India Pondicherry University
Kalapet, Puducherry, India
Moh Sajid Ansari
Department of Botany Kashif Hussain
Aligarh Muslim University School of Pharmacy
Aligarh, India Glocal University
Saharanpur, India
Jay Prakash Awasthi
Department of Life Science and Bioinformatics Touseef Hussain
Assam University Department of Botany
Silchar, India Aligarh Muslim University
Aligarh, India
Bhushan A. Baviskar
Department of Pharmacognosy and Mulla Javed
Phytochemistry Institute of Bioinformatics and Biotechnology
Government College of Pharmacy Savitribai Phule Pune University
Amravati, India Pune, India

Hiren Bhalani C. Jayabaskaran

Department of Biotechnology and Department of Biochemistry
Biochemistry Indian Institute of Science
Junagadh Agricultural University Bengaluru, India
Junagadh, India
Abrar Ahmad Khan
Sourav Bhatacharya Department of Botany
Department of Biotechnology Aligarh Muslim University
Pondicherry University Aligarh, India
Kalapet, Puducherry, India
Anjali A. Kide
Bhaben Chowardhara Department of Pharmacognosy and
Department of Life Science and Bioinformatics Phytochemistry
Assam University Government College of Pharmacy
Silchar, India Amravatima, India

Mudasir Irfan Dar Nataraja Maheshala

Department of Botany Entomology Section
Government Degree College Poonch ICAR-Directorate of Groundnut Research
Poonch, India Junagadh, India

xv
xvi Contributors

Chandrashekar Mootapally Pramod Wasudeo Ramteke

Department of Bioscience Department of Biological Sciences
Institute of Biotechnology Sam Higginbottom University of Agriculture
Saurashtra University Technology and Sciences
Rajkot, India Prayagraj, India

Neelam Nathani Sayan Roy

Department of Bioscience Department of Biotechnology
Institute of Biotechnology Pondicherry University
Saurashtra University Kalapet, Puducherry, India
Rajkot, India
Bedabrata Saha
Sanjib Kumar Panda School of Biological Sciences
Department of Biochemistry National Institute of Science Education and
Central University of Rajasthan Research
Ajmer, India Jatani, India
and
Kishore C.S. Panigrahi Weed Research Department
School of Biological Sciences Newe Ya’ar Research Centre
National Institute of Science Education and Agricultural Research Organization (ARO)
Research Ramat Yishai, Israel
Centre for Interdisciplinary Sciences
Jatani, India Mohd. Shahnawaz
and Department of Botany
Life Sciences Govt. Degree College Paloura
Homi Bhabha National Institute (HBNI) Mishriwala, India
Mumbai, India
Samrin Shaikh
Vishal N. Patil Department of Botany
Post Graduate Department of Botany Savitribai Phule Pune University
Vidya Bharti College Ganeshkhind, Pune, India
Wardha, Seloo, India
Priyanka Sharma
Ashutosh Kumar Rai Department of Biotechnology
Department of Biology Dr. Babasaheb Ambedkar Marathwada
Texas A&M University University
College Station, Texas Aurangabad, India
and
Department of Microbiology and Cell Biology Lata I. Shukla
Indian Institute of Science Department of Biotechnology
Bengaluru, India Pondicherry University
Kalapet, Puducherry, India
Riddhi Rajyaguru
Department of Biotechnology and D. Sruthi
Biochemistry Department of Biochemistry
Junagadh Agricultural University Indian Institute of Science
Junagadh, India Bengaluru, India
Contributors xvii

Yogesh Chandrakant Suryawanshi P. Vivek Vardhan

Department of Yoga & Naturopathy Department of Biotechnology
Vishwakarma University Pondicherry University
Pune, India Kalapet, Puducherry, India

Bilquees Tabasum Zahoor A. Wani

Department of Botany Department of Botany
Savitribai Phule Pune University Government Degree College Kishtwar
Pune, India Kishtwar, India

Rukamsingh Tomar
Department of Biotechnology and
Biochemistry
Junagadh Agricultural University
Junagadh, India
 1 Plant Secondary Metabolites—
The Key Drivers of Plant’s
Defence Mechanisms
A General Introduction
D. Sruthi and C. Jayabaskaran

CONTENTS
1.1 Introduction............................................................................................................................... 1
1.2 Classification of Secondary Metabolites...................................................................................2
1.2.1 Terpenes (Terpenoids).................................................................................................... 2
1.2.1.1 Synthesis of Terpenes..................................................................................... 3
1.2.1.2 Classification of Terpenes...............................................................................4
1.2.1.3 Extraction and Identification of Terpenes....................................................... 5
1.2.1.4 Role and Application of Terpenes................................................................... 5
1.2.1.5 Essential Oil.................................................................................................... 6
1.2.2 Alkaloids........................................................................................................................ 8
1.2.2.1 Synthesis of Alkaloids.................................................................................... 8
1.2.2.2 Classification of Alkaloids..............................................................................8
1.2.2.3 Extraction and Identification of Alkaloids......................................................9
1.2.2.4 Role and Application of Alkaloids ................................................................9
1.2.3 Phenolics...................................................................................................................... 10
1.2.3.1 Synthesis of Phenolics.................................................................................. 10
1.2.3.2 Classification of Phenolics............................................................................ 12
1.2.3.3 Extraction and Identification of Phenolics.................................................... 13
1.2.3.4 Role and Application of Phenolics................................................................ 13
1.3 Current Biotechnological Approaches for the Enhancement of Secondary Metabolite
Contents in Plants.................................................................................................................... 13
1.4 Conclusion............................................................................................................................... 16
Acknowledgements........................................................................................................................... 17
References......................................................................................................................................... 17

1.1 INTRODUCTION
Plants are rich in bioactive phytoconstituents with enormous ethnopharmacological potential and
these high-value phytochemicals are broadly categorized into two classes (primary and second-
ary metabolites) based on their function (Hussein and El-Anssary 2019). The primary metabolites
are distributed widely in all plants and carry out metabolic functions which are crucial for normal
physiological growth and energy requirements of the plants and are normally evident (Hussain
et al. 2012; Wink 2016). In contrast, plants synthesize myriads of organic compounds; most of them
do not involve directly in plant growth and development. These phytochemicals, normally known
as secondary metabolites, are distributed differentially in restricted taxonomic groups of the plant

DOI: 10.1201/9781003034957-1 1
2 Biotechnological Approaches to Enhance Plant Secondary Metabolites

kingdom (Tiwari and Rana 2015). Briefly, secondary metabolites are normally not essential for
plant growth and its development, but they are essential for surviving in the ecosystem through
chemical defence (Iriti and Faoro 2009). Plant secondary metabolites are the chemical constitutes
primarily responsible for the chemical defences to regulate the relationship between the plant and
their ecosystem and further take part in protecting plants from abiotic and biotic stress conditions
(Mazid et al. 2011). Secondary metabolites are not essentially synthesized under all circumstances
but produced undoubtedly for really appreciated reasons, few of such include, toxic materials to
protect the plant from predators, as volatile attractants or as an agent to impart colour and thereby
warn or attract other species and thus, it is evident that all of them play some crucial function for
the safety of their producer (Adeyemi and Mohammed 2014). Plant defence metabolites arise from
the isoprenoid, the alkaloid and the phenylpropanoid pathways which form three major second-
ary metabolite classes viz., terpenes, alkaloids and phenolics, respectively (Iriti and Faoro 2009).
Generally, precursors for secondary metabolite synthesis are products of the primary metabolism
(Iriti and Faoro 2004). Besides their role in plant protection, secondary metabolites are well studied
for various pharmacological and medicinal potentials and many drugs of natural origin against
human ailments have been discovered from different plants (Velu et al. 2018).
Secondary metabolites provide major pharmacologically active high-value natural products used
to treat various diseases since ancient times and it ranges from migraine up to cancer (Hussein and
El-Anssary 2019; Jain et al. 2019). Vinca alkaloids (vinblastine and vincristine), taxanes [e.g. pacli-
taxel (Taxol) and docetaxel (Taxotere)] and combretastatin are some of the secondary metabolites
used to treat cancer (Hartwell 1984; Kingston 2012; Pinney et al. 2012; Roussi et al. 2012). Further,
secondary metabolite drugs have been obtained from plants with anti-diabetic (e.g. galegine alka-
loid from Galega officinalis,), anti-inflammatory (e.g. curcumin and resveratrol) and anti-viral
(e.g. betulinic acid and calanolides) efficacies (Min et al. 1999; Dharmaratne et al. 2002; Furst and
Zudorf 2014; Rios et al. 2015). So there is a huge demand for medicinal plants to isolate pharma-
ceutically active molecules (Dar et al. 2017a). These secondary metabolites are produced in low
quantity in plants; hence, a large amount of plant material is used to extract the drug molecules
(Dar et al. 2017b). To meet the demand of pharmaceutical companies, huge medicinal plants are
being exploited at a mass level. This practice leads to threat the germplasm and various plants were
extinct from the natural environment (Maxted et al. 2020). Hence, it was needed to enhance the
contents of such pharmaceutically active plant secondary metabolites to reduce the exploitation of
the medicinal plants from the natural environment (Jimenez-Garcia et al. 2013). With this regard,
various biotechnological strategies were used by different workers across the globe (Guerriero et al.
2018). So, in this chapter, an effort was made to overview the plant secondary metabolites, to dis-
cuss different classes of plant secondary metabolites, to discuss the biosynthetic pathways that took
part in the production of plant secondary metabolites and to highlight various approaches adopted
to enhance the secondary metabolite contents in plants.

1.2 CLASSIFICATION OF SECONDARY METABOLITES

Secondary metabolites are classified primarily as terpenoids, alkaloids and phenolics depending on
their chemical structure (Shamina and Sarma 2001).

1.2.1 Terpenes (Terpenoids)

Terpenes are ubiquitous in plants and are the largest class of secondary metabolites with more than
22000 compounds (Freeman and Beattie 2008). The high concentration of compounds in turpen-
tine oil has given the alternate name ‘terpenoid’ to these compounds. All terpenes are said to be
the derivative of branched, basic, five-carbon unit isoprene (C5H8) (Bramley 1997; Goodwin and
Mercer 2003). Isoprene is a volatile gas emitted by leaves during photosynthesis and that might
prevent the damage of plant cell membranes due to high temperature or light (Freeman and Beattie
Plant Secondary Metabolites—The Key Drivers of Plant’s Defence Mechanisms 3

2008). Terpenes are the essential metabolite for photosynthesis and also for regulating the plant
metabolic processes (Bramley 1997; Goodwin and Mercer 2003). Even though most of the terpenes
are categorized as secondary metabolites, extremely few among them (e.g. steroids) are included in
the primary metabolite class (Bramley 1997; Goodwin and Mercer 2003).

1.2.1.1 Synthesis of Terpenes

The cytosol and the plastids are the sites of biosynthesis of terpenoids (Aharoni et al. 2006). They
are also known as isoprenoids and synthesized from acetyl-CoA by mevalonate or isoprenoid path-
way (Figure 1.1) (Goodwin and Mercer 2003; Karlic and Varga 2019). Two acetyl-CoA molecules
are combined to form acetoacetyl-CoA and, further form HMG-CoA (β-hydroxy-β-methylglutaryl
CoA) with an additional acetyl CoA molecule. Both of these reactions are catalyzed by HMG-CoA
synthase (Chappell 1995). HMG-CoA reductase further catalyzes the rate-limiting reaction of the
isoprenoid pathway, the synthesis of mevalonic acid from HMG-CoA. From the mevalonate, IPP
(isopentenyl diphosphate), a five-carbon building block for the synthesis of isoprenoid chains is

FIGURE 1.1 Isoprenoid pathway from acetyl-CoA. (Adapted from Iriti and Faoro 2009.)
4 Biotechnological Approaches to Enhance Plant Secondary Metabolites

synthesized and is converted into all the different terpenoids found in nature (Bach 1987; Chappell
1995; Goodwin and Mercer 2003).

1.2.1.2 Classification of Terpenes

Based on the isoprene unit number, terpenes can be classified as ‘hemiterpenes, monoterpenes, ses-
quiterpenes, diterpenes, sesterterpenes, triterpenes, tetraterpenes and polyterpenes with one, two,
three, four, five, six, eight and more than eight isoprene units, respectively’ (Goodwin and Mercer
2003). Table 1.1 represents different classes of terpenoids with their molecular formula.
Hemiterpenes are formed from isopentenyl pyrophosphate and mainly found in plants as signal-
ling defence compounds and are represented by parent isoprene, that is a common volatile com-
pound released from woody trees like willows, oaks, spruce and poplar (Dewick 2009; Osbourn
and Lanzotti 2009). Other examples for hemiterpenes include berry and fruit metabolite prenol and
isovaleric acid, an essential oil constituent (Schnitzler et al. 2012).
Monoterpenes represent the simplest terpenoid class with C10 skeleton and are formed from
geranyl-PP (Croteau 1987; Goodwin and Mercer 2003). Monoterpenes are generally occurring in
secretory glands, as the main constituent of plant essential oil (Goodwin and Mercer 2003). They
are primarily non-volatile and can further be categorized as acyclic, cyclopentanoid, cyclohex-
anoid and irregular monoterpenes based on their structure. Thujene, camphene, pinene, limonene,
myrcene, careen, etc. are some of the monoterpenes (Goodwin and Mercer 2003). Monoterpenes
are widely seen in higher plants and possess a strong characteristic aroma that gives them huge
attention in the perfumery industry (Goodwin and Mercer 2003). They also have applications in
food flavourings and pharmacology (Kang and Lee 2016). Because of their repulsiveness or attrac-
tiveness, monoterpenes involve in plant-plant, plant-phytophagous and plant-microbe interactions
(Croteau 1987).
Sesquiterpenes are the largest terpenes and are synthesized from farnesyl-PP (Cane 1990). They
have a C15 skeleton and their volatility is less compared to monoterpenes (Cane 1990; Stephane and
Jules 2020). However, they co-exist with monoterpenes and found to be the essential part of essen-
tial oil (Goodwin and Mercer 2003; Stephane and Jules 2020). Further, sesquiterpenes possess enor-
mous biological activities as pheromones, insect antifeedant, insect juvenile hormone, plant growth
regulators, mycotoxins and phytoalexines antibiotics (Bowers et al. 1977; Kalsi et al. 1989; Taylor
1990; Morrissey and Osbourn 1999; Dhadialla et al. 2005; Abdelgaleil et al. 2020). Further, sesqui-
terpenes can be classified as acyclic, monocyclic, bicyclic and tricyclic sesquiterpenes (Goodwin
and Mercer 2003). Sesquiterpenes contain a lactone ring that comes under the class sesquiterpene
lactone (Schmidt 2006). Caryophyllene, nerolidol, humulene, elemol, α-cadinene, γ-bisabolene, etc.
are the examples for sesquiterpenes (Goodwin and Mercer 2003).

TABLE 1.1
Classification of Terpenoids
Name of Terpenes Molecular Formula
Hemiterpenes C5H8
Monoterpenes C10H16
Sesquiterpenes C15H24
Diterpenes C20H32
Sesterterpenes C25H40
Triterpenes C30H48
Tetraterpenes C40H64
Polyterpenes (C5H8)n
Plant Secondary Metabolites—The Key Drivers of Plant’s Defence Mechanisms 5

Diterpenes with C20 skeleton are derived from geranylgeranyl-PP and consist of acyclic,
monocyclic, bicyclic, tricyclic or tetracyclic diterpenes (Dogbo and Camara 1987; Goodwin and
Mercer 2003). More than 500 diterpenes are reported. Agathic acid, phytol, α-camphorene, cassaic
acid, steviol, etc. are some of the diterpenes (Goodwin and Mercer 2003). The sesterterpenes have
C25 skeleton and are found together with diterpenoids in lichens, seaweeds, fungi, marine organ-
isms and higher flowering plants and originate from geranylfarnesyl pyrophosphate (Bramley 1997;
Goodwin and Mercer 2003; Dewick 2009; Osbourn and Lanzotti 2009; Gonzalez 2010; Cimmino
et al. 2014). Ophiobolin A and bilosespene A are the examples for sesterterpenes from fungal and
marine metabolites, respectively (Rudi et al. 1999; Au et al. 2000). Leucosesterterpenone and leu-
costerlactone are the tetracyclic sesterterpenes isolated and identified from plant Leucosceptrum
canum (Hussain et al. 2008). Sesterterpenoids have been investigated recently for their chemical,
structural and biological characterizations and possess a wide spectrum of biological properties
against microorganisms and many of them exhibit strong anticancer efficacy (Gonzalez 2010; Wang
et al. 2013; Cimmino et al. 2014; Evidente et al. 2015).
Triterpenes (C30 skeleton) represent another large set of isoprenoid constituents (Goodwin and
Mercer 2003) and are formed from farnesyl-PP (two molecules) through a reductive head-to-head
condensation (Iriti and Faoro 2009). It consists of tetracyclic derivatives and pentacyclic compounds
(Goodwin and Mercer 2003). Lanosterol, euphol, lupeol, cycloartenol, etc. are some of the cyclic
triterpenoids (Goodwin and Mercer 2003). Phytosterols with a cyclopentanoperhydrophenanthrene
ring system are the derivatives of tetracyclic triterpenes, whereas saponins are the water-soluble
glycosidic triterpenes (Goodwin and Mercer 2003). The tetraterpenes (C40) are synthesized from
two geranylgeranyl-PP molecules through head-to-head condensation (Wendt and Schulz 1998) and
contain only one group—the carotenoid pigments (α-and β-carotene, xanthophylls, etc.) (Goodwin
and Mercer 2003). They are mainly seen in chloroplasts and guard it against photodynamic sen-
sitization and further contribute to the light-harvesting process of photosynthesis (Bramley 1997;
Goodwin and Mercer 2003). Polyterpenes (e.g. natural rubber), on the other hand, are the terpenoids
widely seen in plants with higher molecular weight (Bramley 1997; Goodwin and Mercer 2003) and
composed of more than a thousand isoprene units formed through the polymerization of geranyl-
geranyl-PP molecules (Wendt and Schulz 1998). The examples for different terpenoid classes are
shown in Figure 1.2.

1.2.1.3 Extraction and Identification of Terpenes

The non-volatile terpenoids are extracted from the plant samples with organic solvents like hexane
or hexane-ethyl acetate mixture and can be separated from other constituents present in the extract
using column chromatography with a silica stationary phase (Jiang et al. 2016). On the other hand,
the volatile terpenoids from plant tissues can extract directly with traditional extraction techniques
(e.g. hydrodistillation and organic solvent extraction) and more novel techniques like microwave-
assisted extraction and solid-phase micro extraction (Chemat et al. 2012; Pawliszyn 2012; Jiang
et al. 2016). The extracted and purified terpenoids can be identified and quantified using gas chro-
matography-flame ionization detector (GC-FID) and gas chromatography-mass spectrometry (GC-
MS) (Jiang et al. 2016).

1.2.1.4 Role and Application of Terpenes

Terpenoids are highly bioactive and play a key function in plant-plant communication and also
in plant-environment, plant-insect and plant-pathogen interactions (Dudareva et al. 2004; Cheng
et al. 2007; Bouwmeester et al. 2019). Isoprenoids took part in protein prenylation—the synthesis of
differentially lengthened isoprenoid chains anchoring proteins like G-proteins, plastoquinone and
cytochrome-a in membranes (Wendt and Schulz 1998). Terpenoids have great commercial signifi-
cance because of their vast applications as insecticides, flavouring agents and anti-microbial agents
and further due to their role in pharmaceuticals and perfumes (Martin et al. 2003).
6 Biotechnological Approaches to Enhance Plant Secondary Metabolites

FIGURE 1.2 Examples for different classes of terpenoids.

1.2.1.5 Essential Oil

The aromatic plants have characteristic fragrance due to certain compounds which collectively
form oily volatile liquids called essential oil (Joy et al. 2014; Stephane and Jules 2020). Hence, the
essential oil is considered the ‘essence’ of aromatic plants and this volatile liquid is extracted from
their different parts (Joy et al. 2014; Morsy 2017). Each plant species has its characteristic blend of
volatiles which contain species-specific compounds (Janardhanan and Thoppil 2004). Rosaceae,
Lamiaceae, Myrtaceae, Zingiberaceae, Myristicaceae, Acoraceae, Oleaceae, Cupressaceae,
Asteraceae, Umbelliferae, etc. are some plant families that have their essential oil constituents
(Stephane and Jules 2020).
‘Essential oils are synthesized by glandular trichomes and other secretory structures, special-
ized secretory tissues mainly diffused onto the surface of plant organs’ (Sharifi-Rad et al. 2017).
‘It is a heterogeneous, complex mixture composed of terpene hydrocarbons (mainly mono and ses-
quiterpenes), oxygenated compounds, straight-chain non-terpenoid hydrocarbons and their oxygen
derivatives (aldehydes, ketones, alcohols, acids, esters and ethers), phenylpropenes and miscella-
neous compounds (degradation products formed from lactones, unsaturated fatty acids, glycosides,
terpenes, sulphur and nitrogen-containing compounds)’ (Morsy 2017). However, the more volatile
fractions of essential oils are contributed by plant terpenes, especially mono and sesquiterpenes
(Janardhanan and Thoppil 2004).
Considering the physical characteristics of essential oil, most mono and sesquiterpenes have
optical activity, whereas phenylpropenes usually do not. However, the oil as a whole has optical
activity. Refractive index and specific gravity are the other physical parameters of essential oil,
Plant Secondary Metabolites—The Key Drivers of Plant’s Defence Mechanisms 7

which are useful as indicators of oil quality (Waterman 1993). The volatile oils are colourless
or may have a wide range of pale colours. The odour, another important physical parameter of
essential oil, can be classified into six major types viz., woody (odour of wood like sandal wood),
herbaceous (smell of green herbs like dill), fruity (odour of fruits), spicy (smell of spices), green
(smell of cut grass, leaves, etc.) and flowery (odour of flowers) (Janardhanan and Thoppil 2004).
Hydrodistillation is the traditional and most preferable method for essential oil extraction and
can be performed using the Clevenger trap apparatus (AOAC 1975; Rassem et al. 2016; Irshad
et al. 2019). Upon boiling, essential oil escapes along with steam and forms a separate layer in the
Clevenger trap, which can be collected (AOAC 1975). This method is cheap, easy to conduct, suit-
able for field operation and the extracted oil will be free from oil glands/ducts/cells in the plant
tissues (Janardhanan and Thoppil 2004; Irshad et al. 2019). Steam distillation, solvent extrac-
tion, hydrosteam distillation, maceration, enfleurage, expression, microwave-assisted distillation,
microwave hydro diffusion and gravity, high-pressure solvent extraction, solvent-free microwave
extraction, supercritical carbon dioxide extraction, ultrasonic extraction and the phytonic process
are the other essential oil extraction methods (Ranjitha and Vijiyalakshmi 2014; Moghaddam and
Mehdizadeh 2017; Mohammed et al. 2019). Supercritical fluid extraction with supercritical CO2 is
an advanced technique that is a simple, fast, inexpensive, effective and virtually solvent-free sample
pre-treatment method for volatile oil extraction (Pourmortazavi and Hajimirsadeghi 2007). The
selection of extraction method is based on the plant material type and the desired end products
(Janardhanan and Thoppil 2004).
The essential oils are analyzed by various chromatographic techniques like thin-layer chroma-
tography (TLC), high-pressure liquid chromatography (HPLC), column chromatography and gas
chromatography (GC) (Janardhanan and Thoppil 2004). Nuclear magnetic resonance (NMR) spec-
troscopy can elucidate the structure of essential oil constituents, and mass spectroscopy (MS) is
used for direct analysis of mixtures (Janardhanan and Thoppil 2004). Recently, GC coupled with
MS (GC-MS) is extensively used for the separation, identification and quantification of essential oil
constituents (Janardhanan and Thoppil 2004).
Apart from its role as aroma contributors, the essential oil has many other functions in plants.
Essential oils lead to attracting certain insects and are reported to help in pollination (Pichersky and
Gershenzon 2002). Certain volatile oils can protect plants from the attack of animals, microbes and
plant parasites and are also involved in other non-defensive functions like temperature regulation,
decreased water loss and reduced mechanical abrasion (Janardhanan and Thoppil 2004; Sharifi-Rad
et al. 2017). Essential oil and its constituents are also reported for allelopathic potential (EI-Sawi
et al. 2019). Thus, they are involved in the ecology and physiology of plants (Janardhanan and
Thoppil 2004; Sharifi-Rad et al. 2017).
Essential oils are well recognized for their application as perfumery contributors, medicaments
and flavouring agents and hence, they have immense significance in various industries including
perfumery, pharmaceutical and food (Hamid et al. 2011; Chavez-Gonzalez et al. 2016; Sarkic and
Stappen 2018). They are used for perfuming soaps, deodorants, cosmetics, oral care products, for
flavouring food, beverage products, etc. (Sarkic and Stappen 2018). They are also used as labora-
tory reagents, as a solvent in the paint industry, as insecticides, as a component of polishes, pastes,
ink, etc. (Janardhanan and Thoppil 2004; Sarkic and Stappen 2018; Irshad et al. 2019; Kouznetsov
2019). They have also been reported to be used in pharmaceutical industries due to their antiseptic,
stimulative, carminative, expectorant, rubefacient, diuretic, counter-irritant and flavouring prop-
erties (Janardhanan and Thoppil 2004). Essential oils are familiar for their anti-spasmodic, anti-
bacterial, anthelmintic, anti-oxidant, disinfectant, anti-cancer, anti-fungal and anti-inflammatory
activities and also for irritable bowel syndrome treatment (Perez et al. 2011; Valeriano et al. 2012;
Khanna et al. 2013; Cash et al. 2016; Nazzaro et al. 2017; Torres-Martinez et al. 2017; Blowman
et al. 2018; Ferreira et al. 2018; Heghes et al. 2019; Man et al. 2019). They are used for the manufac-
ture of ointments, lotions, creams and various syrups (Dosoky and Setzer 2018; Sarkic and Stappen
2018). Essential oil is also used widely in aromatherapy (Ali et al. 2015).
8 Biotechnological Approaches to Enhance Plant Secondary Metabolites

1.2.2 Alkaloids
Alkaloids represent another vast group of heterogeneous, physiologically active and heterocyclic
secondary metabolites (Goodwin and Mercer 2003). More than 2000 alkaloids are reported from
plants and they are distributed in all plant parts. Alkaloids are majorly basic in nature (Goodwin and
Mercer 2003) and accumulate chiefly in four types of tissues viz., vascular sheaths, actively growing
tissue, epidermal and hypodermal cells, and latex vessels (Goodwin and Mercer 2003). They are
present in vacuoles and thus do not appear in young cells until vacuolation happens (Goodwin and
Mercer 2003). Alkaloids are frequently stored in tissues other than their site of synthesis. Likewise,
secondary structural modifications of alkaloids often occur at sites other than where primary
synthesis took place (Goodwin and Mercer 2003). Dicots are richer in alkaloids than monocots
whereas they have limited distributions in lower plants (Goodwin and Mercer 2003). Apocynaceae,
Rubiaceae, Solanaceae, Papaveraceae, Leguminosae and Fumariaceae are the families rich in
alkaloids, whereas those like Rosaceae and Labiatae contain alkaloids to a lesser extent (Goodwin
and Mercer 2003).

1.2.2.1 Synthesis of Alkaloids

A common pathway that can explain alkaloid biosynthesis does not exist (Iriti and Faoro 2009). The
biosynthetic precursors for alkaloids are almost always amino acids (Goodwin and Mercer 2003).
Other multi-carbon units like nicotinic acid, anthranilic acid, isoprenoids, acetate and purine are
also incorporated into the final structure of some alkaloids (Iriti and Faoro 2009). Even though the
biosynthesis of alkaloids cannot be easily explained, some typical reactions like synthesis of Schiff
bases and the Mannich reaction which take part in the synthesis of different classes of the alkaloid
can be explained (Plemenkov 2001). The Schiff bases are formed within a molecule, by the reaction
of amines with aldehydes or ketones and these reactions are the general method for the produc-
tion of C=N bonds (e.g. synthesis of piperidine alkaloids) (Plemenkov 2001; Dewick 2002). On the
other hand, the Mannich reaction proceeded in both intra and inter molecularly (Plemenkov 2001;
Dewick 2002). In addition to a carbonyl compound and an amine, the Mannich reaction contains
an integral component, a carbanion, which plays the function of the nucleophile in the nucleophilic
addition to the ion formed by the reaction of the carbonyl and the amine (Plemenkov 2001; Dewick
2002). In addition to the biosynthesis of monomeric alkaloids which involved the aforementioned
reactions, there are also dimeric, trimeric and tetrameric alkaloids synthesized through conden-
sation of two, three and four monomeric alkaloids, respectively (Hesse 2003). Dimeric alkaloids
are normally synthesized from the same type of monomers through the following mechanisms:
Michael reaction (e.g. villalstonine), Mannich reaction (e.g. voacamine), condensation of alde-
hydes with amines (e.g. toxiferine), lactonization (e.g. carpaine) and oxidative addition of phenols
(e.g. dauricine) (Hesse 2003).

1.2.2.2 Classification of Alkaloids

The alkaloids formed from amino acids and have nitrogen in a heterocyclic ring are commonly
known as true alkaloids (e.g. nicotine, morphine) and those without such rings are called protoalka-
loids (e.g. ephedrine, cathinone) (Goodwin and Mercer 2003; Wansi et al. 2013). The alkaloids (with
or without heterocyclic rings) which are not formed from amino acids are called pseudoalkaloids,
in which carbon skeleton is derived from isoprenoids (e.g. actinidine, dendrobine) (Goodwin and
Mercer 2003). Besides, steroidal alkaloids are also present and which include, cholestane deriva-
tives, pregnane derivatives and C-nor-D-homosteroidal alkaloids (Goodwin and Mercer 2003).
Alkaloids are also grouped based on their biosynthetic precursors and chemical structure
(Table 1.2) (Iriti and Faoro 2009). The main alkaloid groups are formed from the amino acid pre-
cursors like ornithine (pyrrolidine, tropane and pyrrolizidine alkaloids), leucine (pyrrole alka-
loids), lysine (piperidine, indolizidine and quinolizidine alkaloids), tyrosine (catecholamines,
tetrahydroisoquinoline, isoquinoline and benzyltetrahydroisoquinoline alkaloids), tryptophan
Plant Secondary Metabolites—The Key Drivers of Plant’s Defence Mechanisms 9

TABLE 1.2
Precursors and Different Classes of Alkaloids
Precursors Classes of Alkaloid Examples
Ornithine Pyrrolidine1, Tropane, 1. Cocaine, Atropine
Pyrrozidine, Polyamines2 2. Putrescine, Spermine, Spermidine
Leucine Pyrrole -
Lysine Piperidine, Indolizidine, -
Quinolizidine
Tyrosine Isoquinoline, Tetrahydroisoquinoline, 1. Morphine, Curarines, Papaverine
Benzyltetrahydroisoquinoline1, 2. Adrenaline, Noradrenaline
Catecholamines2
Tryptophan Indole1, Carbolines, Quinoline2, 1. Vindoline, Catharantine
Pyrrolindole, Indolamines3 2. Quinine, Capthotecin
3. Melatonin, Serotonin
Histidine Imidazole -
Phenylalanine Capsaicin, Ephedrine
Anthranilic acid Quinazoline, Quinoline, Acridine -
Purine Theobromine, Caffeine, Theophylline
Geranylgeranyl-diphosphate Terpenoidic -
Cholesterol Steroidal Solanin
Acetate Piperidine -
Nicotinic acid Pyridine Nicotine

Source: Adapted from Iriti and Faoro (2009).

(indolamines, carboline, indole, quinoline, pyrrolindole and ergot alkaloids), histidine (imidazole
alkaloids) and phenylalanine (Cordell et al. 2001; Facchini 2001; Hughes and Shanks 2002).

1.2.2.3 Extraction and Identification of Alkaloids

The alkaloids can be extracted with different methods. The alkaline alkaloids can be extracted
with water or acidic water. The acid extraction method generally uses 0.1% to 1% hydrochloric
acid, sulphuric acid, tartaric acid or acetic acid solution as a solvent (Yubin et al. 2014). Both free
and salt alkaloids can be extracted using organic solvents like methanol or ethanol (Yubin et al.
2014). Further, lipophilic free alkaloids can be extracted using organic solvents like chloroform,
benzene, ether and methylene chloride (Yubin et al. 2014). The extracted alkaloids can be separated
using methods like solvent method, precipitation method, fractional distillation method, gradient
pH method, dialysis method, crystallization method and chromatographic method (Yubin et al.
2014; Feng et al. 2019). The extracted alkaloids can further go for their purity checking using physi-
cal property measurements and chromatographic techniques including TLC, HPLC and GC (Feng
et al. 2019). The purified alkaloids can be further identified through structural elucidation using
different techniques like ultraviolet-visible spectra, infrared spectra (IR), NMR spectroscopy and
mass spectrometry (MS) (Feng et al. 2019).

1.2.2.4 Role and Application of Alkaloids

Alkaloids have many ecological, biochemical and physiological roles in plants (Goodwin and
Mercer 2003). They protect the plants from insects and herbivores, act as N-excretory products in
the same way as uric acid and urea in animals and further as nitrogen reserve and growth regula-
tors (Goodwin and Mercer 2003). They also involve maintaining ionic balance under their che-
lating power (Goodwin and Mercer 2003). Alkaloids represent massive phytoconstituents with
10 Biotechnological Approaches to Enhance Plant Secondary Metabolites

nutritional, pharmacological, toxicological and cosmetic potential (Iriti and Faoro 2009). They
have great importance in pharmaceutical industries because of their vast pharmacological activities
like anti-amoebic, anti-plasmodial, anti-oxidant, anti-cholinergic, anti-malarial, anti-hypertensive,
anxiolytic, anti-microbial, anti-inflammatory, anti-depressant, anti-HIV and anti-tumour properties
(Wright et al. 1991; Cowan 1999; Kupeli et al. 2002; Hesse 2003; Cos et al. 2008; Kulkarni and Dhir
2009; Liu et al. 2013; Yuan et al. 2019).

1.2.3 Phenolics
Phenolics are ubiquitous plant secondary metabolites and these phytoconstituents occur as free
phenols or their glycosides (Marinova et al. 2005; Hussein and El-Anssary 2019). This class of
compounds consists of more than 8000 biologically active constituents which range from simple
phenolic compounds to polymeric structures with a molecular weight of above 30000 Da (Marinova
et al. 2005).

1.2.3.1 Synthesis of Phenolics

Phenolics are the class of phenylalanine derivatives and they possess a basic C6 ─C3 (phenyl-
propane) skeleton. They are synthesized through the phenylpropanoid pathway (Figure 1.3)

FIGURE 1.3 Aromatic amino acids biosynthesis from shikimic acid and phenyl alanine derivatives of phen-
ylpropanoid pathway. (Adapted from Iriti and Faoro 2009.)
Plant Secondary Metabolites—The Key Drivers of Plant’s Defence Mechanisms 11

and are also called phenylpropanoids (Iriti and Faoro 2009; Hyskova and Ryslava 2019).
‘Phosphoenolpyruvate and erythrose 4-phosphate from the glycolysis and pentose phosphate path-
way are the precursors of phenylpropanoid pathway and which leads to the synthesis of two main
intermediates, shikimic and chorismic acid’ (Weaver and Hermann 1997). ‘In the succeeding steps,
after a branch point, phenylalanine and tyrosine are formed from prephenic and arogenic acid,
whereas tryptophan from anthranilic acid’ (Weaver and Hermann 1997). The phenylalanine further
subjected to deamination reaction catalyzed by PAL (phenylalanine ammonia-lyase), which leads to
the synthesis of cinnamic acid and this forms the key step for the synthesis of phenolic constituents
(Iriti and Faoro 2009).
After a series of hydroxylation reactions of the benzene ring, hydroxycinnamates (e.g. coumaric,
sinapic and ferulic acids) are formed from the cinnamic acid precursor (Iriti and Faoro 2009).
Hydroxycinnamates are reduced to their respective alcohols through aldehyde intermediates and
collectively termed monolignols which are further subjected to dimerization or polymerization to
form lignans and lignin, respectively (Iriti and Faoro 2009). Lignification is a complex reaction in
which the polymerization of lignin units is catalyzed by peroxidases, consuming H2O2 (Hahlbrock
and Scheel 1989; Iriti and Faoro 2004). Benzoic and hydroxybenzoic acids (C6 ─C1) like salicylic
acid are another cinnamic acid derivative group, synthesized by the cleavage of C2 fragment from
the phenylpropane structure (Hahlbrock and Scheel 1989; Iriti and Faoro 2004). However, based on
the species and the conditions (e.g. pathogen attack), salicylic acid can also synthesize directly from
chorismate-isochorismate by isochorismate synthase (Wildermuth et al. 2001). Figure 1.4 explains

FIGURE 1.4 Main steps of the phenylpropanoid pathway leading to benzoic acids, hydroxycinnamates,
monolignols and lignin. (Adapted from Iriti and Faoro 2009.)
12 Biotechnological Approaches to Enhance Plant Secondary Metabolites

the phenylpropanoid pathway’s major steps which lead to the synthesis of benzoic acids, hydroxy-
cinnamates, monolignols and lignin.

1.2.3.2 Classification of Phenolics

The phenolics can be classified as simple phenols, phenolic acids, phenylacetic acids, hydroxy-
cinnamic acids, phenylpropenes, coumarins, quinones, stilbenes, xanthones, lignans, neolignans,
melanins, tannins and flavonoids (Goodwin and Mercer 2003).
Simple phenols are not widely distributed and the most common simple phenol is hydroquinone.
The phenolic acids are extensively present throughout the plant kingdom (Goodwin and Mercer
2003). Protocatechuic acid, p-hydroxybenzoic acid, syringic acid and vanillic acid are some of
the phenolic acids (Goodwin and Mercer 2003). Aldehyde or alcohols corresponding to the par-
ent carboxylic acids (e.g. vanillin) are also known (Goodwin and Mercer 2003). Even though the
distribution of phenylacetic acids in plants is not known much, 2- and 4-hydroxyphenylacetic acids
were reported (Goodwin and Mercer 2003). The hydroxycinnamic acids, like p-coumaric acid and
caffeic acid and their methylated derivatives like ferulic acid and synapic acid, are universally pres-
ent in higher plants (Goodwin and Mercer 2003). Phenylpropenes are not widely distributed, but
they occur sporadically in essential oils (e.g. myristicin) (Goodwin and Mercer 2003). Coumarins
are lactones, derived from o-hydroxycinnamic acid and occur abundantly in plants (Goodwin and
Mercer 2003). The quinones mainly include benzoquinones, naphthoquinones and anthraquinones,
whereas lunalaric acid is the most common plant stilbene (Goodwin and Mercer 2003). Over 70
xanthones were reported in plants and their distribution is restricted to Guttiferae and Gentianaceae
families (e.g. mangiferin) (Goodwin and Mercer 2003).
The lignans are naturally occurring phenylpropanoid dimers in which the two C6 ─C3 units
are joined tail-to-tail by the carbon-carbon bond between the middle carbons of their side chains
(e.g. conidendrin) (Goodwin and Mercer 2003). If the C6 ─C3 units are joined head-to-tail, the resul-
tant phenolics are called neolignans (e.g. eusiderin) (Goodwin and Mercer 2003). Melanins are
brown or black, naturally occurring pigments (Goodwin and Mercer 2003). Melanin, characteristics
of plants are called catechol melanins, since they yield catechol on alkali fusion (Goodwin and
Mercer 2003). The tannins are class of high molecular weight chemical constituents with tanning
properties and mainly include hydrolysable and condensed tannins (Goodwin and Mercer 2003).
The flavonoids are the largest group of naturally occurring phenolic compounds and are distrib-
uted in different parts of the plant, both in free form and as glycosides (Sulaiman and Balachandran
2012). These compounds function as co-pigments and are responsible for different colours exhib-
ited by different plant parts (Goodwin and Mercer 2003; Iwashina 2015). The flavonoid glycosides
are found to be occurring as O-glycosides and C-glycosides, where the latter received less atten-
tion (Xiao et al. 2016). The structure of flavonoid consists of 15 carbon atom phenylpropanoid
core, together with 2 six-carbon aromatic rings (ring A and B) joined by a heterocyclic ring which
contains 3 carbon atoms (ring C), which together can be represented as C6 ─C3─C6 (Cheng et al.
2014). The flavonoids can be classified into the following categories. Flavones (contains benzo-
γ-pyrone ring; e.g. apigenin), flavanone (have a 2,3-dihydro skeleton in C6 ─C3─C6 structure of
flavonoid; e.g. naringenin), flavonol (3-hydroxy derivative of flavone; e.g. kaempferol), dihydrofla-
vonol (3-hydroxy derivative of flavanone; e.g. taxifolin), isoflavones (contain benzo-γ-pyrone ring
with phenyl substitution at position 3 of the pyrone ring; e.g. genistein), chalcones (open chain
flavonoids; e.g. butein) and dihydrochalcones (dihydro derivatives of chalcones; e.g. phloretin)
(Goodwin and Mercer 2003; Alihosseini 2016; Goncalves et al. 2018; Wang et al. 2018; Banoth
and Thatikonda 2019; Shao and Bao 2019). They also include the aurones (possess the 2-ben-
zylidene-coumaranone skeleton; e.g. sulphuretin), anthocyanins (glycosides of anthocyanidins;
e.g. pelargonidin), leucoanthocyanidins (have flavan-3,4-diol skeleton; e.g. teracacidin) and biflavo-
noids (dimers of various flavonoids linked by C─C or C─O─C bond; e.g. amentoflavone) (Goodwin
and Mercer 2003; Halbwirth 2010; Mercader and Pomilio 2013; Leicach and Chludil 2014; Antal
et al. 2016).
Plant Secondary Metabolites—The Key Drivers of Plant’s Defence Mechanisms 13

1.2.3.3 Extraction and Identification of Phenolics

Various techniques for extraction of phenolic constituents are developed and which include LLE
(liquid-liquid extraction), UAE (ultrasound-assisted extraction), MAE (microwave-assisted extrac-
tion) and SFE (supercritical fluid extraction) (Jahromi 2019). Maceration, soxhlet extraction and
hydro distillation are the main methods for the extraction of phenolic compounds with the LLE
technique (Garcia-Salas et al. 2010). The main parameters of these methods are the type and polar-
ity of solvents and their ratio, time and temperature of extraction and also, chemical composition and
physical characteristics of the samples (Garcia-Salas et al. 2010). In UAE, using ultrasound waves,
cavitation bubbles are created near the sample tissue and then they break the cell wall, and thereby
the cell content is released (Altemimi et al. 2016). In the other method, the microwave-assisted
extraction, the samples are allowed to interact with the microwave (Sookjitsumran et al. 2016). The
microwave generates heat as a result of molecular motion induction and hence the cell wall will be
ruptured and which will further result in the release of active substances in the cell (Sookjitsumran
et al. 2016). In the supercritical fluid extraction method, the solvent is at a temperature and pressure
above its critical point and there is no surface tension in it (Murga et al. 2000). Hence, it concur-
rently shows the properties of liquid and gas which can be much efficient for phenolic extraction
from plants. High diffusivities and low viscosities of supercritical fluids make them able to extract
different phenolic constituents in less time with more effectiveness (Murga et al. 2000).
Since there are phenolic constituents with various chemical properties and structures, differ-
ent protocols and techniques like UV-visible spectroscopy, column chromatography, near-infrared
reflectance spectroscopy, NMR, HPLC, GC-MS and liquid chromatography-mass spectrometry
(LC-MS) are extensively used for their purification, identification and quantification (Kivilompolo
et al. 2007; Jahromi 2019).

1.2.3.4 Role and Application of Phenolics

The role of phenolic constituents in plant physiology and interactions with biotic and abiotic stress
conditions are difficult to overestimate (Bhattacharya et al. 2010). The phenolic constituents are
in plant tissues as an adaptive response to unfavourable environmental conditions and they play a
pivotal function in the regulation of different environmental stresses, like high light, nutrient defi-
ciency, low temperature and microbial infection (Naikoo et al. 2019). In short, phenolics are synthe-
sized by plants primarily for getting protection from stress and further to get structural integrity and
scaffolding support (Bhattacharya et al. 2010). Phenolics secreted by perturbed or wounded plants
can kill or repel many microorganisms (Bhattacharya et al. 2010). The phenolic compounds have
a broad range of medicinal potentials like anti-oxidant, anti-diabetic, anti-microbial, anti-tumour,
anti-viral, analgesic, anti-inflammatory and anti-pyretic efficacies (Chen and Ho 1997; Narasimhan
et al. 2004; Sun et al. 2007; Adisakwattana et al. 2008; Cheynier 2012).

1.3 CURRENT BIOTECHNOLOGICAL APPROACHES FOR THE ENHANCEMENT

OF SECONDARY METABOLITE CONTENTS IN PLANTS
We are dependent on plants for food, furniture, constructions and fuel since pre-historic times.
Plants in all forms have tendered their services to mankind (Amjad et al. 2013). Humans have
selected plants based on the value and potential for domestication (Ramawat et al. 2009). People
living in different continents of the world also used plants to treat various diseases and various
systems of medicines were generated viz. Ayurvedic, Unani, Chinese system of medicines, etc.
(Nadkarni 1996; Che et al. 2017; Dar et al. 2017a). Due to the advancement of science and tech-
nologies, various other non-conventional methods to treat the diseases were also identified. Each
system of medicine is having advantages and disadvantages. The system which utilized plant-based
natural products for the discoveries of drugs has attracted attention across the globe, due to its least
or no side effects (Dubey et al. 2004; Yuan et al. 2016; Roy et al. 2018). Plant synthesized various
14 Biotechnological Approaches to Enhance Plant Secondary Metabolites

secondary metabolites besides its primary metabolites to resist in the changing environment and to
cross-talk between signalling pathways within the plant and with other biological entities (Jacobo-
Velazques et al. 2015; Martin 2017; Rai et al. 2017). These secondary metabolites are reported to
serve humans across various domains from food to pharmaceutical industries around the globe (Dar
et al. 2017a). To harvest these medicinal principles from the medicinal plants, people have exploited
the plant germplasm from the natural habitat at an alarming rate (Maxted et al. 2020). Many of the
medicinal plants are now considered endangered (Sharma and Thokchom 2014). As per the reports,
the contents of these naturally occurring medicinal components were found low in most of the
plants (Verma and Shukla 2015). So, it was needed to enhance the contents of such key principal
components of the plants. Various people around the globe tried to enhance the contents of the sec-
ondary metabolites in the plants using various methods and were reported significant enhancement
of the targeted secondary metabolites in the tested plants (Hussain et al. 2012). The most common
methods employed to augment the contents of the plant secondary metabolites are as follows:

1. Tissue culture approaches: Plant cell and tissue culture technologies using explants like
leaves, roots, stems and meristems are established for the enhanced in vitro production
of secondary plant metabolites as it offers controlled supply of biochemicals independent
of plant availability and bioreactors are the key process for their commercial production
(Sajc et al. 2000; Hussain et al. 2012). Other advantages of this technique over conven-
tional plant cultivation include the production of a metabolite of interest under controlled
environmental conditions, cultured cells could be devoid of contaminations with micro-
organisms and insects, a cell could be easily multiplied to get their specific metabolite,
automated cell growth control and metabolite process regulation can reduce the labour cost
and further, the productivity could be improved and the metabolite of interest is extract-
able from the cultures (Hussain et al. 2012). Taxol, morphine, codeine, L-Dopa, capsaicin
and berberine are some of the potentially active natural metabolites produced in large
quantities using plant cell and tissue culture techniques (Tam et al. 1980; Yoshikawa et al.
1985; Holden et al. 1988; Morimoto et al. 1988; Wichers et al. 1993; Fett-Neto et al. 1994;
Thengane et al. 2003).
2. Abiotic elicitation: The treatment of plant cells with abiotic elicitors is a useful approach
to increase the secondary metabolite production in cell cultures (Karuppusamy 2009).
Abiotic elicitors represent non-biological origin substances and are grouped into physical,
chemical and hormonal factors (Naik and Al-Khayri 2016). Light, osmotic stress, salin-
ity, drought and thermal stress are the physical elicitors (Naik and Al-Khayri 2016). The
metabolite production can be affected by light and the effect of light on the enhanced pro-
duction of two important secondary metabolites—gingerol and zingiberene—in Zingiber
officinale callus culture is reported (Anasori and Asghari 2008). Osmotic stress, an impor-
tant environmental stress, is proven to alter the physiological and biochemical properties
of plants and increase the secondary metabolite concentrations in plants (Zobayed et al.
2007). Polyethylene glycol is an osmotic agent which can induce water stress in many
plants (Van den Berg and Zeng 2006). Exposure to salinity is also known to stimulate
or induce the production of secondary metabolite (Haghighi et al. 2012). Catharanthus
roseus grown under salt stress showed increased levels of vincristine—an important alka-
loid (Misra and Gupta 2006). Drought stress, which can also greatly reduce plant growth,
can increase secondary metabolite content (Naik and Al-Khayri 2016). Heavy metals rep-
resent chemical elicitors and agents for abiotic stress in living organisms (Cai et al. 2013).
Heavy metal-induced changes in the metabolic activity of plants can alter the production
of photosynthetic pigments, nonprotein thiols, proteins and sugars (Naik and Al-Khayri
2016). Plant hormones are also widely used as elicitors and jasmonic acid, salicylic acid
and gibberellins are the most studied hormones in this category (Naik and Al-Khayri
2016). Jasmonic acid induces the production of different plant secondary metabolites,
Plant Secondary Metabolites—The Key Drivers of Plant’s Defence Mechanisms 15

including rosmarinic acid, terpenoids indoles alkaloids and plumbagin in different cell
cultures (Krzyzanowska et al. 2012; Almagro et al. 2014; Silja et al. 2014). Salicylic acid is
another hormonal elicitor that is reported to enhance the production of various secondary
metabolites like tanshinones (Xiaolong et al. 2015), withanolide A, withanone and witha-
ferin A (Sivanandhan et al. 2013), stilbene (Xu et al. 2015), vincristine and vinblastine
(Idrees et al. 2010). Another phytohormone gibberellin is also known as an elicitor for
secondary metabolite production (Liang et al. 2013).
3. Biotic elicitation: Biotic elicitors represent the substances of biological origin and they
contain polysaccharides of plant cell walls (e.g. pectin, chitin, cellulose and chitosan) and
microorganisms (Naik and Al-Khayri 2016). The treatment of chitosan in Plumbago rosea
cultures enhanced plumbagin content (Komaraiah et al. 2003) whereas chitosan or chitin
application induced the synthesis of fluoroquinolone and coumarins in shoot cultures of
Ruta graveolens (Orlita et al. 2008). Microorganisms also play a role as biotic elicitors
by inducing plant defence response and by triggering the pathways to alter the secondary
metabolite production (Naik and Al-Khayri 2016). Researchers use yeast extract for years
as one of the important biotic elicitors to stimulate the production of various metabo-
lites like ethylene in tomato and tanshinone in Perovskia abrotanoides root culture (Felix
et al. 1991; Arehzoo et al. 2015; Naik and Al-Khayri 2016). The effectiveness of fungal
preparations (pathogenic and nonpathogenic) as elicitors is one of the most effective biotic
strategies to stimulate a plant’s phenylpropanoid and flavonoid biosynthetic pathways
(Dixon et al. 2002; Lattanzio et al. 2006; Naik and Al-Khayri 2016). The production of
indole alkaloids serpentine, ajmalicine and catharanthine has increased up to five times in
C. roseus cell suspensions and raucaffrincine in Rauwolfia canescens using fungal cell
wall fragments (Parchmann et al. 1997; Zhao et al. 2001; Namdeo et al. 2002). Likewise,
fungal mycelia increased the diosgenin content in Dioscorea deltoidea by 72% (Rokem
et al. 1984). Furthermore, bacterial elicitors induce scopolamine biosynthesis in Scopolia
parviflora adventitious hairy root cultures (Jung et al. 2003). Likewise, a significant
increase in the glycyrrhizic acid content was observed in Agrobacterium rhizogenes,
Bacillus aminovorans and Bacillus cereus challenged cultures (Awad et al. 2014). The
secondary metabolite production enhancement in hydroponic cultures using biotic elicitors
was also reported (Vu et al. 2006). Datura innoxia Mill., were cultivated in hydroponic
conditions in which nutrient solution was inoculated with Agrobacterium rhizogenes and
found that there is an increased production of alkaloid content in the plant roots growing
in hydroponic culture (Vu et al. 2006).
4. Hairy root culture: The hairy root system based on inoculation with Agrobacterium rhizo-
genes is another method for the production of secondary metabolites synthesized in plant
roots (Palazon et al. 1997; Karuppusamy 2009).
5. Production of transgenic lines: Genetic manipulation became a promising alternative
for biotechnological exploitation of plant cells and hence the engineered plants will give
an increased enzyme activity and enhanced production of the respective metabolite
(Karuppusamy 2009; Hussain et al. 2012).
6. Biosynthetic metabolic pathway engineering: Metabolic pathway engineering is another
approach for increased secondary metabolite production through the targeted and pur-
poseful alteration of metabolic pathways (Lessard 1996). Metabolic engineering can offer
different strategies to enhance the productivity of the desired metabolite by increasing
the number of producing cells, through over expression of genes, by enhancing the rate-
limiting enzymes or blocking the feedback inhibition mechanism and competitive path-
ways (Hussain et al. 2012).
7. Targeted gene editing using CRISPR CAS9: CRISPR-Cas9-based genome editing sys-
tems are now flashed to the future efforts in the elucidation and engineering of biosyn-
thetic pathways of novel natural products (Nielsen et al. 2017). The CRISPR/Cas9 system
16 Biotechnological Approaches to Enhance Plant Secondary Metabolites

exhibits vast applications with high precision and accuracy compared to other genome
editing tools. Hence, it can probably be used as a common technique for plant metabolic
engineering in the future (Alagoz et al. 2016). The effectiveness of CRISPR-Cas9 genome
editing technology in the manipulation of bioactive alkaloid biosynthetic pathways in
Papaver somniferum L. has been successfully demonstrated (Alagoz et al. 2016).
8. RNAi technology: RNA interference technology (RNAi) possesses the main impact on the
manipulation of secondary metabolites which represents three major groups viz. terpe-
noids, alkaloids and phenylpropanoids (Wagner and Kroumova 2008). RNAi might prove
to be effective for the production of novel natural products from plants, which in turn
can give novel and rapid applications (Borgio 2009). Researchers used RNAi technology
to yield unnatural plant natural products from lengthy and complex metabolic pathways
(Borgio 2009). Runguphan et al. (2009) explained the elimination of monoterpene indole
alkaloids (derived from two starting compounds, tryptamine and secologanin) produc-
tion in C. roseus hairy root culture through RNA mediated suppression of tryptamine
biosynthesis. In this study, they have introduced an unnatural tryptamine analogue to the
production media and established that the silenced C. roseus culture could produce novel
alkaloids derived from this unnatural starting substrate. This provided an advantage that
the novel natural product synthesized this way is not contaminated due to the presence of
normal natural alkaloids present in C. roseus.
9. Induced mutation (gamma radiation), nuclear radiations and mutation breeding are the
other method of enhancement of secondary metabolite production. The effect of gamma
radiation on the formation of biomass and the yield of pharmacologically important second-
ary metabolites in Hypericum triquetrifolium Turra callus cultures induced from different
seedling parts has been demonstrated (Azeeza et al. 2017). Photo-elicitation of bioactive
secondary metabolites using ultraviolet radiation has also been studied (Matsuura et al.
2013). The role of mutation breeding in the alteration of secondary metabolite production
has also been investigated (Kolakar et al. 2018). In periwinkle ‘Dhawal’, a high alkaloid
producing variety has been developed by chemical mutagen treatment of seeds followed by
rigorous selection in widely cultivated variety ‘Nirmal’ (Kulkarni and Baskaran 2003). An
increase in sapogenin (diosgenin) level in Trigonella corniculata following chemical muta-
genesis using dimethyl and diethyl sulphate has also been reported (Kolakar et al. 2018).

These new approaches will help to explore the plants as a potential source of many more phytochem-
icals. Continuation and much deeper efforts in this area will hopefully direct to the improved bio-
technological production of specific, high-value and yet underexplored plant metabolites (Hussain
et al. 2012).

1.4 CONCLUSION
This chapter has dealt with plant secondary metabolites, their synthesis, classification, biological
function, medicinal applications and an overview of the recent advancements for the enhanced
production of novel secondary metabolites in plants. There is a huge range of secondary metabo-
lites present in the plant kingdom, with a very varied distribution. Secondary metabolites are not
required directly for the growth and development of plants but they are necessary for the survival
of plants in adverse conditions. They protect plants from biotic and abiotic stress conditions through
the chemical defence mechanisms. The capability of plants to defend and survive is thus ultimately
because of the ecological functions of their secondary metabolites. Such metabolites are studied
and clinically proven for their ethnopharmacological potential also and which will aid to human
health. However, many of such secondary metabolites are still under-explored and unidentified for
their potential and thus the research area of natural products still requires much more attention and
thorough study. Further, because of the increased commercial significance of high-value secondary
Plant Secondary Metabolites—The Key Drivers of Plant’s Defence Mechanisms 17

metabolites in recent years, there is a great inducement for researchers to focus much on secondary
metabolism, especially in the option of their enhanced production by means of different approaches.
The common approaches mainly include tissue culture approaches, biotic and abiotic elicitation, meta-
bolic engineering, targeted gene editing, RNAi technology, production of transgenic lines, induced
mutation, nuclear radiations and mutation breeding. In future, the aforementioned biotechnological
approaches might be approached as an alternative production method to overcome the less acces-
sibility of biologically potential, commercially important and medicinally valuable plant secondary
metabolites. Advances in bio techniques, especially methods for culturing plant cell cultures, might
give new angles for the commercial processing of even rare medicinal plants and their novel chemicals.

ACKNOWLEDGEMENTS
Dr. D. Sruthi acknowledges the Department of Health Research (DHR), Government of India, New
Delhi, for her award of Young Scientist-HRD scheme (YSS/2019/000035/PRCYSS). Dr. D. Sruthi
is also grateful to Science and Engineering Research Board (SERB), Department of Science and
Technology, Government of India, New Delhi, for her award of National post-doctoral Fellowship
(PDF/2017/000339). Acknowledgement is further extended to Dr. T. John Zachariah, Former Principal
Scientist, and Acting Director, ICAR-Indian Institute of Spices Research, Kozhikode, Kerala, for his
valuable guidance on plant secondary metabolites during the doctoral period of Dr. D. Sruthi.

Contact Information: Please contact the author at: [email protected]

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 Now let us
resume our walk.
The Strand is very
little altered, you
think. Already Exeter
Change is gone;
Exeter Hall is already
built; the shops are
less splendid, and
plate glass is as yet
unknown; in Holywell
Street I can show
you one or two of
the old signs still on
the house walls;
EXETER CHANGE Butcher Row, behind
St. Clement Danes, is
pulled down and the street widened; on the north side there is
standing a nest of rookeries and mean streets, where you will have
your Law Courts; here is Temple Bar, which you will miss; close to
Temple Bar is the little fish shop which once belonged to Mr.
Crockford, the proprietor of the famous club; the street messengers
standing about in their white aprons will be gone in your time; for
that matter, so will the aprons; at present every other man in the
street wears an apron. It is a badge of his rank and station; the
apron marks the mechanic or the serving-man; some wear white
aprons and some wear leather aprons; I am afraid you will miss the
apron.
Fleet Street is much more picturesque than the Strand, is it not?
Even in your day, Eighty-seven, when so many old houses will have
perished, Fleet Street will still be the most picturesque street in all
London. The true time to visit it is at four o’clock on a summer
morning, when the sun has just risen on the sleeping city. Look at
the gables of it, the projecting stories of it, the old timber work of it,
the glory and the beauty
of it. As you see Fleet
Street, so Dr. Johnson
saw it.
There is a good deal
more crowd and
animation in Fleet Street
than in the Strand. That
is because we are nearer
the City, of course; the
traffic is greater; the
noise is much greater. As
for this ring before us, let
us avoid it. A coachman
fighting a ticket-porter is
a daily spectacle in this
thoroughfare; those who
crowd round often get
bloody noses for their
pains, and still more
THE PARISH ENGINE
often come away without
(From a Drawing by George Cruikshank in ‘Sketches by Boz’)
their purses. Look! The
pickpockets are at their
work almost openly. They have caught one. Well, my friend, our long
silk purses—yours will be square leather things—are very easily
stolen. I do not think it will repay you for the loss of yours to see a
poor devil of a pickpocket pumped upon.
You are looking again at the plain windows with the small square
panes. The shops make no display as yet, you see. First, it would not
be safe to put valuable articles in windows protected by nothing but
a little thin pane of glass—which reminds me that in the matter of
street safety you will be a good deal ahead of us; next, an honest
English tradesman loves to keep his best out of sight. The streets
are horribly noisy. That is quite true. You have heard of the roar of
the mighty city. Your London, Eighty-seven, will not know how to
 IN FLEET STREET—PROCLAIMING THE QUEEN.

roar. But you can now understand what its roaring used to be. An
intolerable stir and uproar, is it not? But then your ears are not, like
ours, used to it. First, the road is not macadamised, or asphalted, or
paved with wood. Next, the traffic of wagons, carts, and
wheelbarrows, and hand-carts, is vastly greater than you had ever
previously imagined; then there is a great deal more of porter work
done in the street, and the men are perpetually jostling, quarrelling,
and fighting; the coaches, those of the short stages with two horses,
and the long stages with four, are always blowing their horns and
cracking their whips. Look at yonder great wagon. It has come all
the way from Scotland. It is piled thirty feet high with packages of all
kinds: baskets hang behind, filled with all kinds of things. In front
there sit a couple of Scotch lasses who have braved a three weeks’
journey from Edinburgh in order to save the expense of the coach.
Brave girls! But such a wagon with such a load does not go along
the street in silence. It is not in silence either that the women who
carry baskets full of fish on their
heads go along the street, nor is
the man silent who goes with a
pack-donkey loaded on either
side with small coal; and the
wooden sledge on which is the
cask of beer, dragged along by a
single horse, makes by itself as
much noise as all your carriages
together, Eighty-seven.
And there is nothing, you
observe, for the protection and
convenience of passengers who
wish to cross the road. Nothing
at all. No policeman stands in
the middle of the road to
regulate the traffic; the drivers
pay no heed to the foot CROCKFORD’S FISH SHOP
passengers; at the corner of (From a Drawing by F. W. Fairholt)

Chancery Lane, where the press

is the thickest, the boys and the clerks slip in and out among the
horses and the wheels without hurt: but how will those ladies be
able to get across? They never would but for the crossing-sweeper—
the most remunerative part of the work, in fact, is to convoy the
ladies across the road; if he magnifies the danger of this service,
and expects silver for saving the lives of his trembling clients, who
shall blame him?
There are still left some of the old posts which divided the
footway from the roadway, though the whole is now paved and—
what, Eighty-seven? You have stepped into a dandy-trap and
splashed your feet. Well, perhaps, in your day they will have learned
to pave more evenly, but just at present our paving is a little rough,
and the stones sometimes small, so that here and there, after rain,
these things will happen.
 Here we are at Blackfriars. This is the
Gate of Bridewell, where they used to flog
women, and still flog the ’prentices. Yonder
is the Fleet Prison, of which we have just
read an account in the ‘Pickwick Papers.’
They have cleared away the old Fleet
Market, which used to stand in the middle
of the street, and they have planted it
behind the houses opposite the Prison.
Come and look at it. Let us tread softly over
the stones of Farringdon Market, for
THOMAS CHATTERTON somewhere beneath our feet lie the bones
of poor young Chatterton. No monument
has been erected here to his memory, nor is the spot known where
he lies, but it is somewhere in this place, which is a tragic and
mournful spot, being crammed beneath its pavement with the bones
of the poor, the outcast, the broken down, the wrecks and failures of
life, and littered above the pavement with the wreckage and refuse
of the market. This place was formerly the burial-ground of the Shoe
Lane Workhouse.
We can walk down to the Bridge and look at the river. No
Embankment yet, Eighty-seven. No penny steamers, either. Yet the
watermen grumble at the omnibuses which have cut into their trade.
Here comes the lamplighter, with his short ladder and his
lantern.
Gas, of course, has been introduced for ever so long. They have
blindly followed the old plan of lighting, and have stuck up a gas
lamp wherever there used to be an oil lantern. The theatres and
places of amusement are brilliant with gas, and it is gas which
makes the splendour of the gin-palace. The shops took to it slowly,
but they are now beginning to understand how to brighten their
appearance after dark. Go into any little thoroughfare, however, and
you will see the shops lit with two or three candles still.
 In the small houses and the country towns the candles linger
still. And such candles! For the most part they are tallow: they need
constant snuffing: they drop their detestable grease everywhere—on
the tablecloth, on your clothes, on the butter and on the bread. You,
Eighty-seven, will be saying hard things of gas, but you do not know
from what darkness, and misery of darkness, it saved your
ancestors.
As for the churches, they are not yet generally provided with
gas. There is some strange prejudice against it in the minds of the
clergy. Yet it is not Papistical, or even freethinking. In most of them,
where they have evening service, the pews are provided with two
candles apiece, stuck in tin candlesticks, with four candles for the
pulpit and four for the reading-desk. The effect is not unpleasing,
but the candles continually require snuffing, and the operation is
constantly attended with accidents, so that the church is always
filled with the fragrance of smouldering tallow wicks. The
repugnance to gas is so great, indeed, in some quarters, that one
clergyman, the Rector of Holy Trinity, Marylebone, is going to
commit all his vestrymen to the Ecclesiastical Courts because they
have attempted to light the church with gas.
Here is a City funeral in one of the burial-grounds close to the
crowded street; the clergyman reads the Service, and the mourners
in their long black cloaks stand round the open grave, and the coffin
is lowered into it, and outside there is no cessation at all to the
bustle and the noise; the wagoner cracks his whip, the drover
swears at his cattle, the busy men run to and fro as if the last rites
were not being performed for one who has heard the call of the
Messenger, and, perforce, obeyed it. And look—the mould in which
the grave is dug is nothing but bits of bones and splinters of coffins.
The churchyard is no longer a field of clay: it is a field of dead
citizens. You, friend Eighty-seven, will manage these things better.
Here goes one of the long stages. Saw you ever a finer coach,
more splendidly appointed, with better cattle? Ten miles an hour that
coachman reckons upon as soon as he is clear of London. They say
that in a year or two, when all the railways are opened, the stage
coaches will be ruined, the horses all sold, and the English breed of
horses ruined. We shall travel twenty miles an hour without stopping
to change horses; the accidents will be frightful, but those who meet
with none will get from London to Edinburgh in less than twenty-four
hours. Next year they promise to open the London and Birmingham
Railway.
Here comes a soldier. You find his dress
absurd? To be sure, his tight black stock makes
his red cheeks seem swollen; his queer tall hat,
with the neat red ball at the top, might be
more artistic; the red shoulder roll, not the
least like an epaulette, would hardly ward off a
sword-cut; the coat with its swallow tail is no
protection to the body or the legs; the
whitened belt must cost an infinite amount of
trouble to keep it fit for inspection, and a
working-man’s breeches and stockings would
be more serviceable than those long trousers.
There are always brave fellows, however, ready
to enlist; the soldier’s life is attractive, though 3rd REGIMENT OF BUFFS
the discipline is hard and the floggings are truly
awful.
My friend, it is half-past five, and you are tired. Let us get back
to Temple Bar and dine at the Mitre, where we can take our cut off
the joint for eighteen-pence. About this time most men are thinking
of dinner. Buy an evening paper of the boy.
So: this is cosy. A newly sanded floor, a bright fire, and a goodly
company. James! a clean tablecloth, a couple of candles, and the
snuffers, and the last joint up. What have you got in the paper?
Madagascar Embassy, Massacre in New Zealand—where the devil is
New Zealand?—Suicide of Champion, who made the infernal
machine, Great Distress in the Highlands, Murder of a Process-server
in Ireland, Crossing of the Channel in a Balloon—I hope that some
 day an army may not cross it—
Letter from Syria, concerning the
recent Great Earthquake, Conduct
of the British Legion in Spain,
Seven Men imprisoned for
unlawfully ringing the Bells, Death
of the Oldest Woman in the World,
aged 162 years, said to have been
the Nurse of George Washington—
a good deal of news all for one
evening paper. Hush! we are in
luck. Here is Douglas Jerrold. Now
we shall hear something good.
Here is Leigh Hunt, and here is
Forster, and here—ah! this is
DOUGLAS JERROLD unexpected—here comes none
(From the Bust by the late E. H. Bailey, R.A.) other than ‘Boz’ himself. Of course
you know his name? It is Charles
Dickens. Saw one ever a brighter eye or a more self-reliant bearing?
Such self-reliance belongs to those who are about to succeed. They
say his fortune is already made, though but yesterday he was a
reporter in the House, taking down the speeches in shorthand. Who
is that tall young man with the ugly nose? Only a journalist. They
say he wrote that funny paper called ‘The Fatal Boots’ in Tilt’s
Annual. His name is Thackeray, I believe, but I know nothing more
about him.
Here comes dinner, with a tankard of foaming stout. Is there any
other drink quite so good as stout? After you have taken your dinner,
friend Eighty-seven, I shall prescribe for you what you will never get,
poor wretch—a bottle of the best port in the cellars of the Mitre.
My friend, there is one thing in which we of the Thirties do
greatly excel you of the Eighties. We can eat like ploughboys, and
we can drink like draymen. As for your nonsense about Apollinaris
Water, we do not know what it means; and as for your not being
able to take a simple glass of port, we do not in the least understand
 JOHN FORSTER
(From a Photograph by Elliott and Fry)

it.
No
t
tak
ea
pin
t
of
po
rt?
Ma
n CHARLES DICKENS
ali
ve! we can take two
Leigh Hunt. bottles, and never turn a
-LEIGH HUNT-
hair.
 CHAPTER V.

WITH THE PEOPLE.

When the real history of the people comes to be written—which will

be the History, not of the Higher, but of the Lower Forms of
Civilisation—it will be found that, as regards the people of these
islands, they sank to their lowest point of degradation and corruption
in the middle of the eighteenth century—a period when they had no
religion, no morality, no education, and no knowledge, and when
they were devoured by two dreadful diseases, and were prematurely
killed by their excessive drinking of gin. No virtue at all seems to
have survived among all the many virtues attributed to our race
except a bulldog courage and tenacity. There are glimpses here and
there, when some essayist or novelist lifts the veil, which show
conditions of existence so shocking that one asks in amazement how
there could have been any cheerfulness in the civilised part of the
community for thinking of the terrible creatures in the ranks below.
They did not think of them; they did not know of them; to us it
seems as if the roaring of that volcano must have been always in
their ears, and the smoke of it always choking their throats. But our
people saw and heard nothing. Across the Channel, where men’s
eyes were quicker to see, the danger was clearly discerned, and the
eruption foretold. Here, no one saw anything, or feared anything.
How this country got through without a revolution, how it
escaped the dangers of that mob, are questions more difficult to
answer than the one which continually occupies historians—How
Great Britain, single-handed, fought against the conqueror of the
world. Both victories were mainly achieved, I believe, by the might
and majesty of Father Stick.
He is dead now, and will rule no more in this country. But all
through the last century, and well into this, he was more than a king
—he was a despot, relentless, terrible. He stripped women to the
waist and whipped them at Bridewell; he caught the ’prentices and
flogged them soundly; he lashed the criminal at the cart-tail; he
lashed the slaves in the plantations, the soldiers in the army, the
sailors on board the ships, and the boys at school. He kept
everybody in order, and, truly, if the old violence were to return, we
might have to call in Father Stick again.
He was good up to a certain point, beyond which he could not
go. He could threaten, ‘If you do this, and this, you shall be
trounced.’ Thus the way of transgressors was made visibly hard for
them. But he could not educate—he taught nothing except
obedience to the law; he had neither religion nor morals; therefore,
though he kept the people in order, he did not advance them. On the
other hand, under his rule they were left entirely to themselves, and
so they grew worse and worse, more thirsty of gin, more brutal,
more ignorant. So that, in the long run, I suppose there was not
under the light of the sun a more depraved and degraded race than
that which peopled the lowest levels of our great towns. There is
always in every great town a big lump of lawlessness, idleness, and
hostility to order. The danger, a hundred years ago, was that this
lump was getting every day bigger, and threatening to include the
whole of the working class.
Remember that as yet the government of this realm was wholly
in the hands of the wealthier sort. Only those who had what was
humorously called a stake in the country were allowed to share in
ruling it. Those who brought to the service of their native land only
their hands and their lives, their courage, their patience, skill,
endurance, and obedience, were supposed to have no stake in the
country. The workers, who contribute the whole that makes the
prosperity of the country, were then excluded from any share in
managing it.
It seems to me that the first improvement of the People dates
from their perception of the fact that all have a right to help in
managing their own affairs; I think one might prove that the ideas of
the French Revolution, when they were once grasped, arrested the
downward course of the People—the first step to dignity and self-
respect was to understand that they might become free men, and
not remain like unto slaves who are ordered and have to obey. Then
they began to struggle for their rights, and in the struggle learned a
thousand lessons which have stood them in good stead. They
learned to combine, to act together, to form committees and
councils; they learned the art of oratory, and the arts of persuasion
by speech and pen; they learned the power of knowledge—in a
word, the long struggle whose first great victory was the Reform Act
of 1832 taught the People the art of self-government.
Fifty years ago, though that Act had been passed, the great
mass of the people were still outside the government. They were
governed by a class who desired, on the whole, to be just, and
wished well to the people, provided their own interests were not
disturbed, as when the most philanthropic manufacturers loudly
cried out as soon as it was proposed to restrict the hours of labour.
It is not wonderful, therefore, that the working classes should at that
time regard all governments with hostility, and Religion and Laws as
chiefly intended to repress the workers and to safeguard the
interests of landlords and capitalists. This fact is abundantly clear
from the literature which the working men of 1837 delighted to read.
As regards their religion, there was already an immense advance
in the spread of the Nonconformist sects and the multiplication of
chapels. As for the churches, I am very certain that the working man
does not go much to church even yet, but fifty years ago he
attended service still less often. A contemporary who pretends to
know asserts that nine out of ten among the working men were
professed infidels, whose favourite reading was Paine, Carlile, and
Robert Taylor, the author of ‘The Devil’s Chaplain.’ Further, he
declares that not one working man in a hundred ever opened a
Bible.
I refrain from dwelling upon this state of things as compared
with that of the present, but it appears from a census taken by a
recent weekly newspaper (which, however, omitted the mission
churches and services in school-rooms and other places) that about
one person in nine now attends church or chapel on a Sunday.
As regards drink, a question almost as delicate as that of
religion, it is reported that in London alone three millions of pounds
were spent every year in gin, which seems a good deal of money to
throw away with nothing to show for it. But figures are always
misleading. Thus, if everybody drank his fair share of this three
millions, there would be only a single glass of gin every other day for
every person; and if half the people did not drink at all, there would
be only one glass of gin a day for those who did. Still, we must
admit that three millions is a sum which shows a widespread love of
gin. As for rum, brandy, and Hollands, the various forms of malt
liquor, fancy drinks, and compounds, let us reserve ourselves for the
chapter on Taverns. Suffice it here to call attention to the fact that
there was no blue ribbon worn. Teetotallers there were, it is true,
but in very small numbers; they were not yet a power in the land;
there was none of the everlasting dinning about the plague spot, the
national vice, and the curse of the age, to which we are now
accustomed. Honest men indulged in a bout without subsequent
remorse, and so long as the drink was unadulterated they did
themselves little harm. Without doubt, if the men had become
teetotallers, there would have been very much more to spend in the
homes, and the employers would, also without doubt, have made
every effort to reduce the wages accordingly, so as to keep up the
old poverty. That is what the former school of philosophers called a
Law of Political Economy. The wages of a skilled mechanic fifty years
ago seem to have never risen above thirty shillings a week, while
food, clothes, and necessaries were certainly much dearer than at
present. He had savings banks, and he sometimes put something by,
but not nearly so much as he can do now if he is thrifty and in
regular work. It is quite clear that he was less thrifty in those days
than now, that he drank more, and that he was even more reckless,
if that is possible, about marriage and the multiplication of children.
As for the material condition of the people, there cannot be a
doubt that it has been amazingly improved within the last fifty years.
It is not true, as stated in a very well known work, that the poor
have become poorer, though the rich have certainly become richer.
The skilled working man is better paid now than then, his work is
more steady, his hours are shorter. He is better clad, with always a
suit of clothes apart from his working dress; he is better taught; he
is better mannered; he has holidays; he has clubs; he is no longer
forbidden to combine; he can co-operate; he holds meetings; he has
much better newspapers to read; his food is better and cheaper; he
has model lodging houses. Not only is he actually better; he is
relatively better compared with the richer classes, while for the last
ten years these have been growing poorer every day, although still
much richer than they were fifty years ago. Moreover, it is becoming
more difficult in every line, owing to the upward pressure of labour,
to become rich.
His amusements no longer have the same brutality which used
to characterise them. The Ring was his chief delight, and a well-
fought battle between two accomplished bruisers caused his heart to
leap with joy. Unhappily the Ring fell, not because the national
sentiment concerning pugilism changed, but by its own vices, and
because nearly every fight was a fight on the cross; so that betting
on your man was no longer possible, and every victory was arranged
beforehand. There are now signs of its revival, and if it can be in any
way regulated it will be a very good thing for the country. Then
there was dog-fighting, which is still carried on in certain parts of the
country. Only a few years ago I saw a dozen dog-fights, each with
its ring of eager lookers-on, one Sunday morning upon the sands
between Redcar and Saltburn. All round London, again, there were
ponds, quantities of ponds, all marked in the maps of the period and
now all filled up and built over. Some, for instance, were in the fields
on the east side of Tottenham Court Road. Hither, on Sundays, came
the London working man with ducks, cats, and dogs, and proceeded
to enjoy himself with cat-hunts and duck-hunts in these ponds.
There were also bull-and-bear-baitings and badger-drawings. As for
the fairs, Bartholomew and Greenwich, one is sorry that they had to
be abolished, but I suppose that London had long been too big for a
fair, which may be crowded but must not be mobbed. A real old fair,
with rows of stalls crammed with all kinds of things which looked
ever so much prettier under the flaring lamps than in the shops, with
Richardson’s Theatre, the Wild Beast Show, the wrestlers and the
cudgel-players, the boxers, with or without the gloves, the dwarfs,
giants, fat women, bearded women, and monsters, was a truly
delightful thing to the rustics in the country; but in London it was
incongruous, and even in Arcadia a modern fair is apt to lose its
picturesque aspect towards nightfall. On the whole, it is just as well
for London that it has lost its ancient fairs.
It is not in connection with working men, but with the whole
people, that one speaks of prisons. I do not think that our prison
system at the present day is every thing that it might be. There have
been one or two books published of late years, which make one
uncomfortable in thinking of the poor wretches immured in these
abodes of solitary suffering. Still, if one has to choose between a
lonely cell and the society of the prison birds by day and night, one
would prefer the former. Some attempts had been made in Newgate
and elsewhere to prevent the prisoners from corrupting each other,
but with small success. Those who were tried and sentenced were
separated from those who were waiting their trial; the boys were
separated from the men, the girls from the women. Yet the results
of being committed to prison, for however short a period, were
destructive of all morals and the last shred of principle. Not a single
girl or woman who went into prison modest and virtuous but
became straightway ashamed of her modesty and virtue, and came
out of the prison already an abandoned woman. Not a man or boy
who associated with the prisoners for a week but became a past
master in all kinds of wickedness. In the night rooms they used to
lock up fifteen or twenty prisoners together, and leave them there all
night to interchange their experiences—and what experiences! Only
those who were under sentence of death had separate cells. These
poor wretches were put into narrow and dark rooms, receiving light
only from the court in which the criminals are permitted to walk
during the day. They slept on a mat, and in former days had but
twenty-four hours between sentence and execution, with bread and
water for all their food.
Transportation still went on, with the horrors of the convict ship,
the convict hulks, and the convict establishments of New South
Wales and Tasmania. The ‘horrors’ of the system have always
seemed to me as forming an unessential part of the system. With
better management on modern ideas, transportation should be far
better than the present system of hopeless punishment by long
periods of imprisonment. We can never return to transportation as
far as any colony is concerned, but I venture to prophesy that the
next change of the penal laws will be the re-establishment of
transportation with the prospect of release, a gift of land, and a
better chance for an honest life.
Meantime the following lines belong to Fifty Years Ago. They are
the Farewell of convicts about to sail for Botany Bay:

THE DARBY DAY.

Come, Bet, my pet, and Sal, my pal, a buss, and then farewell—
And Ned, the primest ruffling cove that ever nail’d a swell—
To share the swag, or chaff the gab, we’ll never meet again,
The hulks is now my bowsing crib, the hold my dossing ken.
Don’t nab the bib, my Bet, this chance must happen soon or later,
For certain sure it is that transportation comes by natur;
His lordship’s self, upon the bench, so downie his white wig in,
Might sail with me, if friends had he to bring him up to priggin;
And is it not unkimmon fly in them as rules the nation,
To make us end, with Botany, our public edication?
But Sal, so kind, be sure you mind the beaks don’t catch you
tripping,
You’ll find it hard to be for shopping sent on board the shipping:
So tip your mauns afore we parts, don’t blear your eyes and nose,
Another grip, my jolly hearts—here’s luck, and off we goes!

Debtors’ prisons were in full swing. There were Whitecross

Street Prison, built in 1813 for the exclusive reception of debtors,
who were before this crowded together with criminals at Newgate;
Queen’s Bench Prison, the Fleet, and the Marshalsea. The King’s
Bench Prison was the largest, and, so to speak, the most fashionable
of these prisons. Both at the King’s Bench and the Fleet debtors
were allowed to purchase what were called the ‘Rules,’ which
enabled them to live within a certain area outside the prison, and
practically left them free. They paid a certain percentage on their
debts. This practice enabled the debtor to refuse paying his debts,
and to save his money for himself or his heirs. Lodgings, however,
within the Rules were bad and expensive.
 NEWGATE—ENTRANCE IN THE OLD BAILEY

There was no national compulsory system of education; yet the

children of respectable working men were sent to school. The
children of the very poor, those who lived from hand to mouth by
day jobs, by chance and luck, were not taught anything. If you talk
to a working man of sixty or thereabouts, you will most likely
discover that he can read, though he has very often forgotten how
to write. He was taught when he was a child at the schools of the
National Society, or at those of the British and Foreign Society, or at
the parish schools, of which there were a great many. There were
also many thousands of children who went to the Sunday School.
Yet, partly through the neglect of parents, and partly through the
demand for children’s labour in the factories, nearly a half of the
children in the country grew up without any schooling. In 1837 there
were forty per cent. of the men and sixty-five per cent. of the
women who could not sign their own names.
And there were already effected, or just about to be effected,
three immense reforms, the like of which the nation had never seen
before, which are together working for a Revolution of Peace, not of
war, greater than contemplated by the most sincere and most
disinterested of the French Revolutionaries.
The first was the Reform of the Penal Laws.
In the beginning of the century the law recognised 223 capital
offences. A man might be hanged for almost anything: if he
appeared in disguise on a public road; if he cut down young trees; if
he shot rabbits; if he poached at night; if he stole anything worth
five shillings from a person or a shop; if he came back from
transportation before his time; a gipsy, if he remained in the same
place a year. In fact, the chief desire of the Government was to get
rid of the criminal classes by hanging them. It was Sir Samuel
Romilly, as everybody knows, who first began to attack this
bloodthirsty code. He was assisted by the growth of public opinion
and by the juries, who practically repealed the laws by refusing to
convict.
It was not, again, until the year 1836
that counsel for a prisoner under trial for
felony was permitted to address the jury. In
the year 1834, there were 480 death
sentences; in 1838, only 116. In 1834, 894
persons were sentenced to transportation for
life, and in 1838 only 266. Remember that
this wicked severity only served to enlist the
sympathies of the people against the
Government.
IN THE QUEEN’S BENCH
The second great step was the repeal of
the Acts which forbade combination. Until
the year 1820, the people had been forbidden to combine. Their
only power against employers who worked them as many hours a
day as they dared, and paid them wages as small as they could, who
took their children and locked them up in unwholesome factories,
was in combination, and they were forbidden to combine. When the
law—an old mediæval law—was repealed, it was found that any
attempt to hold public meetings might be put down by force; so
that, though they could not combine, the chief means of promoting
combination was taken from them.
The third great step was the Extension of the Suffrage, so that
now there is no Briton or Irishman but can, if he please, have his
vote in the government of the nation. It is not a great share which is
conferred by one vote, but it enables every man to feel that he is
himself a part of the nation; that the government is not imposed
upon him, but elected and approved by himself.
Considering all these things, have we any reason to be surprised
when we learn that, on the Queen’s Accession, there was among the
people no loyalty whatever? Attachment to the Sovereign, personal
devotion to the young Queen, rallying round the Throne—all these
things were not even phrases to the working class. For they never
heard them used.
There was no loyalty at all, either to the Queen, or to the
institution of a limited Monarchy, or to the Constitution, or to the
Church.
For a hundred and fifty years there had been no loyalty among
the people. Loyalty left the country with James II. Not one of the
Sovereigns who followed him commanded the personal enthusiasm
of the people, not even Farmer George, for whom there had been
some kind of affection with something of contempt. From 1687 until
1837, which is exactly one hundred and fifty years, not one
Sovereign who sat upon the Throne of England could boast that he
had the love of the people. Not one wished to have the love of the
people. He represented a principle: he governed with the assistance
of a few families and by the votes of a small class. As King he was a
stranger. When he drove through the streets, the people hurrahed;
but they did not know him, and they cared nothing for him.
Therefore the sentiment of loyalty had to be re-born. It could
only be awakened by a woman, young, virtuous, naturally amiable,
and resolved on ruling by constitutional methods. Yet in some of the
journals written for, and read by, the working men, the things said
concerning the Queen, the Prince Consort, and the Court were
simply horrible and disgusting. Such things are no longer said. There
are still papers which speak of the aristocracy as a collection of titled
profligates, and of the clergy as a crowd of pampered hypocrites,
but of the Queen it is rare indeed to find mention other than is
respectful. Her life and example for fifty years have silenced the
slanderers. It has been found once more possible for a Sovereign to
possess the love of her people.
The papers read by the working men were not only scurrilous,
but they were Republican and revolutionary. The Republic whose
example they set before themselves was not the American, which is
Conservative, for of this they knew nothing. Let us clearly
understand this. Fifty years ago America was far more widely
separated from England than is China now. The ideal Republic was
then the earlier form of the first French Republic. These people
cared little for the massacres which accompanied the application of
Republican principles. I do not say that they wished to set the heads
of the Queen’s Ladies-in-Waiting on pikes, but they thought the
massacres of innocent women by the French an accident rather than
a consequence. They loved the cry of ‘Liberty, Equality, and
Fraternity,’ and still believed in it. They dreamed of a country which
they thought could be established by law, in which every man was to
be the equal of his neighbour—as clever, as skilful, as capable, as
rich, and as happy. The dream continues, and will always continue,
to exist. It is a generous dream—there never has been a nobler
dream—so that it is a thousand pities that human greed, selfishness,
ambition, and masterfulness will not suffer the dream to be realised.
Those who advocated an attempt to realise it flung hard names at
the Crown, the Court, the aristocracy, the Church, the educated, and
the wealthy. Presently they began to formulate the way by which
they thought to place themselves within reach of their object. The
way was Chartism. They wanted to carry six measures—Universal
Suffrage, Annual Parliaments, Vote by Ballot, Abolition of Property
Qualification, Payment of Members, and Equal Electoral Districts.
Very well; we have got, practically, four out of the six points, and
there are many who think that we are as far off the Millennium as
ever. Yet there are, however, still among us people who believe that
we can be made happy, just, merciful, and disinterested by changing
the machinery. Changing the machinery! The old party of Radicals
still work themselves into a white heat by crying for change in the
machinery.
And now a thing which was never contemplated even by the
Chartists themselves—the really important thing—has been acquired
by the people. They are no longer the governed, but the governors.
The Government is no longer a thing apart from themselves, and
outside them. It is their own—it is the Government of the People of
England. If there is anything in it which they do not like, they can
alter it; if there is anything they agree to abolish, they can abolish it,
whether it be Church, Crown, Lords, wealth, education, science, art
—anything. They may destroy what they please: they may reduce
the English to an illiterate peasantry if they please.
They will not please. I, for one, have the greatest confidence in
the justice, the common-sense, and the Conservatism of the English
and the Scotch. The people do not, as yet, half understand their own
power; while they are gradually growing to comprehend it, they will
be learning the history of their country, the duties and
responsibilities of citizenship, the dangers of revolution, and the
advantages of those old institutions by whose aid the whole world
has been covered with those who speak the Anglo-Saxon speech
and are governed by the English law.
My friends, we are changed indeed. Fifty years ago we were, as
I have said, still in the eighteenth century. The people had no power,
no knowledge, no voice; they were the slaves of their employers;
they were brutish and ill-conditioned, ready to rebel against their
rulers, but not knowing how; chafing under laws which they did not
make, and restraints which kept them from acting together, or from
meeting to ask if things must always continue so. We are changed
indeed.
 We now stand upright; our faces are full of hope, though we are
oppressed by doubts and questions, because we know not which
path, of the many before us, will be the wisest; the future is all our
own; we are no longer the servants; we are the Masters, the
absolute Rulers, of the greatest Empire that the world has ever seen.
God grant that we govern it with wisdom!
 CHAPTER VI.

WITH THE MIDDLE-CLASS.

The great middle-class—supposed, before the advent of Mr. Matthew

Arnold, to possess all the virtues; to be the backbone, stay, and prop
of the country—must have a chapter to itself.
In the first place, the middle-class was far more a class apart
than it is at present. In no sense did it belong to society. Men in
professions of any kind, except the two services, could only belong
to society by right of birth and family connections; men in trade—
bankers were still accounted tradesmen—could not possibly belong
to society. That is to say, if they went to live in the country they
were not called upon by the county families, and in town they were
not admitted by the men into their clubs, or by ladies into their
houses. Those circles, of which there are now so many—artistic,
æsthetic, literary—all of them considering themselves to belong to
society, were then out of society altogether; nor did they overlap
and intersect each other. The middle-class knew its own place,
respected itself, made its own society for itself, and cheerfully
accorded to rank its reverence due. The annals of the poor are
meagre; only here and there one gets a glimpse into their lives. But
the middle-class is much better known, because it has had prophets;
nearly all the poets, novelists, essayists, journalists, and artists have
sprung from it. Those who adorned the Thirties and the Forties—
Hood, Hook, Galt, Dickens, Albert Smith, Thackeray—all belonged to
it; George Eliot, whose country towns are those of the Thirties and
the Forties, was essentially a woman of the middle-class.
 Middle-class life—especially in the
country—was dull, far, far duller than
modern life even in the quietest country
town. The men had their business; the
women had the house. Incomes ran
small; a great deal was done at home
that is now done out of it. There was a
weekly washing-day, when the house
steamed with hot soap-suds, and the
‘lines’ were out upon the poles—they GEORGE ELIOT
were painted green and were square— (Taken from the Drawing in ‘The
Graphic’ by permission)
and on the lines hung half the family
linen. All the jam was made at home; the
cakes, the pies, and the puddings, by the wife and daughters; the
bread was home-made; the beer was home-brewed (and better beer
than good home-brewed no man need desire); all those garments
which are not worn outside were made at home. Everybody dined in
the middle of the day. Therefore, in the society of the country town
dinner-parties did not exist. On the other hand, there were sociable
evenings, which began with a sit-down tea, with muffins and tea-
cakes, very delightful, and ended with a hot supper. Tobacco was not
admitted in any shape except that of snuff into the better kind of
middle-class house; only working men smoked vulgar pipes; the
Sabbath was respected; there was no theatre nearer than the county
town; the girls had probably never seen a play; every man who
respected himself ‘laid down’ port, but there was little drinking of
wine except on Sunday afternoons; no one, not even the ladies,
scorned the glass of something warm, with a spoon in it, after
supper. For the young there was a fair once a year; now and then a
travelling circus came along; there was a lecture occasionally on an
instructive subject, such as chemistry, or astronomy, or sculpture;
there were picnics, but these were rare; if there were show places in
the neighbourhood, parties were made to them, and tea was
festively taken among the ruins of the Abbey.
 Fashion
descends
slowly; it is now
the working
man who takes
his wife into the
country for tea:
fifty years ago
he took his wife
nowhere, and
scorned tea.
Open-air games
and sports there
were none; no
lawn-tennis,
Badminton, or
anything of that
kind in those
days; even
croquet, which
is now so far
lost in the mists
of antiquity that
men of thirty
are too young
to remember
the rage for it,
was actually not
yet invented.
John Galt
Archery
-JOHN GALT-
certainly
existed, and the
comic writers are always drawing pictures of the young ladies
sticking their arrows into the legs of people a hundred feet or so
wide of the target. But archery belonged to a class rather above that
which we are now considering. There was not much sketching and
painting. There was no amateur photography; there was no catching
of strange creatures in ponds for the aquarium—a fashion also now
happily extinct; there was not, in fact, any single pursuit,
amusement, or game which would bring young people together in
the open air. There was no travelling; the summer holiday had not
yet got down in the country. In London, to be sure, everybody down
to Bevis Marks and Simmery Axe went out of town and to the
seaside in July or August; but in the country nobody thought of such
a thing; not the vicar’s daughters, not the solicitor’s wife, not the
family of the general practitioner; the very schoolmaster, who got his
four weeks in the summer and his three at Christmas, spent them at
home in such joy as accompanies rest from labour. With no outdoor
amusements, and with no summer holiday, how much is life
simplified! But the simplicity of life means monotony—faciunt vitam,
balnea, vina, Venus.
In the winter, things were
somewhat different. In some
towns there was the county
ball. At this function one had
the pleasure of gazing upon
ladies and gentlemen of the
highest rank and fashion, and
of observing that they kept to
themselves like a Hindu caste, LA PASTOURELLE
danced with each other at the
upper end of the room, cast disparaging glances at the dresses of
the ladies of the lower end, and sniffed at their manner and
appearance. This was true joy. There were also occasional dances at
home, but these were rare, because people had not learned how to
meet and dance without making a fuss over it, taking up carpets,
putting candles in tin sconces, keeping late hours, and having a
supper, the preparation of which was mainly done by the ladies of
the house, and it nearly killed them, and drove the servants—the
genteel middle-class family often got along with only one—to give
notice. I think that the dances which had gone out in London still
lingered in the country. There were, for instance, the Caledonians as
well as the Lancers; there were country dances without end, the
very names of which are now lost; the gentlemen performed the
proper steps with grace and agility, while the ladies were careful to
preserve an attitude supposed the only one possible for a lady while
dancing, in which the figure was bent forward, the face was turned
up with the chin stuck out, while the hands were occupied in holding
up the dress to the regulation height. The elders, meanwhile, played
long whist at tables lit by candles which wanted snuffing between
the deals. The bashful youth of the party was always covering
himself with shame by his clumsiness in snuffing out the candles, or,
even if he succeeded in taking off the red-hot ball of burnt thread,
he too often neglected to close the instrument with which he
effected the operation, and thereby mightily offended the nostrils of
the company. When there was no dancing the younger members
began with a ‘little music.’ Their songs—how faded and stale they
seem now if one tries to sing them!—turned chiefly on the
affections, and the favourite poet was Felicia Hemans. After the little
music they sat down to a round game, of which there were a great
many, such as Commerce, Speculation, Vingt-et-Un, Limited Loo, or
Pope Joan. The last was played with a board. I remember the board
—it was a round thing, lacquered, and like a punch-bowl, but I think
with divisions; as for the game itself, and what was done with the
board, I quite forget, but both game and bowl lasted quite into the
Fifties. Are there any country circles now where they still play Pope
Joan with mother-o’-pearl counters, and after the game have a
grand settlement, and exchange the counters for silver and copper,
some with chuckles, and others with outward smiles but inward
rage?
People were extremely punctilious on the subject of calls—one
remembers the call in the ‘Mill on the Floss.’ The call was due at
regular intervals, so that even the day should almost be known on
which it was paid or returned. It was a ceremonial which
necessitated a great deal of ritual and make-believe. No one, for
instance, was to be surprised in doing any kind of work. There was a
fiction in genteel families that the ladies of the house never did
anything serious or serviceable after dinner; the afternoon was
supposed to be devoted either to walking, or to making calls, or to
elegant trifling at home. Therefore, if the girls were at the moment
engaged upon any useful work—many of them, poor things, never
did anything but useful work—they crammed it under the sofa, and
pretended to be reading a book, or painting, or knitting, or to be
engaged in easy and fashionable conversation. Why they went
through this elaborate pretence I have not the least idea, because
everybody knew that every girl in the place was always making,
mending, cutting-out, basting, gusseting, trimming, turning, and
contriving. How do you suppose that the solicitor’s daughters made
so brave a show on Sundays if they were not clever enough to make
up things for themselves? Everybody, of course, knew it, and why
the girls would not own up at once one cannot now understand.
Perhaps it was a sort of suspicion, or a faint hope, or a wild dream,
that a reputation for ladylike uselessness might enable them to cross
the line at the County Ball, and mingle with the county people.
Are there still any circles of society in which, if a lady with her
daughters calls upon another lady with her daughters, the
decanters, biscuits, and glasses are placed upon the table, and the
visitors are asked whether they will take port or sherry? This, fifty
years ago, was always done in country towns, and the visitors
always took a glass of port or sherry. In some houses it was not port
and sherry that were placed upon the table, but ‘red’ and ‘white.’ I
do not know whether the red was currant or raspberry, but I think
that the white was generally cowslip. When the visitors were gone,
the ladies got out their work again, threaded their needles, and
spent an enjoyable hour or two in discussing the appearance, the
dress, the manners, and the resources of their visitors. But the visit
did them good, because it compelled company manners, which are
always good for girls, and it dragged them a little out of themselves.
They were too much en famille, these girls; they were never
separated from each other. The boys got out to school or to business
all day; but the poor girls were always together. Side by side they
did their household duties, side by side they sewed and dressmaked,
side by side they walked, side by side they prayed in the church,
side by side they slept. Small chance of happiness was theirs—
happiness is a separate, distinct, individual kind of thing, in which
one can consult one’s own likes—until, in the fulness of time, there
came along the lover—a humdrum, commonplace kind of lover, I
dare say, but his sweetheart was as commonplace as himself—and
she exchanged a house, where she was a better kind of servant, for
one of exactly the same sort, in which she was the mistress. And
when one says mistress, it must be remembered that man was, in
those days, much more of a master in the house than he is now
allowed to be. I speak not at random, but from the evidence of
those who remember and from study of the literature, both that
written by the men and that by the women. I am certain that the
husband, unless he was hen-pecked—a pleasing word, now seldom
used—was always the Master and generally the Tyrant in the house.
Let me, with some diffidence, approach the subject of the
Church in the country town. I never truly understood the Church of
fifty years ago until, in the autumn of 1885, I perambulated with one
who is jealous for Church architecture and Church antiquities the
north-east corner of Norfolk, where there are many churches, and
most of them are fine. In our pilgrimage among these monuments
we presently came upon one at the aspect of which we were fain to
sit down and weep. It was, externally, an old and venerable
structure, which might have been made beautiful within. Plaster
covered the walls, and hid the columns; the interior of the church
was crowded with high pews, painted white, and having along the
top a sham mahogany kind of hand-rail; the chancel was
encumbered with these enclosures, which hid the old brass-work;
that which belonged to the Squire was provided with red curtains on
brass rods to keep the common people from gazing at the Quality.
The reading-desk, pulpit, and altar were covered with a cloth which
had been red, but had long before faded away into an indescribably
shabby brown. The pulpit was not part of the old three-decker, but
was stuck into the wall; the windows had lost their old tracery; the
painted glass was gone; the roof was a flat whitewashed ceiling. The
church, to eyes accustomed to better things, presented a deplorable
appearance. My friend, pointing solemnly to the general shabbiness,
remarked, ‘Donec templa refeceris.’ It was the motto of the journal
started early in the Forties by a small knot of Cambridge men—
among whom was Mr. Beresford Hope, now, alas! no more—who
desired to raise and beautify public worship in the Anglican faith,
and also, I believe, to assert and insist upon certain points of
doctrine. And they clearly perceived that, while the churches
remained in their neglected condition, and church architecture was
at its then low ebb, their doctrine was impossible. How far they have
succeeded not only the Ritualists themselves proclaim, but also
every other party in the Church, and even the Nonconformists, who
have shared in the increased beauty and fitness of public worship.

THE QUEEN RECEIVING THE SACRAMENT AFTER HER CORONATION—WESTMINSTER

ABBEY, JUNE 29, 1838.
(From the Picture by C. R. Leslie, R.A., at Windsor Castle.)
 He who can remember the ordinary Church Services in the early
Fifties very well knows what they were in the Thirties, except that in
the latter there were still some venerable divines who wore a wig.
The musical part of the service was, to begin with, taken slow—
incredibly slow; no one now would, who is not old enough to
remember, believe how slow it was. The voluntary at the beginning
was a slow rumble; the Psalms were very slowly read by the
clergyman and the clerk alternately, the Gloria alone being sung,
also to a slow rumble. The choir was generally stationed in the organ
loft, which has been known to be built over the altar at the east end
—as at St. Mary’s, Cambridge—but was generally at the west end. It
was not a choir of boys and men only, but of women and men. The
‘Te Deum’ was always ‘Jackson’—from my youth up have I loathed
‘Jackson’; there was just one lively bit in it for which one looked and
waited; but it lasted a very few bars; and then the thing dragged on
more slowly than ever till it came to the welcome words, ‘Let me
never be confounded.’ Two hymns were sung—very slowly; they
were always of the kind which expressed either the despair of the
sinner or the doubtful joy of the believer. I say doubtful, because he
was constantly being warned not to be too confident, not to mistake
a vague hope for the assurance of election, and because, with the
rest of the congregation, he was always being told how few in
number were those elect, and how extremely unlikely that there
could be many of those few in that one flock. Read any of the
theological literature of the period, and mark the gulf that lies
between us and our fathers. There were many kinds of preachers,
just as at present—the eloquent, the high and dry, the low and
threatening, the forcible-feeble, the florid, the prosy, the scholarly—
but they all seemed to preach the same doctrine of hopelessness,
the same Gospel of Despair, the same Father of all Cruelty, the same
Son who could at best help only a few; and when any of the
congregation dared to speak the truth, which was seldom, these
blasphemous persons whispered that it was best to live and enjoy
the present, and to leave off trying to save their souls against such
fearful odds, and with the knowledge that if they were going to be
saved it would be by election and by no merit or effort of their own,
while, if the contrary was going to happen, it was no use striving
against fate. Wretched, miserable creed! To think that unto this was
brought the Divine Message of the Son of Man! And to think of the
despairing deathbeds of the careless, the lifelong terror of the most
religious, and the agony of the survivors over the death of one ‘cut
off in his sins’!
What we now call the ‘life’ of the Church, with its meetings,
committees, fraternities, guilds, societies, and organisations, then
simply did not exist. The clergyman had an easy time; he visited
little, he had an Evening Service once a week, he did not pretend to
keep saints’ days and minor festivals and fasts—none of his
congregation expected him to keep them; as for his being a
teetotaller for the sake of the weaker brethren, that would have
seemed to everybody pure foolishness, as, indeed, it is, only people
now run to the opposite belief; yet he was a good man, for the most
part, who lived a quiet and exemplary life, and a good scholar—
scholars are, indeed, sadly to seek among the modern clergy—a
sound theologian, a judge of good port, and a gentleman. But
processions, banners, surpliced choirs, robes, and the like, he would
have regarded as unworthy the consideration of one who was a
Churchman, a Protestant, and a scholar.
To complete this brief study of the Church fifty years ago, let us
remark that out of 11,500 livings which it possessed, 3,000 were
under 100l. and 1,000 under 60l. a year, that there were 6,080
pluralists and 2,100 non-residents, that the Dissenters had only been
allowed to marry in their own chapels and by their own clergy in the
year 1831, that they were not admitted, as Dissenters, to the
Universities, and that the incomes of some of the Bishops were
enormous.