Get Your Free Goodie Box here

Biotechnology for Sustainability by Subhash Bhore, K. Marimuthu and M. Ravichandran [E - HTML preview

PLEASE NOTE: This is an HTML preview only and some elements such as links or page numbers may be incorrect.
Download the book in PDF, ePub, Kindle for a complete version.



Biotechnology for Sustainability

Achievements, Challenges and Perspectives


Subhash Bhore,

K. Marimuthu &

M. Ravichandran

Biotechnology for Sustainability

Achievements, Challenges and Perspectives


Subhash Bhore, K. Marimuthu & M. Ravichandran



Biotechnology for Sustainability

Achievements, Challenges and Perspectives

Subhash Bhore, K. Marimuthu & M. Ravichandran (Editors)

Published by AIMST University


ISBN: 978-967-14475-3-6 (Print version)

eISBN: 978-967-14475-2-9 (e-Book version)


Published by

AIMST University

Printed by

AIMST University


© 2017 by the authors; Licensee, Editors; AIMST University,

Malaysia. This book is an open access book distributed under

the terms and conditions of the Creative Commons Attribution

(CC-BY) license (

CC BY license is applied which allows users to download, copy, reuse and distribute

articles and or data provided the original article and book is fully cited. This open

access aims to maximize the visibility of articles, reviews and or perspectives, much of

which is in the interest of national, regional and global community.

Disclaimer: The information provided in this book is designed to highlight the views,

perspectives, achievements and or research findings of respective contributors. While

the best efforts have been used in preparing this book, Editors and or Publisher make

no representations or warranties of any kind and assume no liabilities of any kind with

respect to the accuracy or completeness of the contents and specifically disclaim any

implied warranties. Neither the Editors nor Publisher of this book shall be held liable or

responsible to any person or entity with respect to any loss or incidental or

consequential damages caused, or alleged to have been caused, directly or indirectly,

by the information highlighted herein. Readers should be aware that the information

provided in this book may change.

All articles and or reviews published in this book are deemed to reflect the individual

views of respective authors and not the official points of view, either of the Editors or of

the Publisher.

Cover image: A diagram showing the 17 Sustainable Development Goals (Credit:

Edited by

Dr. Subhash J. Bhore (Senior Associate Professor)1,

Dr. K. Marimuthu (Professor)1, 2, and

M. Ravichandran (Senior Professor)1, 2

Address for Correspondence:

1Department of Biotechnology, Faculty of Applied Sciences, AIMST University,

Bedong-Semeling Road, 08100 Bedong, Kedah Darul Aman, Malaysia; Telephone

No.: +604 429 8176; e-mail: /

2Chancellery, AIMST University, Bedong-Semeling Road, 08100 Bedong, Kedah Darul

Aman, Malaysia; Tel. No.: +604 429 1054 /8103; e-mail: /


First; July 18, 2017



This book is dedicated to all researchers working in

various domains of biotechnology and to all

stakeholders those are working for the global

sustainable development to improve the health of the

people and planet.



World Environment Day (WED) is a biggest global annual event celebrated each

year on June 5 to create the positive awareness to preserve the environment and planet

earth. This year, the theme for WED-2017 was “Connecting people to nature”. Our

environment should be healthy for our growth, development and to achieve the sustainable

development goals (SDGs) adopted by the international community to transform the world.

Most recently, António Guterres (United Nations Secretary General) precisely

highlighted that “Without a healthy environment we cannot end poverty or build prosperity.

We all have a role to play in protecting our only home: we can use less plastic, drive less,

waste less food and teach each other to care”. In fact, to achieve the SDGs by protecting

environment, everyone needs to do their part.

We strongly believe that biotechnology can play an important role directly or

indirectly in achieving various SDGs. Hence, we had decided to publish a book,

“Biotechnology for Sustainability” to commemorate the WED and to highlight the

achievements, challenges and perspectives in various domains of the biotechnology. In

response to our call for articles, we had received 50 manuscripts. The selected articles

published in this book are highlighting various issues, achievements, challenges and

perspectives for the viable development and sustainability. The World Commission on the

Environment and Development defined sustainability as the “development that meets the

needs of the present without compromising the ability of future generations to meet their

own needs”. The United Nations recent estimate suggest that the world’s food supply needs

to be doubled by the year 2050 to keep up with the growing demand. To achieve this is a

huge challenge; because, the amount of arable land is continuously decreasing as a result of

rising urbanization, saline soils and desertification. Biotechnologists (and plant breeders)

around the world are working persistently to produce crops which will boost the food

production to meet the growing demand. Genetically engineered crop varieties do offer

many promising possibilities to boost nutritive value of the food, sustain farming on

marginal lands, and to minimize the loss by creating pests and disease resistant varieties.

The articles published in this book are going to be useful in creating awareness

about the environmental issues, natural resources, biodiversity conservation, sustainable

development and various biotechnological approaches that could be used to alleviate the

respective challenges.

We would like to express our sincere gratitude and thanks to Dato' Seri Utama Dr.

S. Samy Vellu, Chancellor and Chairman, AIMST University for his support in publishing

this book.

We wish to thank all contributing authors for making a common cause with us. This

book publication project could not have been completed without the courteous cooperation

of the authors to highlight achievements, challenges and or perspectives in using

biotechnological approaches for the sustainability.

We are confident that this book will serve as a reference to various researchers,

scientists, academicians and graduate students involved in biodiversity conservation,

environmental protection and various fields of biology and biotechnology.

It is hoped that a prudent use of biotechnology in the biodiversity conservation,

environmental protection, and production of more and better quality of food, fiber, fuel and

drugs will contribute in accomplishing SDGs and to promote peace in the world.

Subhash J. Bhore

K. Marimuthu

M. Ravichandran

ISBN: 978-967-14475-3-6 ; eISBN: 978-967-14475-2-9 i


Preface ................................................................................................................................ i

Contents ............................................................................................................................ ii

Plant Tissue Culture for Sustainability

C. K. John ....................................................................................................................... 1

Traditional Medicine of the Tribes in Tamil Nadu and Its Sustainable

Use through Biotechnology

Valli Gurusamy, Kavitha Valampuri John, Usha Raja Nanthini

Ayyakkanu, Ramani Bai Ravichandran ......................................................................... 14

Vermitechnology – An Eco-Biological Tool for Sustainable


Mahaly Moorthi, Koilpathu Senthil Kumar Abbiramy, Arumugam Senthil

Kumar and Karupannan Nagarajan................................................................................ 41

Role of Biotechnology in Food Authentication

Shobana Manoharan, Raghavan Kuppu and Ramesh Uthandakalaipandian ................... 51

Management Strategies against Tiny Tigers for Sustainable

Development of Agriculture

Viswa Venkat Gantait ................................................................................................... 58

Designing Greener Pharmaceuticals and Practicing Green Health Is

Required for Sustainability

Sridevi Chigurupati, Jahidul Islam Mohammad, Kesavanarayanan

Krishnan Selvarajan, Saraswati Simansalam, Shantini Vijayabalan and

Subhash Janardhan Bhore ............................................................................................. 68

Clonal Propagation of a High Value Multipurpose Timberline Tree

Species Quercus semecarpifolia Sm. of West Himalaya, India

Aseesh Pandey and Sushma Tamta ............................................................................... 79

Spent Mushroom Substrate of Hypsizygus ulmarius: A Novel

Multifunctional Constituent for Mycorestoration and Mycoremediation

Padmavathi Tallapragada and Ranjini Ramesh .............................................................. 88

Biotechnology for Sustainability of Forests

Kumud Dubey and Kesheo Prasad Dubey ................................................................... 104

Biotechnological Approaches for Conservation and Sustainable Supply

of Medicinal Plants

Sagar Satish Datir and Subhash Janardhan Bhore ........................................................ 117

ISBN: 978-967-14475-3-6 ; eISBN: 978-967-14475-2-9 ii

Making Himalayas Sustainable: Opportunities and Challenges in

Indian Himalayan Region

Harsh Kumar Chauhan and Anil Kumar Bisht ............................................................. 129

Natural Polyphenols and Its Potential in Preventing Diseases Related

To Oxidative Stress as an Alternative Green Nutraceutical Approach

Sreenivasan Sasidharan, Shanmugapriy, Subramanion Lachumy Jothy,

Mei Li Ng, Nowroji Kavitha, Chew Ai Lan, Khoo Boon Yin,

Soundararajan Vijayarathna, Leow Chiuan Herng and Chern Ein Oon ........................ 141

A Review on Green Synthesis of Nanoparticles and Its Antimicrobial


Karthika Arumugam and Naresh Kumar Sharma ......................................................... 171

Production of Secondary Metabolites Using a Biotechnological


Produtur Chandramati Shankar and Senthilkumar Rajagopal ....................................... 187

Potential of Marine Algae Derived Extracts as a Natural Biostimulant

to Enhance Plant Growth and Crop Productivity

Lakkakula Satish* and Manikandan Ramesh ............................................................... 200

Biotransformation of Various Wastes into a Nutrient Rich Organic

Biofertilizer - a Sustainable Approach towards Cleaner Environment

Geetha Karuppasamy, Michael Antony D’Couto, Sangeetha Baskaran and

Anant Achary.............................................................................................................. 212

Bacterial Endophytes as Biofertilizers and Biocontrol Agents for

Sustainable Agriculture

Amrutha V. Audipudi, Bhaskar V. Chakicherla and Shubhash Janardhan

Bhore .......................................................................................................................... 223

Microbial Metabolic Engineering: A Key Technology to Deal with

Global Climate and Environmental Challenges

Meerza Abdul Razak, Pathan Shajahan Begum and Senthilkumar

Rajagopal .................................................................................................................... 248

Biodiesel Production for Sustainability: An Overview

R. Meena Devi, R. Subadevi and M. Sivakumar .......................................................... 262

In vitro Cell Bioassays in Pollution Assessment

Narayanan Kannan, Poorani Krishnan and Ahmad Zaharin Aris ................................. 274

Lipopeptide Biosurfactants from Bioagent, Bacillus as a Weapon for

Plant Disease Management

Sampath Ramyabharathi, Balaraman Meena, Lingan Rajendran and

Thiruvengadam Raguchander ...................................................................................... 287

ISBN: 978-967-14475-3-6 ; eISBN: 978-967-14475-2-9 iii

Biotechnology as a Tool for Conservation and Sustainable Utilization of

Plant and Seaweed Genetic Resources of Tropical Bay Islands, India

Pooja Bohra, Ajit Arun Waman and Anuraj Anirudhan ............................................... 295

Plantibodies for Global Health: Challenges and Perspectives

Prasad Minakshi, Basanti Brar, Manimegalai Jyothi, Ikbal, Koushlesh

Ranjan, Upendra Pradeep Lambe and Gaya Prasad ..................................................... 305

Renewable Energy from Agro-industrial Processing Wastes: An


Sudhanshu S. Behera, Ramesh C. Ray and S. Ramachandran ..................................... 322

Mitigation of Climatic Change by Organic Agriculture

Mohan Mani, Manohar Murugan, Ganesh Punamalai and Vijayalakshmi

Ganesan Singaravelu ................................................................................................... 336

Application of Anti-vibrio and Anti-quorum Sensing Technology for

Sustainable Development in Shrimp Aquaculture

Ramesh Kandasamy, Amutha Raju and Manohar Murugan ......................................... 344

Promiscuous Rhizobia: A Potential Tool to Enhance Agricultural Crops


Ikbal, Prasad Minakshi, Basanti Brar, Upendera Praddep Lambe,

Manimegalai Jyothi, Koushlesh Ranjan, Deepika, Virendra Sikka and

Gaya Prasad ................................................................................................................ 358

Organic Farming and Halalan Toyyiban Foods: An Attempt to Relate


Quamrul Hasan and Zakirah Othman .......................................................................... 376

Biotechnological Approaches: Sustaining Sugarcane Productivity and


Ashutosh Kumar Mall and Varucha Misra .................................................................. 386

Bioremediation: A Biotechnology Tool for Sustainability

Niharika Chandra, Ankita Srivastava, Swati Srivastava, Shailesh Kumar

Mishra and Sunil Kumar ............................................................................................. 398

Sea Urchin - A New Potential Marine Bio-resource for Human Health

M. Aminur Rahman, Fatimah Md. Yusoff, Kasi Marimuthu and Yuji

Arakaki ....................................................................................................................... 417

Marine Pollution and Its Impacts on Living Organisms

Thavasimuthu Citarasu and Mariavincent Michael Babu ............................................. 444

Ecology, Distribution and Diversity of Bioluminescent Bacteria in Palk

Strait, Southeast Coast of India

Srinivasan Rajendran, Ganapathy selvam Govindarasu and Govindasamy

Chinnavenkataraman................................................................................................... 456

ISBN: 978-967-14475-3-6 ; eISBN: 978-967-14475-2-9 iv

Synthesis of Biocompatible Silver Nanoparticles Using Green Alga

( Ulva reticulata) Extract

Ganapathy selvam Govindarasu, Srinivasan Rajendran and Sivakumar

Kathiresan................................................................................................................... 475

Diversity and Ethno-Botanical Potential of Tree Plants of Katarniaghat

Wildlife Sanctuary, Bahraich (UP) India: An Overview

Tej Pratap Mall ........................................................................................................... 486

Free Radical Scavenging Potential and Anticancer Activity of Primula

denticulata Sm. from North-Western Himalayas

Bilal Ahmad Wani, Mohammed Latif Khan and Bashir Ahmad Ganai ........................ 512

Panchakavya: Organic Fertilizer and Its Stimulatory Effect on the Seed

Germination of Abelmoschus esculentus and Solanum melongena

V. Ramya and S. Karpagam ........................................................................................ 525

Increasing Human Interference in Katarniaghat Wildlife Sanctuary

Shiv Pratap Singh ....................................................................................................... 534

ISBN: 978-967-14475-3-6 ; eISBN: 978-967-14475-2-9 v

Biotechnology for Sustainability

Achievements, Challenges and Perspectives

Biotech Sustainability (2017), P1-13

Plant Tissue Culture for Sustainability

C. K. John*

Plant Tissue Culture Division, CSIR-National Chemical Laboratory, Dr. Homi Bhabha

Road, Pune 411008, India;*Correspondence:; Tel.: +91-9822531551

Abstract: The United Nations has placed great emphasis on sustainability. Three of the

most important requirements of sustainable development are: eradicating extreme pov-

erty and hunger, protecting the environment, and conserving biodiversity. Because of human activities the stable functioning of earth‛s life support system – which includes the at-

mosphere, oceans, forests, waterways, biodiversity and biogeochemical cycles, is at risk.

One of the major contributing factors is the large scale destruction of natural forests. Defor-

estation had many adverse effects; most importantly, the effects on climate, environment,

and biodiversity. The three pillars of sustainable development are: sustainable agriculture,

conserving biodiversity, and protecting the environment through reversing the effects of

deforestation by large scale afforestation. Plant Tissue Culture can greatly contribute in all

the three.

Keywords: Afforestation; biodiversity conservation; micropropagation; plant tissue culture;

sustainable agriculture

1. Introduction

variation in plant varieties as possible.

Plant tissue culture can contribute to all

The United Nations Summits and

the three. In this paper I will elaborate on

Commission Reports from the 1987

how Plant Tissue Culture, my area of re-

Brundtland Commission (World Com-

search, can contribute to Sustainable Ag-

mission on Environment and Develop-

riculture, Protecting Forests, and Con-

ment) report onwards have placed added

serving Biodiversity.

emphasis on sustainability of all devel-

opment efforts. Three of the most im-

2. Sustainable development

portant requirements are: 1. Eradicat-

ing extreme poverty and hunger, 2. Pro-

In 1987 it was the Brundtland

tecting the environment, and 3. Conserv-

Commission (World Commission on En-

ing biodiversity. To eradicate extreme

vironment and Development) report “Our

poverty and hunger two things are essen-

Common Future” which brought the con-

tial: first, sustainable agriculture which

cept of “Sustainable Development” into

makes food available/affordable and se-

common use. The World Commission on

cond, creation of jobs which translates to

Environment and Development was set up

purchasing power. One of the major fac-

by the UN General Assembly in 1983.

tors in protecting the environment is re-

Brundtland Commission Report defined

versing the loss of natural forests. Con-

Sustainable Development as “Develop-

serving biodiversity is of great relevance

ment that meets the needs of the present

now than ever before for the reason that

without compromising the ability of the

our world is fast changing. To have crop

future generations to meet their own

varieties suitable for this changing envi-

needs”. According to the Brundtland

ronment is to preserve as much natural

Commission Report, the needs, in particu-

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 1

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John lar the essential needs of the world‟s poor,

the principles of sustainable develop-

to which overriding priority should be

ment into country policies and programs,

given, and the limitations imposed by the

reversing loss of environmental resources,

State of Technology and Social organiza-

and reducing biodiversity loss.

tion on the Environment‟s ability to meet

In 2012, the United Nations

present and future needs should be ad-

Rio+20 summit in Brazil committed gov-

dressed. The Brundtland Commission Re-

ernments to create a set of “Sustainable

port emphasized the need to integrate

Development Goals” (SDGs). On Sep-

economic and ecological factors in deci-

tember 25th 2015, countries adopted a set

sion-making at all levels for sustainable

of goals to end poverty, protect the planet,

development. These factors include, re-

and ensure prosperity for all as part of

viving growth, changing quality of

a 2030 Sustainable Development Agenda.

growth, meeting essential needs for jobs,

Each goal has specific targets to be

food, energy, water and sanitation, ensur-

achieved in 15 years. The 17 Sustainable

ing the resource base, reorienting tech-

Development Goals (SDGs), otherwise

nology and managing risks. In its broadest

known as the Global Goals, are a uni-

sense, the strategy for sustainable devel-

versal call for action to end poverty,

opment aims to promote harmony among

protect the planet and ensure that all

people and between human beings and

people enjoy peace and prosperity al-


ways. The goals are interconnected.

In 1992, at the Earth Summit (Rio,

The key to success on one will involve

1992) there was consensus that environ-

tackling issues associated with anoth-

ment, and economic and social develop-

er. The SDGs work in the spirit of

ment cannot be considered in isolation,

partnership and pragmatism, to make

and in addition to treaties and agreements

the right choices now to improve life,

on climate change, biological diversity,

in a sustainable way, for future genera-

deforestation, and desertification, the Rio

tions. They provide clear guidelines

Declaration contains fundamental princi-

and targets for all countries to adopt in

ples on which nations can base their fu-

accordance with their own priorities

ture decisions and policies, considering

and the environmental challenges of

the environmental implications of socio-

the world at large. The SDGs are an

economic development.

inclusive agenda. They tackle the root

In 2000 the Millennium Sum-

causes of poverty and unite all nations

mit of the United Nations, following the together to make a positive change for

adoption of the United Nations Millenni-

both people and planet (UNDP).The

um Declaration, established the eight Mil-

15th SDG of UN relates to Life on land,

lennium Development Goals (MDGs) to

and involves protecting, restoring and

be achieved by the year 2015. The MDGs

promoting sustainable use of terrestri-

are: 1. to eradicate extreme poverty and

al ecosystems, sustainably managing for-

hunger, 2. to achieve universal primary

ests, combating desertification, and halt-

education, 3. to promote gender equali-

ing and reversing land degradation and

ty and empower women, 4. to re-

halting biodiversity loss.

duce child





The stable functioning of Earth‛s

prove maternal





life support system – which includes the

bat HIV/AIDS, malaria, and other diseas-atmosphere, oceans, forests, waterways,

es, 7. to ensure environmental sustainabil-

biodiversity and biogeochemical cycles, is

ity, and 8. to develop a global partnership

a prerequisite for future human develop-

for development. In the present context

ment. However, as per recent research

Goal 7: Ensuring environmental sustaina-

findings this functioning is at risk (Rock-

bility is very important. Two of the im-

ström et al., 2009). Further human pres-

portant targets of MDG 7 are: Integrating

sure may lead to large-scale, abrupt, and

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 2

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John potentially irreversible changes to Earth‛s

generations depends, stands on three pil-

life support system (Lenton 2011; Bar-


nosky et al., 2012). Likely impacts on


Sustainable agriculture

humanity include: diminishing food pro-


Conserving biodiversity

duction, water shortages, extreme weath-


Protecting the environment

er, ocean acidification, deteriorating eco-

systems, and sea-level rise. In this back-

Increasing food production must

drop Griggs et al. (2013) suggested that

involve, developing/ introducing better

we redefine sustainable development as

(efficient, high yielding, insect-pest re-

“Development that meets the needs of the

sistant) varieties of crop plants, conserv-

present while safeguarding Earth‟s life-

ing biodiversity, and protecting environ-

support system, on which the welfare of

ment. Plant Tissue Culture can greatly

current and future generations depends.”

contribute in all these.

Without economic, technological, and

societal transformations, chances of large-

4. Plant tissue culture

scale humanitarian crises exist. Such cri-

ses could undermine any gains made by

Plant tissue culture is the aseptic

meeting the MDGs. A re-evaluation of the

growing of whole plants or parts (cells,

relationship between people and planet is

tissues/ organs) in/ on defined (synthetic)

necessary (Griggs et al., 2014).

nutrient media under controlled (envi-

ronmental) conditions (temperature, light,

3. Three pillars of sustainable devel-

humidity). Usually in glass vessels (test


tubes, conical flasks, jam bottles etc.) -

for a review see John et al. (1997).

In the second half of the 20th cen-

Plant tissue culture is based on

tury there was intensification of agricul-

cellular „totipotency‟, the inherent poten-

ture in most parts of the world. Intensive

tial of a plant cell to regenerate a whole

agriculture involved: (i) expanding farm

plant. Unlike animal cells, most plant

lands, by removing natural forests, (ii)

cells retain the capacity to regenerate the

better irrigation, by constructing big

whole organism even after undergoing the

dams, which again submerged vast forests

final differentiation. In plants, as long as

in their catchment areas (iii) use of chem-

the cells have an intact membrane system

ical fertilizers and pesticides, to produce

and a viable nucleus, even highly mature

high yields. Destruction of natural forests

and differentiated cells retain the ability

had many adverse effects; most im-

to regenerate to a meristematic state.

portantly, the effects on climate, envi-

Though initially, in the first two decades

ronment, and biodiversity. Extensive use

of the 20th Century progress was slow,

of chemical fertilizers and pesticides also

standardization of universal plant tissue

had their own adverse effects. Excessive

culture media - White‟s (White, 1933),

use of chemical fertilizers has resulted in

Gamborg‟s (Gamborg et al. , 1975) and

nitrate accumulation, increased soil salini-

MS (Murashige and Skoog, 1963)

ty, and water eutrophication. High use of

changed the scene. Plant tissue culture

pesticides has resulted in development of

media contain minerals, growth factors

resistance in many pest species. In recent

and a carbon source (usually sucrose).

years there is much concern about envi-

Controlled environmental factors are light

ronmental contamination by fertilizers

(intensity and length – photoperiod), tem-

and pesticides.

perature, relative humidity. On/ in a cus-

Sustainable Development, that

tom standardized medium, and controlled

meets the needs of the present while safe-

environmental conditions, the explant

guarding Earth‟s life-support system, on

(starting plant material) - usually young,

which the welfare of current and future

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 3

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John undifferentiated tissue, regenerate into

gregates of few cells. These cells/ cell ag-

whole plants.

gregates grow/ divide/ separate as a result

of agitation, and can be continually main-

4.1. Types of cultures

tained in this state. Growth of cells in

Different types of cultures are

suspension culture can be more easily

possible: (i) culture of whole plants, (ii)

manipulated in liquid medium than on

embryo culture (embryo rescue), (iii) or-

semi-solid medium. Slowly agitating the

gan culture (shoot tip culture, root culture,

liquid medium on a rotary shaker is nec-

leaf culture, anther culture etc.), (iv) cal-

essary for the growth of the cultures,

lus culture, (v) cell suspension and single

which can be sub-cultured. Growth in

cell culture, (vi) protoplast culture.

single isolated cells can be induced by

culturing them in hanging drops in micro-

4.2. Callus culture

chambers. Suspension cultures are useful

Callus is an amorphous mass pro-

in plant production by somatic embryo-

duced by cell proliferation, occurring in

genesis form single cells. In regeneration

an unorganized manner. In nature it is a

of plants from callus established on semi-

wound response, or a plant reaction to the

solid media from small cell aggregates,

presence of micro-organisms, insects, or

and for the production of secondary me-

to some kind of stress. Under in vitro

tabolites. Suspension cultures can also be

conditions callusing is a response to en-

initiated from tissue other than callus

dogenous or exogenous growth regula-

(Geile and Wagner, 1980).

tors. The potential for callus formation is

dependent on the tissue (explant) type.

4.5. Protoplast cultures

Meristematic tissues are more suitable for

Protoplasts are plant cells without

callus induction than mature tissues. Cal-

cell walls. In 1882, Klercker isolated pro-

lus cultures can be maintained for long by

toplasts mechanically for the first time.

sub-culturing the primary callus (callus

The yield of protoplasts was very low. In

established originally from the explant),

1960, Cocking using enzymes for the first

at periodic intervals.

time could isolate protoplasts in large

numbers. Protoplasts can be isolated from

4.3. Somaclonal variations

different plant parts, or from tissues al-

Long term callus cultures can

ready in culture. Enzymatic isolation is

however, suffer from spontaneously aris-

now the most commonly used method. A

ing genetic variations, reflected in the

combination of these two can also be

phenotype of plants regenerated from


such calli. These variations are known as

One of the important applications

somaclonal variations. Somaclonal varia-

of protoplasts is in somatic hybridization.

tions are reported in many species. The

Many agents like NaNO3 (Power et al. ,

basis of somaclonal variations is not well

1990), a higher pH, and a higher concen-




tration of calcium ions in the medium

ments, activation of endogenous trans-

(Melchers and Labib, 1974), polyethylene

posons, and changes in the status of DNA

glycol (Kao and Michayuluk, 1974;

methylation, are considered to be the con-

Wallin et al. , 1974), and a high strength

tributing factors.

electric field (Zimmermann and Scheu-

rich, 1981), are used for obtaining fusion

4.4. Suspension cultures

between protoplasts. Protoplasts, when

Culture of unorganized plant cells,

placed in appropriate media regenerate

as single cells/ cell aggregates, in liquid

cell walls and form calli, from which

medium. Friable callus when cultured in

plants can be regenerated. Protoplasts are

agitated liquid medium, the cells separate

used for producing somatic hybrids (para-

and form a suspension of single cells/ ag-

sexual hybrids), for genetic manipulation,

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 4

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John and for basic studies on a variety of as-shoot tips of virus infected plants are also

pects. Regenerating plants from proto-

virus-free. Morel and Martin (1950; 1955)

plasts is difficult in some species. Con-

could produce healthy plants from virus-

ventional hybridization depends on affini-

free plants through shoot-tip culture from

ty of gametes. Wide crosses are not pos-

infected mother plants. This is possible

sible because of well-established cross

because the pathogen concentration is not

breeding barriers. Protoplast fusion makes

uniform in the infected plants, and apical

such hybridizations possible.

buds of rapidly growing shoots are often

not invaded by the virus. Morel (1960)

4.6. Anther (isolated microspore) cultures

used shoot apices of orchids to obtain

Guha and Maheshwari (1964) ob-

their rapid clonal multiplication. Shoot tip

tained haploid embryos, directly from an-

culture has two important practical appli-

ther cultures of Datura innoxia. The

cations: (i) virus eradication and (ii) mi-

origin of these embryos was traced to the




pollen grains. The potential of anther cul-

were followed by in vitro propagation of

ture for obtaining haploid plants, and

plants from shoot tip culture. Initially

from them by chromosome doubling of

most of the species micropropogated were

homozygous diploid plants was apparent.

herbaceous (Morel, 1964; Murashige,

In 1974, Nitsch had reported regeneration

1974). Now methods are available for the

of haploids and homozygous diploids by

micropropagation of a large number of

chromosome doubling, from isolated mi-

species belonging to a wide range of plant

crospore culture (Nitsch, 1974a; 1974b).


Culturing the microspores along with an-

ther wall is essential for success. In iso-

4.8. Embryo culture

lated microspores, pollen embryogenesis

Very young to mature embryos

is induced only rarely.

can be cultured in vitro. Embryo culture is

This technique has great potential

one of the oldest applications of plant tis-

in plant breeding. Normally it takes self-

sue culture in plant breeding. It has many

ing for many generations to obtain homo-

practical applications, and very useful in

zygosity in parental lines required in

obtaining hybrid plants from crosses in

breeding programmes. This time can be

which post-zygotic incompatibility exists.

considerably reduced by haploid culture

In post zygotic incompatibility, fertiliza-


tion and zygote formation occur on cross

pollination. The zygote grows, but is not

4.7. Meristem culture and shoot tip cul-

accepted by the endosperm. This results


in embryo abortion at some stage of de-

When growing points (meristems)

velopment before maturing of the seed. In

of shoots are cultured they continue their

such instances when the ovary/ ovule/

organized growth. The shoots/multiple

embryo with a part of the maternal tissue

shoots produced can be rooted to produce

is excised and cultured on a suitable me-

plantlets. This capacity has practical ap-

dium and under optimum culture condi-

plication and economic significance for

tions, it matures to produce a seedling.

plant propagation.

This procedure is hence called embryo

Culture of the meristemic zones

rescue. Sharma et al. (1980) obtained few

/extreme shoot tip is known as meristem

hybrids between Solanum melongena and

culture, and culture of small segments (5-

S. khasianum by this method. Embryo

10 mm in size) from the shoot tip is

culture is useful also in overcoming seed

known as shoot tip culture. It was known

dormancy and for obtaining seed germi-

that meristems of virus infected roots are

nation in some vegetatively propagating

free of the pathogen (White, 1933; 1934).

species in which seeds are produced but

Limasset and Cornuet (1949) found that

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 5

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John normally do not germinate (e.g. some

shoots or leaves. These organs may arise

wild bananas).

out of pre-existing meristems or out of

differentiated cells. Indirect pathway in-

4.9. Invitro pollination and fertilization

cludes a callus stage.

Pre-zygotic incompatibility is one

Direct pathway bypasses a callus

of the major limitations in obtaining hy-

stage. The cells in the explant act as direct

bridization between many plant species

precursors of a new primordium, an organ

and varieties. In pre-zygotic incompatibil-

or a part in its most rudimentary form or

ity, the zygote is not formed on cross pol-

stage of development.

lination. The pollen either do not germi-

nate on the stigma of the female parent or

4.12. Somatic embryogenesis

the pollen tube gets arrested at some point

In plants, embryo-like structures

of its growth on the stigma/ in the style.

can be generated from non-germ cells

A variety of methods used in vivo to

(somatic cells), by circumventing the

overcome this barrier.

process of normal fertilization. As somat-

Kanta et al. (1962) developed an

ic embryos are formed without fertiliza-

in vitro technique for overcoming pre-

tion event, they are genetically identical

zygotic incompatibility. In this method,

to the parent tissue, and are therefore

the mature/nearly mature ovaries/ovules


are cultured on suitable media and polli-

Somatic embryogenesis may be

nated in vitro with cross pollen to obtain

direct or indirect. Indirect somatic embry-





ogenesis involves a callus phase prior to


embryo production. Direct somatic em-

bryogenesis involves production of em-

4.10. Root cultures

bryos from organized tissue without an

Tip portions from primary and

intervening callus phase. Irrespective of

secondary roots of many plants can be

the mode of production, anatomical and

cultured. In 1922, Kotte and Robbins in-

physiological features of somatic embryos

dependently postulated that true in vitro

are highly comparable to zygotic embry-

cultures could be raised from meristemat-

os. The morphological and temporal de-

ic cells from root tips and shoot tips.

velopments of somatic embryos are very

Kotte (1922) could cultivate root tips of

similar to that of zygotic embryos. They

pea and maize in nutrient media for long,

both proceed through a series of distinct

but no sub-culturing were done. Robbins

stages, namely, globular, heart, torpedo

(1922) could subculture his maize root

and cotyledon or plantlet stages for dicot-

cultures. White (1934) obtained unlim-

yledons, and globular, elongated, scutellar

ited growth of tomato roots, using the

and coleoptilar stages for monocotyle-

same medium as Robbins (1922), with

dons. These stages typically span a period

yeast extract. Root cultures are useful in:

of several days. In dicots initially small

(i) secondary metabolite production, and

globular embryos form which undergo

(ii) in basic studies on nematode infec-

isodiametric growth and establish bilat-

tions, mycorrhizal associations, and root

eral symmetry. In monocots, especially in

nodulation by Rhizobium bacteria.

grasses, the transition from globular stage

follows a series of events occurring sim-

4.11. Organogenesis

ultaneously; such as the development of

Organogenesis is the process of

scutellum, initiation of the coleoptilar

initiation and development of a structure

notch, tissue differentiation with the de-

that shows natural organ form and/or

velopment of embryogenic vascular sys-

function. It is the ability of non-

tem and accumulation of intracellular

meristematic plant tissues to form various

storage substances. Somatic embryogene-

organs de novo; the production of roots,

sis is used for: large-scale clonal propaga-

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 6

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John tion of elite cultivars, as an alternative to

Rapid and large-scale clonal (ge-

conventional Micropropagation, produc-

netically uniform) propagation of plants

ing synthetic (artificial) seeds. Indirect

(micropropagation) may allow faster pro-

somatic embryogenesis (via callus) or

duction of plants that are slow to propa-

secondary embryogenesis is used in gene

gate in vivo.

transfer. Somatic embryogenesis also of-

The time required for bulking-up

fers potential model for the study of mo-

of new cultivars before they are commer-

lecular, regulatory and morphogenetic

cially introduced can be drastically de-

events in plant embryogenesis.

creased. Storage of germplasm, e.g. Cry-


4.13. Micropropagation

Micropropagation is the tissue cul-

4.13.2. The process of micropropagation

ture method of clonal propagation of

a. A small piece of the plant to be cloned

plants. Plant tissue culture is rapidly be-

(the explant) is removed from a

coming a commercial method for propa-

healthy, well-maintained stock plant

gating difficult-to-propagate plants, new

and surface sterilized (explant varies

cultivars (selections, hybrids, transgenic),

with species, but shoot tips, leaves,

rare/endangered species. Micropropaga-

stem pieces, lateral buds, and young

tion is usually achieved by the release

flowers or floral parts are used).

(from dormancy), and growth of pre-

b. Surface sterilized explants are rinsed

existing (axillary/ lateral) meristems in

with sterile water, and placed aseptical-

the initial culture. This is followed by re-

ly in/ on specially formulated and steri-

peated enhanced formation of axillary

lized medium in culture vessels.

shoots by sub-culture on medium supple-

c. The explant may proliferate directly by

mented with plant growth regulators. The

enhanced lateral branching, or the tis-

shoots produced are rooted either in vitro

sue may undergo a certain period of

or ex vitro (out of culture).

unorganized growth (callus) prior to

There are many advantages of Mi-

shoot differentiation.

cropropagation. Shoot production is relia-

d. The growth of the cultures is principal-

ble and consistent. Multiplication rates

ly determined by the plant growth reg-

can be three-fold to eight-fold a month.

ulator (PGR) content of the culture

Plants produced via shoot culture are usu-

medium (the auxin and cytokinin alone

ally true-to-type and uniform. Allows

or in combination and concentration/s).

propagation of rare/ endangered/ hybrid/

Most cultures are established within 4

induced mutant/ genetically transformed

to 12 weeks depending on the species/

plants. There also are few disadvantages.


PGRs do not release apical dominance in

e. A proliferating shoot culture can be

all species. There may be a difference in

sub-cultured to produce divisions

results between juvenile and mature tissue

which will multiply rapidly.

of perennial species; shoot cultures may

f. Rate of multiplication vary and are af-

require a reversion to juvenility. Rooting

fected by many factors. Production of

of the micro-shoots may be difficult. Get-

thousands, and in some cases millions

ting uniform shoot production in vitro,

of plants a year from a single explant

which is very important in commercial

has been demonstrated

operations, may not be possible in some

instances. The procedure is relatively la-

5. Role of plant tissue culture in sus-

bor intensive, with high upfront costs to

tainable agriculture

get started.

Sustainable agriculture requires

4.13. 1. Applications of micropropagation

efficient, biotic and abiotic stress resistant

crop varieties. Germplasm collection and

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 7

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John storage, simple crop improvement meth-jor driver of loss of biodiversity. This puts

ods such as selection and bulking by rapid

in jeopardy the sustainability of agricul-

and large scale cloning by micropropaga-

ture and ecosystem services and their

tion can be of great use. Plant tissue cul-

ability to adapt to changing conditions.

ture techniques such as: anther culture,

This also poses serious threat to food and

dihaploid production, embryo rescue, and

livelihood security.

in vitro pollination and fertilization can be

very useful in developing crop varieties

6.4. Plant tissue culture methods for con-

through hybridization. Callus culture, cell

serving biodiversity

suspension culture, organogenesis and

Plant Tissue Culture offers novel

somatic embryogenesis are essential for

options for collection, multiplication and

crop improvement through transgenics

medium/ long-term ex situ conservation

(various genetic engineering techniques).

of plant biodiversity. By plant cell, tissue,

and organ culture techniques, rapid and

6. Role of plant tissue culture in con-

large scale multiplication and season in-

serving biodiversity

dependent production of planting material

is possible. This has helped in the conser-

6.1. Biodiversity

vation of many endangered species. Me-

"Biological diversity means the

dium-term conservation is achieved by

variability among living organisms from

slow growing cultures. Cryopreservation

all sources including, inter alia, terrestrial,

(at −196 °C, in liquid nitrogen) allows the

marine and other aquatic ecosystems and

safe and cost-effective long-term conser-

the ecological complexes of which they


are part; this includes diversity within

species, between species and of ecosys-

6.5. In vitro collection

tems.” - Definition of Biodiversity by

Potential advantages of in vitro


methods are: (i) Less space requirement,

(ii) Pathogen-fee plants, (iii) No need for

6.2. Importance of biodiversity

transfer (under storage conditions), (iv)

Biodiversity is essential to: (i) en-

Stored cultures can be used as stock for

sure the production of food, fibre, fuel,

vegetative preservation, and (v) Interna-

fodder, etc., (ii) maintain other ecosystem

tional exchange of plant material made

services, (iii) allow adaptation to chang-

easy because, no use of soil, and no path-

ing conditions - including climate change,


and (iv) sustain rural peoples' livelihoods

Basic goals of an in vitro storage

(Convention of Biological Diversity).

system are: to maintain genetic stability,

to keep in indefinite storage without loss

6.3. Threats to biodiversity

of viability, and most importantly, to be

Biodiversity is




ous threat as a result of human activities.

Three types of Plant Tissue Culture sys-

The main dangers worldwide are: (i) In-

tems are available. They are: (i) Normal

vasion by alien species, (ii) Environmen-

growth, (ii) Slow growth, and (iii) Cryo-

tal degradation, (iii) Climate change and


global warming, (iv) Urbanization and

habitat conversion, (v) Population growth

6.5.1. Normal growth

and ever-increasing demand for resources,

Normal Growth is achieved either

(vi) Unsustainable over-exploitation of

on semi solid media or in liquid media.

natural resources.

Normal growth is similar to multiplication

Agriculture contributes signifi-

stage in micro-propagation, and requires

cantly to conservation and sustainable use

frequent sub-culture. Considered as genet-

of biodiversity. However, it is also a ma-

ically stabile when achieved through di-

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 8

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John rect organogenesis from apical buds / ax-Cryopreservation is the storage of

illary buds as explants.

living tissues at ultra-low temperatures

(˗196°C). It is useful in conservation of

6.5.2. Slow growth storage

plant germplasm of vegetatively propa-

By deliberate slowing down of

gated species, recalcitrant seed species

growth cultures can be stored at least for

(coconut palm etc.), conservation of tissue

6 months and maximum up to 6 years

with specific characteristics, cell lines

without sub-culturing. There are many

producing secondary metabolites, genet-

ways to achieve slow growth. First, by

ically transformed tissues, tissues compe-

manipulating storage temperature (cold

tent to transformation/ mutagenesis, path-

storage at 1-9°C) and light (low light in-

ogen (virus) eradicated tissue for future

tensity). Second, increasing osmotic po-

multiplication (as is done in Banana).

tential of the media [by using osmotically

Cryopreservation procedures are

active compounds such as sucrose (at

available only for limited number of plant

higher ~6%), mannitol etc.]. Third, by

species. Each species/ variety/ tissue type,

addition of inhibitors or retardants, for

needs standardization for: explant size

e.g. mineral oil overlay (callus), reduced

and type, water content, and natural freez-

oxygen tension etc.

ing resistance. Most studies on cryopres-

Plant Growth Retardants are

ervation of plants involve only one or a

chemicals that slow cell division and






elongation in shoots. They cause plants to

germplasm collections stored in liquid

be shorter and more compact, interrupt

nitrogen currently exist (with a relatively

cell division, stem elongation, and inflo-

limited number of accessions).

rescence / flower formation. But roots

continue to grow. Plant growth retardants

7. Role of plant tissue culture in pro-

may reduce the natural Gibberellic acid,

tecting the environment

or may produce more ethylene.

Forests are complex ecosystems,

6.5.3. Cold storage

predominantly composed of trees and

Storage at non-freezing temps,

shrubs, and usually have closed canopies.

from 1-9° C dependent on species. Stor-

There is nearly 4 billion hectares of forest

age of shoot cultures (stage I or II) works

in the world (this is about 30% of the total

well for strawberries, grapes, may be for

land cover). Depending on the physical,

many more spp. Transferred to fresh me-

geographical, climatic and ecological fac-

dium every 6 months/ annually/ or longer

tors, there are different types of forests

periods basis. Advantages of cold storage

like evergreen forest (mainly composed of

are: (i) simple, (ii) high rates of survival,

evergreen tree species) and deciduous

and (iii) useful in micropropagation (es-

forest (mainly composed of deciduous

pecially in periods of low demand). The

tree species). India‟s recorded forest area

disadvantages are: (i) may not be suitable

is 76.52 million hectares. This is 23.28%

for some tropical, subtropical species be-

of the country‟s total geographical area.

cause of susceptibility to cold injury, (ii)

Over 90% of the forest area is under gov-

requires refrigeration, which is more ex-

ernment ownership and is managed by the

pensive than storage at ultra-low tempera-

forest departments of the state govern-

tures (in cryopreservation). An alternative

ments (State of Forests Report, 2009).

to cold storage is the use of a medium

Forests are important both economically

with reduced nutrients and lacking su-

and ecologically, and render many ser-

crose (as reported in coffee).

vices to the life support system of earth.

Forests are the primary source of

6.5.4. Cryopreservation

wood. Wood is used to fulfil three basic

needs: (i) energy, (ii) construction materi-

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 9

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John al, and (iii) industrial raw material. About

due to conversion to agriculture land and

2 billion people in the developing world

urbanization. Deforestation has severe

are dependent on forests for their basic

consequences for the environment and

energy needs (fuel for cooking food). Un-

climate. More than 2000 times the total

til recently wood has been the chief con-

energy consumption of the world popula-

struction material. High strength to

tion, of solar energy reaches the earth‟s

weight ratio and availability in many

surface. Because of deforestation, not on-

kinds, ease of cutting and shaping with

ly this natural source of energy is wasted,

simple tools, and insulation to heat, make

but also has a serious negative impact on

wood an ideal construction material for

the environment, by way of surface heat-

many purposes. Major industrial uses of

ing, and desertification.

wood are: (i) paper and pulp, (ii) rayon,

In the tropical and sub-tropical re-

and (iii) plywood. Besides wood, forests

gions of the world, receiving about 600

are a source of a variety of non-wood for-

mm rainfall and above, plantation forestry

est products (NWFPs). Thus forests con-

of economically important tree species

tribute greatly to the economy. Around

(say teak for timber and eucalypts for

1.6 billion people depend on forests for

pulp) can take away pressure for forestry

their livelihood. This includes some 70

resources from natural forests and can add

million indigenous people.

to the forest cover. For this, large num-

Forests play an important role in

bers (in millions) of plating material of

maintaining ecological balance. Forests

superior varieties (fast growing, better

are atmospheric filters. They are the ma-

adapted, disease and insect-pest resistant

jor suppliers of oxygen. In photosynthesis

etc.) are necessary. But forest tree species

they fix atmospheric carbon dioxide into

are difficult to breed, because of their

carbohydrates, sugars, proteins, and many

long generation cycles, highly heterozy-

forms of biomass, thus playing a signifi-

gous natural populations, openly cross

cant part in the global carbon cycle. For-

pollinated nature, and lack of knowledge

ests contribute large quantities of mois-

about their genetics.

ture to the atmosphere, thus regulating

Clonal propagation of „superior‟

climate. Forests also conserve soil and

genotypes (identified for desirable traits)

water resources.

through tissue culture has been used very

The term forest implies „natural

profitably in case of many tree species.

vegetation‟ of the area, existing from

Eucalypts have been multiplied and used

thousands of years and supporting a varie-

for plantation forestry for a long time

ty of biodiversity. More than half of the

(FAO Report, 1981). Eucalyptus wood

known terrestrial plant and animal species

from plantation forestry has been used as

live in forests (Millennium Ecosystem

timber, industrial raw material and fuel.

Assessment, 2005). The forest ecosystem

Plantation forestry using eucalypts may

has two components - biotic and abiotic.

not be suitable for some places because of

The living component includes plants

high water demand. Teak ( Tectona gran-

(trees, shrubs, herbs etc.), animals and

dis Linn. f.) is another success story. In

microorganisms. Forests are home to

early 1980s, scientists from CSIR-

more than 80 per cent of all terrestrial

National Chemical Laboratory, Pune for

species of animals, plants and insects.

the first time regenerated complete plant-

Globally, deforestation is the major cause

lets from an 80 year old „elite‟ tree (Gupta

of loss of biological diversity, and is a

et al., 1983). Poplars ( Populus spp.) are

matter of great concern (Laurance, 2007).

another tree species which clonally prop-

Worldwide, the area of natural

agated through plant tissue culture tech-

forests decreases by some 13 million ha

niques, and widely cultivated in planta-

annually (this is about 3% of the total for-

tion forestry in many parts of the world.

est area). This loss of forest area is mostly

In India, poplars ( Populus spp.) are the

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 10

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John most popular tree species in agro-forestry

Steffen, W. and Shyamsundar, P.

production systems. Poplars are usually

(2014). An integrated framework for

intercropped with agricultural crops like

sustainable development goals. Ecol-

wheat, rice and sugar cane. Poplars are

ogy and Society 19, 49.

well known for their fast growth, out-

Guha, S. and Maheshwari, P. (1964). In

standing properties and quick and high

vitro production of embryos from an-

financial returns. Timber from poplars

thers of Datura. Nature 204, 497-98.

often forms the backbone of match, paper,

Gupta, P. K., Mehta, U. and Masca-

sports goods, plywood, and composite

renhas, A. F. (1983). A tissue culture

board industries.

method for rapid multiplication of

Eucalyptus camaldulensis. Plant Cell


Rep. 2, 296-99.

Hartley, M. J. (2002). Rationale and

Barnosky, A. D., Hadly, E. A., Bas-

methods for conserving biodiversity

compte, J., Berlow, E. L., Brown, J.

in plantation forests. Forest Ecology

H., Fortelius, M., Getz, F. M.,

and Management 155, 81–95.

Harte, J., Hastings, A., Marquet, P.

John, C. K., Nadgauda, R. S. and Mas-

A., Martinez, M. D., Mooers, A.,

carenhas, A. F. (1997). Tissue Cul-

Roopnarine, P., Vermeij, G., Wil-

ture of Economic Plants, Center for

liams, J. W., Gillespie, R., Kitzes,

S&T of Non-Aligned and other De-

J., Marshall, C., Matzke, N., Min-

veloping Countries, New Delhi, and

dell, D. P., Revilla, E. and Smith, A.

Commonwealth Science Council,

B. (2012). Approaching a state shift

London. pp. 3-34.

in Earth‟s biosphere. Nature 486, 52-

Kanta, K., Rangaswamy, N.S. and Ma-


heshwari, P. (1962). Test tube ferti-

Cocking E. C. (1960). A method for the

lization in a flowering plant. Nature

isolation of plant protoplasts and

194, 1214-1217.

vacuoles. Nature 187, 962-63.

Kao, K. N. and Michayluk, M. R.

FAO (1999). State of world‟s forests

(1974). A method for high frequency

1999, United Nations Food and Agri-

intergeneric fusion of plant proto-

cultural Organization, Rome, 154 pp.

plasts. Planta 115, 355-67.

FAO (2007). State of world‟s forests

Kotte, W. (1922). Kulturversuchemitiso-





United Nations Food and Agricultur-

Allgem. Bot. 2, 413-434.

al Organization, Rome.

Klercker, I. A. F. (1892). Eine method

Gamborg, O. L., Shyluk, J. and Kar-


tha, K. K. (1975). Factors affecting

Oefvers K. Vetensk. Akad.Foerh. 9,

the isolation and callus formation in


protoplasts from shoot apices of Pi-

Laurance, W. F. (2007). Have we over-

sum sativum L. Plant Sci. Lett. 4,

stated the tropical biodiversity crisis?


Trends Eco. Evol. 22, 65-70.





Lenton, T. M. (2011). Beyond 2°C: rede-

Gaffney, O., Rockström, J., Öh-

fining dangerous climate change for

man, M. C., Shyamsundar, P.,

physical systems. Wiley Interdiscipli-

Steffen, W., Glaser, G., Kanie, N.

nary Reviews: Climate Change 2(3),

and Noble, I. (2013). Sustainable


development goals for people and

Limasset, P. and Cornuet, P. (1949).

planet. Nature 495, 305-7.

Recherche du virus de la mosaique

Griggs, D., Stafford Smith, M., Rock-

du Tabacdans les meristemes des

ström, J., Öhman, M. C., Gaffney,

plantesinfectes. C. R. Ac. Sc. 228,

O., Gisbert Glaser, G., Noble, I.,


ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 11

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John Melchchers, G. and Labib, G. (1974).

lated plant protoplasts. Nature 225,

Somatic hybridization in plants by


fusion of protoplasts - I. Selection of

Reid, W. V. and Miller, K. R. (1989).

light resistant hybrids of a haploid

Keeping options alive: the scientific

light sensitive varieties of tobacco.

basis for conserving biodiversity.

Mol. Gen. Genet. 135, 277-94.

World Resources Institute, Washing-




ton, DC. 128 p.

(2005). Ecosystem and human well-

State of Forests Report (2009). Ministry

being: current state and trends. Find-

of environment and forests, Govern-

ings of the conditions and trends

ment of India, New Delhi.

working group, In: Hassan, R.,

Raghavan, V. (1980). Embryo culture,

Scholes, R. and Ash, N. (eds.) Mille-

In: Vasil, I. K. Ed. Perspectives in

nium ecosystem assessment series.

plant cell and tissue culture, Int. Rev.

Island Press, Washington, U.S.A.

Cytology Suppl. 11B, Academic

Morel, G. (1950). Sur la culture des tis-

Press, New York, USA, pp. 209-240.

sue de deux Monocotyledones. C. R.

Robbins, W. J. (1922). Effect of auto-

Ac. Sc. 230, 1099-1101.

lyzed yeast and peptone on the

Morel, G. (1960). Producing virus free

growth of excised corn root tips in

cymbidium. Bulletin of American Or-

the dark. Bot. Gaz. 74, 59-79.

chid Society 29, 495-497.

Rockström, J. et al. (2009). A safe oper-






ating space for humanity. Nature 461,

l‟acidepantotheninesur la croissance


des tissues d‟Aubepinecultives in

Sharma, D. R., Chowdhury, J. B., Ahu-

vitro. C. R. Ac. Sc. 243, 166-168.

ja, U., and Dhankhar, B. S. (1980).

Morel, G. and Martin, C. (1950). Gue-

Interspecific hybridization in genus






Solanum. A cross between S.

teintesd‟unemaladie a virus. C. R. Ac.

melongena and S. khasianum through

Sc. 235, 1324-25.

embryo culture. Zeitschrift fur Pflan-

Morel, G. and Martin, C. (1955). Gue-

zenzuchtung, 85(3), 248-253.

rison de pommes de terreatteintes de

Shindell , D. Kuylenstierna, J. C. I.,

maladies a virus. C. R. Ac. Agri. 41,






R., Amann, M., Klimont, Z., Anen-

Murashige, T. (1974). Plant propagation

berg, S. C., Muller, N., Janssens-

through tissue cultures. Ann. Rev.

Maenhout, G.,Raes, F., Schwartz,

Plant Physiol. , 24, 135-65.

J., Faluvegi, G., Pozzoli, L., Ku-

Murashige, T. And Skoog, F. (1963). A




revised medium for rapid growth and

L., Emberson,



bioasays with tobacco tissue cultures.

D., Ramanathan, V., Hicks, K.,

Physiol Plant. 15, 473-497.




T., Milly,

Nitsch, C. (1974a). La culture de pollen

G., Williams, M., Demkine, V. and

isolesurmihensynthetique. C. R. Ac.

Fowler, D. (2012). Simultaneously

Sc. 278, 1031-34.

mitigating near-term climate change

Nitsch, C. (1974b). Pollen culture – a

and improving human health and

new technique for mass production of

food security. Science 335, 183–189.

haplid and homozygous plants. In:

UNEP (1994). Convention on biological

Kasha, K. J. Ed. Haploids in Higher

diversity, UNEP/CBD, Switzerland,

Plants – Advances and Potential.

November 1994, 34 p.

University of Guelph, Guelph, Swe-

United Nations (2005). 2005 world

den. pp. 123-35.

summit outcome [adoption resolu-

Power, J. B., Cummins, S. E., and

tion, 60th session].United Nations,

Cocking, E. C. (1990). Fusion of iso-

New York, New York, USA. [online]

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 12

Biotech Sustainability (2017)

Plant Tissue Culture for Sustainability John URL:

SDSN) (2013). An action agenda for


sustainable development. Report for

United Nations (2012). The future we

the UN Secretary-General. Sustaina-

want: outcome document adopted at

ble Development Solutions Network,

Rio+20. United Nations, New York,

New York, New York, USA. [online]










United Nations (2012 a). The millennium

United Nations Environment Pro-

development goals report 2012. Unit-

gramme (UNEP) (2013). Embedding

ed Nations, New York, New York,

the environment in sustainable devel-



opment goals. UNEP Post-2015 Dis-


cussion Paper 1.UNEP, Nairobi,






United Nations (2013). A new global


partnership: eradicate poverty and

Wallin, A., Glimelius, K. G. and Eriks-

transform economies through sus-

son, T. (1974). The induction of ag-

tainable development. The report of

gregation and fusion of Daucus caro-

the High-Level Panel of eminent per-

ta protoplasts by polyethylene glycol.

sons on the post-2015 development

Zeitschrift fur Planzenphysiol. 74,

agenda.United Nations, New York,




White, P. R. (1933). Plant tissue culture:


Results of preliminary experiments


on the culturing of isolated stem tips


of Stellaria media. Protoplasma 19,

United Nations Open Working Group


(UN OWG) (2013). Interim progress

White, P. R. (1934). Multiplication of the

report to UN General Assembly.

viruses of tobacco and ancuba mosaic

United Nations, New York, New

in growing excised tomato roots.




Phytopathol. 24, 1003-11.

URL: http://sustainabledevelopment.

White, P. R. (1933). Potentially unlim-

ited growth of excised tomato root tip


in a liquid medium. Plant Physiol, 9,

United Nations Open Working Group


(UN OWG) (2014). Outcome docu-

Zenktler, O. M. (1980). Intra-ovarian

ment - Open Working Group on Sus-

and in vitro pollination. In: Vasil, I.

tainable Development Goals (19th

K. Ed. Perspectives in plant cell and

July 2014).United Nations, New

tissue culture, Int. Rev. Cytology

York, New York, USA. [online]

Suppl. 11B, Academic Press, New

URL: http://sustainabledevelopment.

York, USA, pp. 137-156.

Zimmerman, U. and Scheurich, P.


(1981). High frequency fusion of

United Nations Sustainable Develop-

plant protoplasts by electric fields.





Planta 151, 26-32.

© 2017 by the author. Licensee, Editors and AIMST University, Malay-

sia. This article is an open access article distributed under the terms and

conditions of the Creative Commons Attribution (CC BY) license


ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 13

Biotechnology for Sustainability

Achievements, Challenges and Perspectives

Biotech Sustainability (2017), P14-40

Traditional Medicine of the Tribes in Tamil Nadu and Its

Sustainable Use through Biotechnology

Valli Gurusamy1, Kavitha Valampuri John2, Usha Raja Nanthini Ayyakkanu2,

Ramani Bai Ravichandran3,*

1Vice Chancellor, Mother Teresa Women’s University; 2Department of Biotechnology,

Mother Teresa Women’s University; 3Department of Zoology, University of Madras, India;

*Correspondence:; Tel: +91 9444020828

Abstract: India is a land of mega biodiversity representing about 7% of the world‟s flora

and 6.5 per cent of world‟s fauna. Tamil Nadu, one of the southern most states of India, is

rich in forest cover and cultural diversity. Genomic evidence supports the peopling of Tamil

Nadu from the 1st wave of migration of humans from the „Out of Africa‟ exodus and points

out that the tribes of state were among the earliest settlers in the region. The tribal popula-

tion of Tamil Nadu represents 1.02% of the total population of the state. Living in close as-

sociation with the forest, they have accumulated a treasure trove of ethno botanical

knowledge in the form of traditional medicine. The future of sustainable use of renewable

forest product lies with the molecular tools of Biotechnology. We present here an analysis

of the documented literature of the medicinal plants used by the tribes of Tamil Nadu for

treatment of common disorders. We also present the challenges and prospects within the

scope of Biotechnology to ensure sustainable use of traditional medicine for the betterment

of mankind and environment.

Keywords: Biotechnology; sustainable; tribes; traditional medicine; Tamil Nadu.

1. Introduction

along with their 1st wave of migration out

of Africa. Traditional Medicine is the sum

India is a land of enormous cultur-

total of long-standing information on the

al, linguistic and religious diversity pre-

knowledge, skills, and health practices

sumably because of Man‟s long stay, for

based on the theories, beliefs, and experi-

the past 50-70,000 years in this continent.

ences indigenous to different cultures or

This is an outcome of various migrations

local communities. Traditional medicine

that took place into India, serving as a

incorporates plant, animal and mineral

major corridor for the dispersal of modern

based medicines and encompasses spir-

humans out of Africa (Cann, 2001). Ar-

itual therapies, manual techniques and

chaeological evidences indicated that the

exercises which can be applied singularly

Indian subcontinent was peopled by vari-

or in combination for the maintenance of

ous migrations since Palaeolithic (300-

health through the prevention, diagnosis,

400,000 BCE), starting with the Late

improvement or treatment of physical and

Pleistocene (Misra, 2001). „The Castes

mental illness. Traditional knowledge has

and Tribes of Southern India‟ was an at-

been well preserved and orally passed

tempt to catalogue these populations

from one generation to the next in the

(Thurston, 1909).The knowledge of the

form of stories, legends, folklore, rituals,

medicinal value of plants, animals and

songs, art, and even laws. Since there is

other substances and their uses goes back

no written script the exchange of know-

to the time of the earliest settlers probably

how between diverse communities is a

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 14

Biotech Sustainability (2017)

Traditional Medicine of the Tribes in Tamil Nadu… Gurusamy et al.

process of evolution through trial and er-

cient lineage from the 1st wave of the „out

ror which makes documentation and rec-

of Africa‟ exodus and thus in vast ethno-

ord-keeping almost impossible. This pro-

botanical knowledge. According to the

cess of exchange and assimilation is con-

2011 census, 104 million tribal people

tinuous, and today there is a growing

speaking over 227 linguistic groups in-

awareness among the medical community

habit varied geographic and climatic

about the intrinsic value of traditional

zones of the Indian subcontinent. Ethno-

medicine, and as a result in India Ayurve-

medicine includes plants, animal products

da, Unani and Siddha have entered the

and minerals used by tribal communities

mainstream to compliment biomedicine.

of a particular region or country for me-

Contemporary Indian society faces the

dicinal purposes other than those men-

challenge of integrating the best of the

tioned in classical streams of the respec-

different healing traditions to provide a

tive cultures. Tribal people have been us-

holistic health care.

ing a large number of wild plants as doc-

umented by ethnobotanical investigations.

2. Traditional knowledge

The application of most of the plants rec-

orded are either lesser known or hitherto

Even before classical medical

unknown to the outside world. India has

knowledge of ancient India was codified

been the country most concerned about

into the canonical texts of Ayurveda in

the conservation of its medicinal plants.

the 6th century BC, there were abundant

There are over 45,000 species of vascular

sources of medical knowhow in the sub-

plants reported from India of which as

continent from prehistoric times. Tradi-

many as 15,000 may be used medicinally.

tional healers can be either folk or tribal

The folk medicine system of India use

healers and have worked in intimate rela-

about 5,000 plant species with about

tion with their environment. Traditional

25,000 formulation, whereas the tribal

healing ranges from simple home reme-

medicine involves the use of over 8,000

dies related to nutrition and treatment for

plant species with about 1,75,000 specific

minor illnesses, to more sophisticated

preparations (Pushpangadan and George,

procedures such as midwifery, bone set-

2010). More than 90% of the raw material

ting, blood-letting (therapeutic phleboto-

for traditional medicine comes from wild

my) and treatment of snake bites and

harvesting as this the common method

mental disorders. Some healing practices

used for collecting them (Tandon, 1996;

were considered to be sacred and were

Gupta 1998; Ved et al., 1998). About 71

associated with rituals that helped safe-

medicinal plant species are classified as

guard them for there is a substantial over-

“rare”, and of this 92% are in active trade,

lap between healing plants and sacred

and 74% are traded nationally. It has been

plants. Categories of traditional healers

estimated that between 4,000 and 10,000

are traditionally trained healers, old indi-

medicinal plant species in India face ex-

viduals of the community, educated indi-

tinction in the local, regional and national

viduals acquiring certain knowledge from

levels (Hamilton, 2004). In an effort to

their predecessors, ancient inscriptions in

create leadership in affordable and holis-

the form of copper plate/palm leaf writ-

tic health care, India is committed to

ings, old and recent publications in re-

promoting traditional medicines like

gional language.

Ayurveda which remained untapped due

to inadequate scientific scrutiny. Steps

3. Indian ethnobotany

are being taken to bring in regulatory

amendments in research and effective en-

India is one of the richest coun-

forcement for integration of quality prod-

tries in the world not only in biodiversity

ucts, practices and practitioners into the

but also in different ethnic groups of an-

AYUSH (Ayurveda, Yoga and Naturopa-

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 15

Biotech Sustainability (2017)

Traditional Medicine of the Tribes in Tamil Nadu… Gurusamy et al.

thy, Unani, Siddha and Homoeopathy)

in moderate altitude of the district name-

system at central and state level.

ly: New Kotagiri (Aggal), Kil-Kotagiri,

Kundah, Kallimalai, Gudalur, Trichigadi

4. Traditional knowledge of the tribes

and Sholur Kokal.Their settlement is

of Tamil Nadu

called the “Kokkal” with linear row of

houses in streets, „Keri‟. Each village has

According to the 2001 census,

three Keri known as Kizhkeri, Nadukeri

tribal population in Tamil Nadu is 6,

and Melkeri. Keri, clan, exogamy is note-

51,321 which constitute 1.02% of the to-

worthy among Kotas (Kavitha V.J.,

tal population. There are 36 tribes and sub

2008). Their chief diety is Kambattrayan.

tribes in Tamil Nadu. Out of the 36

Their population was 1,894 in 2001 cen-

Scheduled Tribe communities in the


state, about six tribal populations Todas,

Irulas: They are also called as Iu-

Kotas, Krumbas, Paniyas, Irulas, Kat-

van,Villiar. The Irulas are distributed in

tunayakas have been classified as primi-

the lower altitudes of the Nilgiri hills (dis-

tive tribes with incredibly high anthropo-

trict). They are negrotoid in appearance

logical significance. The primitive tribes

whose chief occupation is as plantation

occupy the length and breadth of the Nil-

labourers in the estates. Their settlements

giri district in the Western Ghats of Tamil

are called “Aral”. Their dialect is Tamil

Nadu. One sixth of the land mass of Tam-

mixed with Malayalam. Their community

il Nadu is covered by forests. The tribes

is divided into seven exogamous clans

of Tamil Nadu live in and around the re-

(Kuems): Kupper, Sambe, Kalkatti, Ku-

served forests and have gained immense

runagar, Devanan, Peradar and Punger

knowledge on the use of forest produce to

(Rajan and Sethuraman, 1991). They are

treat common disorders.

basically hunter gatherers. Their popula-

Todas: They are called by other

tion was 6,700 in 2001 census.

names like Tudas, Thuduvans, and Todar

Kurumbas: The Kurumbas prac-

(Kavitha V.J., 2008). They are profes-

tice hunting food gathering economy,

sional pastoralists and dairy men, a purely

well-versed in honey collection tech-

pastoral economy in India today, living in

niques. They are plain dwelling people

the higher altitudes in the traditional

living in the interior forests of the district.

houses called “Munds” that are half barrel

Their staple foods are wild tubers (Di-

shaped and are vegetarians. Their dialect

oscorea bulbosa), wild fruits and other

is independent form of Dravidian Tamil-

minor forests produces. Their settlements

Malayalam. They are fair skinned and

are called “Mottam”. They are dark

wear ornaments and their dress is akin to

skinned and speak the Kurumba dialect.

the Roman „toga‟. They have two exoga-

Kurumbas are a heterogenous population

mous divisions called Tarthar and Teivali.

having divisions such as Halu Kurumbas,

There are five socially distinguishable

Betta Kurumbas, Mullu Kurumbas, Jess

sects (clans) such as Pelki, Pekkan,

kurumbas and Urali Kurumbas. Their

Kuttan, Kenna and Jodi (Rajan and Sethu-

population was 6,872 in the 2001 census.

raman, 1993). Their population was 1,600

Paniyas: The Paniyas are negro-

in the 2001 census.

toid people living in bamboo huts at the

Kotas: Their other names are Ko-

junction of Kerala and Tamil Nadu bor-

ter, Kothewars, and Kohatur. The Kotas

der. They work as labourers with Waya-

are musicians and excellent craftsmen

nad Chettis though they were basically

having mastery over ironworking. They

hunter gatherers. Their settlements are

are light skinned, with copper hair. They

called “Paddi”. They possess excellent

speak the “Kota” a Dravidain language.

skills in the art of fishing by employing

Their distribution in the Nilgiri district is

certain plant parts like bark of Eugenia sp.

confined only to seven villages inhabiting

and leaves of Aibizzia sp. as stupefying

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 16

Biotech Sustainability (2017)

Traditional Medicine of the Tribes in Tamil Nadu… Gurusamy et al.

agents. They speak the Paniya dialect,

dicinal knowledge of the tribes of Tamil

practice Illam, patilineage, and their in-


heritance is by “Marumakkathayam”.

They numbered 6,700 in 2001 census.

5. Ethnomedical wealth of tribes of

Kattunayakas: They are also

Tamil Nadu

called as Shola Nayakans, Jenu or Teen

Kurumbas. They are another group of for-

A total of 229 medicinal plants

est dwellers who are nomadic in nature,

used by the tribes of Tamil Nadu belong-

their staple foods are honey, wild fruits

ing to 79 families for the treatment of

and tubers. Their settlements are called

more than 40 disorders were documented.

“Paadi”. They are short and black with

The percent representation of the families

protruding forehead. They have curly hair

of plants used as medicine by the tribes of

and speak Kannada language. Eating bi-

Tamil Nadu is represented in Figure 1.

son flesh is a cultural taboo with them.

Euphorbiaceae is the largest family repre-

The social customs and religious practices

sented by 18 species at 12% followed by

of Kattunayakas are akin to Kurumbas in

Fabaceae by 14 species (9%), Lamaceae

many respects. They population was

by 13 species (8%), Asteraceae by 11

1,425 in the 2001 census.

species (7%), Solaneaceae and Rutaceae

Paliyars: They are found in the

by 10 species each (6%) and Ascelpiada-

hilly regions of Madurai, Dindigul, Theni,

cea by 8 species(5%). This data marks a

Tirunelveli and Virudhunagar districts. It

direction for scientific researchers as to

is believed that Paliyars are indigenous

which of the families to search for to

people of Palani hills of Kodaikanal and

identify bio active compounds.

speak Tamil. Physically they are similar

Of the primary parts of the plant

to the Semong of Malaya and other Indian

used the leaves formed 40% of usage in

tribal communities. They can be grouped

traditional medicine followed by the root

into three categories based on their life

and bark at 11%, whole plant and fruit at

styles, namely, nomadic, semi nomadic

8%, stem at 7% and seed at 5% (Figure

and settled. Nomadic Paliyars don't build

2). It is of utmost importance to see this

houses; they live temporarily in rock

data in the light of the major families rep-

caves called 'Pudai'.

resented for medicinal use by the tribes of



Tamil Nadu. It is also important to test

knowledge for the tribes of Tamil Nadu

these bioactive compounds from different

was retrieved using Pubmed and Google

parts of the plant as a combination and to

using the keywords ethnomedicne, tribes,

use bioactive compounds different fami-

Tamil Nadu. The ethnobotanical data pre-

lies in conjunction for any particular dis-

sented here included knowledge form


seven tribal groups of Irula, Pani-

The data when analysed for major

ya,Kurumba, Kota, Thoda, Kattunayak-

remedies against diseases it revealed that

kans and Paliyars (Table 1). These pub-

the tribes of Tamil Nadu had traditional

lished research articles were then ana-

remedies for wounds (31) and skin prob-

lysed manually to ascertain the traditional

lems (29), followed by stomach aches

knowledge of these communities related

(17), diahorrea and headache (13), Cold,

to the study area and the usage pattern of

cough, fever (12) and rheumatic diseases,

the medicinal plant species. The data was

gastric disorders and toothache (9). It is

further analysed using graphical represen-

interesting to note that the ethnomedicine

tations for summarizing and interpreting

of the tribals has 9 remedies for women to

for the major families of plant species

ease labour pain and 3 to induce lactation

represented, part of the plant used and

(Figure 3). Tribal ethnomedicine also has

cure for disorder from the traditional me-

remedies for diabetes and jaundice (5).

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 17

Biotech Sustainability (2017)

Traditional Medicine of the Tribes in Tamil Nadu… Gurusamy et al.

Anacardiaceae Aplaceae


Menispermaceae Myrataceae



2% Meliaceae



2% Mimosaceae

2% Pandanaceae




































Figure 1: Percent representation of major plant families used by Tamil Nadu tribes as medi-




Nuts Res in

Fruits Flowers





0% 1%




2% Latex










Whole Plant






Figure 2: Parts of the Plant used in traditional healing among the tribes of Tamil Nadu.

ISBN: 978-967-14475-3-6; eISBN: 978-967-14475-2-9 18

Biotech Sustainability (2017)

Traditional Medicine of the Tribes in Tamil Nadu… Gurusamy et al.

Table 1: Ethnomedical traditional knowledge of the tribes in Tamil Nadu


Botanical Name

Local name