Protocols for In Vitro Cultures and Secondary Metabolite Analysis of Aromatic and Medicinal Plants

481 Pages·2016·20 MB·English

by  S. Mohan Jain

Methods in
Molecular Biology 1391
S. Mohan Jain Editor
Protocols for In Vitro
Cultures and Secondary
Metabolite Analysis
of Aromatic and
Medicinal Plants
Second Edition
M M B
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life and Medical Sciences
University of Hertfordshire
Hatfield, Hertfordshire , AL10 9AB, UK
For further volumes:
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Protocols for In Vitro Cultures
and Secondary Metabolite
Analysis of Aromatic
and Medicinal Plants
Second Edition
E dited by
S. Mohan Jain
Applied Biology, University of Helsinki, Helsinki, Finland
Editor
S. Mohan J ain
Applied Biology
University of Helsinki
Helsinki, Finland
ISSN 1064-3745 ISSN 1940-6029 (electronic)
Methods in Molecular Biology
ISBN 978-1-4939-3330-3 ISBN 978-1-4939-3332-7 (eBook)
DOI 10.1007/978-1-4939-3332-7
Library of Congress Control Number: 2016934450
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Prefa ce
It is well known that plants consistently synthesize, accumulate, and use a bewildering
range of secondary metabolites as a part of their overall defense strategy. Many of these
metabolites have been used around the world as medicines for various human health prob-
lems. In recent years the quest for quality of life and a common belief that plants are “natu-
ral and therefore safe” have paved the way for a wider acceptance of plant-based medicines
worldwide. International trade in medicinal plants has become a major force in the global
economy, and the demand is increasing in both developed and developing countries. Thus,
the continued rise in consumer demand for plant-based medicines and the expanding world
population have resulted in the indiscriminate harvest of wild species of medicinal plants.
As well, a reduction of natural habitats for medicinal plants has placed many wild species in
danger of extinction. The impact of rapid climate changes may also have an adverse effect
on wild plant species leading to the loss of useful genetic material. Most medicinal plants
are harvested from the wild, and the traditional agricultural and horticultural practices have
not been developed even for most commonly used medicinal plant species.
The quality and consistency of the products are most challenging issues facing the
plant-based medicines. The production of medicinal metabolites in plants is affected by
plant genotype, cultivation, harvesting, processing, and distribution. Medicinal plant prep-
arations may also be contaminated with microbes and soil contaminants such as heavy met-
als, herbicides, pesticides, and other agricultural chemicals which can cause qualitative and
quantitative changes in the levels of medicinal metabolites. The widespread occurrence of
chemical variability and compromised quality of medicinal plants remain the major factors
in inconsistent results of clinical trials of plant-based medicines. New regulations are cur-
rently being developed internationally to ensure consistency, safety, and effi cacy of plant-
based medicines as well as how they are developed, manufactured, and marketed. Clearly,
there is an imminent need for the development of new technologies and production
approaches to improve the overall strategy of medicinal plant production to comply with
upcoming legal regulations.
In vitro cell culture and controlled environment production systems offer an excellent
opportunity for the selection and seasonal independent propagation of elite lines with spe-
cifi c, consistent levels of medicinal metabolites with minimum contamination. Additionally,
the plant materials produced by in vitro techniques allow effi cient application of the emerg-
ing analytical methods. The impact of these techniques perhaps greatest in the improvement
of medicinal plants since the resulting genetic diversity may open avenues for the discovery
of new medicinal metabolites and treatments. This book provides a detailed step-b y-s tep
description of protocols for the establishment of in vitro cultures of important medicinal
plants, their mass multiplication in controlled environment, and stepwise secondary metabo-
lite analysis. In addition many of these protocols will provide a basis for much needed efforts
of in vitro germplasm conservation or cryopreservation of medicinal plant species at the
brink of extinction as well as to protect them from the adverse impact of rapid climatic
changes. This book will certainly appeal to graduate and postgraduate students, researchers,
biotechnologists, industry, and the Government agencies and could be used as a textbook.
v
vi Preface
This book contains 30 book chapters dealing with in vitro propagation of medicinal
plants. Each manuscript has been peer reviewed and revised accordingly.
We greatly appreciate all reviewers who have contributed their time to review manu-
scripts that certainly helped in improving the quality of contributory manuscripts. Our
sincere thanks are due to the Humana Publishers for giving us the opportunity to edit
this book.
Helsinki, Finland S. M ohan Jain
Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . x i
1 Protocols for In Vitro Mass Multiplication and Analysis
of Medicinally Important Phenolics of a Salep Orchid,
Satyrium nepalense D.Don (“Salam Mishri”). . . . . . . . . . . . . . . . . . . . . . . . . . 1
Shashi B. Babbar and D eepak K . Singh
2 I n Vitro Culture and Phytochemical Analysis of P assiflora tenuifila
Killip and P assiflora setacea DC (Passifloraceae). . . . . . . . . . . . . . . . . . . . . . . . 13
Jenny Sumara Sozo , D aniel C uzziol C ruz , Ana Flavia Pavei ,
Isadora M edeiros da Costa Pereira , M arcia W olfart , F ernanda Ramlov ,
Daiane Fiuza Montagner , Marcelo Maraschin , and Ana Maria V iana
3 S hoot Tip Meristem Cryopreservation of H ypericum Species. . . . . . . . . . . . . . 31
Katarína Bruňáková and Eva Č ellárová
4 P rotocols for Biotechnological Interventions in Improvement
of Vanilla (V anilla planifolia Andrews.). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Minoo Divakaran , K . Nirmal Babu, and K.V. Peter
5 A ssessment of In Vitro Biological Activities of Anthocyanins-Rich
Plant Species Based on P linia cauliflora Study Model . . . . . . . . . . . . . . . . . . . 6 5
Heloisa S . Pitz , A driana C.D. T revisan , F austo R. C ardoso ,
Aline P ereira , E duardo L .G. M oreira , Manuel A. d e Prá ,
Letícia Mazzarino , Maria B . V eleirinho , Rosendo A . Yunes ,
Rosa M.R. d o Valle , and Marcelo M araschin
6 B iotechnological Approaches for Biomass and Cardenolide
Production in D igitalis purpurea L. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 1
Naivy P érez-Alonso , Borys C hong-Pérez , A lina C apote , A nabel Pérez ,
André Gerth , Geert A ngenon , and Elio J iménez
7 In Vitro Regeneration of Endangered Medicinal Plant
Heliotropium kotschyi (Ramram) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 03
Manal Ahmed S adeq , Malabika R oy P athak , A hmed Ali S alih ,
Mohammed Abido , and Asma Abahussain
8 M icropropagation and Biomass Production of True-to-Type
Stevia rebaudiana Bertoni. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
Arpan R. Modi , Vikas Sharma , Ghanshyam Patil ,
Amritpal S . S ingh , N. S ubhash , and Nitish Kumar
9 P anax ginseng Adventitious Root Suspension Culture:
Protocol for Biomass Production and Analysis
of Ginsenosides by High Pressure Liquid Chromatography. . . . . . . . . . . . . . . 125
Hosakatte N iranjana Murthy and K ee Y oeup P aek
vii
viii Contents
10 Immobilization of R ubia tinctorum L. Suspension Cultures
and Biomass Production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 41
Pınar N artop
11 I n Vitro Propagation and Conservation of B acopa monnieri L. . . . . . . . . . . . . 153
Neelam S harma , Rakesh Singh , and Ruchira Pandey
12 Establishment, Culture, and Scale-up of B rugmansia candida
Hairy Roots for the Production of Tropane Alkaloids . . . . . . . . . . . . . . . . . . . 173
Alejandra B eatriz C ardillo , Julián R odriguez Talou ,
and Ana M aría G iulietti
13 Micropropagation, Acclimatization, and Greenhouse
Culture of V eratrum californicum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
Sarah A . W hite , J effrey A delberg , Jacqueline N aylor-Adelberg ,
David A . Mann , J u Y eon S ong , and Youping Sun
14 I n Vitro Propagation of W ithania somnifera (L.) Dunal . . . . . . . . . . . . . . . . . 2 01
Pritika Singh , Rupam Guleri , and Pratap Kumar P ati
15 In Vitro Propagation of Sambong (B lumea balsamifera Linn.) . . . . . . . . . . . . 2 15
Thelma L. Soriano and Evangelina C . C angao
16 P roduction of Gymnemic Acid from Cell Suspension Cultures
of Gymnema sylvestre . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229
Praveen N agella , Vijayalaxmi S . D andin ,
and Hosakatte N iranjana M urthy
17 Scale-Up of A grobacterium rhizogenes- Mediated
Hairy Root Cultures of Rauwolfia serpentina:
A Persuasive Approach for Stable Reserpine Production. . . . . . . . . . . . . . . . . . 241
Shakti Mehrotra , Vikas S rivastava , M anoj K . G oel ,
and Arun K . K ukreja
18 In Vitro Shoot Cultures and Analysis of Steroidal
Lactones in W ithania coagulans (Stocks) Dunal. . . . . . . . . . . . . . . . . . . . . . . . 2 59
Rohit Jain , S umita Kachhwaha , and S. L . Kothari
19 In Vitro Propagation of C annabis sativa L. and Evaluation
of Regenerated Plants for Genetic Fidelity and Cannabinoids
Content for Quality Assurance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 75
Hemant L ata , Suman Chandra , Ikhlas A . Khan ,
and Mahmoud A . E lSohly
20 T ranscript Quantification of Genes Involved in Steviol Glycoside
Biosynthesis in S tevia rebaudiana Bertoni by Real-T ime Polymerase
Chain Reaction (RT-PCR) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 89
Arpan Modi , N itish K umar , and Subhash N arayanan
21 In Vitro Propagation and Conservation of W ithania somnifera (Dunal) L. . . . 3 03
Nigar Fatima , N aseem Ahmad , and Mohammad Anis
22 Construction of Hypericin Gland-Specific cDNA Library via
Suppression Subtractive Hybridization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 17
Rupesh Kumar Singh , W eina H ou , and Gregory Franklin
Contents ix
23 In Vitro and Cryopreservation Techniques for Conservation
of Snow Mountain Garlic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 35
Ritu M ahajan
24 I n Vitro Callus Induction and Plant Regeneration
from Stem Explants of C eropegia noorjahaniae,
a Critically Endangered Medicinal Herb . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 47
Jaykumar J . Chavan and Mahendra L. A hire
25 S omatic Embryogenesis of Date Palm (P hoenix dactylifera L.)
Through Cell Suspension Culture. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357
Poornananda M . N aik and Jameel M. A l-Khayri
26 P rotocols for Improvement of Black Pepper (P iper nigrum L.)
Utilizing Biotechnological Tools. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 367
K. Nirmal Babu , Minoo D ivakaran , G . Y amuna ,
P. N . Ravindran , and K. V . P eter
27 Protocols for In Vitro Propagation, Conservation,
Synthetic Seed Production, Microrhizome Production,
and Molecular Profiling in Turmeric ( Curcuma longa L.) . . . . . . . . . . . . . . . . 3 87
K . Nirmal Babu, Minoo Divakaran, Geetha S. Pillai,
V. Sumathi, K. Praveen, Rahul P. Raj, H.J. Akshita,
P.N. Ravindran, and K.V. Peter
28 P rotocols for In Vitro Propagation, Conservation,
Synthetic Seed Production, Embryo Rescue,
Microrhizome Production, Molecular Profiling,
and Genetic Transformation in Ginger ( Zingiber officinale Roscoe.) . . . . . . . . 4 03
K. Nirmal Babu, K. Samsudeen , Minoo D ivakaran ,
Geetha S . P illai , V. Sumathi, K. Praveen,
P.N. Ravindran, and K.V. Peter
29 Hairy Root Cultures of G ymnema sylvestre R. Br.
to Produce Gymnemic Acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 27
J. Rajashekar, Vadlapudi Kumar, V. Veerashree, D.V. Poornima,
Torankumar Sannabommaji, Hari Gajula, and B. Giridhara
30 In Vitro Mass Propagation of C ymbopogon citratus
Stapf., a Medicinal Gramineae. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 45
Elisa Q uiala , R aúl B arbón , Alina Capote , N aivy P érez ,
and Elio J iménez
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 59