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

402 Pages·2009·7.98 MB·English

by  Kee-Yoeup Paek

M M B ™
ETHODS IN OLECULAR IOLOGY
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John M. Walker
School of Life Sciences
University of Hertfordshire
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Protocols for In Vitro Cultures
and Secondary Metabolite
Analysis of Aromatic
and Medicinal Plants
Edited by
S. Mohan Jain and Praveen K. Saxena
Department of Applied Biology, University of Helsinki, Helsinki, Finland
Department of Plant Agriculture, University of Guelph, Guelph, ON, Canada
Editor
S. Mohan Jain Praveen K. Saxena
Department of Applied Biology Department of Plant Agriculture
University of Helsinki University of Guelph
Helsinki Guelph, ON
Finland Canada
ISSN: 1064-3745 e-ISSN: 1940-6029
ISBN: 978-1-60327-286-5 e-ISBN: 978-1-60327-287-2
DOI: 10.1007/978-1-60327-287-2
Library of Congress Control Number: 2008942394
© Humana Press, a part of Springer Science+Business Media, LLC 2009
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Preface
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 problems. In fact,
more than 80% of the world’s population relies on plants for principle health care. Nearly
half of the medical prescriptions in the developed world are of plant origin.
In recent years the quest for quality of life and a common belief that plants are “natu-
ral and therefore safe” has paved the way for a wider acceptance of plant-based medi-
cines 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 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 the most challenging issues facing the
plant-based medicines. The production of medicinal metabolites in plants is affected by
plant genotype, growth environment cultivation, harvesting, processing, and distribution.
Medicinal plant preparations may also be contaminated with microbes and soil contami-
nants such as heavy metals, 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 medici-
nal plants remain the major factors in inconsistent results of clinical trials of plant-based
medicines. New regulations are currently being developed internationally to ensure con-
sistency, safety and efficacy 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 up-coming legal regulations.
In vitro cell culture and controlled environment production systems offer an excellent
opportunity for the selection and season-independent propagation of elite lines with specific,
consistent levels of medicinal metabolites with minimum contamination. Additionally, the
plant materials produced by in vitro techniques allow efficient application of the emerging
analytical methods—such as metabonomics—for complete chemical profiling which has
enormous potential for the discovery of new medicinal compounds.
Traditional breeding programs for medicinal plants are generally difficult to establish
primarily due to lack of defined chemistry of medicinal components. Little knowledge of
the genetic regulation of pathways of potential bioactive molecules further compounds
this problem. In vitro techniques such as somaclonal variation, chemical mutagenesis,
haploidy, protoplast fusion, and genetic transformation are applicable to create novel
v
vi Preface
germplasm. The impact of these techniques is 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. Furthermore, the genetic manipulation of
plant cells and organs has great advantages for producing secondary metabolites and other
bioactive natural products. Together with the tools of chemical and genomic analyses,
the in vitro culture methods hold the key to fundamental research on the biochemical
and molecular basis of the mode of action of plant-based medicines. This book provides
a detailed step-by-step description of protocols for the establishment of in vitro cultures
of important medicinal plants, their mass multiplication in a controlled environment, and
step-wise secondary metabolite analysis, genetic transformation, large-scale metabolite
production in a bioreactor, and molecular markers. The role of altered microgravity in
plant metabolite production is also described. In addition, many of these protocols will
provide a basis for much needed efforts of in vitro germplasm conservation or cryopreser-
vation of medicinal plant species at the brink of extinction as well as efforts to protect
them from the adverse impact of rapid climatic changes. This book will certainly appeal to
graduate and post graduate students, researchers, biotechnologists, industry, government
agencies, and could be used as a text book.
This book contains 31 book chapters, divided into five sections. Section I contains
16 chapters describing step-wise protocols for micropropagation and chemical analysis
of secondary compounds of different medicinal plants. Section II contains five chapters
which address the transgenic approach for producing secondary metabolites. Section III
contains two chapters which cover molecular markers/microsatellites. Section IV has six
chapters which address biotransformation, bioreactors, and metablomics. Finally, section
V contains two review chapters describing plant secondary metabolites in altered micro-
gravity and role of biotechnology in producing anti-cancer compounds. Each chapter has
been peer reviewed and revised accordingly.
We appreciate the time and effort that all reviewers have put into the development of
these chapters which aided in improving the quality of the material presented herein. We
extend our most sincere thanks to the staff of Humana Press for giving us the opportunity
to present this book to our audience.
Helsinki, Finland S. Mohan Jain
Guelph, Canada Praveen K. Saxena
Contents
Preface ........................................................................................................................... v
Contributors .................................................................................................................. xi
PART I: MICROPROPAGATION AND CHEMICAL ANALYSIS
1. Establishment of Adventitious Root Cultures of Echinacea
purpurea for the Production of Caffeic Acid Derivatives ........................................ 3
Kee-Yoeup Paek, Hosakatte Niranjana Murthy, and Eun-Joo Hahn
2. In Vitro Propagation of Rauwolfia serpentina Using Liquid Medium,
Assessment of Genetic Fidelity of Micropropagated Plants,
and Simultaneous Quantitation of Reserpine, Ajmaline, and Ajmalicine ................. 17
M.K. Goel, S. Mehrotra, A.K. Kukreja, K. Shanker, and S.P.S. Khanuja
3. Medicinal Properties, In Vitro Protocols
and Secondary Metabolite Analyses of Scots Pine .................................................. 35
Hely Häggman, Anna Maria Pirttilä, Karoliina Niemi,
Tytti Sarjala, and Riitta Julkunen-Tiitto
4. Saussurea medusa Cell Suspension Cultures
for Flavonoid Production ...................................................................................... 53
Chun-Zhao Liu and Praveen K. Saxena
5. Large-scale In Vitro Multiplication of Crataeva nurvala ....................................... 61
Shashi B. Babbar, Neetika Walia, and Amandeep Kaur
6. Bilberry In Vitro Protocols and Analyses
of Phenolic Compounds ....................................................................................... 71
Laura Jaakola, Kaisu Riihinen, Hely Häggman, and Anja Hohtola
7. In Vitro Propagation of Two Tuberous Medicinal Plants:
Holostemma ada-kodien and Ipomoea mauritiana .................................................. 81
S. Pillai Geetha, A.V. Raghu, Gerald Martin, Satheesh George,
and Indira Balachandran
8. In Vitro Production of Gymnemic Acid from Gymnema
sylvestre (Retz) R. Br. Ex Roemer and Schultes Through
Callus Culture Under Abiotic Stress Conditions .................................................... 93
Abdul Bakrudeen Ali Ahmed, Adhikarla Suryanarayana Rao,
and Mandali Venkateswara Rao
9. Establishment of Plant Regeneration and Cryopreservation
System from Zygotic Embryo-Derived Embryogenic Cell
Suspension Cultures of Ranunculus kazusensis ...................................................... 107
Suk Weon Kim and Myung Jin Oh
10. In Vitro Culture and Secondary Metabolite Isolation in Bryophytes ...................... 117
Aneta Sabovljevic, Marko Sabovljevic, and Nebojsa Jockovic
vii
viii Contents
11. Micropropagation and In Vitro Conservation of Vanilla
(Vanilla planifolia Andrews) ................................................................................. 129
Minoo Divakaran and K. Nirmal Babu
12. Protocol for In Vitro Regeneration and Marker Glycoside
Assessment in Swertia chirata Buch-Ham ............................................................. 139
Sushma Koul, K. A. Suri, P. Dutt, M. Sambyal,
A. Ahuja, and M.K. Kaul
13. Protocols for Establishment of an In Vitro Collection
of Medicinal Plants in the Genus Scutellaria ......................................................... 155
Ian B. Cole, Faisal T. Farooq, and Susan J. Murch
14. Protocols for In Vitro Culture and Phytochemical Analysis
of Phyllanthus Species (Euphorbiaceae) ................................................................. 167
Elizabete Catapan, Fábio Netto Moreno, Márcio Luís Busi da Silva,
Michel Fleith Otuki, Rivaldo Niero, Valdir Cechinel Filho,
Rosendo Augusto Yunes, and Ana Maria Viana
15. In Vitro Clonal Propagation of Asparagus racemosus,
a High Value Medicinal Plant ................................................................................ 179
Sanjay Saxena and Nishritha Bopana
16. Micropropagation of Penthorum chinense Through
Axillary Buds ........................................................................................................ 191
Jun Yang and Zheng-song Peng
PART II: TRANSGENIC APPROACHES
17. Spontaneous Plant Regeneration and Production
of Secondary Metabolites from Hairy Root Cultures
of Centaurium erythraea Rafn .............................................................................. 205
Angelina Subotic´ , Slad¯ana Jevremovic´, Dragoljub Grubišic´ ,
and Teodora Jankovic´
18. Transgenic Hypericum perforatum ........................................................................ 217
G. Franklin, Margarida M. Oliveira, and Alberto C.P. Dias
19. Agrobacterium-Mediated Transformation
of Ruta graveolens L. ............................................................................................ 235
Karine Lièvre, Thi Lê Minh Tran, Sébastien Doerper,
Alain Hehn, Paul Lacoste, Brigitte Thomasset,
Frédéric Bourgaud, and Eric Gontier
20. Gene Expression Profiling in Taxus baccata L. Seedlings and Cell Cultures ........... 249
Katarína Brunˇáková and Ján Košuth
21. Catapol Production in Chinese Foxglove (Rehmannia
glutinosa Libos.) Hairy Roots Transformed
with Agrobacterium rhizogenes ATCC15834 ......................................................... 263
Sung Jin Hwang
PART III: MOLECULAR MARKERS AND MICROSATELLITES
22. Identification of Medicinal Plants and Plant Sequences:
A Multiplexed MLPA Assay .................................................................................. 277
Roger A. Barthelson
Contents ix
23. Isolation of Microsatellites from Catharanthus roseus (L.)
G. Don Using Enriched Libraries. ......................................................................... 289
Sabhyata Bhatia and Bhumika Shokeen
PART IV: BIOTRANSFORMATION, BIOREACTORS, AND METABOLOMICS
24. Production of Cinnamyl Glycosides in Compact Callus
Aggregate Cultures of Rhodiola rosea Through
Biotransformation of Cinnamyl Alcohol ................................................................ 305
Zsuzsanna György and Anja Hohtola
25. Spearmint Plantlet Culture System as a Means
to Study Secondary Metabolism ............................................................................ 313
Brent Tisserat, Mark Berhow, and Steven F. Vaughn
26. Bioreactor Production of Secondary Metabolites
from Cell Cultures of Periwinkle and Sandalwood ................................................. 325
Jagan V. Valluri
27. Camptothecin Production by In Vitro Cultures and Plant Regeneration
in Ophiorrhiza Species .......................................................................................... 337
Takashi Asano, Hiroshi Sudo, Mami Yamazaki, and Kazuki Saito
28. Metabolomic Analysis of Ocotea odorifera Cell Cultures:
A Model Protocol for Acquiring Metabolite Data.................................................. 347
Marcelo Maraschin, Paulo Fernando Dias, Ênio Luiz Pedrotti,
Hiliana Nunes Ferreira Morais, Ana Maria Viana,
and Karl Vernon Wood
29. The Production of 9-methoxycanthin-6-one from Callus
Cultures of (Eurycoma longifolia Jack) Tongkat Ali ............................................... 359
Mahmood Maziah and Noormi Rosli
PART V: ALTERED GRAVITY AND BIOTECHNOLOGY
30. Plant Secondary Metabolism in Altered Gravity ..................................................... 373
Lindsey K. Tuominen, Lanfang H. Levine, and Mary E. Musgrave
31. The Role of Biotechnology in the Production
of the Anticancer Compound Podophyllotoxin ..................................................... 387
Hemant Lata, Cassia S. Mizuno, and Rita M. Moraes
Index ........................................................................................................................... 403
Contributors
ABDUL BAKRUDEEN ALI AHMED (cid:129) Department of Plant Science, School of Life Sciences,
Bharathidasan University, Tiruchirappalli, Tamil Nadu, India
A. AHUJA (cid:129) Biodiversity and Applied Botany Division, Indian Institute of Integrative
Medicine, Jammu, India
TAKASHI ASANO (cid:129) Graduate School of Pharmaceutical Sciences, Chiba University,
Chiba, Japan, CREST, Japan Science and Technology Agency, Saitama, Japan
SHASHI B. BABBAR (cid:129) Department of Botany, University of Delhi, Delhi, India
K. NIRMAL BABU (cid:129) Indian Institute of Spices Research, Kerala, India
INDIRA BALACHANDRAN (cid:129) Centre for Medicinal Plants Research, Arya Vaidya Sala,
Kerala, India
ROGER A. BARTHELSON (cid:129) Plant Sciences, University of Arizona, Tucson, AZ
MARK BERHOW (cid:129) New Crops and Processing Technology Research, National Center for
Agricultural Utilization Research, Agricultural Research Service-USDA, Peoria, IL
SABHYATA BHATIA (cid:129) National Institute of Plant Genome Research, Aruna Asaf Ali
Marg, New Delhi, India
NISHRITHA BOPANA (cid:129) TERI University, New Delhi, India
FRÉDÉRIC BOURGAUD (cid:129) UMR Nancy-Université-INRA Agronomie et Environnement
Nancy-Colmar, Vandoeuvre-Lès-Nancy, France
KATARÍNA BRU N˘ÁKOVÁ (cid:129) P. J. Šafárik University in Košice, Faculty of Science,
Institute of Biology and Ecology, Košice, Slovakia
ELIZABETE CATAPAN (cid:129) Departamento de Botânica, Universidade Federal de Santa
Catarina, Florianopolis, SC, Brazil
IAN B. COLE (cid:129) Chemistry, I.K. Barber School of Arts & Sciences, University of British
Columbia, Okanagan, Kelowna, BC, Canada
MÁRCIO LUÍS BUSI DA SILVA (cid:129) Departamento de Botânica, Universidade Federal de
Santa Catarina, Florianopolis, SC, Brazil
ALBERTO C.P. DIAS (cid:129) Departamento de Biologia, Universidade do Minho,
Campus de Gualtar, Braga, Portugal
PAULO FERNANDO DIAS (cid:129) Plant Morphogenesis and Biochemistry Laboratory,
Federal University of Santa Catarina, Florianopolis, SC, Brazil
MINOO DIVAKARAN (cid:129) Providence Women’s College, Kerala, India
SÉBASTIEN DOERPER (cid:129) UMR Nancy-Université-INRA Agronomie et Environnement
Nancy-Colmar, Vandoeuvre-Lès-Nancy, France
P. DUTT (cid:129) Natural Product Chemistry Division, Indian Institute of Integrative
Medicine, Jammu, India
FAISAL T. FAROOQ (cid:129) Chemistry, I.K. Barber School of Arts & Sciences, University of
British Columbia, Okanagan, Kelowna, BC, Canada
VALDIR CECHINEL FILHO (cid:129) Núcleo de Investigações Químico-Farmacêuticas,
Universidade do Vale do Itajaí, Itajaí, SC, Brazil
G. FRANKLIN (cid:129) Departamento de Biologia, Universidade do Minho, Campus de
Gualtar, Braga, Portugal
xi