CSIR-CIMAP

Central Institute of Medicinal and Aromatic Plants

Council of Scientific & Industrial Research (CSIR-CIMAP)
Kukrail Picnic Spot Road; PO CIMAP, Lucknow-226015 (India)

Dr. (Mrs) ARCHANA MATHUR 04th August, 2019

Ex-Chief Scientist &
Presently CSIR Emeritus Scientist
Department of Plant Biotechnology

CERTIFICATE
This is certified that Ms. Pooja Chauhan D/o Shri. Chandra Bhushan Chauhan, carried
out her dissertation cum project work under my supervision for partial fulfillment of her
M.Sc (Biotechnology) from Dr. Ram Manohar Lohia Avadh University, Ayodhya (U.P.).
The ensuing dissertation report entitled “Biotechnological Interventions in Some Medicinal
and Aromatic Plants” embodies the details pertaining to the lay out and salient findings of
the experiments with which she was associated during her training period at CSIR-CIMAP
from 4th February to 4th August, 2019. During this period Pooja was provided with in depth
exposure to all basic tools of Plant Biotechnology like plant morphogenesis, in vitro tissue
banking, genomic DNA isolation, Synthetic seed Technology etc.
During her project work Pooja has shown keen interest in all the task assigned to her. She
is very hardworking and blessed with a strong desire to learn more. She has shown a
systematic approach towards her laboratory experimentations. I have always found her
very sincere towards her assigned work. I am confident that Pooja will be able to
independently plan, execute and interpret her experiments in her future research career.
I wish her all the best in life.

(Archana Mathur)

Note: No part of this report can be reproduced or otherwise used without prior permission of Director, CSIR-
CIMAP, Lucknow

Phone: ++91-522-2718673; ++91-522-2359623; ++91-522-2718631; FAX: ++91-522-2342666; Email:

a.mathur@cimap.res.in; archnacimap@cimap.res.in; website: www.cimap.res.in
 CERTIFICATE

This is to certify that the thesis, entitled “BIOTECHNOLOGICAL INTERVENTIONS IN

SOME MEDICINAL AND AROMATIC PLANTS” submitted to Dr. Ram Manohar
Lohia Awadh University, Ayodhya, in partial fulfillment of the requirement for the Degree of
M.Sc. Biotechnology, is a record of original research work done by Ms. Pooja Chauhan during
the period of 04/02/2019 to 04/08/2019 of her study in the department of plant biotechnology,
at CIMAP Institute under my supervision and guidance and the project report has not formed
the award of any Degree/Diploma/Associateship/Fellowship or other similar title to any
candidate of any university.

Signature of H.O.D Signature of Supervisor

BIOTECHNOLOGICAL INTERVENTIONS IN SOME
MEDICINAL AND AROMATIC PLANTS

A Dissertation Submitted To
Dr RML Avadh University, Ayodhya
For partial fulfillment
For the M. Sc. Biotechnology Degree

CSIR-CIMAP
Division of Plant Biotechnology
CSIR-Central Institute of Medicinal and Aromatic Plants,
Lucknow - 226015, India

Under The Guidance Of

Dr. ARCHANA MATHUR
CSIR- EMERITUS SCIENTIST
CSIR – CIMAP, LUCKNOW

SUBMITTED BY
Ms Pooja Chauhan
Department of biotechnology
Ram Manohar Lohia Avadh University, Ayodhya , (U.P.)
 DECLARATION

I hereby declare that the project entitled “Biotechnology Interventions in some Medicinal
and Aromatic Plants” is being submitted for the partial fulfillment of award of degree of
Master of Science in biotechnology, Dr. Ram Manohar Lohia Avadh University, Ayodhya,
(U.P.).

It is an authentic record of the work carried out under the kind guidance of Dr.
Archana Mathur, Ex-chief Scientist in CSIR- CIMAP, Lucknow (U.P.)

I have not submitted matter embodied in this dissertation for any other degree or deploma.

Date:
Pooja Chauhan
Place: Lucknow M.Sc. Biotechnology(4th semester)
Dr. R.M.L.A. University, Ayodhya

(Utter Pradesh)
 ACKNOWLEDGEMENT

The successful completion of any task would be incomplete without thanking the people who
made it possible. With all my submissiveness I thank God for his constant presence throughout
my work. I take pleasure in expressing a few words of gratitude and respect to all those who
helped me in completing my dissertation work.

I express my deepest sense of gratitude to my guide Dr. (Mrs.) Archana Mathur, Ex-chief
Scientist for her meticulous guidance, support, expert supervision, keen interest, constant
involvement and encouragement throughout the period of my dissertation work.

I am very much thankful to Dr. Abdul Samad, Director, CIMAP, Lucknow, for giving me
an opportunity to carry out my project work and also for allowing me to use relevant facilities
in this esteemed Institution throughout the course of my project.

I sincerely express my heartfelt thanks to Dr. A.K Mathur, Ex-chief Scientist for his constant
support throughout the work.

I would like to express my thanks to all my seniors Dr. Archana, Mamta, and Dr. Neha for
their enthusiastic expert advice and input into the project. The technical help rendered by Mr.
Chandra pal Kumar (lab attendant)

I would also like to thank Dr. Raksha pal Singh and Dr. Rajendra patel (Biology Central
Facility) for providing the instrumental facilities. I am also thankful to Head of Library ‘Gyan
Surbhi” in CIMAP, for providing useful and important literature. I immensely thank all faculty
members of my university for giving me an opportunity to carry out my project work in this
esteemed Institution and also for their cordial support

Thanks to all my friends for their support and co-operation and for the fun we shared while at
the work.

My acknowledgement will be incomplete if I do not mention my parents, with whose

blessings I was able to achieve my goal successfully. There are no words to express my
feelings towards them. I silently acknowledge my debt to them.

POOJA CHAUHAN
 CONTENTS Page No.

Acknowledgements

1. Abbreviations 1

2. Introduction 2-9

3. Review of literature 10-17

4. Objectives 18

5. Materials and methods 19-25

6. Observation and results 26-35

7. Plant tissue culture techniques learnt 36-46

8. References 47

9. Appendices 48-51
INDEX OF TABLE

S. No. Title Of Table

1. Some important elements for plant nutrition and their
physiological function.
2. Composition of MS Media used during establishment of in
vitro cultures.
3. Plant growth regulators used with the nutrient medium

4. Effect of growth regulators on seed germination of Uraria

picta
5. Shoot Induction percentage from nodal explants of D.
gangeticum
6. The genomic DNA yield of M-mother plant and 11 randomly
selected plantlets using nano-drop spectrophotometer.
7. List of 2 ISSR primers used in the genetic fidelity testing of
Asparagus adscendens
8. Effects of different substrates on Solanum surattanse
synthetic seed germination .Observations were taken after
20 days
9. List of plants in which different explants have been
encapsulated.
10. List of plants showing potential for synthetic seed
technology
11. PCR components (Genei)

12. Composition of reaction mixture

13. PCR conditions

INDEX OF FIGURES

S.No Title of figures

1. Maintainence and multiplication of ongoing in vitro-growing (A) Stevia

rebaundiana, (B) Paris polyphylla, Valeriana wallichii, (C) Centella asiatica,and (D)
Ocimum sanctum, multiple shoot cultures in the in vitro tissue bank
2. Seed germination and seedling development in Uraria picta under in-vitro
conditions. A) Germination of inoculated seeds B) Shoot initiation; C) shoot
elongation; D) shoot multiplication
3. Shoot bud induction and proliferation from nodal explants of D. gangeticum. A)
Nodal explants, B) Shoot bud initiation, C) & D) Shoot proliferation
4. Multiplication and acclimatization of Asparagus adscendens A) Established in-vitro
shoots; B) Proliferated shoots after subculture; C)Transferred plants in the soil
after rooting in culture room; D)Acclimatized plant in the glasshouse
5. Multiplication and acclimatization of S. surattense .A) subcultured in-vitro shoots;
B) Plantlets with rooting; C)Transferred plants in the soil after rooting in culture
room; D)Acclimatized plant in the glasshouse
6. 0.8% Agarose gel electrophoresis showing quality and quantity of DNA extracted
by modified CTAB method of M mother and C1-C11 clones of in-vitro cultured
Asparagus adscendens plant
7. 1.2 % agarose gel electrophoresis of ISSR-PCR showing Genetic fidelity testing of 1
mother and 11 clone plantlets of Asparagus adscendens
8. Synthetic seed production from nodal explants of S. surattense.
A)synthetic seeds ; B)&C) Shoot emergence & elongation; D) Rooting from
elongated shoot (arrow)
9. Flow chart of Artificial seed technology