In-vitro Propagation of Rosa Centifolia, a Plant of

Ornamental and Medicinal Value

Dissertation thesis submitted to

H.N.B Gahrwal (Central) University, Dehradun

In partial fulfilment of the requirement for the degree of

Master of Science in Biotechnology
by
DAWOOD YOUSUF 4TH SEMESTER
Under the supervision of
VIKAS MISHRA
(Asst. Prof. & Head Dept. of Biotechnology)
At

Sai Biological Research Centre

A Unit of Sai Group of Institution, Dehradun

2019
 Declaration by the Candidate

I, DAWOOD YOUSUF, hereby declare that the thesis entitled “In Vitro propagation of Rosa
Centifolia a plant of ornamental and medicinal value”, submitted to the Department of
Biotechnology, HNB Garhwal University, Srinagar in partial fulfillment of the requirements for
the Degree of Master of Technology in Biotechnology, is an independent research work carried
out by the undersigned during a period of six months, under the supervision and guidance of
Mr. VIKAS MISHRA, (Asst. Prof. & Head Department of Biotechnology) and to the best of
my knowledge and belief this thesis has not formed on the basis of award of any Degree/
Diploma/Associate ship/Fellowship or other similar title to any candidate of any University.

Place: Signatureof candidate

Date:
 ACKNOWLEDGEMENT

Successful completion of project involves interest and efforts of many people and so this
Becomes obligatory on my part to record thanks to them . by the grace of the al-mighty and help
of a lot many people , I was able to accomplish the task ahead of me. Thus , it becomes
immensely important for me to thank all the people who helped me in completing this task.
Therefore, on this was first of all I would like to thank from the core of my heart to Mr. Vikas
Mishra (Asst. Prof. &Head Dept. of Biotechnology SIPAS,Dehradun) under the supervision of
whom my dissertation work has been accomplish. I heartily wish to pay my deep regards to Dr.
Sandhya Dogra (Principal) Sai institute of paramedical and allied sciences, Dehradun for her
guidance , help and encouragement .
I wish to convey my sincere thanks to Mr. Harish Arora and Mrs. Rani Arora (Honorable
chairperson) SIPAS, DEHRADUN for providing the all facilities for accomplishment of this
training.
I would like to extend my grateful thanks to my friends Vandana, Nassirudin, Priyanka,Anupriya
and Anshdeep) . At last but not least I want to thanks my Parents of them I am able to carry my
studies in a graceful manner.

DAWOOD YOUSUF
 Table of Contents
Chapter Contents Page No.
Abbreviations
1. Introduction
1.1 Plant Tissue Culture
1.1.1 Uses of tissue culture
1.1.2 Types of tissue culture
1.2 ROSE(Rosa Centifolia)
2. Review Literature
3. Materials and Methods
3.1 Laboratory requirements
3.2 Sterlization
3.3 Media constituents
3.4 Composition of Ms media
4 Stages of Micropropagation
4.1 stage 0
4.2 stage 1
4.3 stage 2
4.4 stage 3
4.5 stage 4
5 Methodology of work done
5.1 plant material
5.2 sterlization of explants
5.3 culture media
5.4 culture conditions
6 Results and discussion
6.1 Rosa Centifolia showing shoot initiation
on ms media after 1 week
6.2 Multiple shoot formation of Rosa
Centifolia in Ms media

7 Summary and conclusion

7.1 conclusion
8 Literature cited
 List of Tables
Table No. Content of table Page No.
1 3.4 Shows composition of murashige and skoog
medium.
2 6.1 Shows initiation of shoot from stem of rose.

3 6.2 Pathway subculturing of rose shoots.

List of Figures
S. No. Content of figure Page No.
1 6.1 Fig Rosa Centifolia showing shoot initiation on
MS media after 1 weeek.
2 6.2 Fig Multiple shoot formation of Rosa Centifolia
on MS media 0.5 mg/1 2,4 d,1 mg/1 BAP after 2 and
3 weeks.
 Abbreviations
IAA INDOLE 3 ACETIC ACID
IBA INDOLE 3 BUTYRIC ACID
NAA NAPTHALENE ACETIC ACID
B5 GAMBORG’S MEDIUM
BAP BENZYLE AMINO PURINE
2IP ISO PENTENYLE ADENINE
CaCl₂ CALCIUM CHLORIDE
2,4D 2,4 DI CHLOROPHENOXY ACETIC ACID
Fe EDTA FERROUS ETHYLENE DI AMINO TETRA
ACETIC ACID
FeSO₄ FERROUS SULPHATE
GA₃ GIBBRELC ACID
HgCl₂ MERCURIC CHLORIDE
H₃BO₃ BORIC ACID
KIN KINETIN (FURFURYL AMINO PURINE)
KNO₃ POTASSIUM NITRATE
KH₂PO₃ POTASSIUM HYDROGEN PHOSPHATE
MS MURASHIGE SKOOG’S MEDIUM
Myo-inositol MEMBER OF VIT. B-COMPLEX
NH₄NO₃ AMONIUM INTRATE
NH₄HPO₄ AMONIUM HYDROGEN PHOSPHATE
(NH₄)₂SO₄ AMONIUM SULPHATE
NaHPO₄ SODIUM HYDROGEN PHOSPHATE
TDZ THIDIAZURON