morphological and molecular characterization in custard apple

149 Pages·2017·4.89 MB·English

MORPHOLOGICAL AND MOLECULAR
CHARACTERIZATION IN CUSTARD APPLE

BY
SAITWAL YOGESH SUDHAKAR
(Reg. No. 10/74)

DEPARTMENT OF HORTICULTURE
POST GRATUATE INSTITTUTE,
MAHATMA PHULE KRISHI VIDYAPEETH,
RAHURI - 413 722 (M.S.) INDIA.

2013
i

MORPHOLOGICAL AND MOLECULAR
CHARACTERIZATION IN CUSTARD APPLE

A thesis submitted to the
MAHATMA PHULE KRISHI VIDYAPEETH,
RAHURI – 413 722, AHMEDNAGAR,
MAHARASHTRA, INDIA

in the partial fulfilment of the requirements for the degree

of

DOCTOR OF PHILOSOPHY (AGRICULTURE)

in

HORTICULTURE (FRUIT SCIENCE)

by

SAITWAL YOGESH SUDHAKAR
(Reg. No. 10/74)

DEPARTMENT OF HORTICULTURE,
POST GRADUATE INSTITUTE,
MAHATMA PHULE KRISHI VIDYAPEETH,
RAHURI – 413 722, DIST. - AHMEDNAGAR,
MAHARASHTRA, INDIA
2013
ii

MORPHOLOGICAL AND MOLECULAR
CHARACTERIZATION IN CUSTARD APPLE

by
SAITWAL YOGESH SUDHAKAR
(Reg. No. 10/74)

A thesis submitted to the
MAHATMA PHULE KRISHI VIDYAPEETH,
RAHURI – 413 722, AHMEDNAGAR,
MAHARASHTRA, INDIA

in the partial fulfilment of the requirements for the degree
of

DOCTOR OF PHILOSOPHY (AGRICULTURE)

in

HORTICULTURE (FRUIT SCIENCE)

approved by

Dr. A. M. Musmade
(Chairman and Research Guide)

Dr. S. S. Mehetre Dr. V. S. Supe
(Committee member) (Committee member)

Dr. A. A. Kale Dr. V. R. Joshi
(Committee member) (Committee member)

DEPARTMENT OF HORTICULTURE,
POST GRADUATE INSTITUTE,
MAHATMA PHULE KRISHI VIDYAPEETH,
RAHURI – 413 722, DIST. - AHMEDNAGAR,
MAHARASHTRA, INDIA
2013
iii

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iv

CANDIDATE’S DECLARATION

I hereby declare that the dissertation or a part
there of has not been previously
submitted by me to any other
university or institute
for degree or
diploma

Place : M.P.K.V., Rahuri
Date: / /2013 (Y.S.Saitwal)
v

Dr. A. M. Musmade
Professor,
Department of Horticulture,
Mahatma Phule Krishi Vidyapeeth
Rahuri-413 722, Dist. Ahmednagar,
Maharashtra State, INDIA.

CERTIFICATE

This is to certify that the thesis entitled, “MORPHOLOGICAL AND
MOLECULAR CHARACTERIZATION IN CUSTARD APPLE” submitted to the
Faculty of Agriculture, Mahatma Phule Krishi Vidyapeeth, Rahuri Dist. Ahmednagar (M.S.)
in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY
(AGRICULTURE) in HORTICULTURE (FRUIT SCIENCE), embodies the results of a
piece of bona fide research work carried out by Saitwal Yogesh Sudhakar, under my
guidance and supervision and that no part of this thesis has been submitted for any other
degree or diploma.
The assistance and help received during the course of this investigation have been
duly acknowledged.

Place: MPKV, Rahuri (A. M. Musmade)
Date : / /2013 Research Guide
vi

Dr. B.R. Ulmek
Associate Dean,
Post Graduate Institute,
Mahatma Phule Krishi Vidyapeeth,
Rahuri-413 722, Dist. Ahmednagar,
Maharashtra State, INDIA.

CERTIFICATE

This is to certify that the thesis entitled, “MORPHOLOGICAL AND
MOLECULAR CHARACTERIZATION IN CUSTARD APPLE” submitted to the
Faculty of Agriculture, Mahatma Phule Krishi Vidyapeeth, Rahuri Dist. Ahmednagar (M.S.)
in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY
(AGRICULTURE) in HORTICULTURE (FRUIT SCIENCE), embodies the results of a
piece of bona fide research work carried out by Saitwal Yogesh Sudhakar, under the
guidance and supervision of Dr. A. M. Musmade, Professor, Department of Horticulture,
Mahatma Phule Krishi Vidyapeeth, Rahuri and that no part of this thesis has been submitted
for any other degree.

Place : M.P.K.V., Rahuri
Date : / /2013 (B.R. Ulmek)
vii

A C K N O W L E D G E M E N T

Formal words cannot carry the fragrance of emotions with them; still they
are only available means of expressing emotions. My acknowledgements are much
more than what i am expressing here.
Nothing can severe the spiritual relation between guru and his disciple. It is
something eternal. Success depends on the sincerity and effort on the part of
disciple.
The culmination of research work is an indispensable corner stone in the life
of any student with the research guide being the driving force behind. I feel it is
indeed my great privilege and immense pleasure in availing this golden opportunity
to express my wholehearted sense of gratitude and ineffable indebtedness to my
honourable Research Guide Dr. A.M. Musmade, Professor, Department of
Horticultur, M.P.K.V., Rahuri. Being Chairman of my Advisory Committee, I am
incapable to express the debt of thanks to him for his learned counsel, intellectual
inspiration, keen interest, constructive criticism, valid suggestions, arduous and
meticulous guidance, constant generous encouragement and willing help during the
entire course of this study and for the painstaking efforts for finalizing the
manuscript that stepped up to see the present form. I must confess that, really,
very fortunate I am, for I got an opportunity to work under his versatile, scholastic
and intellectual guidance during my Ph.D. (Hort.) programme.
I am highly grateful to the Members of my Advisory Committee, Dr.S.S.
Mehetre, Ex-DOR, M.P.K.V., Rahuri, Dr. V.S. Supe, Associate Professor,
Department of Horticulture, Dr. A.A. Kale, Associate Professor of Biochemistry,
State Level Biotechnology Centre and Dr. V.R. Joshi, Assistant Professor,
Department of Horticulture, M.P.K.V., Rahuri for their valuable comments and
constructive suggestions in enlightening my views at various stages of this
investigation and Scientist of Maharashtra State visited to my research trial and
given valuable guidance and inspiration for conducting this unique research work.
I would like to acknowledge with high respect the gratefulness of Dr. S.A.
Ranpise, Head, Department of Horticulture, Dr. A.S. Jadhav, Professor and
Incharge, State Level Biotechnology Centre, M.P.K.V., Rahuri for their constant
viii

inspiration, talented and well versed advice by providing necessary facilities to
carry out this research work.
I accolade my highest respect with reverence to Dr. T.A. More Sir,
honourable Vice-Chancellor, M.P.K.V., Rahuri, Dr. R.S. Patil, DOR, M.P.K.V., Rahuri,
Dr. B. R. Ulmeck, Dean and Associate Dean (PG), M.P.K.V., Rahuri, without whom I
would not be a Scholar.
Everything in my life is nothing of I ignore the all time encouragement
evergreen affection and latent love of my Pappa Shri. Sudhakar Saitwal and Aai
Sau. Mangala Saitwal which elevate my morals during whole educational career.
The words with me are insufficient to express the feeling to my heart to
acknowledge them for their difficult job of educating me and keeping comfort
without which this work would not have seen the light of the day at all.
I can not eschew to express special thanks to my beloved brother Pankaj,
Vahini Sau. Arti, Sister Sau. Punam Tai and Jijaji Shri. Pravin.
One needs sincere friend’s at all major junctures in life to bear strains and
fatigue carefully. I have been more than lucky in this respect and my cardiac sense
of gratitude towards my intimate friends Chandrakant, Prashant2, Suhas, Ram,
Vishal, Mahesh, Dhanaji, Mohan, Aditya, Mahendra, Pramod and Prasad for their
encouragement and kindly co-operation.
No words of gratitude can equate the tremendous encouragement with love
and affection that have been bestowed on me by all the evangelists and apostles
who constantly counseled me through their soul prayers and blessings with lot of
ethical convictions that exalted my aspirations up to this stage and I must pray for
persisting their obligations with my soul till the eternity.
I ever rest thanks to all of them who have directly and indirectly helped me
for this fruitful outcome. Nevertheless, I beg pardon politely to those who have
helped me but their names are not appearing in this acknowledgement.
Place : Rahuri
Date : (Y.S.Saitwal)
ix

CONTENTS

CANDIDATES DECLARATION iv
CERTIFICATES
1. Research Guide v
2. Associate Dean (PGI) vi
ACKNOWLEDGEMENTS vii
LIST OF TABLES xii
LIST OF FIGURES xiii
LIST OF PLATES xiv
LIST OF ABBREVATIONS xv
ABSTRACT xvii
1. INTRODUCTION 1-4
2. REVIEW OF LITERATURE 5-19
2.1 Morphological and biochemical characterization 5
2.1.1 Morphological characterization 5
2.1.2 Biochemical characterization 8
2.2 Molecular characterization 10
2.2.1 DNA extraction 10
2.2.2 ISSR analysis 11
2.2.3 SSR analysis 16
3. MATERIAL AND METHODS 20-38
3.1 Morphological and biochemical characterization 20
3.1.1 Experimental material 20
3.1.2 Experimental site 23
3.1.3 Experimental methods 23
3.1.4 Morphological and biochemical observations 24
3.2 Molecular characterization 31