(Solanum nigrum L.) GENOTYPES NANDHINI, TMSc.

200 Pages·2013·4 MB·English

GENETIC DIVERSITY STUDIES IN MAKOI (Solanum nigrum L.)
GENOTYPES

Thesis submitted in part fulfillment of the requirement for the degree of
Doctor of Philosophy in Horticulture (Spices, Plantation, Medicinal and Aromatic
crops) to Tamil Nadu Agricultural University, Coimbatore – 641 003

By

NANDHINI, T. M.Sc., (Hort.)
ID. No. 10- 633- 005

DEPARTMENT OF MEDICINAL AND AROMATIC CROPS
HORTICULTURAL COLLEGE AND RESEARCH INSTITUTE
TAMIL NADU AGRICULTURAL UNIVERSITY
COIMBATORE - 641 003

2013
Acknowledgement
I bow my head with all reverence to the Lord, who kindly imbued the energy and enthusiasm
through ramifying paths of thick and thin of my efforts.
With deep respect and esteem regents, I owe my inexpressible personal indebtedness to my
chairman Dr.P.Paramaguru, Professor and Head, Coconut Research Station, Veppankulam for his
exemplary guidance, constant encouragement and support throughout the course of the study.
I am highly obliged to the members of the advisory Committee Dr. R.M.Viyakumar, Professor
and Head, Department of Medicinal and Aromatic crops, HC & RI, Coimbatore. Dr. S.Robin, Professor
and Head, Department of Rice, CPBG, Coimbatore, and Dr. R.Kasthuri, Professor (Biochemistry),
Coimbatore
for their valuable and constant encouragement offered during the course of study.
I offer my sincere thanks to Dr.P.Jansirani, Professor and Head, Department of Spices and
Plantation crops, HC & RI, TNAU, Coimbatore for her valuable advices during my course work and
Research.
I deem it is a special privilege to put forth my sincere thank to Dr.Mariappan, Dean
Horticulture and Dean Former Dean Dr. N. Kumar, Horticultural College and Research Institute, Tamil
Nadu Agricultural University, Coimbatore.
I enroll my deep sense of gratitude to Dr. Natarajan, Professor of Horticulture, for his
instinctive encouragement, help and guidance.
I am happy to express my immense thanks to Dr. Durga Devi, Dr. L.Nalina, Dr. Chitra,
Dr. M. Ganga, Dr. Suganthy, Dr. Mohanalakshmi, Dr. Meena, Dr. P. Jegadheswari. and Dr. Saravanan
for their encouragement, care, timely help and proper guidance during the course of my study.
I am very thankful to Dr.Thondaiman, Mr.Ramchander, Dr.Ashok, Dr.Krishnamoorthy,
Dr.Mahalingam, Dr.Anandhi, Dr.Karthikeyan, Dr.Sugapriya and Mr.Swaminathan for their timely
help rendered for accomplishment of my thesis work.
I wish to express the sense of love, heartfelt thanks and affection to my dear friends Mekala,
Priya, Ramya, Narmi, Goms akka, Janani, Ranchana, Vaidehi akka, Selva, Allirani akka, Dhivya,
Tamil, Deepa, Sajitha akka, Anushiya, Visalakshi akka, Karthi anna, MuthuKumar, Jagadesh,
Rajamanigam sir, Ravanachander, Vasu sir, Suresh sir, Ganesh and Dinesh sir for their unreserved help,
constant encouragement and extraordinary support have helped me to succeed in all my endeavours.
I express my sincere thanks to my beloved Juniors Ammaan, Sumitha, Arthi, Sangeetha,
Imbarasi, Princy, Murali, Mani, Anok, Govind and Suresh for their constant motivation and affection
during these years of my studies.
My special thanks to farm workers of Medicinal plant Unit for their assiduous help rendered in
carrying out the research work.
Mere words cannot express the gratitude owe to my lovable husband Mr.S.Mohandass, who
stood by me at tough times and whose persuasion, patience and encouragement was the inspiration of
this research work.
Words are ineffable to express my indebtedness to my loving father (Mr.M.Thirumoorthy and
Mr.R. Sunderajan), mother (Mrs.T.Ponnamaal and S.Subulakshmi), brothers Dinesh and Jai for their
insisting inspiration, constant patting and everlasting love bestowed on me at every stage of my
endeavour.

(T.Nandhini)
ABSTRACT

GENETIC DIVERSITY STUDIES IN MAKOI (Solanum nigrum L.) GENOTYPES
By
T. Nandhini, M.Sc., (Hort)
Degree : Doctor of Philosophy in Horticulture
Chairman : Dr. P. Paramaguru, Ph.D.,
Professor and Head,
Coconut Research Station,
Veppankulam - 614 906.
2013
An investigation was carried out with 45 makoi (Solanum nigrum L.) genotypes at
the Horticultural College and Research Institute, Coimbatore during 2011 - 2013 to study
the extent of variability, genetic divergence through morphological characters and
molecular markers and association among yield and its component characters.
Morphological characterization of 45 genotypes based on 14 IPGRI descriptors was
done and the cluster analysis grouped the genotypes into four major clusters. Ploidy analysis
and DNA barcoding analysis revealed that TN Sn 10 was S.nigrum L. (hexaploid) and TN
Sn 30, TN Sn 32, TN Sn 44 and TN Sn 47 were S.villosum Mill. (tetraploids) while
genotypes TN Sn 08, TN Sn 12, TN Sn 23, TN Sn 38, TN Sn 52 and TN Sn 53 were
S.americanum Mill.(diploids). The genotypes TN Sn 10 and TN Sn 30 recorded highest
herbage and alkaloid yield and can be used in breeding programme to get high alkaloid
yielding varieties. Though TNSn 19 and TN Sn 22 recorded lesser yield than TN Sn 10 and
TN Sn 30, they showed high nutritive value and palatable characters and hence they can be
used for evolving varieties that can be used as green leafy vegetable.
In Mahalanobis D2 analysis, the genotypes were grouped into eight clusters. Among
the eight clusters, the cluster V was the largest with 15 genotypes followed by cluster I with
11 genotypes. The cluster IV and III has maximum inter cluster distance which showed
maximum divergence between the two clusters. Fresh and dry herbage yield per plant
contributed maximum towards divergence.
The genetic relationship and variation of 45 accessions were examined by SSR
analysis using 10 primer pairs. A total of 22 fragments from ten SSR markers were formed
of which seven primers were polymorphic. PIC was highest for the SSR primers STWIN 12
G (0.698). The Jaccard‟s similarity coefficient ranged from 0.36 to 1.00. Based on the
banding pattern, the cluster analysis was done using UPGMA and the dendrogram was
constructed which resulted in eight clusters.
In all the three species, the maximum phenotypic and genotypic coefficient of
variations was observed for protein content, dry herbage yield per plant, fresh herbage yield
per plant. Heritability estimates were higher for quality characters than the biometric
characters. Ascorbic acid content, protein content, iron content and days to flower bud
initiation recorded high estimates of heritability coupled with high genetic advance.
Correlation studies indicated that total alkaloid content expressed highly significant
and positive correlation withdry herbage yield, crude fibre, number of leaves and number of
primary branches. It also had significant negative association with protein content, number
of berries per plantand days to fifty per cent flowering.
In a nutshell, wide diversity existed among the genotypes in morphological and
biochemical characters and are beneficial for developing superior varieties through selection
or hybridisation programme. Correlation revealed earlier flowering with lesser berry yield
can be considered for higher herbage yield.
CONTENTS

CHAPTER TITLE PAGE NO.
1 INTRODUCTION
2 REVIEW OF LITERATURE
3 MATERIALS AND METHODS
4 EXPERIMENTAL RESULTS
5 DISCUSSION
6 SUMMARY
REFERENCES
PLATES
ANNEXURE
PUBLICATIONS
LIST OF TABLES
Table No. Title Page No.
1. List of forty five Genotypes
2. List of morphological characters observed
List of morphologically distinct genotype used for ploidy and barcode
3.
analysis
4. List of SSR primer pairs used in the study
5. Morphological characterization of Genotypes
Variations in stomatal, pollen, flower and berry characters in distinct
6.a.
Genotypes
6.b. List of genotypes subjected to DNA barcode analysis
Analysis of variance for different characters of Solanum nigrum L.
7.
genotypes
Analysis of variance for different characters of Solanum villosum Mill.
8.
genotypes
Analysis of variance for different characters of Solanum nigrum Mill.
9.
genotypes
10. Mean performance of genotypes for plant height (cm)
11. Mean performance of genotypes for plant spread (N-S) (cm)
12. Mean performance of genotypes for plant spread (E-W) (cm)
13. Mean performance of genotypes for number of primary branches
14. Mean performance of genotypes for stem girth (cm)
15. Mean performance of genotypes for number of leaves per plant
16. Mean performance of genotypes for leaf length (cm)
Table No. Title Page No.
10. Mean performance of genotypes for leaf breadth (cm)
11. Mean performance of genotypes on days to flower bud initiation
12. Mean performance of genotypes for days to fifty per cent flowering
13. Mean performance of genotypes for number of berries per plant
14. Mean performance of genotypes for fresh herbage yield per plant (g)
22. Mean performance of genotypes for dry herbage yield per plant (g)
23. Mean performance of genotypes for fresh herbage yield per plot (Kg)
24. Mean performance of genotypes for dry herbage yield per plot (Kg)
Mean performance of genotypes for Fresh herbage yield per hectare (t)
25.
(estimated)
Mean performance of genotypes for dry herbage yield per hactare (t)
26.
(estimated)
27. Mean performance of genotypes for total leaf area (cm2) per plant
28. Mean performance of genotypes for total chlorophyll (mg g-1)
29. Mean performance of genotypes for total soluble proteins (mg g-1)
30. Mean performance of genotypes for total phenols (mg g-1)
31. Mean performance of genotypes for total alkaloid content (%)
32. Mean performance of genotypes for protein content (mg 100g-1)
33. Mean performance of genotypes for crude fibre (%)
34. Mean performance of genotypes for iron content (mg 100g-1)
Table No. Title Page No.
35. Mean performance of genotypes for ascorbic acid (mg g-1)
36. Organoleptic evaluation of high yielding genotypes
37. Distribution of genotypes into different clusters based on D2 statistics
38. Inter and intra cluster D2 and D (with in parentheses) values
39. Relative contribution of eleven agronomic traits towards divergence
40. Cluster Mean values for eleven characters
SSR primers showing total and polymorphic amplicons generated pattern for
41.
45 genotypes
42. Jaccard‟s similarity coefficient among genotypes from SSR data
43. Clustering of 45 genotypes based on SSR markers
44. Estimates of variability and genetic parameters of Solanum nigrum L. genotypes
Estimates of variability and genetic parameters of Solanum americanum Mill.
45.
genotypes
Estimates of variability and genetic parameters of Solanum villosum Mill.
46.
genotypes
Estimates of simple correlation coefficients among 12 biometric traits in
47.
Solanum nigrum L. genotypes
Estimates of simple correlation coefficients among 12 biometric traits in
48.
Solanum americanum Mill. genotypes
Estimates of simple correlation coefficients among 12 biometric traits in
49.
Solanum villosum Mill. genotypes
LIST OF FIGURES

Figure
Title Page No.
No.
1.
Diversity in growth habit and branching
2.
Diversity in stem and leaf pubescence
3.
Diversity in stem color
4.
Diversity in berry color
5. Dendrogram based on morphological characters of forty five genotypes
Phylogenetic analysis of the ITS sequences of the Solanum species using
6.
MEGA5 software
7. Fresh herbage yield per hectare of high yielding genotypes
8. Biochemical contents in high yielding genotypes
9. Organoleptic evaluation of high yielding genotypes
10. Relative contribution of characters towards divergence in makoi genotypes
11. Dendrogram based on SSR markers of forty five genotypes