12

1. SYNOPSIS
ON
MOLECULAR DIVERSITY IN FOXTAIL MILLET
CORE GERMPLASM [Setaria italica (L.) Beauv.] FOR
YIELD AND NUTRITIONAL CHARACTERS
STUDENT
D. Grace Beulah
ID.No.18PHGPB103
ADVISOR
Dr. G. Roopa Lavanya
Associate Professor
Department of Genetics and Plant Breeding
Naini Agriculture Institute
Sam Higginbottom University of Agriculture, Technology & Sciences Prayagraj
(Allahabad) -211007, U. P., India

2. STUDENTS ADVISORY COMMITTEE
1
Dr. G.Roopa Lavanya
Advisor
Associate Professor
Department of Genetics and Plant
Breeding
2 Dr. C. V. Chandra Mohan Reddy
( Co-advisor)
Senior Scientist
Genetics and Plant Breeding
ANGRAU-RARS, Nandyal
Andhra Pradesh, India
3 Dr. Vaidurya Pratap Sahi
(Member)
Head, Department
Genetics and Plant Breeding
4 Dr. Eapen P. Koshy
( Member)
Assistant Professor
Department of Molecular and Cellular
Engineering, JIBB
5 Dr. Joy Dawson
( Member)
Professor and Head
Department of Agronomy.

3. Topics to be discussed
1. Introduction
2. Research gap
3. Objectives
4. Materials and methods
a) List of genotypes
b) Experimental details
c) Plan of work
d) Observations to be recorded
5. Expected outcome
6. References

4. Introduction
Foxtail millet, scientific name Setaria
italica (synonym Panicum italicum L.), is an
annual grass grown for human food. It is the
second-most widely planted species of millet, and
the most grown millet species in Asia.
Foxtail millet is an annual grass with slim, vertical,
leafy stems which can reach a height of 120–
200 cm (3.9–6.6 ft).The seed head is a dense,
hairy panicle 5–30 cm (2.0–11.8 in) long.
Foxtail millet is well recognized as a short duration
and drought tolerant crop. In India it is cultivated
over an area of 0.87 lakh ha with
total production of about 0.66 lakh tones and with
productivity of 762 kg/ha .

5. At present, in India the crop is cultivated on a
very limited area of around 0.5 million
hectares in sporadic patches in the states of
Andhra Pradesh, Karnataka, Tamil Nadu,
Maharashtra, Rajasthan, Madhya Pradesh,
Uttar Pradesh and North Eastern states.
The foxtail millet grain is (per 100g) rich in
protein (11.2g) and iron (2.8mg) as compared
to rice (7.9 g protein and 1.8 mg Fe) and rich
in fat 4.0g per 100g which is superior to rice
and wheat.

6.  The SSR markers are now the markers of choice in
most areas of molecular genetics as they are highly
polymorphic even between closely related lines,
require low amount of DNA, can be easily automated
for high throughput genotyping, can be exchanged
between laboratories and are highly transferable
between populations. The SSR markers are co-
dominant markers and good for studies of population
genetics and mapping.

7. Research gap
At present there is a need to increase the
area under foxtail millet being cultivated in
India. It is essential to improve the production
of foxtail millet with varietal differences for
identification of genotype with high yielding
and high nutrient content for the exploitation
the value addition and its nutritional benefits to
diabetes patients. Not much work has been
done in Foxtail Millet as it is under utilized
crop.

8. Objectives
1. To select genotypes for high grain nutrient
content
2. To assess the extent of diversity among Foxtail
millet accessions for quantitative and
qualitative traits
3. To study association between yield and grain
yield component traits
4. To asses genetic diversity in the foxtail millet
core collection using SSR markers

9. .
Materials and methods
a) List of genotypes
Checks
1. 3156
2. 3085
3. 3222
4. Prasad
5. Suryanandi
.

10. SiA 3842 SiA 3852 SiA 3862 SiA 3872 SiA 3882 SiA 3892 SiA 3902 SiA 3913 SiA 3923 SiA 3937
SiA 3843 SiA 3853 SiA 3863 SiA 3873 SiA 3883 SiA 3893 SiA 3903 SiA 3914 SiA 3924 SiA 3938
SiA 3844 SiA 3854 SiA 3864 SiA 3874 SiA 3884 SiA 3894 SiA 3904 SiA 3915 SiA 3925 SiA 3941
SiA 3845 SiA 3855 SiA 3865 SiA 3875 SiA 3885 SiA 3895 SiA 3905 SiA 3916 SiA 3926 SiA 3942
SiA 3846 SiA 3856 SiA 3866 SiA 3876 SiA 3886 SiA 3896 SiA 3906 SiA 3917 SiA 3928 SiA 3943
SiA 3847 SiA 3857 SiA 3867 SiA 3877 SiA 3887 SiA 3897 SiA 3907 SiA 3918 SiA 3930 SiA 3944
SiA 3848 SiA 3858 SiA 3868 SiA 3878 SiA 3888 SiA 3898 SiA 3908 SiA 3919 SiA 3932 SiA 3945
SiA 3849 SiA 3859 SiA 3869 SiA 3879 SiA 3889 SiA 3899 SiA 3909 SiA 3920 SiA 3933 SiA 3946
SiA 3850 SiA 3860 SiA 3870 SiA 3880 SiA 3890 SiA 3900 SiA 3910 SiA 3921 SiA 3934 SiA 3947
SiA 3851 SiA 3861 SiA 3871 SiA 3881 SiA 3891 SiA 3901 SiA 3912 SiA 3922 SiA 3936 SiA 3948

11. SiA 3949 SiA 3962 SiA 3972 SiA 3982 SiA 3992 SiA 4003 SiA 4015 SiA 4026 SiA 4036 SiA 4046
SiA 3951 SiA 3963 SiA 3973 SiA 3983 SiA 3993 SiA 4004 SiA 4016 SiA 4027 SiA 4037 SiA 4047
SiA 3952 SiA 3964 SiA 3974 SiA 3984 SiA 3994 SiA 4005 SiA 4017 SiA 4028 SiA 4038 SiA 4048
SiA 3953 SiA 3965 SiA 3975 SiA 3985 SiA 3995 SiA 4006 SiA 4019 SiA 4029 SiA 4039 SiA 4049
SiA 3954 SiA 3966 SiA 3976 SiA 3986 SiA 3996 SiA 4007 SiA 4020 SiA 4030 SiA 4040 SiA 4051
SiA 3955 SiA 3967 SiA 3977 SiA 3987 SiA 3997 SiA 4008 SiA 4021 SiA 4031 SiA 4041 SiA 4052
SiA 3956 SiA 3968 SiA 3978 SiA 3988 SiA 3998 SiA 4009 SiA 4022 SiA 4032 SiA 4042 SiA 4053
SiA 3958 SiA 3969 SiA 3979 SiA 3989 SiA 3999 SiA 4010 SiA 4023 SiA 4033 SiA 4043 SiA 4054
SiA 3959 SiA 3970 SiA 3980 SiA 3990 SiA 4000 SiA 4011 SiA 4024 SiA 4034 SiA 4044 SiA 4055
SiA 3960 SiA 3971 SiA 3981 SiA 3991 SiA 4002 SiA 4013 SiA 4025 SiA 4035 SiA 4045 SiA 4056

12. SiA 4057 SiA 4067 SiA 4077 SiA 4087 SiA 4098 SiA 4108 SiA 4118 SiA 4128 SiA 4138 SiA 4158
SiA 4058 SiA 4068 SiA 4078 SiA 4088 SiA 4099 SiA 4109 SiA 4119 SiA 4129 SiA 4139 SiA 4160
SiA 4059 SiA 4069 SiA 4079 SiA 4089 SiA 4100 SiA 4110 SiA 4120 SiA 4130 SiA 4140 SiA 4161
SiA 4060 SiA 4070 SiA 4080 SiA 4090 SiA 4101 SiA 4111 SiA 4121 SiA 4131 SiA 4141 SiA 4162
SiA 4061 SiA 4071 SiA 4081 SiA 4091 SiA 4102 SiA 4112 SiA 4122 SiA 4132 SiA 4142 SiA 4163
SiA 4062 SiA 4072 SiA 4082 SiA 4092 SiA 4103 SiA 4113 SiA 4123 SiA 4133 SiA 4143 SiA 4164
SiA 4063 SiA 4073 SiA 4083 SiA 4094 SiA 4104 SiA 4114 SiA 4124 SiA 4134 SiA 4144 SiA 4165
SiA 4064 SiA 4074 SiA 4084 SiA 4095 SiA 4105 SiA 4115 SiA 4125 SiA 4135 SiA 4145 SiA 4166
SiA 4065 SiA 4075 SiA 4085 SiA 4096 SiA 4106 SiA 4116 SiA 4126 SiA 4136 SiA 4147 SiA 4167
SiA 4066 SiA 4076 SiA 4086 SiA 4097 SiA 4107 SiA 4117 SiA 4127 SiA 4137 SiA 4157 SiA 4168

13. 1 -20
5 checks
21-40
5 checks
41-60
5 checks
61-80
5 checks
81-100
5 checks
101 -120
5 checks
121-140
5 checks
141-160
5 checks
161-180
5 checks
181-200
5 checks
201 -220
5 checks
221-240
5 checks
241-260
5 checks
261-280
5 checks
281-300
5 checks
Block -1 Block -2 Block -3
Layout

14. Experimental details
• Location : Regional Agricultural Research
Station, ANGRAU, Nandyal
• Design : Augmented design
• Plot size : 2.25 m x 3.0 m
• Fertilizers : 20 kg/ha N as basal & 20 kg/ha as top
dressing
• Season : Kharif-2021 and Rabi 2021-2022
• Entries : 300+ 5 checks
• Spacing : 22.5 x 10 cm

15. Plan of work
1) First year: Kharif -2021
Screening and evaluation of 300 foxtail millet genotypes
available in the Regional Agricultural Research Station
(ANGRAU) Nandyal, Andhra Pradesh for the selection
of high yielding genotypes.
The details are as follow:
• Site of experiment: Central Farm, RARS, Nandyal
• Number of Genotypes: 300
• Experimental Design: Augmented design

16. 2) Second year: Rabi 2021-2022
Screening and evaluation of 300 foxtail millet
genotypes. The details are as follow;
• Site of experiment : Central Farm, RARS, Nandyal
• Number of Genotype: 300
• Experimental Design: Augmented design.
3) Third year: Kharif, 2022
• Nutritional characters analysis at RARS, Nandyal.
• Molecular diversity analysis will be done at Yogi
Vemana University, Kadapa.

17. 3.c. Observations to be recorded
In the present investigation, the data on the following pre-harvest,
post-harvest and quality components characters will be recorded.
Pre – Harvest Observations:
1.Days to 50 % flowering
2. Days to maturity
3. Plant height (cm)
4. Flag Leaf length (cm)
5. Flag Leaf width (cm)
6. Inflorescence length
7. Leaf area index
8. Panicle length (cm)
9. Peduncle length (cm)
10. No. of productive tillers
11. Fruit color seed color

18. Post –Harvest Observations:
1. Panicle weight(g)
2. Biological Yield per Plant (g):
3. Harvest index (%)
4. Seed yield per plant (g)
Nutritional characters: Micro Kjeldhal Distillation Method
(A.O.A.C, 1070)
1. Protein content (%)
2. Iron content (mg/100g)
3. Zinc content (mg/100g)
4. Calcium content (mg/100g)
Molecular Diversity Analysis
1. DNA extraction (CTAB method )
2. PCR amplification

19. 3. d. Methodology
The observations will be recorded on five randomly selected plants
in each entry of each replication for all the characters except days
to emergence, days to maturity which will be recorded on plot
basis. Observations on various traits will be recorded as described
below.
a. Pre -Harvest Observations:
1. Days to 50 per cent flowering:
The total number of days taken for flowering from sowing to
opening of first flower will be calculated in 50 per cent of the
plants.

20. 3.Plant height (cm):
The plant height will be observed by measuring total height from
ground level to the tip of the main inflorescence at dough stage and
expressed in centimeters.
4.Flag Leaf length (cm):
Flag Leaf length will be observed by measuring total flag leaf size
from base to tip of the leaf.
5.Leaf width (cm):
Leaf width will be observed by measuring at widest point of the
leaf.
6.Leaf area index:
It is defined as ratio of total projected leaf area per unit ground
area.

21. 7. Panicle length (cm): It will be measured from base to the tip of
inflorescence on the main tiller at dough stage.
8. Peduncle Length (cm):
It will be measured from top most nodes to the base of
inflorescence.
9. No. of Productive tillers:
Number of basal panicle bearing tillers are counted at harvest.
10. Fruit colour Seed colour:`
Observed by seeing the seed we can decide yellow, Red.
11.Inflorescence length:
Observed by seeing the inflorescence we can decide size of bristles
is ranges from small, medium, long.

22. b. Post-Harvest Observations:
1. Panicle Weight (g):
Total panicle weight of each plant after harvest will be taken and
mean of five plants was computed.
2. Biological Yield per Plant (g):
Dry fodder yield will be taken at the time of harvesting in five
randomly selected plants and the average showed the dry fodder
yield per plant.
3.Seed Yield per Plant (g):
Weight of total grain yield of tagged plants will be recorded and the
mean yield per plant will be calculated.
4. Harvest Index:
It is the ratio of ecological yield to the biological yield is harvesting
index.

23. C. Grain Nutrients Estimation:
The for estimation of grain nutrients content viz., protein content
and grain micronutrients (calcium, iron and zinc) content. Total protein
was estimated in selected genotypes using Micro Kjeldhal
Distillation Method (A.O.A.C, 1070).
About 3-5 gram of dry seed was randomly drown from each of the
genotype were powdered.
One gram of powder was accurately weighed using electronic
balance and subjected to digestion.
Total nitrogen in the seed obtained after titration was multiplied
with the factor 6.25 to arrive at protein content and expressed in
percentage.
Grain micronutrients (calcium, iron and zinc) were estimated using
Atomic Absorption Spectrophotometer (AAS) using mineral solution
which was made using diacid mixture.

24. D.DNA extraction and purification:

25. PCR amplification
The volume of the reaction mixture will be 25 μL which
consisted of 1 μL (20 ng) DNA sample, 0.25 μL (40μmol/L)
primers, 2.5 μL 10× PCR buffer, 0.5 μLdNTPs (10 mmol/L), 2
μL MgCl2 (20 mmol/L), 0.1μLTaq (5 U/ μL) and 17.65 μL
ddH2O.
The PTC-200 Peltier Thermal Cycler was used and
programmed for35 cycles of 94(1 min), 55(1 min), 72(2 min),
then followed by post-extension at 72for 10 min.
PCR products (7.0–7.5 μL) were used for electrophoresis on
2% agarose gels stained with ethidium bromide at 100–120 V/cm
for 100–200 min,and photographed under UV light using Gel
Doc 2000system.

27. 3.e. Statistical Analysis :The data will be subjected to statistical
and biometrical analysis as detailed below:
1.Coefficient of Variation (Burton, 1952)
1.Genotypic Coefficient of Variation (GCV)
2.Phenotypic Coefficient of Variation (PCV)
2.Heritability(Broad sense) (Burton and Devane, 1953)
3.Genetic advance (Johnson et al. 1955)
4.Genetic divergence (Mahalanobis, 1936)
5.Correlation co-efficient analysis (Al-Jibouriet al., 1958)
6.Nutrition and grain quality analysis
7.Molecular Marker Analysis
8.Analysis of molecular diversity will be done by using NTSYS-pc
ver. 2.2

28. Expected outcome
• Identification of genotypes with high grain
yielding and its component characters and
grain nutrients content.
• Identification of diverse genotype at molecular
level.

29. References
Anju, T., and S. Sarita 2010. Suitability of foxtail millet (Setaria
italica) and Barnyard millet (Echinochloa frumentacea) for development
of low glycemic index biscuits. Mal. J, utr. 16(3): 361-368.
Austin, D.F. 2006. Foxtail millets (Setaria: Poaceae) - Abandoned food
in two hemispheres, Econ Bot., 60(2): 143-158
Burton, G.W. 1952. Quantitative inheritance in grasses. Proc. 6th Int.
Grassland Cong., 1: 277-283.
Chin Yun Chiang, Heng-Sheng Lin, Song-Bin Chang, Gwo-Ing Liao,
and Chang – Sheng Kuoh, (2012).Genetic Diversity in the Foxtail
Millet (Setaria italica) Germplasm as Determined by Agronomic Traits
and Microsatellite Makers, Australian Journal of Crop Science,
6(2):342-349
D.Ratna Babu, Md Ayesha, J. Dayal Prasad Babu, V.SrinivasaRao,
(2017), Genetic Parameters for Grain yeild and Nutritional Quality Traits
in Foxtail millet .International Journal of Current Microbiology and
Applied Sciences ISSN: 2319-7706 Volume 8 Number 02.

30. Johnson, H.W., H.F. Robinson, and R.E. Comstock. 1955. Estimation
of genetic variability and environmental variability in soybean. Agron. J.
47: 314-318
Kottapalli, K.R., M.D. Burow, G. Burow, J. Burke, and N. Puppala.
2007. Molecular characterization of the U.S. peanut mini core collection
using microsatellite markers. Crop Res. 47: 1718-1727
Mahalanobis, P.C. 1928. A statistical study at Chinese head
measurement. Journal of Asiatic Society of Bengal. 25: 301-307.
Sandhu, T,S., Arora, B.S. and Yashvir Singh, (1974), Interrelationships
between yeild and yeild components in Foxtail millet. Indian Journal of
Agricultural Sciences.,44(9):563 566.
Winter, P. and G. Kahl. 1995. Molecular marker technologies for plant
improvement. World J. of Microbiology and Biotechnology 11: 438–448.
http://www.fao.org/docrep/t0818e/T0818E0a.htm
http://www.aicrpsm.res.in
https://thebiologynotes.com/polymerase-chain-reaction-pcr/