Research Journal of Biotechnology

Vol. 9(2) February (2014)

Res. J. Biotech

Molecular Characterization in Grain Amanranthus

(Amaranthus spp.) using RAPD marker
Patel K. S.1*, Parihar A.2, Chaurasia Pratibha1, Patel P. J.1 and Pachchigar Karen2
1. N.V.Patel College of Pure and Applied Sciences, S.P. University, V.V. Nagar - Anand-388120, Gujarat, INDIA
2. Dept. of Molecular Biology and Biotechnology, C.P.College of Agriculture, S.D.Agricultural University, S.K.Nagar, INDIA
*drakarsh@gmail.com

acids. Grain Amaranth are widely scattered in Asia. The

crop is important and widely spread in India. For an
effective breeding programme, information concerning the
extent and nature of genetic diversity within a crop species
is essential. It is particularly useful for characterizing
individual accessions and cultivars and as a general guide
in the selection of parents for hybridization.

Abstract
RAPD assessment on genetic variation of total 58
accessions comprising 15 parental genotypes and
their 43 F1 hybrids of grain Amaranthus were
analyzed. Among the 40 tested primers, four primers
viz. OPA-06, OPA-10, OPA-14 and OPA-15 were able
to discriminate all the genotypes sufficiently. In RAPD
analysis, at the intraspecic level, the percentages of
RAPD polymorphism were found to be 71.43 (A.
hypochondriacus), 100 (A. cruentus), 100 (A.
caudatus), 75.76 [IC-1733 (A. edulis)] and 84.85
[SKGPA-144 (A. tricolor)] and hybrids showed varied
degree of polymorphism ranging from 42.86 to 71.43
per cent.

There are various techniques for studying the genetic

variability of crop germplasm including morphological
traits, total seed protein, isozymes and various types of
molecular markers. However, DNA-based markers provide
powerful and reliable tools for discriminating variations
within crop germplasm and for studying evolutionary
relationships5. Several molecular approaches have been
employed to assess genetic diversity and relationships but
isozyme or random amplified polymorphic DNA (RAPD)
data can be generated faster and with less labor than other
methods such as RFLP (restriction fragment length
polymorphism) and the use of microsatellites. With the
development of polymerase Chain Reaction (PCR) based
RAPD and SSR; most of the problem associated with
RFLP was overcome.

The population matrix showed that allele at SKGPA144 (Amaranthus tricolor) and F1 hybrids have
maximum genetic distance of 0.742 to 0.962 in all
populations while population IC-1733 (Amaranthus
edulis) has minimum genetic distance among all the
germplasm. The lowest and the maximum genetic
distance of IC-1733(Amaranthus edulis) was 0.663
and 0.720 respectively to all populations. It further
matches the pattern of allelic frequency distribution
where SKGPA-144(Amaranthus tricolor) and F1 have
remarkable high polymorphism.

Random amplified polymorphic DNA (RAPD) markers

offer quick screening of different regions of the genome for
genetic polymorphisms. The technique of RAPD gained
importance in genetic research due to its simplicity,
speed15, efficiency, relative ease to perform and nonrequirement of sequence information9. RAPD analysis has
been used to document the genetic variation in Amaranth.
The PCR based fingerprinting techniques are very efficient
both in cost and time to identify markers associated with a
trait. The RAPD markers are easier and quicker to use and
are preferred in application where the relationships between
closely related breeding lines are of interest7. RAPD
analysis enables the detection of informative genetic
markers at a large number of loci in both coding and noncoding regions of the genome16.

Keywords: RAPD, Genomic DNA, polymorphism and

genetic variation.

Introduction
Amaranth is a neglected pseudocereal crop belonging to
family Amaranthaceae (Dicotyledons, Order Caryophyllales). The genus Amaranthus includes five major
species of grain type Amaranth viz. Amaranthus
hypochondriacus L (2n=32), A. cruentus, (2n=34), A.
caudatus L. (2n=32 and occasionally 2n=34 also), A. adulis
Spegazzini (2n=32) and A. tricolor (2n=34). The grain
Amaranth is thought to be of American origin. Sauer13 has
presented several evidences which include archaeological,
historical and folklore evidences pointing to the new world
as the centre of origin and domestication. Grain Amaranth
(Amaranthus spp.) is one of the underutilized crops,
offering a lot of potentiality to be explored as future food
crop by virtue of high quality of protein rich grain and
carotene rich leaves. The grain has high protein with high
lysine content and good source of other essential amino

Material and Methods

The experiment was conducted at Plant Biotechnology
Laboratory, Department of Genetics Plant Breeding, C.P.
College of Agriculture, S.D. Agricultural University, S.K.
Nagar, during year 2012-13.
Plant material: The leaf tissues were harvested from one
to two week old plant at initial two to three leaf stage when
leaves have limiting amount of polysaccharides8. A total of
58 accessions used were comprised of 15 parental

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Research Journal of Biotechnology

Vol. 9(2) February (2014)

Res. J. Biotech

genotypes and their 43 F1 hybrids. The leaves were

lyophilized by freeze drying and then stored in a 80oC
freezer until use.

Amaranth genotypes grouped into three clusters i.e. A

cluster, B cluster and C cluster (Fig.1). The cluster B has
Amaranthus caudatus as main parent and other included
under cluster B were hybrids while cluster A further was
divided into two sub-clusters i.e. A1, A2 and A3 where A1
has Amaranthus hypochondriacus as main parent and A2
further divided into two sub clusters as A2-1 and A2-2 has
remaining F1 and cluster C consisting of IC-1733
(Amaranthus edulis Spegazzini), SKGPA-144 (Amaranthus
tricolor) and Amaranthus cruentus as main parent.

Genomic DNA Extraction: Two hundred milligram leaf

tissue was washed in distilled water and ground into wash
buffer (1 ml/100 mg) with -marcaptoethanol (60
l/100mg) by using autoclaved pre-chilled mortar and
pestle. The DNA was isolated using the Cetyltrimethyl
Ammonium Bromide (CTAB) method4 with some
modification. The extracted DNA was resuspended in
250l TE buffer and restored at -20oC for further use.

The RAPD-based clustering (Fig. 1) revealed that A.

caudatus is closely related to A. hypochondriacus. The
similar results were also observed in previous RAPD-based
analysis3,14 and hybrid fertility data6. Pal and Khoshoo12
reported A. tricolor as one of the common progenitors of
A. cruentus and A. hypochondriacus. The present study
revealed low genetic similarities between A. tricolor and
other grain Amaranth supporting the previous finding.

RAPD Analysis and PCR amplification: The molecular

characterization through RAPD analysis was carried out by
method given by Mathews et al10 with minor modification.
The amplification was performed in 200 l thin walled
PCR tube containing a 25l reaction mixture (2.5l taq
buffer, 16l sterile DDH2O, 2.0 l MgCl2, 1.0l dNTP,
2.0l Primers, 1.0l Taq DNA polymerase and 0.5l
sample DNA). The reaction volume of 25l was subjected
to amplification through PCR in thermal cycler along with
the control (without template DNA). The PCR product was
resolved on 1.5 per cent agarose gel in 1X TAE buffer. The
gel was visualized under UV using gel documentation
system (Alfa Innotech Corporation, USA)

At the intra specic level, the percentages of RAPD

polymorphism were found to be 71.43 (A.
hypochondriacus), 100 (A. cruentus), 100 (A. caudatus),
75.76 [IC-1733 (A. edulis)] and 84.85 [SKGPA-144 (A.
tricolor)] and hybrids showed varied degree of
polymorphism ranging from 42.86 to 71.43 per cent. Using
GenAlex 6.5, all data were analyzed where statistical
analysis for allele frequency and its overall distribution
between and inter population suggested that in each
population, loci two, three and four have shown maximum
frequency with mutation of 0.667, 0.5 and 0.393
respectively. Estimated allele frequency for dominant allele
ranged from 1.0 to 0.293 exhibiting large scale scattering of
dominant nature of allele in population. In addition to it,
strikingly IC-1733 (Amaranthus edulis Spegazzini) and
SKGPA-144 (Amaranthus tricolor) at loci 5, 6 and 2, 3
respectively, have shown complete recessive nature of
expression (Table 2).

Data Scoring and Analysis: DNA finger prints were

scored for the presence=1 or absence=0 of band of various
molecular weight size in the form of binary matrix. The
polymorphism percentage was calculated as per the
formula suggested by Blair et al1:
The polymorphism (%) = a-b/a X 100
where a = Total number of bands and b= Number of
Monomorphic bands. The Polymorphism Information
Content (PIC) value was calculated as per equation of
Botstein et al2:
PICj =1

pi2

In most cases, allelic frequency has shown two no. of

different alleles (Na) while number of maximum effective
alleles in Amaranthus cruentus and Amaranthus caudatus
were 1.625 and 1.619 respectively where as IC1733(Amaranthus edulis Spegazzini) was found to possess
the least no. of effective alleles as 0.169 indicating
distribution of alleles in IC-1733 (Amaranthus edulis
Spegazzini) was not stable (Table 3). This finding is similar
to previous data where IC-1733 (Amaranthus edulis
Spegazzini) at loci five and six has complete recessive
expression.

where i = The ith allele of the Jth marker, b = The number

of alleles at the Jth marker and p= allele frequency.

Results and Discussion

PCR amplification of DNA, using four different primers for
RAPD analysis, produced 33 DNA fragments. All the
selected four primers amplified DNA fragments across 58
genotypes were studied. The total number of amplified
fragments varied from seven to ten for all the primers viz.
OPA-06, OPA-10, OPA- 14 and OPA- 15 with size ranging
from 63 to 400 bp. Out of 33 DNA fragments, 33 were
polymorphic giving a 100 per cent polymorphism. The
average polymorphic band per primer was 8.25 and percent
polymorphism displayed 100 per cent for all primers. The
PIC value varied from 0.61 (OPA-10) to 0.97 (OPA-15)
(Table 1). Dendrogram was constructed with the data
generated by all four primers and their amplicons of 58

Shannons information index showed the maximum

relatedness between Amaranthus cruentus and Amaranthus
caudatus among all the populations with mean values of
0.531 and 0.589 respectively.The least similar populations
were found to be IC-1733(Amaranthus edulis Spegazzini)
and SKGPA-144(Amaranthus tricolor) with mean value of
0.236 and 0.351 respectively.

30

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Vol. 9(2) February (2014)

Res. J. Biotech

Table 1
RAPD analysis showing Number of bands, Total Loci, Polymorphic Loci, Percentage Polymorphism, PIC value and
Molecular weight range (bp)
S.
N.
1
2
3
4

Locus
name
OPA-06
OPA-10
OPA-14
OPA-15
Total

No. of
bands
116
177
202
197
692

Total
Loci
8
10
8
7
33

Polymorphic
Loci
8
10
8
7
33

Percentage
Polymorphism
100
100
100
100
100

PIC
value
0.93
0.61
0.88
0.97
3.39

Molecular weight range

(bp)
63-414
234-1347
156-2227
237-3025
-

Table 2
Band Frequency(P), Estimated Allele Frequency (p and q), Samples Size(N), No. of Alleles (Na), No. of Effective
Alleles (Ne), Shannon Information Index(I), Expected Heterozygosity (He) and Unbiased Expected
Heterozygosity(uHe)
Pop
Pop1

Pop2

Pop3

Pop4

Pop5

Pop6

Locus
Locus1
Locus2
Locus3
Locus4
Locus5
Locus6
Locus7
Locus1
Locus2
Locus3
Locus4
Locus5
Locus6
Locus7
Locus1
Locus2
Locus3
Locus4
Locus5
Locus6
Locus7
Locus1
Locus2
Locus3
Locus4
Locus5
Locus6
Locus7
Locus1
Locus2
Locus3
Locus4
Locus5
Locus6
Locus7
Locus1
Locus2
Locus3
Locus4
Locus5
Locus6
Locus7

Band Freq.
1.000
0.333
0.333
0.333
1.000
0.167
0.333
0.333
0.667
0.667
0.500
0.167
0.667
0.167
0.167
0.667
0.667
0.333
0.333
0.667
0.333
0.833
0.000
0.167
0.667
0.000
0.000
1.000
0.333
0.000
0.000
0.500
0.333
0.167
0.500
0.036
0.250
0.393
0.179
0.250
0.286
0.250

P
1.000
0.184
0.184
0.184
1.000
0.087
0.184
0.184
0.423
0.423
0.293
0.087
0.423
0.087
0.087
0.423
0.423
0.184
0.184
0.423
0.184
0.592
0.000
0.087
0.423
0.000
0.000
1.000
0.184
0.000
0.000
0.293
0.184
0.087
0.293
0.018
0.134
0.221
0.094
0.134
0.155
0.134

Q
0.000
0.816
0.816
0.816
0.000
0.913
0.816
0.816
0.577
0.577
0.707
0.913
0.577
0.913
0.913
0.577
0.577
0.816
0.816
0.577
0.816
0.408
1.000
0.913
0.577
1.000
1.000
0.000
0.816
1.000
1.000
0.707
0.816
0.913
0.707
0.982
0.866
0.779
0.906
0.866
0.845
0.866

N
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
6.000
28.000
28.000
28.000
28.000
28.000
28.000
28.000

31

Na
1.000
2.000
2.000
2.000
1.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
0.000
2.000
2.000
0.000
0.000
1.000
2.000
0.000
0.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000
2.000

Ne
1.000
1.428
1.428
1.428
1.000
1.189
1.428
1.428
1.953
1.953
1.707
1.189
1.953
1.189
1.189
1.953
1.953
1.428
1.428
1.953
1.428
1.935
1.000
1.189
1.953
1.000
1.000
1.000
1.428
1.000
1.000
1.707
1.428
1.189
1.707
1.037
1.302
1.525
1.205
1.302
1.355
1.302

I
0.000
0.477
0.477
0.477
0.000
0.296
0.477
0.477
0.681
0.681
0.605
0.296
0.681
0.296
0.296
0.681
0.681
0.477
0.477
0.681
0.477
0.676
0.000
0.296
0.681
0.000
0.000
0.000
0.477
0.000
0.000
0.605
0.477
0.296
0.605
0.090
0.394
0.528
0.311
0.394
0.431
0.394

He
0.000
0.300
0.300
0.300
0.000
0.159
0.300
0.300
0.488
0.488
0.414
0.159
0.488
0.159
0.159
0.488
0.488
0.300
0.300
0.488
0.300
0.483
0.000
0.159
0.488
0.000
0.000
0.000
0.300
0.000
0.000
0.414
0.300
0.159
0.414
0.035
0.232
0.344
0.170
0.232
0.262
0.232

uHe
0.000
0.327
0.327
0.327
0.000
0.174
0.327
0.327
0.532
0.532
0.452
0.174
0.532
0.174
0.174
0.532
0.532
0.327
0.327
0.532
0.327
0.527
0.000
0.174
0.532
0.000
0.000
0.000
0.327
0.000
0.000
0.452
0.327
0.174
0.452
0.036
0.236
0.350
0.173
0.236
0.266
0.236

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Table 3
Data of mean and gross mean value of allelic frequency for different parameter
Pop
N
Na
Ne
I
He
uHe
Pop1 Mean 6.000 1.714 1.272 0.315 0.194 0.212
SE
0.000 0.184 0.077 0.085 0.054 0.059
Pop2 Mean 6.000 2.000 1.625 0.531 0.357 0.389
SE
0.000 0.000 0.134 0.067 0.057 0.062
Pop3 Mean 6.000 2.000 1.619 0.538 0.360 0.393
SE
0.000 0.000 0.122 0.056 0.049 0.053
Pop4 Mean 6.000 1.000 1.297 0.236 0.161 0.176
SE
0.000 0.378 0.169 0.121 0.087 0.094
Pop5 Mean 6.000 1.429 1.351 0.351 0.227 0.247
SE
0.000 0.369 0.113 0.099 0.067 0.073
Pop6 Mean 28.000 2.000 1.290 0.363 0.215 0.219
SE
0.000 0.000 0.056 0.052 0.036 0.037
Grand Mean and SE over Loci and Pops
N
Na
Ne
I
He
uHe
Total Mean 9.667 1.690 1.409 0.389 0.252 0.273
SE
1.280 0.105 0.051 0.036 0.026 0.028
Table 4
Data of polymorphism percentage in population and mean population polymorphism percentage
with its standard deviation
Population Polymorphism (%)
Pop1
71.43
Pop2
100.0
Pop3
100.0
Pop4
42.86
Pop5
71.43
Pop6
100.0
Mean
80.95
SE
9.52
Table 5
Total banding pattern among populations
Population
Pop1 Pop2 Pop3 Pop4
No. Bands
7
7
7
4
No. Bands Freq. >= 5%
7
7
7
4
No. Private Bands
0
0
0
0
No. LComm Bands (<=25%)
0
0
0
0
No. LComm Bands (<=50%)
0
0
0
0
Mean He
0.194 0.357 0.360 0.161
SE of Mean He
0.054 0.057 0.049 0.087
Mean uHe
0.212 0.389 0.393 0.176
SE of Mean uHe
0.059 0.062 0.053 0.094

Pop5
5
5
0
0
0
0.227
0.067
0.247
0.073

Table 6
Genetic distance matrix based on Nei genetic distance
Pop1 Pop2 Pop3 Pop4 Pop5 Pop6
Pop1 0.000
Pop1
Pop2 0.416 0.000
Pop2
Pop3 0.411 0.009 0.000
Pop3
Pop4 0.411 0.331 0.328 0.000
Pop4
Pop5 0.298 0.107 0.103 0.138 0.000
Pop5
Pop6 0.373 0.050 0.039 0.251 0.031 0.000 Pop6

32

Pop6
7
6
0
0
0
0.215
0.036
0.219
0.037

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Table 7
Genetic distance matrix based on Nei genetic identity
Pop1 Pop2 Pop3 Pop4 Pop5 Pop6
Pop1 1.000
Pop1
Pop2 0.660 1.000
Pop2
Pop3 0.663 0.991 1.000
Pop3
Pop4 0.663 0.718 0.720 1.000
Pop4
Pop5 0.742 0.899 0.902 0.871 1.000
Pop5
Pop6 0.689 0.951 0.962 0.778 0.970 1.000 Pop6
Table 8
Summary of Genic Variation Statistics for All Loci11
Locus
Sample Size
na*
ne*
h*
I*
OPA10-1
58
2.0000 1.5234 0.3436 0.5273
OPA10-2
58
2.0000 1.3960 0.2837 0.4576
OPA10-3
58
2.0000 1.5278 0.3455 0.5295
OPA10-4
58
2.0000 1.4382 0.3047 0.4826
OPA10-5
58
2.0000 1.5098 0.3377 0.5206
OPA10-6
58
2.0000 1.4194 0.2955 0.4717
OPA10-7
58
2.0000 1.5882 0.3704 0.5572
OPA10-8
58
2.0000 1.4728 0.3210 0.5016
OPA10-9
58
2.0000 1.4876 0.3278 0.5094
OPA10-10
58
2.0000 1.7291 0.4217 0.6126
OPA06-1
58
2.0000 1.4813 0.3249 0.5061
OPA06-2
58
2.0000 1.8742 0.4664 0.6592
OPA06-3
58
2.0000 1.5973 0.3740 0.5612
OPA06-4
58
2.0000 1.3430 0.2554 0.4232
OPA06-5
58
2.0000 1.4662 0.3180 0.4980
OPA06-6
58
2.0000 1.3389 0.2531 0.4204
OPA06-7
58
2.0000 1.3918 0.2815 0.4551
OPA06-8
58
2.0000 1.4576 0.3139 0.4934
OPA15-1
58
2.0000 1.9463 0.4862 0.6793
OPA15-2
58
2.0000 1.9553 0.4886 0.6817
OPA15-3
58
2.0000 1.6190 0.3823 0.5704
OPA15-4
58
2.0000 1.8800 0.4681 0.6609
OPA15-5
58
2.0000 1.9025 0.4744 0.6673
OPA15-6
58
2.0000 1.5453 0.3529 0.5378
OPA15-7
58
2.0000 1.5901 0.3711 0.5581
OPA14-1
58
2.0000 1.7760 0.4369 0.6287
OPA14-2
58
2.0000 1.6779 0.4040 0.5938
OPA14-3
58
2.0000 1.9912 0.4978 0.6909
OPA14-4
58
2.0000 1.9767 0.4941 0.6872
OPA14-5
58
2.0000 1.7324 0.4228 0.6138
OPA14-6
58
2.0000 1.4887 0.3283 0.5099
OPA14-7
58
2.0000 1.2138 0.1762 0.3198
OPA14-8
58
2.0000 1.3447 0.2564 0.4244
Mean
58
2.0000
St. Dev
0.0000
* na = Observed number of alleles
* ne = Effective number of alleles
* h = Nei's (1973) gene diversity
* I = Shannon's Information index

33

1.5964
0.2140

0.3630
0.0825

0.5458
0.0922

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Fig. 1: Dendrogram showing clustering of different species of Amaranth and their hybrids and their using MEGA
software based on Nei11 genetic distance with NEIGHBOUR method obtained from different primers

Figure 2: Band patterns across population

The overall likeliness among population mean is 0.398
(Table 3) which showed population is moderately
dissimilar from its origin which matches to our selection of
genotypes and cross combination tested.

Expected heterozygosity (He) revealed genetic diversity

within population. Amaranthus caudatus and Amaranthus
cruentus are highly diverse with He mean of 0.357 and
0.360 respectively, as matching to data of per cent

34

Research Journal of Biotechnology

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Res. J. Biotech
2. Botstein D., White R. L., Skolnick M. and Davis R. W.,
Construction of a genetic linkage map in man using restriction
fragment polymorphisms, Am J Hum Genet, 32, 314-331 (1980)

polymorphism in both the populations, The IC-1733

(Amaranthus edulis Spegazzini) has shown least per cent
polymorphism as 42.86, while overall mean per cent
polymorphism of loci in all populations is 80.95 9.52
(Table 4) which exhibited that all genotypes of population
are quite diverse from its origin and again confirming our
previous result of Shannons information index.

3. Chan K. F. and Sun M., Genetic diversity and relationships

detected by isozyme and RAPD analysis of crop and wild species
of Amaranthus, Theor. Appl. Genet., 95, 865-873 (1997)
4. Doyle J. J. and Doyle J. L., Isolation of plant DNA from fresh
tissue, Focus, 12, 13-15 (1990)

Banding pattern graph (Fig. 2) for allelic data depicted that

among all populations, no. of bands and its frequency was
greater than 5 per cent in all the cases where as none of the
populations showed unique allele loci to single population
and showed inter cross nature of population with its overall
distribution among population.

5. Gepts P., The use of molecular and biochemical markers in

crop evolution studies, In Hecht M. K., ed., Evolutionary
Biology, 27, 5194, Plenum Press, New York (1993)
6. Gupta V. K. and Gudu S., Interspecific hybrids and possible
phylogenetic relations in grain amaranths, Euphytica, 52, 33-38
(1991)

The overall population banding pattern showed that in

Amaranth tricolor and F1 hybrid, mean heterozygosity
remained similar as 0.247 and 0.219 while it decreased to
lowest at 0.161 at IC-1733 (Amaranthus edulis Spegazzini)
and among most cases, average mean heterozygosity was
found to be 0.55(Table 5) which matched to our previous
Shannons index with 0.398.

7. Hallden C., Nilsson N. O., Ebding L. M. and Sdl T., Evalution

of RFLP and RAPD markers in a comparision of Brassica napus
breeding lines, Theor. Appl. Genet., 123-128 (1994)
8. Hyam R., Field collection: Plants, In Karp A., Isaac P. G.,
Ingram D. S., eds., Molecular tool for Screening biodiversity,
Chapman and Hall, London, 49-50 (1998)

Looking at polymorphism among populations, calculation

of genetic distance was carried out by using Nei11 genetic
distance method and pair wise analysis of each allele is
done at all loci for accurate distance analysis (Table 6). The
pair wise population of Nei11 genetic distance formed
genetic distance matrix based on biased allelic distribution.
The population matrix showed that allele at SKGPA144(Amaranthus tricolor) and F1 hybrids have maximum
genetic distance of 0.742 to 0.962 at all populations while
population IC-1733 (Amaranthus edulis) has minimum
genetic distance among all the germplasm.

9. Karp A., Reproducibility testing of RAPD, AFLP and SSR

markers in plants by a network of European laboratories, Mol.
Breed., 3, 381390 (1997)
10. Mathews M. D., Srinivasachary Sujatha, JeVrey R., Mike L.
B., Gale D. and Katrien M. D., The genetic map of Finger millet
Eleusine coracana, Theor. Appl. Genet., 114, 321-332 (2007)
11. Nei M., Molecular evolutionary genetics, Columbia
University Press, New York (1987)
12. Pal M. and Khoshoo T. N., Evolution and improvement of
cultivated amaranths, VI. cytogenetic relationships in grain types,
Theor. Appl. Genet, 43, 242-251 (1973)

The lowest and the maximum genetic distance of IC-1733

(Amaranthus edulis) were 0.663 and 0.720 respectively to
all populations. It further matches the pattern of allelic
frequency distribution that SKGPA-144(Amaranthus
tricolor) and F1 have remarkable high polymorphism
(Table 7). Analysis of population polymorphism as multi
descriptive statistic using program popgene 32 has been
carried out effectively for all possible 33 loci from four
primers of RAPD.

13. Sauer J. D., The grain amaranths and their relatives a revised
taxonomic and geographic survey, Ann. Missouri Bot., 54, 103137 (1967)
14. Transue D. K., Fairbanks D. J., Robison L. R. and Andersen
W. R., Species identification by RAPD analysis of grain amaranth
genetic resources, Crop Sci, 34, 1385-1389 (1994)

Results showed that loci OPA 15-1, OPA14-3, OPA14-4

and OPA 06-2 have maximum effective number of allele
i.e. alleles distributed equally in ideal population while
OPA 06-4 (1.34), OPA 06-6 (1.33), OPA14-7(1.21) and
OPA14-8 (1.34) have lower distribution (Table 8) which
suggested that all loci remained quite scattered and
polymorphic among all populations as cumulatively.

15. Welsh J. and McClelland M., Fingerprinting genomes using

PCR with arbitrary primers, Nucleic Acids Res, 18, 72137218
(1990)
16. Williams J. G. K., Kubelik A. R., Livak K. J., Rafalski J. A.
and Tingey S. V., DNA polymorphisms amplied by arbitrary
primers are useful as genetic markers, Nucleic Acids Res, 18,
6531-6535(1990).

References

(Received 15th August 2013, accepted 20th November

2013)

1. Blair M. W., Panaud O. and McCouch S. R., Inter simple

sequence repeat (ISSR) amplification for analysis of
microsatellite motif frequency and finger printing in rice (Oryza
sativa L.), Theor. Appl. Genet., 98, 780792 (1999)

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