American Journal of Plant Sciences, 2011, 2, 629-635
doi:10.4236/ajps.2011.25074 Published Online November 2011 (http://www.SciRP.org/journal/ajps)
629
Genetic Variability in Germplasm Accessions of
Capsicum annuum L
Shrilekha Misra1*, Raj Kishori Lal2, Mahendra Pandurang Darokar2, Suman Preet Singh Khanuja2
1Banasthali University, Rajasthan, India; 2Central Institute of Medicinal and Aromatic Plants, Lucknow, India.
Received December 13th, 2010; revised March 14th, 2011; accepted May 1st, 2011.
ABSTRACT
Capsicum annuum is the most widely cultivated species of peppers (chilies) in the world. For culinary purposes, its
fruits are used for pungency (capsaicin) and also color (capsanthin). Capsaicin is also used for medicinal purposes
particularly in anti-inflammatory formulations. Genetic divergence among 38 accessions collected from diverse loca-
tions in India (28 from Uttar Pradesh, 5 from Assam, 3 from Maharash tra and 2 from Uttaranc hal), was estimated from
the data pooled over 3 consecutive years for 15 morphological, growth and chemotypic characters that included days to
first and second flowering, fruit onset, plant height, primary, secondary and tertiary branches, leaf surface area, fruit
length and diameter, fruit surface area, fresh and dry fruit weight, capsaicin and capsanthin content. Based on this
characterization the plants could be grouped into 7 clusters wherein substantial diversity among accessions was indi-
cated by the wide range of 2
D values (752.901 - 1918683.00). Accessions with distinct identity were marked, which
are likely to be quite suitable for breeding through hybridization combining desirable traits. The accessions labeled
number 38, 27, 26, 14 and 24 to high capsaicin content (%); 35, 23, 3, 16, 29 and 11 for high capsanthin content (%)
and 26 and 27 for dual purpose had characteristics de sirable. Above accessions could be utilized in hybridiza tion pro-
gramme for C. annuum crop improvement.
Keywords: Genetic Diversity, Capsicum Annuum, Capsaicin and Capsanthin Content, Recombination Breeding,
Geographic al Di st ri but i o n
1. Introduction
Capsicum is the unique genus of the family Solanaceae
finding diverse uses from nutritional and culinary to
pharmaceutical uses. In this genus, native to the Ameri-
cas, more than 30 species have been described, but only
five of them, Capsicum annuum var. annuum, C. chi-
nense, C. frutescens, C. baccatum var. pendulum, and C.
pubescens are considered to be domesticated (Pandey,
Pozzobon et al., 2006; Moscone et al., 2007) [1-3]. Pun-
gent peppers commonly known as chilies in India be-
long to the species C. annuum L. which is the most
widely cultivated species in the world. Both hot and
sweet peppers have been reported to have originated
from C. annuum [4].
The pungent chemical principle of Capsicum is cap-
saicin, which is synthesized and accumulated specifically
in fruits. The capsaicin has found use in a wide range of
pain problems, including post-mastectomy syndrome,
urticaria, psoriasis, diabetic neuropathy, arthritis, pruritis,
contact allergy, post-surgical neuromas etc. [5]. The sweet
pepper on the other hand is one of the richest sources of
carotenoids. The ketoxanthophylls, capsanthin and cap-
sorubin are the major pigments contributing to the red
color of Capsicum, while β-carotene and zeaxanthin are
responcible for the yellow-orange colors [6].
The characterization and the evaluation of the Capsi-
cum domesticated species are particularly interesting for
gene bank curators, since a wide variability, not yet fully
known and exploited, is available in these species (Guz-
mán et al. [7], 2005; Sudré et al. [8], 2006; Ince et al. [9],
2009). In this study, genetic variability for diverse traits
in available germplasm collections assembled from dif-
ferent places in India were evaluated as a prelude to crop
improvement.
2. Materials and Methods
Genetic divergence among 38 indigenous accessions of
Capsicum annuum collected from various geographical
locations (Tabl e 1) in India was studied. The plants were
grown in randomized block design at the research farm

Genetic Variability in Germplasm Accessions of Capsicum annuum L 630
Table 1. Accessions/collections of Capsicum annuum and
their geographic origin.
Accession No. Pl ace of collection/acquisition
1 Lucknow, U.P.
2 "
3 "
4 "
5 "
6 "
7 "
8 "
9 "
10 "
11 "
12 "
13 "
14 "
15 "
16 "
17 "
18 Ramnagar, Uttranchal.
19 Pantnagar, Uttranchal.
20 Lucknow, U.P.
21 "
22 "
23 Nagpur, Maharashtra.
24 Assam.
25 Assam.
26 Assam.
27 Assam.
28 Assam.
29 Lucknow, U.P.
30 "
31 "
32 "
33 "
34 "
35 "
36 "
37 Pune, Maharashtra.
38 Pune Maharashtra.
of CIMAP, Lucknow, India during 1998, 1999 and 2000
with two replicates in each year. Each treatment con-
sisted of single row 45 cm long and 60 cm apart. The
plant received normal intercultural operations, irrigations
and fertilizer applications (20 kg N, 30 kg P2O5 and 30
kg K2O per hectare).
Observations were recorded for the 15 characters for
days to first flowering, days to second flowering, days to
fruit initiation, plant height, primary branches/plant,
secondary branches/primary branch, tertiary branches/
secondary branch, leaf surface area, fruit length, fruit
diameter, fruit surface area, fruit weight fresh and dry,
capsaicin and capsanthin content. Mean values for all the
characters (pooled over three years) were subjected to D2
and canonical analysis of genetic divergence based on
the procedures outlined by Mahalanobis [10] and Rao
[4].
3. Results
Highly significant differences (P < 0.01) among the col-
lection of C. annuum for all the studied 15 characters
indicated the presence of considerable genetic diversity.
Tremendous variation in shape and size of fruits were
observed (Table 2, for the mean, ranges and C.D.).
Hence D2 values for 703 pairs of genotypes were deve-
loped. Based on these values, all 38 accessions/genotypes
could be grouped into seven clusters such that the geno-
type within the clusters had smaller 2
D-values among
themselves than those belonging to different clusters
(Table 3). Divergence for the pooled characters (in terms
of D2 values) ranged from 752.901 to 1918683 and clus-
ter mean (for 164 to 241 days—first flowering, 000 - 243
days to second flowering, 259 - 336 days of fruit initia-
tion, 50 - 106 cm plant height, 5 - 8 primary branches, 7 -
10 secondary branches, 5 - 9 tertiary branches, 20.06 -
45.39 cm2 leaf surface area, 2.332 - 9.960 cm fruit length,
0.802 - 1.830 cm fruit diameter, 4.312 - 27.400 cm2 fruit
surface area, 0.76 - 4.17 mg fruit weight fresh, 0.20 -
0.75 mg fruit weight dry, 0.000% - 0.0869% capsaicin
content and 0.063% - 0.517% capsanthin content). It is
evident from the results, Table 2 and Figure 1 (in Fig-
ure 1, D = 2
D), That the mean inter-cluster 2 D -
values and the cluster mean indicated the highest diver-
gence between clusters designated III and VII (2
D=
1665293.0), corroborating the proposed variation of the
germplasm into different gene complexes based on cyto-
genetic aspects and botanical characters. Divergence was
also noted between clusters designated I and VII (2
D =
1661982.0). II and VII (2
D = 1167484.0) and between
IV and VII (2
D = 1119514.0). The lowest inter-cluster
distance was noticed in between cluster I and cluster III
(2 D = 76941.01) (Table 3). Considerable differences in
economic traits, growth and other parameters were noted
(Tables 2 and 4). The comparative characteristic features
of Capcicum annuum plant collected from various places

Genetic Variability in Germplasm Accessions of Capsicum annuum L 631
Table 2. Mean performance (X), ranges and critical difference of 38 accessions of Capsicum annuum.
Accession
No.
Days to
first
flowering
Days to
second
flowering
Days to
fruit
initiation
Plant
height
(cm)
No. of
primary
branches/
plant
No. of
secondary
branch es/
primary
branch
No. of
tertiary
branch es/
secondary
branch
Leaf
surface
area (cm2)
Fruit
length
(cm)
Fruit
diameter
(cm)
Fruit
surface
area (cm2)
Fruit
weight
fresh (g)
Fruit
weight
dry (g)
Capsaicin
(%)
Capsanthin
(%)
1 173 233 271 100 4 9 5 29.98 5.98 0.66 6.65 0.81 0.23 0.018 0.036
2 180 243 275 103 7 7 6 34.66 3.01 1.09 5.18 2.81 0.50 0.036 0.052
3 156 000 207 53 7 6 5 11.41 2.23 0.65 2.64 0.48 0.16 0.090 0.307
4 184 237 267 63 8 7 6 19.68 5.18 0.81 7.52 0.93 0.32 0.102 0.046
5 177 242 272 106 5 8 6 24.00 2.62 1.84 10.94 3.81 0.88 0.002 0.015
6 177 241 253 91 8 9 6 18.26 2.16 0.68 3.23 0.54 0.23 0.049 0.008
7 178 237 253 79 5 9 8 40.19 3.90 1.35 10.56 4.44 0.96 0.002 0.140
8 144 000 174 41 8 8 6 10.62 1.61 1.08 3.58 0.72 0.24 0.004 0.186
9 174 237 275 91 9 9 6 16.59 4.38 0.68 5.35 0.74 0.17 0.075 0.183
10 178 238 267 96 8 8 7 23.04 4.19 0.67 4.67 1.26 0.23 0.043 0.071
11 180 244 261 82 8 8 6 14.8802.56 0.71 3.04 0.36 0.14 0.011 0.204
12 168 221 251 98 8 7 4 16.63 4.18 0.77 4.69 0.53 0.23 0.014 0.062
13 175 232 255 92 7 8 6 14.19 3.11 0.78 3.32 0.45 0.15 0.086 0.106
14 181 237 256 90 10 7 7 24.79 4.74 0.76 5.52 1.21 0.25 0.140 0.042
15 189 240 281 92 11 6 8 38.75 3.06 0.79 4.28 0.53 0.20 0.100 0.079
16 178 241 267 84 9 10 6 13.10 3.75 0.74 5.21 0.85 0.25 0.089 0.291
17 179 240 266 81 11 10 7 8.53 3.96 0.97 6.32 0.52 0.19 0.049 0.067
18 152 000 179 58 8 10 8 8.83 4.52 0.82 6.60 1.00 0.25 0.045 0.035
19 168 000 207 50 6 7 5 14.18 3.50 0.81 4.07 0.61 0.21 0.060 0.078
20 166 237 279 85 8 8 6 37.37 3.24 1.51 8.42 2.26 0.49 0.036 0.149
21 175 242 250 98 8 6 6 27.69 4.06 1.31 9.75 2.05 0.44 0.082 0.101
22 171 248 310 91 9 10 6 19.55 3.95 1.21 8.31 1.83 0.42 0.083 0.145
23 176 240 297 79 8 7 7 25.27 3.72 1.21 9.52 2.55 0.54 0.044 0.334
24 183 237 303 110 10 10 5 22.50 1.62 1.35 4.98 1.21 0.20 0.128 0.043
25 161 241 268 96 8 11 6 9.40 2.13 1.03 4.01 2.56 0.51 0.082 0.017
26 179 237 302 111 11 10 5 43.28 1.45 1.35 4.97 1.31 0.32 0.144 0.191
27 183 240 265 72 7 7 5 6.62 2.65 0.74 3.84 0.48 0.20 0.145 0.145
28 180 237 305 42 9 9 5 44.01 2.78 0.76 4.13 0.77 0.22 0.131 0.045
29 157 000 187 59 8 10 6 17.66 3.80 0.76 4.24 0.69 0.23 0.011 0.214
30 177 232 266 50 5 7 5 23.23 5.84 1.43 14.98 2.01 0.49 0.001 0.051
31 179 244 278 74 8 7 5 29.07 3.32 0.74 4.38 0.80 0.22 0.001 0.111
32 159 224 252 83 7 9 5 20.77 2.97 1.02 4.28 0.65 0.25 0.068 0.180

Genetic Variability in Germplasm Accessions of Capsicum annuum L 632
33 180 225 271 78 8 7 5 24.45 2.38 0.99 2.91 0.47 0.16 0.096 0.014
34 181 226 270 84 6 9 6 25.45 3.38 0.74 4.47 1.30 0.28 0.026 0.193
35 176 243 271 106 6 8 6 45.39 3.61 1.82 14.68 2.75 0.70 0.000 0.478
36 218 000 294 92 8 10 8 44.42 9.96 1.59 27.40 3.18 0.82 0.104 0.086
37 178 245 254 102 10 11 6 28.03 4.65 0.70 5.54 1.27 0.24 0.085 0.053
38 160 000 187 41 6 7 5 9.45 4.30 1.09 8.60 1.05 0.31 0.219 0.100
Mean (X) 174 237 258 81 7 8 5 23.31 3.64 1.00 6.64 1.36 0.34 0.065 0.123
Range 144 - 218 221 - 248 174 - 31041 -
111 4 - 11 6 - 114 - 8 6.62 -
45.39
1.45 -
9.96
0.65 -
1.84
2.64 -
27.40
0.36 -
4.44
0.14 -
0.96
0.00 -
0.22 0.008 - 0478
CD (1%) 5.133 4.228 4.559 2.608 1.740 1.591 1.363 0.567 4.508 4.159 0.084 0.036 3.069 0.004 0.003
Table 3. Average intra- (bold) and inter cluster divergence (D2 value) in Capsicum annuum acce ssions.
Clusters I II III IV V VI VII 2
D Accession included in
clusters
I (21) 43480.89 91363.24 76941.01 219655.20 303668.90298298.301661982.002695389.50
3, 6, 9, 10, 11, 12, 13, 14,
15, 16, 17, 19, 26, 27, 28,
29, 31, 32, 33, 34, 37
II (7) 54333.34 118249.60 141860.60166808.30126699.801167484.001866798.80 1, 4, 18, 21, 22, 23, 38
III (5) 60444.35 133822.00257733.40331515.701665293.002643999.00 2, 8, 20, 24, 25
IV (2) 32932.59 84986.18 195033.901119514.001927804.40 5,7
V (1) 00000.00 90935.49 758072.301662204.50 35
VI (1) 00000.00 604546.801647029.90 30
VII (1) 00000.00 6976892.10 36
2
D = Average D2 value.
Table 4. Cluster means and other allied genetic parameters in Capsicum annuum accessions.
Clusters Days to Ist
Flower Ing
Days to Iind
Flowering
Days of
Fruit
Initiation
Plant
Height
(cm)
Prmary
Bran ches
/Plant
Sec.
Br./Pri.
Br.
Teri. Br./
Sec. Br.
(Sq.cm)
Leaf
Surface
Area
(Sq.cm)
Fruit
Length
(cm)
Fruit
Diameter
(cm)
Fruit
Surface
Area
(Sq.cm)
Fruit Wt.
(Fresh)
(Mgs)
Fruit
Wt.
(Dry)
(Mgs)
Capsaicin
Content
(%)
Capsanthin
Content
(%)
I 175 231 262 80 8 8 5 21.59 3.250.80 4.31 0.76 0.22 0.08690.156
II 164 171 267 76 7 8 6 20.06 4.531.01 8.14 1.46 0.33 0.074 0.120
III 167 239 259 87 8 8 6 22.90 4.531.01 8.14 1.46 0.33 0.074 0.120
IV 177 240 262 92 5 8 7 32.09 3.261.59 10.75 4.17 0.75 0.000 0.111
V 176 243 336 106 6 8 6 45.39 3.621.83 14.68 2.75 0.71 0.000 0.517
VI 177 232 265 50 5 7 5 23.23 5.851.43 14.99 2.01 0.50 0.004 0.019
VII 241 000 293 91 8 10 9 44.42 9.961.59 27.40 3.19 0.71 0.039 0.148
Z 1 vector 1.7709 –0.1702 –4.1256 4.8078 7.4035 –6.4503–2.99240.24950.33070.15730.79110.1059 0.3597 –5.30541.1607
Ranks III XI XIII II I VX XII VIII VII IX V X VI IVX IV
Z 2 vector –0.0115 –9.8341 –1.6872 0.1167 5.6000 0.1705–0.22910.5147–0.40220.3234–8.66040.5715 –0.1568 –2.7722–3.7760
Rank VII VX XI VI I V IX III X IV IVX II VIII XII XIII

Genetic Variability in Germplasm Accessions of Capsicum annuum L 633
that largest cluster grouping was comprised of accessions
collected from Lucknow (Uttar Pradesh); Pantnagar and
Ramnagar (Uttaranchal); Assam and Pune (Maharashtra)
followed by cluster II comprised of accessions originated
from Uttar Pradesh, Uttaranchal and Maharashtra, and
cluster III comprised the accessions collected from Uttar
Pradesh and Assam states of India.
Notwithstanding the fact that, based on 2
D-values, all
38 accessions/genotypes could be grouped into seven
clusters such that the genotype within the clusters had
smaller 2
D-values among themselves than those be-
longing to different clusters (Table 3). It is evident from
the results, Table 3 and Figure 1, that the mean in-
ter-cluster 2
D-values and the cluster mean indicated the
highest divergence between clusters designated III and
VII (2
D= 1665293.0), corroborating the proposed varia-
tion of the germplasm into different gene complexes by
Moscone et al. (2007) [3] in investigation based on cy-
togenetic aspects and botanical characters. Divergence
was also noted between clusters designated I and VII
(2
D = 1661982.0). II and VII (2 D = 1167484.0) and
between IV and VII (2 D = 1119514.0). The lowest inter-
cluster distance was noticed in between cluster I and
cluster III (2
D = 76941.01) (Table 3). The highest intra
cluster 2 D values depicted by clster III (2 D =
60444.35) having 5 accessions followed by cluster II
(2
D = 55333.340 having 7 and cluster I (2 D =
43480.89) having 21 accessions. The 0.00 intra-cluster
value depicted by clusters V to VII having unique indi-
vidual and highly diverse accession (Figure 1 in terms of
D = 2
D).
Figure 1. Genetic diversity (D = 2
D
) among 38 acces-
sions of C. annuum.
of India are presented in Table 1. Days to first flowering
varies from 174 - 175, days to 2nd flowering 237 - 247,
days to fruit initiation 265 - 267, plant height 81 - 82 cm,
primary branches/plant 7 - 8, secondary branches/pri-
mary branch 7 - 8, tertiary branches/ secondary branch 5
- 6, leaves surface area 23.11 - 23.30 per sq.cm., fruit
length 3.6 - 3.63 cm , fruit diameter 0.998 - 1.00 cm fruit
surface area 6.642 - 6.65/sq.cm, fruit weight fresh 1.37 -
1.38 mg, fruit weight dry 0.309 - 0.311 mg, capsaicin
content 0.072% - 0.077%, capsanthin content 0.140 -
0.142% (Table 5). Other morphological characters were
also exhibited tremendous variation such as color of leaf
flower and fruits, plant type, shape and morphology of
fruit etc. (Table 5).
Accession number 5, 7, 30, 35 and 36 from Lucknow
(Uttar Pradesh) appeared highly divergent being repre-
sented within four clusters. The clusters V to VII were
highly divergent and unique including only one accession
each (35-V, 30-VI and 36-VII cluster). All belonging
from Lucknow (Uttar Pradesh) represented enormous
variation with respect to their morphology, growth be-
havior, ecological requirement and genetic diversity. The
primary branches per plant (7.4035, rank I) followed by
plant height (4.8078, rank II) and days to first flowering
(1.7709, rank III) at the primary axis (Z1) and the pri-
mary branches per plant (5.6000, rank I) followed by
fruit weight fresh (0.5715, rank II) and leaf surface area
(0.5147, rank III) at the secondary axis (Z2), respectively
were the largest contributor to genetic divergence. The
least contributor to genetic divergence were secondary
branches per primary branch (–6.4503) followed by cap-
saicin content (–5.3054) and days of fruit initiation
(–4.1256) at primary axis, respectively. Least contribut-
ing traits fruit surface area (–8.660), capsanthin content
(–3.776) and capsaicin content (–2.7722) at the secon-
dary axis, respectively (Table 4). The distribution pattern
4. Discussion
In the Capsicum annuum accessions, 55.263% of the
accessions could be grouped within one cluster I (21 ac-
cessions); in cluster II, 18.421% (7 accessions); cluster
III 13.158% (5 accessions), followed by cluster IV 5.263%
(2 accessions). The cluster V, VI and VII were found
unique included only one accession in each namely, 35,
30 and 36, which were in 7.896% of the total accessions
(Table 3). An analysis of cluster composition revealed

Genetic Variability in Germplasm Accessions of Capsicum annuum L 634
Table 5. Average intra and inter cluster D value in Capsicum annuum accessions.
Clusters I II III IV V VI VII
I (21) 208.52 302.26 277.38 468.67 551.06 546.16 1289.17
II (7) 283.09 343.87 376.64 408.42 355.94 1080.50
III (5) 245.85 365.81 507.67 575.77 1290.46
IV (2) 181.47 291.52 441.62 1058.07
V (1) 000.00 301.55 870.67
VI (1) 000.00 777.52
VII (1) 000.00
Where D = 2 D.
of accessions of diverse origin in the clusters indicates
that genetic diversity observed within C. annuum was not
related to geographic origin. Noted difference in plant
characters probably occurred over time with free move-
ment of plant material from location to location, mutation,
hybridization, gene recombination and selection. Our
findings were also in consonance of results of Pandey
and Dobhal [1] and Varalakshmi and Haribabu [11] in C.
annuum crop. Selection of promising accessions based
on genetic divergence and capsaicin and capsanthin con-
tent quality are useful in chili crop improvement. The
accessions labeled number 38, 27, 26, 14 and 24 to high
capsaicin content (%); 35, 23, 3, 16, 29 and 11 for high
capsanthin content (%) and 26 and 27 for dual purpose
had characteristics desirable. Among the above, the ac-
cession no. 35 (0.000% capsaicin content) should be
utilized for the study of genetics of capsaicin content,
making hybrids between the parents 0.000 X high cap-
saicin content.
Mainly due to the versatility of application, C. annuum
plants have become important from an economic point of
view in several countries since their fruits can be used for
different purposes, such as in cooking, in industry (e.g.,
production of “pepper spray”) and for medicinal and or-
namental purposes (Pickersgill, [12] 1971; [13] Moscone
et al. [3], 2007). In Brazil this vegetable is of great im-
portance, ranking second in the generation of foreign
currency among exported vegetables (Embrapa, [6,14]
2009). Therefore, the above listed accessions of the C.
annuum could be utilized, as parents for hybridization in
recombination breeding programme followed by repeated
selection to obtain maximum gain.
5. Acknowledgements
This research work was supported by a grant from CSIR
(Council of Scientific and Industrial Research) and con-
ducted at CIMAP (Central Institute of medicinal and
Aromatic Plants) Lucknow, India.
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