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Assessment of natural cross-pollination levels in chili pepper (Capsicum

annuum L.)
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FORUM IPIMA IOP Publishing
IOP Conf. Series: Earth and Environmental Science 196 (2018)
1234567890 ‘’“” 012008 doi:10.1088/1755-1315/196/1/012008

Assessment of natural cross-pollination levels in chili pepper

(Capsicum annuum L.)

A W Ritonga1, M Syukur1, Rahmi Yunianti1, and Sobir1

1
Department of Agronomy and Horticulture, Faculty of Agriculture, Bogor Agricultural
University, Bogor, West Java, Indonesia.

Email : [email protected]

Abstract. Inconsistency of the current information on the natural cross-pollination (NCP)

stimulated this research to update the natural cross-pollination rates of chili pepper. The
objective of this study were to determine the level of natural cross pollination and the various
factors that influence of the NCP in chili pepper. The experiment used four genotypes of chili
peppers, “IPBC2, IPBC5, IPBC120” (green hypocotyl) and IPBC20 (purple hypocotyl). The
experiment utilized the dominance of purple hypocotyl colors over green hypocotyl colors to
determine NCP. The experimental field consisted of three blocks of trial. Each blocks was
planted by purple hypocotyl genotype in the central plot and green hypocotyl genotypes in the
surrounding four plots. Seeds were harvested from each plant and were examined for the levels
of NCP in the next generation. The result showed that there were the differences on percentages
of NCP on different chili pepper genotypes. The results also showed that the levels of NCP in
chili pepper could be associated with by stigma exertion length and the level of successful
artificial cross-pollination.
Keywords: Artificial cross-pollination, heterosis, inbreeding depression, purple hypocotyl, and
stigma exertion.

1. Introduction
Information on natural cross-pollination (NCP) rates of a crop species in specific environment is
important when choosing a breeding method for crop improvement, germplasm maintenance and
commercial seed production. Chili pepper is known as self-pollinated crops [1]. Self-pollinated crops
are plants which have NCP rates under 10%. Chili pepper has a perfect flower. The time of anthers
dehisce is almost the same time with the receptive stigma. Pollination occurs after the flower open in
chili pepper.
There were variations in NCP in chili pepper. [2], [3], and [3] reported that the NCP rates in chili
pepper could reach over 50% in Italy and Spain while [5] reported the NCP rate in chili pepper ranged
from 0.26% to 3.56%. This inconsistencies prompted several investigators to suggest that Capsicum
should be considered as facultative cross-pollination species [6,7]. This variation depends on the
characteristics of the genotypes, the regions, or the environments within the region. Observations on the
flower of Serrano pepper verified that NCP in chili pepper occurs on the flower whose pistil in longer
than the stamen.
The variation of NCP in crop species can influence method and purpose of crops improvement.
Crops which have high levels of NCP are generally bred as hybrid varieties, while crops with low level
of NCP are generally bred and released as pure line varieties. The objective of this study was to

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 FORUM IPIMA IOP Publishing
IOP Conf. Series: Earth and Environmental Science 196 (2018)
1234567890 ‘’“” 012008 doi:10.1088/1755-1315/196/1/012008

determine the NCP rates of some genotypes of chili pepper in Indonesia and the various factors that
influence the NCP.

2. Materials and methods

This experiment was conducted from September 2011 until March 2012 at the IPB experimental field
Leuwikopo and the Plant Breeding Laboratory, Department of Agronomy and Horticulture, Faculty of
Agriculture, Bogor Agricultural University. Four genotypes of chili peppers were used in this
experiment. IPB C20 genotype, which has purple hypocotyl, was used as pollen source for NCP
identification. IPB C2 and IPB C5 genotypes are female and male parents for the hybrid variety of IPB
CH3, whereas IPB C120 is an inbreed line variety (local variety) of chili pepper were used NCP
evaluation.
The experimental field consisted of three blocks of trial in which each of blocks consisted of 9 m2
pollen source plot in the central block and 10.5 m2 receptor pollen plot in the surrounding the pollen
source plot. The pollen source plot was planted by purple hypocotyl genotypes (IPB C20) with spacing
radius 0.5m x 0.5m, whereas receptor pollen plot was planted by green hypocotyl genotypes (IPB C2,
IPB C5 or IPB C120) with radius 0.5, 1, 1.5, 2.5 and 3.5 m from pollen source plot (figure 1).

Figure 1. The experimental field.

All seeds were harvested from each plant in pollen receptor plots and were examined for NCP in the
next generation. The dominance of purple hypocotyl colors over green hypocotyl colors was utilized to
identify NCP. Seeds were sown in transparent plastics box, seeds from NCP produced purple hypocotyl
H
colors after one week. The percentage of NCP was determined by calculating as t = p x 100%, where H
is the frequency of seedling with purple hypocotyl color; p is number seeds were sown. In addition, the
observations also were made on anther length, carpel length, stigma exertion, and the percentage of
successful of artificial cross-pollination. Artificial cross-pollination was done by hand pollination at the
first node to twelve nodes on each plant. IPB C2, IPB C5 and IPB C120 genotypes were used as the
female parents, whereas IPB C20 genotype was used male parent. The percentage of successful artificial
A
cross-pollination was determined by calculating as T = b x 100%, where A is a success number of
artificial pollination; b is number artificial pollination was done.
Data were analyzed using Minitab 17. T-testing was employed when necessary to detect significant
differences among means. A probability level of p < 0.05 was considered statistically significant. A
linier regression correlation test was performed to investigate correlation between percent natural cross-
pollination and stigma exertion.

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IOP Conf. Series: Earth and Environmental Science 196 (2018)
1234567890 ‘’“” 012008 doi:10.1088/1755-1315/196/1/012008

3. Result and discussion

The result showed that there were significant differences in the levels of natural cross-pollination among
the genotypes of chili pepper (table 1). This is consistent with [6] that there were differences in the
levels of NCP in different types of chili pepper. IPB C2 genotype had the highest percentage NCP with
24.53% while the lower percentage was found on IPB C120 and IPB C5 genotypes with less than 10%
(table 1).

Table 1. Percent natural cross-pollination percentages of three genotypes of chili pepper on different
planting radius.
Radius Genotypes
Means
(m) IPB C2 IPB C5 IPB C120
0.5 22.11 6.75 6.69 11.85
1 25.75 3.73 15.86 15.11
1.5 17.46 3.03 10.42 10.30
2.5 34.20 1.08 14.86 16.71
3.5 23.16 6.25 0.00 9.80
a b b
Means 24.53 4.17 9.57
Values followed by same letters within a row are not significantly different at 5% level of probability

These results were consistent with the result of field experiment. There were different levels of NCP
on different blocks. The highest percentage of NCP was found on IPB C2 block. A total of 60.87% of
the plants on IPB C2 block were contaminated by IPB C20 pollen (table 2).

Table 2. Percentages of plants contaminated by IPB C20 pollen.

Percentages of contaminated plants
Genotypes Number of plants Contaminated plants
(%)
IPB C2 115 70 60.87a
IPB C5 120 11 9.17c
IPB C120 120 24 20.00b
Values followed by same letters within a column are not significantly different at 5% level of probability
Percent natural cross-pollination ranged from 9.80% to 16.71% depending on different planting
radius. However, there was no significant difference of NCP among different planting radius in this
experiment (table 1). It was indicated that wind was not potential factor for chili pepper pollinator. This
was reinforced by the result which showed that the pattern of pollen contamination were random and
irregular (figure 2). [2] reported that the extension of vegetation, infrequent winds and large population
of insect pollinator could be responsible for the high levels of NCP in chili pepper.

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IOP Conf. Series: Earth and Environmental Science 196 (2018)
1234567890 ‘’“” 012008 doi:10.1088/1755-1315/196/1/012008

Table 3. Anther length, carpel length and stigma exertion in three genotypes of chili pepper.
Anther length Carpel length Stigma exertion
Genotype
(mm) (mm) (mm)
IPB C2 2.36c 4.54b 1.01a
IPB C5 3.17a 3.32c -0.91b
IPB C120 2.90b 5.50a 0.99a
Values followed by same letters within a column are not significantly different at 5% level of probability

Figure 2. Pattern of natural cross-pollination on three genotypes of chili

peppers.

Observations on flower morphology showed significant differences on anther length, carpel length
and stigma exertion length among the chili pepper genotypes. IPB C5 genotype had the longest anther
length whereas IPB C120 genotype had the longest carpel length. The highest stigma exertion was
found on IPB C2 and IPB C120 genotypes (table 3). Stigma exertion length was probably one of the
factors which responsible for chili pepper NCP. There was significant positive correlation between
stigma exertion length and NCP percentages (figure 3). This caused IPB C2 genotype which had the
highest stigma exertion length also had the highest NCP percentages while IPB C5 genotypes which had
the lowest stigma exertion length also had the lowest NCP percentage. These results were closely
parallel with the findings from the previous studies. The positive correlation between stigma exertion
length and NCP percentage was also had been reported by [8] and [2] in chili pepper, [9] in Mimulus
ringens, and [10] in Datura stramonium.

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IOP Conf. Series: Earth and Environmental Science 196 (2018)
1234567890 ‘’“” 012008 doi:10.1088/1755-1315/196/1/012008

S 21.2341
100
R-Sq 6.0%
R-Sq(adj) 5.4%

80
Natural cross-pollination (%)

y = 13.96 + 4.03 x

60

40

20

-3 -2 -1 0 1 2 3
Stigma Exertion (mm)

Figure 3. Relationship between percent natural cross-pollination and

stigma exertion in chili pepper.

In this experiment, the observations were also made on levels of successful artificial cross-
pollination. There were significant differences on the percentage of successful artificial pollination
among the different genotypes of chili pepper. IPB C2 genotype had the highest percentage with 50.57
% successful artificial pollination, while the lowest percentage was resulted by IPB C5 genotype (table
4). This indicated that IPB C2 genotype had cross ability better than IPB C5 and IPB C120 genotypes.
This probably caused IPB C2 genotype which had the same stigma exertion length with IPB C120
genotype, but had the higher NCP percentages than IPB C120 genotype.

Table 4. Artificial-pollination success percentages on three genotypes of chili peppers.

Pollination success
Genotypes Number of pollination Pollination success percentages
(%)
IPB C2 176 89 50.57 a
IPB C5 133 19 14.29 c
IPB C120 193 59 30.57 b
Values followed by same letters within a column are not significantly different at 5% level of probability
Variation in natural cross-pollination has been attributed to a variety of factors, including:
asynchrony in flowering times [11,12]; distance to nearest neighbor [13,14]; pattern of pollinator
movement [13,11]; aspects of floral morphology [15,10,16]; and genetic differences in the level of self-
fertility [14]. However, stigma exertion length and level of cross ability were two main factors that were
responsible on chili peppers NCP in this experiment.
The levels difference of NCP will affect the breeding method of chili peppers. Chili pepper
genotypes which had a high level of NCP will likely to behave like cross-pollinated crops. It was also
likely to have high heterosis and inbreeding depression so it is very possible to be released as hybrid
varieties. [17] reported that hybrid variety IPB CH3 (using IPB C2 as female parent) produced higher of
fruit weight and fruit yield per plant than some commercial hybrids whereas [19] reported that IPB C2
genotype had higher heterosis and general combining ability for fruit weight character than the other

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IOP Conf. Series: Earth and Environmental Science 196 (2018)
1234567890 ‘’“” 012008 doi:10.1088/1755-1315/196/1/012008

genotypes. A good crossability in this genotype will also facilitate the hybrid seed production activities.
In contrast to this genotypes, chili pepper genotypes with a low level of NCP will likely to behave like
self-pollinated crops. So, it was also likely to have low heterosis and inbreeding depression and it is
very possible to be released as inbred line varieties. It was no wonder that IPB C120 genotype (local
variety of chili pepper) had long been used as inbreed line variety in Indonesia.

References
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Acknowledgement
Ministry Research, Technology, and Higher Education Republic of Indonesia and Center for Tropical
Horticulture Studies, Bogor, Agricultural University for providing full funding of the experiment and
genetic materials.