KEPROK (Citrus reticulata) GENETIC DIVERSITY ASSESSMENT BASED ON

MORPHOMETRIC STUDIES IN INDONESIA

Amelia Tridiptasari, Jamilatus Sa’diyah, Nurfitri Arfani, Nia Kurniawan

Ecology Laboratory of Mathematics and Natural Science
Brawijaya University, Malang
Email: atridiptasari@gmail.com

ABSTRAK
Indonesia is a tropical country that is known as a producer of citrus. Some areas in
Indonesia was known as a producer of citrus, especially keprok, namely in Kalimantan,
Sumatra, and Madura. The number keprok producing areas in Indonesia with different
geographical conditions can influence the diversity of the keprok. This study aims to determine
the genetic diversity of keprok (Citrus reticulata) from several regions in Indonesia based on
morphometric characteristics and the influence of the environment. Keprok used from
Situbondo, Jember, Madura, and other three areas from Bengkulu, Borneo, Pacitan were
sampled from Balitjestro, Batu, East Java. The result shows two groups of keprok based
morphometric proximity genetic basis, i.e. keprok group I of Pacitan, Situbondo, and Madura
and group II of Bengkulu, Jember, and Kalimantan.

Keywords: Citrus reticulata, Morphometric, Diversity, Environment

INTRODUCTION
Keprok orange (Citrus reticulata)
included in the family Rutaceae (Etebu &
Nwauzoma, 2014) is a horticultural crop
most traded in the world (Turner & Betty,
2013). This plant has an important role in
the world market and within the country,
either in the form of fresh or processed.
The development of citrus cultivation in
Indonesia has reached about 70-80%
annually. Development opportunities citrus
cultivation Indonesia at the ASEAN level
is quite good considering Indonesia
become Country with the largest harvested
area and production for citrus in ASEAN
(the Ministry of Agriculture, 2015).
Almost the entire territory of the 33
provinces in Indonesia has developed a
citrus. There are five areas that dominate
the production of oranges (70%) of the
overall production Indonesia, among
others: North Sumatra, East Java, South
Sumatra, Sulawesi Selatan, and West
Kalimantan (Morey, 2007). Based on
information from the Ministry of
Agriculture (2015) note that this type of
local oranges are cultivated in various
regions in Indonesia covers keprok (Citrus
reticulata / Nobilis L.), tangerine (C.
microcarpa L. and C.sinensis L.)
consisting of Siam Pontianak, Siam Garut,
Siam Lumajang, as well as a large citrus
(C. maxima Herr.) consisting of orange
Nambangan-Madison and Bali.
Citrus crop diversity in Indonesia is
influenced by various factors. These
factors include the different geographic
circumstances in various parts of
Indonesia, local citrus harvest unbalance
with needs of the domestic market
throughout the year, causing the entry of
imported oranges, and genetic factors that
plants (Hanif, 2015). All these factors
caused changes in the diversity of citrus
spread in Indonesia. In addition, the
identification of citrus plant taxonomy is
still difficult because of the many crosses
spontaneously and commercial (Turner,
2013).
 Identification of the citrus crop
diversity can basically be done in various
ways. One way is to use morphometric
analysis. Morphometric analysis is a way
to assess the phenotypic variation that is
due to biotic and abiotic factors. A
morphometric method performed by using
the size and the ratio of the size of an
organism to measure variations of a
biological specimen, and comparing the
genetic variations, environmental and
phenotype (Paris et.al, 2016). This method
has also been applied in plants and plant
organs for years (Cope, et al, 2012).
Morphometric methods, in general, has
been widely used to determine the kinship.
METHODS
Study Area
The citrus sample used in this study was
from six locations in Indonesia. These
locations include Bengkulu, West
Kalimantan, Pacitan, Jember, Situbondo,
6
5
3 4
Figure 1. Origin site of C. reticulata sampled (1) Bengkulu, (2) Kalimantan Barat, (3) Pacitan, (4) Jember,
(5) Situbondo, (6) Madura
The study was conducted in October-
December 2016. The samples of citrus fruit
Bengkulu, Pacitan, and Kalimantan
conducted at the Research Institute for
Citrus and Subtropical Fruit (Balitjestro),
of a particular species, the differentiation
of the various species, to determine the
variation of species, and for the
identification of a species (Soladoye et al,
2010). Based on the foregoing, the
research on "Keprok (Citrus reticulata)
Assessment Based On Genetic Diversity
Studies morphometric In Indonesia" needs
to be implemented.
The purpose of this study was to
determine the genetic diversity of citrus
fruits based morphometric and to explain
the influence of the environment on the
genetic diversity of citrus fruits based
morphometric.
and Madura. Keprok Bengkulu, Pacitan,
and West Kalimantan were used in this
study were taken from the Balitjestro,
Batu, East Java. Map location of the origin
of citrus fruits can be seen in Figure 1.
Batu, East Java. Keprok Jember, Situbondo,
and Madura were taken from the area of
origin of citrus fruits. Geographical
conditions of the area of origin of citrus
presented in Table 1.
Table 1. Geographically characteristics of C. reticulata
No Region Elevation Altitude Air Rainfall Fertility
(mdpl) temperate (mm/year)
(oC)
1 Bengkulu High 800-1000 19 2.500 Thriving
2 Pacitan Low 700 22 1000 Clay sanding
3 West Kalimantan Low < 500 22 2.800 Lightly thriving
4 Jember Low 100-500 23 1.969 Arid
5 Situbondo Low 250 24,7 994 Arid
6 Maduta Low 200-500 28 1.328 Arid

Measurement Data Morphometric

Citrus Fruits
This study uses descriptive
exploratory with quantitative and
qualitative approaches. Characters
measurement morphometric includes fruit
width (LB), fruit length (PB) (Bercu,
2016), the amount of flesh (SDB), the
width of flesh (A), the width rind
(mesocarp / albedo) (B), broad beans
(LBJ), and the number of seeds (SBJ). Part
of morphometric measured citrus fruit is
shown in Figure 2.
 reducing its dimensions. This is done by
eliminating the correlation between
independent variables through the
a
transformation of the independent variable
origin to new variables that are not
correlated at all. Based on the analysis of
the PCA, obtained a major component that
c
is able to retain most of the information
b which was measured using the total
diversity by using a bit of the main
components only. In this study,
Figure 2. (a) Length, (b) width, (c) mesocarp /
albedo
Abiotic factors data used in this
study include altitude, temperature, and
rainfall at each sampling locations keprok.
Measurement morphometric performed
using calipers and gauges while data from
literature abiotic Ministry of Agriculture
Agricultural Research and Development
Agency, 2015. The results of
morphometric measurements keprok in
millimeters (mm).
Data analysis
Citrus fruit morphometric
measurement results were tabulated and
compiled using Microsoft Excel.
Furthermore, the data used to determine
the kinship keprok by using software
PAST 3 with Principal Component
Analysis (PCA) includes the analysis with
dendrogram and Biplot analysis. PCA
analysis with dendrogram with Bray-
Curtis similarity scale can determine
kinship with Biplot keprok while analysis
to determine the character that supports the
grouping of keprok.
RESULT AND DISCUSSION
Comparison of morphometric characters in
this study was determined by Principal
Component Analysis (PCA) to see
morphometric characters that have
relevance to other characters. PCA aims to
simplify the observed variables by
the PCA is done by using biplot analysis similarities closest, and so on until it forms
and dendrogram analysis. a kind of trees where there is a hierarchy
Dendrogram Analysis (levels) which clearly between objects,
Dendrogram analysis is a from the most similar to least similar
multivariate analysis that aims to classify (Santoso, 2004).
the objects of the data based on similar Based on the dendrogram analysis
characteristics examined its (Mattjik, obtained measurement results
2011). The similarity of these morphometric (Citrus reticulata) were get
characteristics is usually measured using from different regions (Figure 3), which
the size of the proximity between objects showed that the index of similarity Bray-
that can be a similarity measure or Curtis, there are two clusters with
unsimilarity. similarity of 85% where cluster 1 consists
One technique of grouping is of keprok from Pacitan, Situbondo, and
engineering hierarchy, that began the Madura while the second cluster consists
grouping of two or more objects with of Kalimantan, Bengkulu, and Jember.

Figure 3. Dendrogram analysis of Citrus reticulata (1,2,3: Bengkulu; 4,5,6: Pacitan; 7,8,9:Kalimantan;
10,11,12: Madura; 13,14,15: Situbondo; 16,17,18: Jember)
Biplot Analysis
In cluster 1, showed that keprok Pacitan Grouping formed on dendrogram
and Situbondo have 97% similarity. Both analysis can be seen in two dimensions
of this keprok have similar characteristics using biplot analysis. Biplot analysis is
between the flesh of the fruit, altitude, and used to describe the line (object) and
also rainfall. In addition, in cluster 2, columns (variables) that exist in the data
between Kalimantan and Bengkulu has matrix together in a low-dimensional
93% similarity with the character of graph (usually two or three) (Ariawan, et
measurement similar to the albedo and al., 2013). This depiction includes
altitude. diversity and correlation between
variables, as well as the proximity between
objects that will be able to identify the
object grouping.
Biplot analysis is a statistical
technique that can descriptive multiple
dimensions simultaneously serving a
cluster object of observation and variables
in a graph on a plane two dimension so
Figure 4. Biplot analysis of Citrus reticulata
Keprok from Madura and Situbondo has
character temperatures were positively
correlated. While the individual variation
of keprok in Madura look bigger than the
individual variation in the area of
Situbondo. Madura and Situbondo are an
area with an average temperature of 24-
28oC. The difference in temperature at the
source of the keprok used in the study
essentially influence the differences in skin
color and flesh color areas that have
differences in the temperature range even
if the fruit of the same age (Figure 5), it is
due to the presence of pigment in the skin
and flesh that changes with changes in
temperature. Iglesias (2007) states that
changes chloroplasts to chromoplast led to
a decline of chlorophyll and carotenoid
pigments additions and changes the color
of skin and flesh from green to orange.
This happens because the ambient
temperature is low, the availability of
that the characteristics of the variable and
the object of observation and the variables
can be analyzed (Diyarti, 2003).
A group of keprok in various areas
based on biplot analysis in this study
showed variations of different groups in
various areas studied (Figure 4).
carbohydrates, the hormone ethylene,
causing greening occurrence. While the
high environmental temperature, nitrogen
availability, and hormone gibberellin cause
regreening. Character heights in this study
correlated inversely with the temperature
factor, ie the higher the temperature of an
area in the lower area.
Discoloration of the skin and flesh
of the fruit in each region in this study had
a parallel correlation. It supported the
statement Alquezar et al. (2008) that there
is a correlation parallels between changes
in skin color and flesh color keprok is
caused by the difference in carotenoid
pigments that are specific to the skin and
fruit, on citrus fruits are ripe, the
carotenoid content β, β-xanthophylls can
reach 98% of total carotenoids in flavedo
and albedo (Rodrigo in Alquezar et al.,
2008).

Figure 5. (a,c) peel color and flesh of Keprok
from Madura (280C), (b,d) peel color and flesh
of Keprok from Bengkulu (190C).
source: private documentation
Pacitan keprok characterized by
mesocarp (albedo), altitude, fruit length
and fruit width contrasting with Jember
keprok. Jember keprok characterized by
the amount of fruit flesh contrasting with
Pacitan. In this study note that the most
conservative character is the amount of
flesh.
Character fruit length, fruit width,
and the amount of flesh on each region
may be associated with nutritional
imbalance citrus that affects the size of a
keprok. Kays research results (1999) states
that the lack of intake of nitrogen,
potassium, zinc, and iron in citrus caused
the small size of keprok.
Bengkulu keprok seeds are
characterized by wide characters. This
characterization can be seen from the
length of the vector of the character that is
longer than the vector formed by other
groups. In addition to differences in seed
width characters, different characters skin
texture and grain of each region in the
observation were also observed. This
occurs because the character differences
citrus fruit maturation process that led to
the decline in the production of ethylene,
sensitivity, and respiration which gradually
led to changes in skin texture and
grapefruit (Iglesias, 2007).
Environmental factors that affect
the quality of the next keprok soil as a
growing medium and provider of plant
nutrients. Kalimantan and Bengkulu have
the character of rainfall with rainfall 2500-
2800 with fertile soil conditions. Iglesias
(2007) explains that a good ground for the
development of citrus is soil water content
and nutrients is always fulfilled all the
time with the availability of abundant
humus. With these findings, it can be seen
various characters from different areas of
keprok in Indonesia which can then be
used for the development of local keprok
in Indonesia.
CONCLUSION
Based on morphometric
measurements can be seen that there is a
genetic diversity of keprok (Citrus
reticulata) from various regions in
Indonesia with biplot analysis and
dendrogram analysis. In this study, we
obtained two clusters with 85% similarity
index where the first cluster consists of
Pacitan, Situbondo, and Madura while the
second cluster consists of Borneo, Madura,
and Bengkulu. Environmental influences
that cause the genetic diversity of citrus
fruits in various areas including air
temperature, altitude, and rainfall.
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