REGULATION OF FRUIT REIPENING USING PLANTS

GROWTH REGULATORS

By:
Dion Satrio Pambudi B1B015018
Hasnadhiazahra Rohadi B1B015028
Regita Antlia Putri B1B016037
Group :I
Subgroup :2
Assistant : Ryan Bagus Saputra

PRACTICAL REPORT OF PLANT PHYSIOLOGY II

MINISTRY OF RESEARCH, TECHNOLOGY, AND HIGHER EDUCATION

JENDERAL SOEDIRMAN UNIVERSITY
FACULTY BIOLOGY
PURWOKERTO

2017
 I. INTRODUCTION

A. Background

In the last few years, fruit-ripening research has challenged the classical
definitions of climacteric and non-climactericfleshy fruits (Paul et al., 2012). A
unique example of this controversy is the fig, Ficus carica L. The ripening process in
fig fruit is categorized as climacteric, showing a rise in respiration rate and ethylene
production at the onset of the ripening phase (Marei and Crane, 1971). Surprisingly,
ripening-related ethylene production increases following pre- or postharvest 1-
methylcyclopropene (1-MCP) application in an unexpected auto-inhibitory manner.
Moreover, postharvest 1-MCP treatment does not affect the ripening parameters of
the treated fruit (unlike other climacteric fruits), while application to fruit on the tree
improves fruit-storage abilities, inhibiting deterioration with minor effects on fruit
growth and ripening.
Based on their mode of ripening, fleshy fruits are divided into two categories,
climacteric and nonclimacteric, depending on the presence or absence of the
climacteric rise in respiration and of autocatalytic ethylene production (Lelivre et
al., 1997). In climacteric fruit, the plant hormone ethylene is the major cue that
controls most aspects of ripening. By contrast, the ripening of nonclimacteric fruit
does not strictly depend on ethylene, and the nature of the triggers of ripening in this
type of fruit remains yet to be elucidated. Since the upstream components of the
ethylene transduction pathway are common to all ethylene responses, the apparent
simplicity of the ethylene signaling pathway cannot account for the wide diversity of
ethylene responses. A plausible hypothesis is that differential responses to ethylene
are directed at the level of ethylene response factor (ERF) transcription factors,
which are encoded by one of the largest families of plant transcription factors, and
therefore, are most su ted to conferring such a large diversity and specificity of
ethylene responses (Mingchun et al., 2015).
The involvement of ethylene in fruit ripening was initially reported a long time
ago (Burg and Burg, 1962), and since then, direct evidences have accumulated to
demonstrate that ethylene mediates fruit ripening at the physiological, biochemical,
andmolecular levels. Altering ethylene at the level of its biosynthesis, perception,
signal transduction, or gene transcription was shown to impact fruit ripening (Liu et
al., 2014). According to the currently accepted model, ethylene signaling relies on a
linear transduction pathway where the hormone is perceived by a specific receptor,
which initiates a signaling cascade by releasing the block exerted by CTR1 on EIN2.
This activates a transcriptional cascade, involving EIN3/EIL1 as the primary
transcription factor and then ERFs, which in turn regulate genes underlying ripening-
related traits, such as color, firmness, aroma, taste, and postharvest shelf life (Chang
et al., 2013).
B. Purpose

To determine the concentration of growth regulators that able to accelerate the

rippening of fruit.
 II. REVIEW OF LITERATURE

As a developmental process, fruit ripening is coordinated by a complex network

of endogenous and exogenous cues. Indeed, the making of a fruit is a genetically
regulated process unique to plants involving three distinct stages: fruit set,
development, and ripening. Fruit development is characterized by a series of
developmental transitions tightly coordinated by a network of interacting genes and
signaling pathways. Among these, ripening has received the greatest attention from
both geneticists and breeders. From the scientific point of view, fruit ripening is seen
as a process in which the biochemistry and physiology of the organ are
developmentally altered to influence the appearance, texture, flavor, and aroma
(Giovannoni, 2004). Since most of the fruit sensory and nutritional quality traits are
elaborated at the ripening stage, deciphering the key genetic and molecular factors
regulating ripening becomes a major task toward improving overall fruit quality
(Carrari and Fernie, 2006). In addition, the cont ol of fruit ripening is also
instrumental to maintain the quality attributes of the fruit during the postharvest shelf
life (Liu et al., 2014).
According Abidin (1985), based on their amylum content, the fruit can be
divided into climacteric fruit and non-climacteric fruit. Climacteric fruit is the fruit
that contains a lot of amylum, such as banana, mango, apple, avocado and the
ripeness can be triggered by ethylene. Endogenous ethylene produced by fruits that
have ripened by itself can stimulate the ripening of the fruit ripened bunch. Non
climacteric fruit is a fruit that contains a little bit of amylum, such as oranges,
grapefruit, watermelon and pineapple. Provision of ethylene in the fruit can stimulate
respiration rate, but does not stimulate endogenous ethylene production and ripening
fruit.
Ethylen is a hydrocarbon is a saturated at room temperature in the form of gas.
These compounds can cause the important changes in the process of growth and
maturation results of agriculture. Ethylen is hormone grow in general is different
with auxin, giberelin and sitokinin. When normal conditions, ethylen will be in the
form of gas and the structure of the chemical is very simple once. In nature ethylen
role will be when the change occurs physiologically on a plants. This hormone will
play a role in the process of maturing fruit in phase klimaterik (Abidin, 1985).
Klimaterik is a sudden period is unique to a specific fruit where during this process
occurs making ethylen accompanied with the commencement of the process of
maturing fruit and fruit showed an increase of CO 2 that suddenly during the process
of maturing (Kusumo, 1990).
According to Abidin (1985), ethylen is growth hormone which in normal
conditions in the form of gas and have a very simple chemical structure that consists
of 2 carbon atoms and 4 hydrogen atoms. Ethylen classified as hormones that are
active in the process of maturing. There are two hypothesis about the relationship
between ethylen and maturing fruit. The first hypothesis, maturation is process that
resulted in the organization between the cells become impaired. This disorder is a
pioneer hidrolisa pati, chlorophyll, pektin and tannin by the enzymes in it that will
produce the ingredients such as ethylen, pigments, energy and polypeptide. The
second hypothesis, maturing in it as a phase of the end of the process of the
decomposition of substrate and the process which is needed by the ingredients to
synthesize specific enzymes, of which will be used in the process of whitered.
The concentration of ethylene used, ettilen types can be divided into three forms:
solid, liquid, and gas. Ethrel is one type of hormone with a liquid form in
concentrations below 50% whose role is to help speed up the ripening of fruit, when
the concentration used is too low, the effect of Ethrel itself will be low so as not so
affect the ripening of fruit, as fruit ripening is aided by the Ethrel. Another type of
other fruit ripening hormone is a solid form with a concentration above 50%, such
that carbide is commonly known by the public, although it is solid but actually when
objects and react to a chemical reaction occurs meal will form gases, function like
Ethrel. Another form is a gas, for example ethylene gas concentrations below 10%, it
is generally able to solve chlorophyll ethylene work on young fruit to cause the red
or orange, because chlorophyll has been reduced by ethylene gas. As a result of the
excess ethylene will hinder the growth of plants (inhibits the elongation of plant),
inhibits the growth and development of roots, leaves, stems, and flowers (Wang et
al., 2002).
The research of Annona squamosa L.is one of the perishable climateric fruit. The
aims of this research were to study the ethylene levels, physiological and
biochemical characters with application of temperatures and chitosan during storage.
The treatment of temperature 25 and chitosan 0% was used as controls. The
parameters observed included in respiration rate, weight loss, levels of ethylene,
pigments, fruit hardness, ascorbic acid content and sugar reduction. Sugarapple fruit
were taken from the same tree. The results showed that the treatment of temperature
and chitosan affects the carotenoid levels in skin of sugar apple fruit. The treatment
of temperature and chitosan is able affect the vitamin C which content in the sugar
apple fruit. The higher of the temperature, the vitamin C content of fruit sugar apple
also higher. While the results of this research show that levels of ethylene at 25 C,
ethylene levels higher than the temperature. The higher levels of chitosan, also makes
the lower levels of ethylene. Ethylene regulate fruit ripening by coordinating the
expression of genes responsible for various processes including respiration rate
increase, autocatalytic production of ethylene, the degradation of chlorophyll,
carotenoid synthesis, the conversion of starch into sugar, and increased activity of
enzymes breaking the cell wall (Mudyantini et al., 2015).
The color change is most notable changes during maturation, occurs synthesis of
certain pigments, such as carotenoids and flavonoids in addition occur revamp
chlorophyll. The color of ripe fruit is caused by the synthesis of carotenoids and
anthocyanins. In the mature period marked by the reduction carotene. For most fruits,
the first sign of maturity is the loss of green color. Chlorophyll content of fruit that
was ripe gradually decreased, in general, a number of permanent green dye contained
in fruits, especially in tissue sections in fruit. His or her fruit caused by the reshuffle
protopektin insoluble into soluble pectin or hydrolysis of starch (such as pumpkin) or
fat (like the avocado.) Synthesis wood substance on some vegetables such as fruit
can also adversely affect the texture (Mudyantini, 2015).
 III. MATERIALS AND METHODS

A. Materials

The tools used in this laboratory activity are measuring glass, stir bar, beaker
glass and analytical scale.
The materials used in this laboratory activity are banan kapok, ethrel (2
chloroetylphosponic acid), papers and distilled water.
B. Methods

1. Make ethrel solution with a concentration of 300 ppm, 600 ppm and 900
ppm.
2. Dip the fruits in each concentration for 5 minutes.
3. Wrap the fruit with paper and put it into the storage area.
4. Observe each day during 7 days and note the change that occur (color,
texture, taste and aroma.
 IV. RESULT AND DISCUSSION

A. Result

Table 4.1.1 Observation of Bananas Ripening Entourage I group 1

concentratio Color Aroma

n 1 2 3 4 5 6 7 1 2 3 4 5 6 7
0 ppm + + + + + + + + + + + + + +
+ + + + + + + + + + + + + +
300 ppm
+ + + + + + + + +
+ + + + + + + + + + + + + +
600 ppm
+
+ + + + + + + + + + + + + +
900 ppm
+ + + + + + + + +

concentratio Texture Taste

n 1 2 3 4 5 6 7 1 2 3 4 5 6 7
0 ppm + + + + + + + + + + + + + +
+ + + + + + + + + + + + + +
300 ppm
+ + + + + + + +
600 ppm + + + + + + + + + + + + + +
+ + + + + + + + + + + + + +
900 ppm
+ + + + + + + +

Interpretation :
+ : Change of color, texture, and aroma low
++ : Change of color, texture, and aroma medium
+++ : Change of color, texture, and aroma high
++++ : Change of color, texture, and aroma very high

Picture 1. Banana 0, 300, 600, 900 ppm Picture 2. Banana 0 ppm (18/3/2017)
Picture 3. Banana 300 ppm (18/3/2017) Picture 4. Banana 600 ppm (18/3/2017)

Picture 5. Banana 900 ppm (18/3/2017) Picture 6. Banana 0 ppm (22/3/2017)

Picture 7. Banana 300 ppm (22/3/2017) Picture 6. Banana 600 ppm (22/3/2017)

Picture 6. Banana 900 ppm (22/3/2017)

 B. Discussion

Based practicum can be concluded that the ethylene concentration of 300 and
900 were the most effective. Due on day 7 pieces with a concentration of 300 and
900 already experiencing changes in color and texture that turns into soft. In addition
to the flavor of the fruit is already sweet and feasible to eat. According Andre (2012)
that the pace of work on fruit ripening ethylene will significant when the
concentration used enhanced and storage process suitable enough oxygen. Reference
data in accordance with Andre (2012) for the parameters of color and flavor, where
the entire treatment the concentration levels showed significant results and nice.
Although external factors such as the content of oxygen in the storage environment
affecting ethilen working speed can affect the results, but overall there is a parameter
which indicates a positive value for this ethylene work.
Mechanism of action of ethylene in ripening fruit that is by adding ethylene from
outside. Among the many changes caused by ethylene are changes in cell membrane
permeability, resulting in the destruction of chlorophyll in chloroplasts by enzymes.
With the destruction of pigment chlorophyll in the cells of the fruit is not protected
so that the fruit ripening color appeared. Ethylene biosynthesis occurs in the plant
tissue that is a change of the amino acid methionine starting compound for the help
of light and FMN (Flavin Mono Nucleotides) becomes methionil. The compound
was taken over by the aid of light and FMN become Ethylene, methyl disulphide and
formic acid. (Sumarjono, 1981).
In the process of maturation of the fruit organoleptic changes occur that include
texture, flavor, and the flavor and color. Changes in texture to the fruit occurs due to
degradation of hemicellulose and protopektin. Softening the texture of the fruit due
to a change protopektin on fruit unripe into a water-insoluble water-soluble pectin
during fruit ripening. In fruits are phenolic substances, phenolic substance that is the
determinant of flavor. Reduced levels of phenolics in fruits cause astringent taste and
masamnya was reduced. The first sign of maturity of fruit color on the fruit is the
loss of green color. The color change is most notable changes during fruit ripening.
In addition to going on an overhaul of chlorophyll in this process occurred synthesis
of certain pigments, such as the carotenoid pigments that already exist but have not
yet appear to be visible at the time of ripening fruit and the fruit becomes yellow.
Color changes in fruits due to the loss of chlorophyll and other pigments cause seems
colors contained in the fruit (Kevany et al., 2007).
In the research of Liu et al., (2015) which is using tomato (Solanum
lycopersicum) as a reference species, the literature aims to revisit the mechanisms by
which ethylene regulates fruit ripening by which this hormone coordinates the
ripening process and emphasizes the interplay between ethylene and ripening-
associated developmental factors and the link between epigenetic regulation and
ethylene during fruit ripening. Altering ethylene at the level of its biosynthesis,
perception, signal transduction, or gene transcription was shown to impact fruit
ripening. It has long been considered that other plant hormones besides ethylene are
likely required for climacteric fruit ripening such as ABA is known to promote
ripening, whereas auxin seems to have anantagonistic effect. Down-regulation of the
key ABA biosynthesis enzyme 9-cis-epoxycarotenoid dioxygenase 1 in tomato fruit
resulted in altered rmness and color but surprisingly higher ethylene production,
indicating the complexity of the ABA/ethylene interplay during ripening. The
expression of ethylene biosynthesis and signaling genes is regulated by auxin in
tomato and other eshy fruits, such as peach.
Factors that affect the activity of ethylene gas by Ridhyanty et al., (2015), that
the stimulation of the maturation are often performed with ethylene gas, carbide, and
Ethrel / ethepon. Antigens will trigger the maturation of the fruit to ethylene work
then triggers the ripening process in the fruit. Ethrel or better known as ethepon is a
chemical compound that serves to trigger growth. Its use varies from one type of
plant or fruit, the concentrations used, it is also a time to use. However, the use
ethepon among farmers or banana traders are still very rare compared with the use of
acetylene. Ethepon very quickly converted into ethylene in plants / fruits, but it also
has a very low toxicity level, so that the residue is not harmful to humans.

V. CONCLUSION AND SUGGESTIONS

A. Conclusion

1. The effective concentration in the ripening of bananas in this lab showed

ethylene concentrations of 300 and 900 is better than 300 concentration.
2. The changes that occur in the fruit during the ripening, seen constantly
increasing every day.
B. Suggestions

1. Itll better if the student can learn how to make ethylene concentration from
0, 300, 600, 900 ppm.
 2. Or if the student cant make ethylene, the assistant can demonstrated how to
make ethylene hormones.

REFERENCES

Abidin, Z. 1985. Dasar-Dasar knowledge about Organizing Grow. Space: Bandung.

Andre, Veliarry. 2012. Kelebihan dan Kekurangan Hormon pada Tanaman.

Grammedia: Jakarta.

Burg SP, Burg EA (1962) Role of ethylene in fruit ripening. Plant Physiol 37: 179
189.

Chang KN, Zhong S, Weirauch MT, Hon G, Pelizzola M, Li H, Huang SS, Schmitz
RJ, Urich MA, Kuo D, et al . 2013. Temporal transcriptional response to
 ethylene gas drives growth hormone cross-regulation in Arabidopsis. eLife 2:
e00675.

Kevany BM, Tieman DM, Taylor MG, Cin VD, Klee HJ. 2007. Ethylene receptor
degradation controls the timing of ripening in tomato fruit. The Plant Journal
51, 458467.

Kusumo, S. 1990. Organizing the plant crops. Yasaguna: Jakarta.

Lelivre JM, Latch A, Jones B, Bouzayen M, Pech JC. 1997. Ethylene and fruit
ripening. Physiol Plant 101: 727739.

Liu M, Diretto G, Pirrello J, Roustan JP, Li Z, Giuliano G, Regad F, Bouzayen M .

2014. The chimeric repressor version of an Ethylene Response Factor (ERF)
family member, Sl-ERF.B3, shows contrasting effects on tomato fruit ripening.
New Phytol 203: 206218.

Marei N, Crane JC. 1971. Growth and respiratory response of fig (Ficus carica L. cv.
Mission) fruits to ethylene. Plant Physiology 48, 249254.

Mingchun Liu, Julien Pirrello, Christian Chervin, Jean-Paul Roustan, and Mondher
Bouzayen. 2015. Ethylene Control of Fruit Ripening: Revisiting the Complex
Network of Transcriptional Regulation. Plant Physiology: Vol. 169, pp. 2380
2390.

Mudyantini, Widya, Anggarwulan, E., Rahayu, P. 2015. Ripening inhibition of sugar-

apple fruit (Annona squamosa L.) with low temperature and chitosan coating.
AGRIC Journal: Vol. 27: 23-29.

Paul V, Pandey R, Srivastava G. 2012. The fading distinctions between classical

patterns of ripening in climacteric and non-climacteric fruit and the ubiquity of
ethylenean overview. Journal of Food Science and Technology:49, 121.
Ridhyanty, S. P., Julianti, E., Lubis, L. M. 2015. Pengaruh Pemberian Ethepon
Sebagai Bahan Perangsang Pematangan Terhadap Mutu Buah Pisang
Barangan (Musa paradisiaca L). Program Studi Ilmu dan Teknologi Pangan
Fakultas Pertanian USU Medan. J.Rekayasa Pangan dan Pert., Vol.3 No.1
Th. 2015.

Wang K, Li H, Ecker J; Li; Ecker. 2002. Ethylene Biosynthesis and Signaling

Networks. Plant Cell 14: S13151. doi:10.1105/tpc.001768.