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Ethylene Biosynthesis in Tropical

Fruits
Controlling the Ripening Process
Via Postharvest Technology
Presenter K D Golden
Objectives

1.To determine ethylene production at the


various stages of ripening.

2.To enhance the shelf life of bananas using


postharvest technology
Introduction

Effects of ethylene on plants include:-


a) Ripening
b) Senescence (aging)
c) Abscission
d) Promotion or inhibition of cell elongation
e) Promotion or inhibition of flowering
f) Stress related responses (wounding &
flooding)
Introduction contd

Controlling the ripening process in fruits has been a problem for


many years. In recent times techniques such as genetic
engineering and post harvest technology had been used to control
the ripening process.
In this study we looked at the respiration rate and ethylene
biosynthesis in bananas (cv giant Cavendish), and how postharvest
technology can enhance the shelf life of bananas. Ethylene
synthesis in Carica papaya and Artocarpus altilis (breadfruit) was
also determined during the ripening process.

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How is Ethylene synthesized in plants

a) Methionine 1 SAM
b) SAM 2 ACC
c) ACC 3 ethylene
1 = s- Adenosyl Methionine Transferase
2 = ACC Synthase
3 = ACC Oxidase
ACC = 1-amino cyclopropane 1-carboxylic acid
SAM = S-adenosyl methionine
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The conversion of ACC into ethylene

ACC Oxidase
COOH Ethylene +
NH2 + O2 + Fe 2+ HCN + CO2

ascorbate

Ethylene ethylene oxidase Ethylene oxide

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Inhibitors of ethylene biosynthesis

ACC synthase is inhibited by:-


a) Amino oxyacetic acid
b) Amino ethyloxy vinylglycine
ACC oxidase is inhibited by:-
a) Cobaltous ions
b) N propyl gallate
c) Dintophenol
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Postharvest technology

Controlled atmosphere is most commonly used to


slow the ripening of fruits and to slow the decay of
vegetables
This can be done by modification of the
oxygen/carbon dioxide concentration of the
atmosphere
Low oxygen concentration (3-4%) slows ripening
because it reduces the autocatalytic production of
ethylene.
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Postharvest technology contd

For controlled atmosphere experiments, the use of a


semipermeable membrane was developed by
Marcelli and Leteinturier (1966-1967).
The membrane is an elastomer of silicone which
has the ability to exchange gases at different rates
depending on their chemical and physical
properties.
At 1 atm its permeability to CO2 is 1750 dm3 .day
-1.M -2 ; O is 320 dm3. day -1.M-2; ethylene is 700
2
dm3 .day -1.M -2.
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Postharvest technology contd

Procedures to calculate the membrane area (cm2) required for a


particular storage chamber are based on the level of CO 2 desired in the
chamber. They are summarized by the following equation:-
RR x 24h DAY 1 x M
Area = ---------------------------
P CO2 x %O2 x 1.977
RR= respiration rate (mg CO2 evolved kg 1 h 1)
M= mass of stored product (kg)
P CO2 = permeability of silicone membrane to CO2
%O2 = O2 gradient across membrane(%), 1.977 = conversion factor

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Typical chamber

Silicone Septum for gas


membrane sampling

PVC pipe. I.D. =


0.25M, L=50 cm

Threaded rod

Fruit
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Methods

Ethylene production was measured by a Pye Unicam (204)


gas chromatograph (GC) fitted with a flame ionization
detector. GC parameters were as follows: column temp =
70 oC, injector temp = 120 oC, detector temp= 250 oC and
carrier gas flow rate (N2) = 30 ml / min.
The respiration rate (RR) was determined by measuring the
amount of O2 consumed or the amount of CO2 evolved (mg
kg -1 h - 1 ) by a Hewlett Packard GC fitted with a thermal
conductivity detector.

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Ethylene Standard Curve
250
Area under curve
(counts / V)
200
150
100
50
0
0 5 10 15 20 25
Ethylene ( nmol)

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Silicone Membrane System
The use of a silicone membrane that is fitted to a chamber (50
cm x 25 cm i.d.) can be used to modify the composition of the
air (3% O2 and 5% CO2) and prevent ethylene - stimulated ripening
of Giant Cavendish bananas. Experiments were carried out in a cold
room at 15C and terminated after 6 weeks.

Silicone
membrane

Chamber
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Diffusion Channel System
Diffusion channels, installed on laboratory scale chambers, were
used to provide and maintain controlled atmosphere (CA) conditions
(3% O2 and 5% CO2) to prevent ethylene - stimulated ripening of
Giant Cavendish bananas. Experiments were carried out in a cold
room at 15C and terminated after 6 weeks.

Diffusion
channel

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Results I

Ethylene biosynthesis during the


ripening stages of Carica papaya and
Artocarpus altilis (breadfruit)
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Results II

Results showings the enhancement of the shelf-life


of bananas using controlled atmosphere (CA)
Respiration rate and ethylene production at 15 0C

60 8
Res piration rat e

Ethylene production
g .h )
-1 -1

g .h )
-1
45 6

-1
A
2.k

l .k
gCO

d(
30 4

e
te(m

rodc
u
nra

nep
tio

le
ira

15 2

thy
p

E
Rs
e

0 0
0 4 8 14 25 32 37

Time (days)

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Respiration rate and ethylene production at 25 0C

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Respiration rate B
120
.h )
-1 Ethylene production

Ethyleneproduced( l. kg .h )
-1
12
-1
g

-1
2.k
espirationrate (mgCO

80
8

40
4
R

0 0
0 3 5 7 10 12 14 16 18

Time (days)

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Bananas stored at CA using diffusion channel

Figure 2 Green bananas after storage Figure 3 After CA storage in diffusion channels,
using diffusion channels for 6 weeks fruit in Figure 2 were induced to ripen at room
temperature

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Bananas stored at CA using silicone membrane

Figure 3 Green bananas after CA storage Figure 4 After CA storage using silicone membranes, fruit in Figure 4 above
with silicone membranes for 6 weeks were induced to ripen at room temperature.

Results showed that an atmospheric composition of 3.5% CO2 and 3% O2 was most suitable for preserving banana quality. Fruits stored in
this atmosphere remained in an unripe state for 6 weeks and were of superior quality to controls and fruits stored in different CO 2/ O2 atmospheres.
Fruit stored at RA were fully ripe after 29 days.

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Thiabendazole Treatment

Figure7. Bananas treated with Figure 8. Bananas that were not


thiabendazole (100 ppm) treated with thiabendazole
before storage at CA before storage at CA.

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Discussion
1. The primary benefit of the CA/MA storage system is that the fruit will maintain its freshness
and quality for a longer period of time than it would if stored under regular atmosphere.
Hence more foreign exchange can be earned due to less spoilage. Indeed this technique can
be a good alternative to genetically engineered fruits, taking into account the ever growing
resistance to genetically engineered products in the UK.

2. There is also considerable use of chemicals in postharvest storage, particularly to protect the
fruit from insects, bacterial and fungal pathogens and to reduce or inhibit ripening. The
widespread use of these chemicals should be restricted, due to cost and health implications.

3. The main objective was achieved. The shelf-life of the fruit was enhanced, while maintaining
optimum quality of the fruit. Ethylene production was implicated in the ripening process of
fruits.

4. The techniques used for enhancing the shelf-life of bananas were successful:-
a) silicone membrane, which is differentially permeable to gases.
b) the diffusion channel system which is based on the principle of diffusion of gases through
channels 25
Future work
1. To apply the technology to
other fruits such as naseberry,
mangoes, starapples etc.
2. To scale-up the chambers to
store larger quantity of fruits.

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Acknowledgement

Thanks to:-
Opal Williams -Stewart ( UWI Mona)
Prof V Raghavan (Chair, Dept of Agriculture &
Biosystems engineering McGill university)
Yvan Gariepy (McGill University)
Valerie Orsat (McGill University)
F. McGilchrist (Physiology work shop, UWI Mona)
W. Campbell (Physiology work shop, UWI Mona)
References

H M Dunkley & K D Golden (1998). ACC oxidase from Carica papaya, isolation
and characterization. Physiologia Plantarum. 103. 225-232.

W J Lipton (1980). Controlled atmospheres for vegetables and fruit- Why and
When. In Postharvest biology & handling of fruits & vegetables. Editors N F Haard
& D K Salunke. The AVI Publishing company, INC. Westport, Connecticut.

O J Williams & K D Golden (2002). ACC oxidase from Artocarpus altilis


(breadfruit). Plant Physiology & biochemistry. 40. 273-278.\

O J Williams, G S Raghavan, K D Golden and Y Gariepy. 2003. Postharvest storage


of giant Cavendish bananas using ethylene oxide and sulphur dioxide. Journal of
Science Food and Agriculture. 83. 180 186.

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