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Agar-agar a promising edible coating agent for management of postharvest
diseases and improving banana fruit quality
Article in Journal of Plant Protection Research · October 2018
DOI: 10.24425/122938
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Jour nal o f P la n t Pro t e ct io n R e s e a rc h ISSN 1427-4345
ORIGINAL ARTICLE
Agar-agar a promising edible coating agent for management
of postharvest diseases and improving banana fruit quality
El Sayed Hussein Ziedan1*, Hassan Mohamed El Zahaby2,
Hanafey Farouk Maswada2, El Hassan Abd El Rafh Zoeir2
1
Department of Plant Pathology, National Research Centre, Dokki, Cairo, Egypt
2
Department of Agricultural Botany, Faculty of Agriculture, Tanta University, Tanta, Egypt
Vol. 58, No. 3: xx–xx, 2018
DOI: 10.24425/122938
Abstract
This study was executed to investigate the potential of agar-agar, a nontoxic and non-de-
Received: March 21, 2018 gradable gelling agent, as a promising coating agent to improve and protect banana fruit
Accepted: August 1, 2018 against fungal postharvest diseases i.e., crown, finger, neck and flower end rots which are
caused by fungal isolates of Colletotrichum musae and Fusarium moniliforme. Coated-ba-
*Corresponding address: nana fruit samples with different concentrations of agar-agar suspension particularly at
ziedanehe@yahoo.com 2.0 g · l−1 exhibited a significant reduction in incidence and severity of postharvest diseases
compared to untreated fruit. Banana fruits dipped in agar suspension at 2.0 g · l−1 for 5, 10
and 15 min showed significant reduction in disease incidence and severity. Moreover,
application of agar suspension as a coating agent at 2.0 g · l−1 significantly decreased
weight loss (%), firmness loss (%), and soluble solid concentration of banana fruit for
15 days at 25 ± 2°C. Scanning electron microscopy observation confirmed that the fruit
coated with agar colloid at 2.0 g · l−1 had significantly fewer cracks and showed smoother
surfaces than untreated fruit. This explains the quality improvement in agar-coated fruit
compared to uncoated fruit. Overall, agar colloid, a safe coating agent, could be used to
protect banana fruit against postharvest rot diseases and extend fruit storage life during
ripening and storage.
Key words: agar, banana, disease, fruit, fungi, storage
Introduction
Banana (Musa spp.) is one of the most popular fruits Iqbal 2016; Zoeir et al. 2017b). Natural coating agents
worldwide. Ripening banana fruit has a relatively short are composed of polysaccharides, proteins, lipids, and
postharvest shelf-life due to fungal infection (Hossain composites. The most common edible coating agents
and Iqbal 2016). Colletotrichum musae and Fusarium include waxes, chitosan, gelatin and gums which help
moniliforme are the major fungi causing postharvest suppress decay during postharvest storage and improve
diseases of banana fruit i.e., crown rot, finger rot, neck fruit quality (Ali et al. 2010; Cháfer et al. 2012; Dhall
rot and flower end rot (Zoeir et al. 2017a). Applica- 2013). Agar is a food additive of universal use consid-
tion of fungicides is expensive, has harmful effects on ered in the USA as GRAS (Generally Recognized as
human health and the environment, and becomes less Safe) by the FDA (Food and Drug Administration)
effective after prolonged use (Lassois et al. 2008). Fun- (FDA 1972). Agaro-polysaccharides can be used as
gicide alternatives such as essential oils, salts, organ- a functional food to prevent inflammatory diseases
ic acids and chitosan have been used for controlling (Enoki et al. 2010). Moreover, enzymatic hydrolysis
postharvest rot diseases of banana fruit (Hossain and is not relevant since there are few agarases (enzymes
2 Journal of Plant Protection Research 58 (3), 2018
that break down agaroses), found only in marine bac- served as a control. Fruit samples were incubated at
teria, in a few bacilli that are not normally found in 23 ± 2°C for 15 days. Postharvest disease incidence
food products (Armisen and Galatas 2000). Since it was calculated of diseased fruits showing symptoms
is non-degradable by microorganisms it is used in the of crown rot, neck rot, finger rot and flower end rot
preparation of microbiological and plant tissue culture according to Zoeir et al. (2017a, b) as follows:
media. The aim of this study was to evaluate agar-agar
Disease [%] Number of diseased fruits
suspension as an edible coating agent for the manage- u 100.
ment of postharvest rot diseases of banana fruit while incidence Total number of banana fruit
maintaining fruit quality. Disease severity was ranked by observing the per-
centage of rotten symptoms based on a linear scale
from (0−4) as follows:
0 = healthy fruit, free of rot and discoloration,
Materials and Methods
1 = 1−25% rotten and discoloration area,
2 = 26−50% rotten and discoloration area,
Plant material and pathogenic fungi 3 = 51−75% rotten and discoloration area,
Healthy ripe uniform banana fruits (Musa acumina- 4 = 76−100% rotten and discoloration area.
ta cv. Balady), the most susceptible cultivar to post-
harvest rot diseases were obtained from orchards in
El-Gharbiya Governorate, Egypt. Highly aggressive Determination of some physicochemical
isolates of Colletotrichum musae and Fusarium mon- properties of banana fruits
iliforme, which cause major postharvest diseases of For weight loss determination, five fruit samples in
banana fruit (Zoeir et al. 2017a), were provided by the each replicate for each treatment were marked be-
Agricultural Botany Department, Faculty of Agricul- fore storage and weighed using a digital balance. The
ture, Tanta University, Tanta, Egypt. same fruit samples were weighed at the beginning of
the experiment and after storage period s of 0, 10 and
15 days. The fruit weight loss percentage was calculat-
Source and preparation of agar suspensions ed using the following formula (El-Sharony and Amin
Agar-agar, a polymer made up of a subunit of galactose 2015):
sugar extracted from red-purple marine algae, mainly W1 W2
Weight loss [%] u 100,
Gelidium amansii, was provided by the Chemical In- W1
dustrial Development Company (CID), Egypt. Differ- where: W1 and W2 − initial weights and weight at spe-
ent concentrations of agar colloid at 0.5, 1.0 and 2 g · l−1 cific intervals (5, 10 and 15 days), respectively.
(w/v) were prepared by dissolution of each amount of
agar in boiling distilled water for 1−2 min, then cooled Peel tissues from one side of banana fingers were
to 50°C. removed and pulp firmness measurements were taken
at three different points using a fruit firmness tester
(Fruit Pressure Tester). Firmness values were the force
required (kg) for complete penetration of 1 cm (Ra-
Effect of agar colloid on postharvest diseases
nasinghe et al. 2005). Firmness loss (%) was calculated
of banana fruits
as follows:
Banana fruits cv. Balady were disinfected by double F1 F2
Firmness loss [%] u 100,
immersion in 2% of 70% ethanol for 5 min and allowed F1
to dry at room temperature under sterile conditions.
where: F1 and F2 − initial firmness and firmness at spe-
Fruit samples were placed separately in polyethylene
cific intervals (5, 10 and 15 days), respectively.
bags that had been previously disinfected with 70%
ethanol and exposed to UV light for 20 min. Fruit Soluble solid concentration (SSC) content of ba-
samples were dipped in different concentrations of nana fruit pulp was estimated using Abbe’s refractom-
agar colloids, 0.0, 0.5, 1.0 and 2.0 g · l−1 for 5 min at eter. According to Akter et al. (2013), a drop of banana
40−50°C. In addition, different dipping times (5, 10 juice squeezed from the fruit pulp on the prism of the
and 15 min) of banana fruit in agar suspension (2 g · l−1) refractometer and the percent of soluble solid content
at 40−50°C were tested. Banana fruit samples were were recorded as % Brix from direct reading of the in-
infested by mixtures (1 : 1) of 106 · ml−1 spore suspen- strument. Temperature corrections were made using
sions of either C. musae or F. moniliforme. Ten fruits the temperature correction chart that accompanied
were used as replicates and ten fruit free treatments the instrument.
El Sayed Hussein Ziedan et al.: Agar-agar a promising edible coating agent for management of postharvest diseases… 3
Scanning electron microscopy (SEM) Results
observations
Discs (4 mm) of the cortex of banana fruit that had Effect of agar concentrations on banana
been previously dipped in agar suspension (2 g · l−1 postharvest diseases
for 10 min) and artificially infested with the causal or-
ganisms, then incubated at 23−25°C for 5 days were Three concentrations of agar suspension i.e., 0.5, 1.0
scanned by Scanning Electron Microscopy quanta and 2.0 g · l−1 were tested against postharvest disease
FEG250 field emission at the National Research Cent- incidence (%) by dipping banana fruit cv. Balady for
er unit, Egypt as follows: cortex discs were fixed in 5 min before artificial infestation by causal pathogen
buffered osmium tetroxide (2%), then dehydrated by inocula of C. musae and F. moniliforme. Data (Fig. 1)
a graded series of ethanol (25, 50, 75, and two 100%) indicated that all agar concentrations significantly sup-
once for 10 min at each step, then coated with gold and pressed postharvest rot diseases of banana fruit com-
viewed by SEM (Pathan et al. 2010). pared to control 10 days after infestation at 25 ± 2°C.
In addition, all agar concentrations suppressed disease
Statistical analysis incidence (%) and severity of crown, neck, finger and
The experiment was conducted using a completely flower end rot compared to control 15 days after in-
randomized design with three replications. Data set was festation (Table 1). It was observed that the reduction
statistically analyzed by analysis of variance (ANOVA) of postharvest disease incidence and severity was in-
technique using computer software SAS program (SAS creased with increasing agar concentrations. An agar
Institute, Cary, NC, USA). Duncan’s multiple range tests suspension at 2 g · l−1 was the best treatment for reduc-
were used to compare differences between treatments at ing incidence and severity of all postharvest diseases
p ≤ 0.05 of each variable (Snedecor and Cochran 1980). of banana fruit.
Control 0.5 g · l–1 1.0 g · l–1 2.0 g · l–1
Fig. 1. Effect of different concentrations of agar suspensions on postharvest diseases of banana fruit after 10 days of storage
at 25 ± 2°C
Table 1. Effect of different agar suspension concentrations on postharvest disease incidence (%) and severity (DS) of banana fruit
15 days after infestation at 25 ± 2°C
Postharvest rot disease incidence [%] and severity (DS)
Treatment crown rot neck rot finger rot flower end rot
[%] DS [%] DS [%] DS [%] DS
Control 100.0 a 4.0 a 100.0 a 4.0 a 100.0 a 4.0 a 100.0 a 4.0 a
Agar [0.5 g · l ]
−1
100.0 a 4.0 a 60.0 b 3.0 b 100.0 a 4.0 a 70.0 c 3.0 b
Agar [1.0 g · l ]
−1
80.0 b 4.0 a 60.0 b 3.0 b 100.0 a 4.0 a 60.0 b 3.0 b
Agar [2.0 g · l ]
−1
40.0 c 2.0 b 20.0 c 1.0 c 30.0 b 2.0 b 20.0 d 1.0 c
Values in each column followed by the same letter are not significantly different at p ≤ 0.05 according to Duncan’s multiple range
4 Journal of Plant Protection Research 58 (3), 2018
times of banana fruit in agar colloid (2 g · l−1) signifi-
Effect of dipping time of banana fruit in agar
cantly reduced all postharvest rot diseases of banana
suspension on banana postharvest diseases
fruit compared to control 15 days after artificial in-
Different dipping times of banana fruit i.e., 5, 10 and festation by causal pathogens. Moreover, the disease
15 min in agar suspension at 2 g · l−1 were tested (Ta- incidence (%) and severity decreased with increasing
ble 2). Data obtained indicated that different dipping dipping times (Table 2).
Table 2. Effect of different dipping times in agar suspension (2 g · l−1) on postharvest rot disease incidence and severity of banana fruit
15 days after infestation at 25 ± 2°C
Dipping Postharvest rot disease incidence [%] and severity (DS)
Treatment time crown rot neck rot finger rot flower end rot
[min] [%] DS [%] DS [%] DS [%] DS
Control – 100 a 4.0 a 70.0 a 3.0 a 100 a 4.0 a 80.0 a 4.0 a
Agar colloid 5 70.0 b 3.0 b 50.0 b 2.0 b 80.0 b 4.0 a 30.0 b 2.0 b
[2 g · l ]
−1 10 50.0 c 2.0 c 40.0 c 2.0 b 0.0 c 0.0 b 0.0 c 0.0 c
15 50.0 c 2.0 c 30.0 d 2.0 b 0.0 c 0.0 b 0.0 c 0.0 c
Values in each column followed by the same letter are not significantly different at p ≤ 0.05 according to Duncan’s multiple range
Fig. 2. Effect of agar suspension [2 g · l−1] on (A) weight loss [%], (B) firmness loss [%] and (C) soluble solid
concentration (Brixº) of banana fruit during different storage time (5, 10 and 15 days) at 25 ± 2°C (Values are
means ± SE. Values followed by the same letter are not significantly different at p ≤ 0.05 according to Duncan’s
multiple range)
El Sayed Hussein Ziedan et al.: Agar-agar a promising edible coating agent for management of postharvest diseases… 5
Fig. 3. Scanning Electron Microscopy (SEM) observation on the surface of (A) uncoated banana fruit
and (B) coated banana fruit with agar suspension at 2 g · l−1 by dipping for 10 min
Effect of agar coating on physiochemical to studies on safe, eco-friendly and cost-effective fruit
properties of banana fruits preservation techniques to maintain quality and ex-
tend the shelf-life of fruit (Maqbool et al. 2010). Over
Weight loss (%), firmness loss (%) and soluble solid
the last two decades, the development and use of ed-
concentration (SSC) of coated and uncoated banana
ible coatings to prolong the shelf-life and improve fruit
fruit gradually increased with increasing storage peri-
ods (Figs. 2A−C). Banana fruit coated with agar colloid quality has been intensively researched (Dhall 2013).
at 2 g · l−1 for 10 min had less weight loss and firmness Agar-agar, a hydrophilic colloid extracted from mari
loss percentages and SSC than control (uncoated) fruit ne red algae (Davidson 2006), has multiple applica-
during storage periods (10 and 15 days). At the end tions particularly in the human food industry. Since
of storage, agar-coated fruit clearly showed the low- it is non-degradable by microorganisms, agar-agar is
est weight loss and firmness loss percentages and SSC commonly used as a solidifying agent in the prepara-
compared to control fruit (Figs. 2A−C). tion of microbiological and plant tissue culture media
(Armisen and Galatas 2000). The present study dem-
onstrated that agar suspensions at different concentra-
Scanning electron microscopy (SEM) tions, especially at 2 g · l−1, applied by dipping banana
observations fruit for 10 min could be a promising protective coat-
Banana fruit coated with agar colloid at 2 g · l−1 for ing agent for management of postharvest rot diseases.
10 min at 40−50°C, which had been artificially inocu- The lowest concentrations of agar suspension (0.5 and
lated with pathogenic fungi as well as control fruit, were 1 g · l−1) had less efficacy than 2 g · l−1. This could be
investigated by scanning electron microscopy (SEM) as because low concentrations of agar coating were less
shown in Figure 3. Results indicated that the surface of thick and did not completely cover fruit surfaces. The
banana fruit had significantly fewer cracks and smooth- same results were obtained by Ali et al. (2010) for to-
er surfaces as well as limited mycelia of pathogenic fungi mato fruit coated with gum Arabic. Agar as a polysac-
compared with the untreated fruit. Thus, agar could be charide coating agent resembles chitosan and gum
used as a protective agent against postharvest diseases. Arabic for controlling postharvest diseases. Chitosan
was effective in controlling postharvest diseases in li-
tchi (Zhang and Quantick 1997), sweet cherries (Ro-
manazzi et al. 2012) and banana fruits (Hossain and
Discussion
Iqbal 2016). Fruit weight loss during storage is attrib-
uted to increased water loss (transpiration) and respi-
Increasing awareness of health problems and environ- ration (Yaman and Bayoindirli 2002). The weight loss
mental issues due to application of pesticides has led reduction of agar-coated banana fruit compared to
6 Journal of Plant Protection Research 58 (3), 2018
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