Detection of the fungal infection in post-harvest

onions by an electronic nose

Malgorzata Labanska Sascha Jenkins Sarah van Amsterdam
The Plant Breeding and School of Life Sciences, School of Life Sciences,
Acclimatization Institute - National University of Warwick University of Warwick
Research Institute Warwick, UK Warwick, UK
Bonin, Poland sascha.jenkins.1@warwick.ac.uk sarah.van-amsterdam@warwick.ac.uk
m.labanska@ihar.edu.pl
John Clarkson James Covington
School of Life Sciences, School of Engineering,
University of Warwick University of Warwick
Warwick, UK Warwick, UK
john.clarkson@warwick.ac.uk j.a.covington@warwick.ac.uk

Among emerging novel trend streams in agriculture, sometimes be seen in the field, a major problem for the
2022 IEEE International Symposium on Olfaction and Electronic Nose (ISOEN) | 978-1-6654-5860-3/22/$31.00 ©2022 IEEE | DOI: 10.1109/ISOEN54820.2022.9789676

techniques for the analysis of the volatile organic compounds industry is the rotting of bulbs in store post-harvest. Here we
emitted from plants are gaining increasing interest. In this demonstrate that electronic nose can discriminate between
work, the electronic nose abilities to detect onions infected with samples with varying proportions of infected onion bulbs and
Fusarium oxysporum f. sp. cepae as well as to discriminate show that storage temperature affects the rate of
between samples with varying proportion of infected onion development of Fusarium basal rot.
bulbs were evaluated. It was demonstrated that changes in the
samples related to the development of the disease can be
observed on the PCA plot. It was also proved that storage of II. EXPERIMENTAL
the onions at a low temperature inhibits the development of the
infection. Presented findings of the preliminary studies A. Sample preparation
revealed that electronic nose is a promising analytical tool for Twenty onion bulbs were inoculated with Fusarium
the detection of the infection of the onion bulbs. oxysporum f. sp. cepae (FOC) isolate FUS2. The outer dry
scales of the bulbs were removed, and a 2-3 mm slice of the
Keywords—electronic nose, volatile organic compounds, gas basal plate was removed with a scalpel, before spraying
analysis, onions, plant disease detection
bulbs with 70% ethanol. Once dry, an 8 mm plug of potato
dextrose agar (PDA) containing the actively growing edge of
I. INTRODUCTION FOC isolate FUS2 was inverted and placed on the cut basal
Plant diseases caused by pathogens and pests lead to plate of each bulb. Non-inoculated bulbs were also prepared
numerous short- and long-term consequences, including in the same way, except no agar plug was placed on the basal
economic loss, reduced availability of food, decreased food plate. Bulbs were placed in damp boxes in sealed plastic bags
quality and possible accumulation of toxins during storage. and incubated at 20˚C.
Early plant disease detection enables a quick response to After initial incubation the non-inoculated and inoculated
these issues, by informing application of pesticides, different bulbs were placed in the sealed 3 L plastic boxes with inlet
management of the infected crops (e.g. elimination or selling and outlet fittings added at both ends (Fig 1.). Each box
into the animal feed market), or as in the case of stored crops contained healthy and inoculated bulbs (a total of five bulbs)
such as onion or potato, increasing ventilation or changing at varying proportions resulting in six samples, i.e. 0%, 20%,
storage conditions to minimise further disease development 40%, 60%, 80% and 100% infected bulbs. Samples were
or spread [1,2]. Among emerging novel trends in agriculture prepared and stored for 5 weeks at 25ºC. To maintain high
methods based on the analysis of the volatile organic humidity throughout the experiment, bulbs were suspended
compounds (VOCs) released by plants are gaining increasing on mesh over some water.
attention. The idea of analysing VOC profiles lies in the
ability of plants to release various compounds according to
their health status and the biochemical response to pests or
pathogens [3,4]. The electronic nose systems are potentially
useful tools for the plant disease detection and recognition of
infected plants, before occurring visual symptoms of the
disease. So far, the electronic nose systems have been
applied for the disease detection of a wide range of crops
including onions [5], potatoes [6], rice [7], apples [8] and
wheat [9].
The aim of this work was to evaluate the ability of
electronic nose technology to detect onion bulbs infected
with the soilborne fungal pathogen Fusarium oxysporum f.
sp. cepae, which causes Fusarium basal rot disease.
Although above-ground symptoms of this disease can
This work has been supported by the Polish National Agency for Academic Fig. 1. A measurement setup with PEN 3 electronic nose.
Exchange within a framework of the Bekker programme
(PPN/BEK/2020/1/00383).

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Additionally, two more samples with 0% and 100% infected W1C
100%
20
onion bulbs were stored at 4ºC to determine the influence of W3S W5S 80%
60%
temperature storage on infection development. 15 40%
20%
B. Electronic nose analysis 10 0%
The PEN 3 electronic nose system (Airsense Analytics W2W 5 W3C
GmbHm Germany) equipped with 10 MOS sensors was used
to analyse the volatiles released by the onion bulbs. Each 0
sensor exhibits sensitivity towards a different class of
volatile compounds (Tab. 1) [10]. A wire resistor generated
high temperature that induce interactions between volatiles W2S W6S
from the sample and the metal-oxide semiconductor surface
resulting in changes in the conductance of the sensor.
Recorded sensor’s signal is the ratio of the resistance of a
sensor exposed to a sample and resistance of the sensor W1W W5C
exposed to a clean air. The measurements were performed
twice every week, for 5 weeks starting from the week after W1S

inoculation. Prior to the measurements the boxes stored at
4ºC were placed at 25ºC to reduce influence of the
temperature on the sensors signals. The headspace from the
each box was pumped in the sensor chamber at a constant
rate of 400 mL/min and sensors signals were recorded for
180 s with 2 s intervals. Before each measurement, a 60 s
cleaning phase was applied, and the baseline was set for 10 s.
The average values of the differential sensors’ responses
recorded between 170-174 s formed the datasets that were
subjected to analysis using Principal Component Analysis
(PCA). The raw sensors signals were autoscaled in columns.
Data analysis was conducted employing MATLAB software
(Math Works Inc., Natick, MA).
III. RESULTS AND DISCUSSION
First, responses of the sensors were evaluated based on
the radar plots. A radar chart of the sensors responses
towards samples with varying proportion of the infected
onion bulbs recorded two weeks after inoculation is
presented on Fig. 2. It was found that patterns of the sensors
responses vary depend on the proportion of the infected
onion bulbs. The greatest contribution to the differences in
those patterns was the sensor selective towards sulphuric -
organic substances (i.e. sulphides). Furthermore, sensors
generally described as selective towards alcohols, methane
and broad range of organic substances demonstrated
significant changes in responses. Generally, the responses of
the sensors increased with an increase in the proportion of
the infected onion bulbs. The smallest responses were
recorded for samples consisting of non-inoculated onions
(0%), whereas the highest signals were observed for samples
with 80% and 100% infected onions.
Fig. 2. The gas sensors responses towards samples with varying disease
rate stored at 25ºC. Measurements made two weeks after inoculation.Onion
bulbs stored in the plastic boxes.
These results are in good accordance with a GC-MS
analysis from the literature [4] which revealed that various
VOCs are released during development of the Fusarium
infection in onions including sulphuric organic compounds
such as methyl propyl sulphide, and methyl propyl
disulphide, and dimethyl sulphide, alcohols and ketones. It
can be seen that several sensors contribute to the volatile
patterns of the samples, and therefore multivariate data
approach is required to discriminate between samples with
varying proportion of the infected and non-infected bulbs.
Over time, visual symptoms of the disease were
observed. Red-brown discoloration and rot were developed
on the base of the bulbs, and the tissue became soft.
Significantly earlier symptoms have been observed in the
case of bulbs stored at 25 C than 4C.
The responses of the sensors towards samples consisting
of 100% and 0% infected bulbs kept under different storage
conditions were processed by PCA to determine the
influence of storage temperature on the disease development.
The resulting PCA score plot is presented in Fig 3. Each
point represents a different storage duration (the lighter
colour the longer storage). The first two principal
components capture 90% of the variance of the dataset. Two
clusters of the samples are visible on the plot. The first one
consisting of the non-infected samples and samples at the
early stage of infection (low PC1 values and high PC 2
values), and the second one with infected samples.
3

TABLE I. DESCRIPTION OF THE PEN 3 ELECTRONIC NOSE SYSTEM

SENSORS 2
PC 2 (13.59%)

Sensors General description 1

W1C aromatic compounds
W5S broad range of compounds 0

W3C ammonia, aromatic compounds

-1
W6S hydrogen 100%, 25°C
W5C alkens, aromatic compounds 0%, 25°C
-2
W1S methane, broad range of compounds 100%, 4°C
0%, 4°C
W1W sulphur - organic compounds -3
-4 -2 0 2 4 6 8 10
W2S alcohols, broad organic compounds
PC 1(76.71%)
W2W aromatic compounds, sulphur organic compounds
W3S alkens, aliphatic organic compounds Fig. 3. The PCA score plots of the samples stored at different
temperatures

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 The greatest differences in the location of the samples on
the PCA plot were observed in the case of the sample
consisting of 100% infected onion bulbs stored at 25ºC. The
correlation between location on the PCA (especially towards
PC1) and the development of the infection is noticeable. The
higher value of the PC1 the more severe disease. Minor
differences between samples kept at 4ºC suggest that little
disease development occurred at this storage temperature.
Low temperature is commonly used in commercial storages
to prevent development of disease. The least differences
between samples were noticed in the case of the non-
inoculated onion bulbs stored at 4ºC.
Finally, sensors signals recorded during four weeks of the
experiment were subjected to PCA processing score plot of
the samples consisted of varying ratios of inoculated to non-
inoculated onion bulbs stored at 25ºC is shown in Fig 4. In
this case, prior to the data processing, a feature extraction
step was implemented that involved the elimination of the
insignificant sensors’ signals from the dataset. Therefore, the
dataset was formed only by sensors signals that differed
significantly depending on the sample tested. This led to
better discrimination between samples. The first two
principal components captured 95% of the variance of the
dataset. It was found that samples with lower proportion of
the infected onion bulbs are placed close to each other and
are characterized by low PC1 and PC2 values, whereas as the
ratio of inoculated to non-inoculated onions increases, the
samples are more dispersed in the plot and have significantly
higher value of PC1. Analysis of the loading plot (data not
shown) revealed that all six sensors significantly contribute
to presented discrimination. Furthermore, as the storage time
increases, the samples are placed further from each other and
have higher values of PC1 indicating the development of the
infection. On the other hand, samples slightly overlap each
other suggesting that a more advanced pattern recognition
method might improve discrimination between samples.
Nevertheless, PCA results have shown that the location of
the sample on the plot is related to the disease severity.
IV. CONCLUSIONS
In this work, the PEN 3 electronic nose system was
applied to discriminate between onions infected with
Fusarium oxysporum f. sp. cepae and those non-inoculated
as well as to differentiate between samples with varied
proportion of infected onion bulbs based on the volatiles
released by the onions. Additionally, the influence of
temperature storage on the development of Fusarium
infection was also determined. The sensors responses
showed significant differences between volatiles released by
onions non-inoculated and those infected with Fusarium
species with varying disease ratios. Sensor sensitive towards
sulphuric organic compounds was found to have the highest
contribution to differentiating samples. It was shown that
PCA analysis of the sensors’ responses provided clear
discrimination between diseased samples and uninfected
controls. Moreover, changes in the samples related to the
development of the disease were observed on the PCA plot.
It was also shown that storage of the onions at low
temperature inhibits the development of the disease. The
findings of this preliminary study suggest that electronic
nose is a promising analytical tool for the detection Fusarium
basal rot in onion bulbs.
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1
time
PC 2(4.12%)
0
100%
80%
60%
-1 time
40%
20%
0%
-4 -2 0 2 4 6
PC 1(91.08%)
Fig. 4. PCA score plot of the samples with different disease rate and
with varying storage time.
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