Moving beyond the molecular mechanism of superficial scald in apple fruit

M. Michailidis1*, C. Skodra1, Ε. Karagiannis1,2, M. Samiotaki3, I. Ganopoulos4,5, G. Tanou5,6, C. Bazakos4,5,7,

A. Dalakouras 8, Α. Molassiotis1
1Aristotle University of Thessaloniki, Department of Ηοrticutlure, Laboratory of Pomology, 54124 Thessaloniki, Greece, msmichai@agro.auth.gr,
2University of Western Macedonia, Department of Agriculture, 53100 Florina, Greece, 3Institute for Bioinnovation, Biomedical Sciences Research

Center “Alexander Fleming”, 16672, Vari, Greece, 4Institute of Plant Breeding and Genetic Resources, ELGO-DIMITRA, Thessaloniki-Thermi, 570001,
Greece, 5Joint Laboratory of Horticulture, ELGO-DIMITRA, Thessaloniki-Thermi 57001, Greece, 6Institute of Soil and Water Resources, ELGO-DIMITRA,
Thessaloniki-Thermi, 57001, Greece, 7Max Planck Institute for Plant Breeding Research, Department of Comparative Development and Genetics,
Carl-von-Linn, 8Institute of Industrial and Forage Crops, Department of Plant Production, 1 Theofrastou Str., 41335, Larisa, Greece

1. INTRODUCTION
Although low temperature storage can delay apple (Malus domestica (A) EH cold Vs LH cold (B)
Borkh.) fruit ripening and senescence, it may cause the superficial scald
disorder under subsequent ripening at room temperature, which
seriously affects fruit quality. The aim of this research was the in-depth
characterization of the molecular mechanisms underlying scald
development, using high-throughput sequencing technologies
(transcriptomics, methylomics), coupled with mass spectrometry-derived
proteomics and metabolomics.
2. MATERIALS AND METHODS
‘Granny Smith’ fruits were harvested at two harvest (early and late)
stages, and after 3 months of cold storage (0 °C), they were exposed to Figure 4. (A) Global and (B) target
room temperature (20 °C) for scald development (Fig. 1). methylation of AFS.
(EH) (Cold)
Early Harvest

Right Epigenomics Finally, RNAi-based silencing of AFS and PPO genes through pump-infiltration
after cold
storage and injection in apple peel (Fig. 5) influenced the scald phenotype. In
0oC
particular, PPO alleviated superficial scald in comparison to GFP and
(LH) 3 months cold
Transcriptomics
storage untouched fruit, whereas in AFS scald symptoms were affected but at a lower
Late Harvest

(RT)
level (Fig. 6).
Scald assessment
Shelf life Proteomics
20oC
Commercial orchard in Imathia region
(North Greece)

Figure 1. Experimental design to study superficial scald in ‘Granny Smith’

3. RESULTS AND DISCUSSION

Data indicated that scald symptoms were mostly observed in early
harvested apples, which is highly linked to their enhanced transcriptional
and translational activity during and just after cold storage (Fig. 2).

Figure 5. Experimental design of AFS and PPO during scald development.

4 months cold storage plus Scald score

10 days at 20 oC

AFS PPO GFP UNT

0 1-25 26-50 51-75 76-100%

4
hpAFS a a
a
a
Superficial scald index

hpPPO
hpGFP a
3 a b
H2 O b
a
a
a
a b
2

b c
b
c c
1
c
c

1d 4d 6d 8d 10d

Figure 6. AFS and PPO scald development after agroinfiltration.

Figure 2. Physiological traits and superficial scald determination.
4. CONCLUSION
Analysis of post-translational protein
The combination of large scale -omics like
modifications (PTMs) also revealed
epigenomics and functional characterization
strong oxidation events following cold
approaches like RNAi-based functional
exposure (Fig. 3). Global and target
characterization of AFS and PPO using different
methylation analysis identified several
agroinfiltration approaches (Fig. 7) may contribute to
genes, including α-farnesene synthase
understand the molecular mechanism of superficial
(AFS) and polyphenol oxidase (PPO),
scald development in apple fruit.
that subjected to cold-derived
epigenetic alternations (Fig. 4). 5. FUNDING
This project has received funding from the Hellenic
Foundation for Research and Innovation (HFRI) and
the General Secretariat for Research and Technology
Figure 3. Categories of post- (GSRT), under grant agreement No. 148.
translational protein modifications.
Figure 7. Agroinfiltration via pump and injection in apple peel .

V Balkan Symposium on Fruit Growing, Zagreb, Croatia, June 18-21, 2023