Calcium signature in kiwifruit ripening through multi-omics integration

M. Michailidis 1,2,3*, C. Polychroniadou1,2,3, I.-D. S. Adamakis 4, I. Ganopoulos2,5, G. Tanou2,3, C. Bazakos2,3,6,

E. Karagiannis7, A. Molassiotis1
1AristotleUniversity of Thessaloniki, Department of Ηοrticutlure, Laboratory of Pomology, 54124 Thessaloniki, Greece, msmichai@agro.auth.gr, 2Joint
Laboratory of Horticulture, ELGO-DIMITRA, Thessaloniki-Thermi 57001, Greece, 3Institute of Soil and Water Resources, ELGO-DIMITRA, Thessaloniki-
Thermi, 57001, Greece, 4University of Athens, Department of Biology, National and Kapodistrian GR- 15784 Athens, Greece, 3Institute for
Bioinnovation, Biomedical Sciences Research Center “Alexander Fleming”, 16672, Vari, Greece, 5Institute of Plant Breeding and Genetic Resources,
ELGO-DIMITRA, Thessaloniki-Thermi, 570001, Greece, 6Max Planck Institute for Plant Breeding Research, Department of Comparative Development
and Genetics, Carl-von-Linn, 7University of Western Macedonia, Department of Agriculture, 53100 Florina, Greece

1. INTRODUCTION
Calcium is an important macronutrient in plants that fulfills a structural (A)
role in cell wall and membrane stability as well as it is the most versatile
second messenger operating in a variety of physiological, developmental
and stress-related processes. Calcium in fruit affect composition, and
structure of different metabolites, genes and proteins but our knowledge (B)
in climacteric fruit metabolism such as kiwifruit is still limited. The aim of
Ca.L
this research was to understand the physiological and molecular aspect CN.L
of calcium during kiwifruit postharvest ripening. CN.E
Ca.E
‘Hayward’ kiwifruits
2. MATERIALS AND METHODS
‘Hayward’ kiwifruit harvested at Figure 4. (A) Metabolic analysis and (B) RNAseq analysis of calcium treatment in
the commercial stage (Brix above early (E) and late (L) response.
6.25%) and immediately dipped in
2% CaCl2. Afterwards, fruit cold Ca-exposed fruit was characterized by elevated abundance of numerous (>890)
stored at 0 °C for 3 months and the proteins, notably at late period. Weighted correlation and network analysis based
H2O 2% CaCl2 subsequent ripening traits were on the integration of transcriptome, metabolome and proteome datasets
evaluated at 20°C for 7 days (Fig. identified candidate modules involved in the Ca action in kiwifruit (Fig. 5).
1). (A) (C)

Figure 1. Experimental design to

Cold storage (0 °C, RH 95%) study calcium in postharvest fruit
for 3 months
physiology

3. RESULTS AND DISCUSSION

Calcium treated kiwifruits exhibited higher Ca concentration and
intracellular calcium (Ca2+) fluoresce signals in outer pericarp and
placenta that accompanied by a reduction in softening and ethylene
arp and
production. (Fig. 2).
tissue Epidermis and Inner pericarp and Control Calcium
outer pericarp vascular tissue

0.3
Calcium content

Figure 5. (A) Venn diagram of proteins, (B) WGCNA modules construction and
Fluoro 3AM
Control

(%, DW)

0.2

0.3 (C) networking illustration.

Calcium content

0.1
✱
Fluoro 3AM

0.0
(%, DW)

CN Ca 0.2
4. CONCLUSION
0.1
We summarize our findings in a model that highlights the main metabolic
Calcium

0.0
events regulated by calcium application and include cell wall formation, protein
CN Ca ubiquitylation, ethylene, ascorbic acid and L-cysteine biosynthesis (Fig. 6).

Figure 2. Calcium content and physiological

traits determination.

Fluorescent microscopy with cell wall

primary antibodies showed that de-
esterified homogalacturonans (HG)
(antibody LM19) was increased while
arabinogalactan proteins (AGPs) (antibodies
LM19 and LM30) were depressed in Ca-
treated fruit. (Fig. 3). Metabolomic analysis
revealed that 27 primary metabolites were
altered by Ca in pericarp tissue (i.e. glucolic
acid, oxoproline and galactinol) (Fig. 4A).
Figure 6. Schematic presentation of calcium metabolic regulation in kiwifruits.
RNAseq analysis performed either at early or
late period following Ca dipping identify 89 5. FUNDING
and 370 DEGs (Fig. 4B). This work was supported by the Operational Program Competitiveness,
Entrepreneurship and Innovation, under the call RESEARCH–CREATE–INNOVATE
Figure 3. Fluorescent microscopy. (project code: Τ2ΕΔΚ-03007; Premium Kiwi).

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