The document discusses using biotechnology to increase the storability and shelf life of fruit. It notes that fruit currently has limited storability and shelf life, resulting in losses for growers and less food availability. Biotechnology can help by developing cultivars with better storage and shelf life through techniques like gene editing. Specific genes involved in ethylene production and cell wall degradation impact ripening and shelf life. New breeding techniques have the potential to precisely modify these genes in popular cultivars to extend shelf life while maintaining taste. This could help tropical crops like mango that currently have limited export opportunities due to short shelf life.

1. International Symposium Feb15-17, Rome
Use of biotechnologies to increase the
storability and shelf life of fruit
2016.02.16 Eric van de Weg

2. International Symposium Feb15-17, Rome
Problem
 Fruit has a limited storability & shelf-life
● Loss of produce
● Decay, diseases
● Short selling window
● Over-supply -> low prices
 Impact
● Growers
● Less food to consume
● Less food to sell
-> Reduced income & financial sustainability
● Global population
● Less food
● Inefficient use scarce resources
http://www.gardenguyhawaii.com/2011/11/mango-anthracnose.html
http://www.123rf.com/photo_
702968_hand-of-overripe-and-
decaying-bananas-on-white-
background-isolated.html
M. J. Mahovic
ttp://edis.ifas.ufl.edu/LyraEDISServlet?command=getImageDetail&image_soid=FIGURE
4&document_soid=HS131&document_version=51786

3. International Symposium Feb15-17, Rome
Biotechnology can help
 Making available of cultivars with better
● Storage & shelf-life
● Harvest window
Classical Breeding
+ Molecular markers
New Breeding
Technologies
Decrease post harvest loss: Pre-harvest measures

4. International Symposium Feb15-17, Rome
Two types of fruit
Climacteric fruit
Non-climacteric fruit
Tomato, Apple, Banana, Mango, Peach, Pears, Avocado, Melon
● Needs ethylene for ripening
Citrus, Grape, Watermelon, Strawberries
● does not need ethylene

5. International Symposium Feb15-17, Rome
Ethylene
 Gas
 Signalling hormone
 Initiates ripening
 Ethylene from ripe fruit  accelerates ripening unripe fruit
Pictures taken from http://glutenfreehomemaker.com/how-to-ripen-green-fruit-kitchen-tip/ http://www.realfoods.co.uk/article/the-real-foods-guide-to-ripening-bananas

6. International Symposium Feb15-17, Rome
Ripening & Taste
 Ripe fruit:
● Good texture, juicy
● Excellent flavour, aroma, color
● Most delicious
 Harvested very unripe: No full maturation is feasible
● Delicate balance
Harvest date / storability / taste
 Harvested unripe
● Better storability and shelf-life
● Less taste !

7. International Symposium Feb15-17, Rome
Harvest date  Targeted market
Image: https://www.youtube.com/watch?v=fGLK_vBUymA
Long storage: stages 1 & 2

8. International Symposium Feb15-17, Rome
Biodiversity: Apple cultivars
 Storability, Temp controlled: 0.5 – 4 months
 Shelf life: 2 – 4 wks
 Storability: Temp + air controlled: 0.7 – 13 months

9. International Symposium Feb15-17, Rome
Biodiversity: Tomato cultivars
 Shelf life: 7 days – 2 months
 Large extension: Thanks to classical breeding &
Biotechnology !

10. International Symposium Feb15-17, Rome
Biodiversity: underlying genes
 Natural variants of a gene (mutant)
 Tomato:
● Combining 3 natural genes
● Ripening blocked
● Extreme long shelf-life
● Taste: “to be repaired” by stimulating other
pathways
 Apple:
● Joint effect of ~7 natural genes
● Ripening process slowed down
● Taste maintained

11. International Symposium Feb15-17, Rome
Experiences Tomato, apple......
 Some specific genes involved in
1. Production and perception of ethylene in fruit
2. Production/ activity cell-wall degrading enzymes
ACS
ACO
Ethylene
ETR
Effects
ACO-Gene
2 genes for
production
1 gene for
perception
- Other functions maintained !
- On-off switches
On/
Off
On/
Off
One for fruit maturation
- On-switch corrupted
- No ethylene production
- Deletion of 2 nucleotides

12. International Symposium Feb15-17, Rome
Prospects Tropical crops: Mango?
 Harvested fruit: Limited storability & shelf-life
 ~96% production for home-market, ~4% export
 Storability & shelf improved:
● Harvesting unripe fruit
● Forced ripening close to selling:
● Ethylene + Temperature treatment
● Less tasty
http://www.fruitnet.com/americafruit/article/164144/first-aussie-mangoes-shipped-to-the-ushttp://www.dawn.com/news/1182818 http://www.skymetweather.com/content/agriculture-and-economy/mango-exports-from-india-
face-a-hard-time/
http://www.ctvnews.ca/business/why-the-eu-alphonso-mango-ban-s-sweet-news-in-india-1.1809895

13. Natural biodiversity Mango
King of the mangoes: Rich, creamy, tender pulp
texture
 Export
‼ Old cultivar, before 1515
cv "Alphonso”
https://en.wikipedia.org/wiki/Alphonso_(mango)

14. International Symposium Feb15-17, Rome
New Australian cultivar: CalypsoTM
Breeding
Average
Australia
Calypso
Production ton/ha 10 30
% 1st grade 60 85
Source: http://www.hin.com.au/__data/assets/pdf_file/0006/9591/Mangos.pdf
Production & Shelf-life increased
Currently: Successful in export
x Less tasty
http://www.freshplaza.com/article/135404/Calypso%C2%AE-Aussie-mangoes-hit-in-US
First shipment to USA

15. International Symposium Feb15-17, Rome
Biotechnology, storability & shelf life
 Need
● More cvs with good storability & shelf life
● Maintaining excellent taste
 New Breeding Techniques ready to go
● Gene-editing
● No adding of new DNA
● Improvement existing, popular cultivars
● Turn-around shorter than classical breeding
 Target genes known
● Ethylene related
● Cell-wall degradation

16. International Symposium Feb15-17, Rome
GM-fruit: Current status
 Approved
● Apple No flesh browning 2015 USA
● Eggplant Insect resistance 2013 Bangladesh
● Plum Virus resistance 2007 USA
● Melon Storability, shelf-life 1999 USA
● Papaya Virus resistance 1998 USA
● Tomato Storability, shelf-life 1992-9 USA,Canada,
Mexico, China
 Field tests
● Banana Black Sigatoka fungus USA
● Orange Citrus greening USA

17. International Symposium Feb15-17, Rome
Storability & shelf life: Current GMO’s
 Derived from two approaches:
1. Insertion foreign gene
● Redirecting pathways
● New products
● Additional antibiotic gene added to trace
GMO-cells
2. Silencing:
● Blocking genes = pathways
● No new proteins produced
● Methods not specific: similar genes blocked too
● Additional gene added to trace GMO-cells

18. International Symposium Feb15-17, Rome
Storability & shelf life: Innovation
 Precision Biotechnology
● Editing : Removal of a tiny DNA segment
● Promotor : Switch off !
Changes only in
 specific tissue(s)
 Specific development stage(s)
 e.g. only Mature Fruit
 Advantage:
● Identical / close to natural genes (mutations)
 Disadvantage:
● More expensive: Additional research needed on
promotors
ACO-Gene
On/
Off
On/
Off

19. International Symposium Feb15-17, Rome
Who benefits?
Local: benefits directly
Export: May takes longer to build a market
Different turnaround, different prospects ?
https://www.youtube.com/watch?v=XTphqTbP8iM https://www.youtube.com/watch?v=XTphqTbP8iMhttps://www.youtube.com/watch?v=6MALFk56jPE
http://www.abc.net.au/news/2015-10-20/washing-mangoes/6870578 http://www.abc.net.au/news/2013-09-30/mango-packing-shed/4989350
http://www.abc.net.au/news/2015-09-22/mangoes-in-bin-to-be-inspected/6795344

20. International Symposium Feb15-17, Rome
Time-line: Action now  impact 15-20 yrs
 Application new technology
 In vitro plantlet -> fruit bearing trees
 Evaluation different new plants
● Overall agronomic performance
● Shelf life <> taste
0-3
3-6
6-8
9
9-12
13
~15
 Admission procedure
 Best plant: Multiplication few -> thousands
 Plantation of orchards
 First production
en.wikipedia.
http://baguioheraldexpress.com/wp-content/uploads/2015/01/Banana-Plantation.jpg

21. International Symposium Feb15-17, Rome
Biotechnology on tropical crops
 New Breeding Techniques ready to go
● Gene-editing
● No adding of new DNA
● Improvement existing, popular cultivars
● Turn-around shorter than classical breeding
 Genes of interest known from other crops
 Info on New Technologies:
● Brochure: http://edepot.wur.nl/357723
● Site event: today 13.15-14.15
● Venue: Sheikh Zayed Centre
● “New breeding technologies for
smallholders’ challenges”

22. International Symposium Feb15-17, Rome
Questions to be adressed
Level of precision breeding ?
● More specificity -> less undesired site-effects ?
● Higher investments, Delayed return on investment
Who controls cultivar development ?
● Private? Public? Public Private Partnerships
Theme for development aid ?
● 90% of GM-growers = small farmers!
● Food security, jobs & income
● Vision infrastructure development  long term effects

23. International Symposium Feb15-17, Rome
Use of biotechnologies to increase the
storability and shelf life of fruit
Eric.vandeweg@wur.nl,
Plant Breeding Wageningen UR, The Netherlands
Thanks
Questions ?

24. International Symposium Feb15-17, Rome
Biotechnology on tropical crops
 New Breeding Techniques ready to go
● Gene-editing
● No adding of new DNA
● Improvement existing, popular cultivars
● Turn-around shorter than classical breeding
 Genes of interest known from other crops
 Info on New Technologies:
● Brochure: http://edepot.wur.nl/357723
● Site event: today 13.15-14.15
● Venue: Sheikh Zayed Centre
● “New breeding technologies for
smallholders’ challenges”