Deep Dive

1. Green Gold the Tech-Challange
Dr. Elene Kakabadze
015730668427
elene.kakabadze@gmail.com
19.04.2021

2. “If I were a teenager today, I'd be hacking biology.” - Bill Gates
Transforming
biological innovation

3. Our experiences from Bio.Kitchen suggest a clear set of values
enabling lean development of life science innovation.
Rapid Prototyping
Transdisciplinarity approach
Hacker Community
Freedom to operate
Founder’s Mindset
Its all about YOUR project!

4. Bio.Kitchen is an open community lab for tech-talents
5 workstations: micro- and molecular biology, chemistry, lab-on-Chip and lab automation
Rüdiger
Trojok
Lab Lead
Leo
Sprengel
Lab Assistant
Andreas
Jefimowicz
Lab Assistant
Dr. Elene
Kakabadze
Freelancer

5. Biocommons
Synbio Oath
Read & Write DNA
printing DNA
Biological
Transformation
cradle to cradle
economics
Personalised
Medicine
future regulation
Fluid.Bio
SLA 3D printing
Lego Biochips
Future lab
Bio-Computer
workshop
Biohacking
pharma bioethics
seminar
biological rapid
prototyping
Syn.bio
cell free system
workshop
Biosensors
station
Internet of Living
Things workshop
Lab.Bio
fungi cultivation
plant cultivation
insect cultivation
Microbiologyfor
beginners
Micro.Bio
sequencing DNA
Bio.Kitchen
lab intro
workshop
yeast cultivation
Sprunginnovation
open source
licencing
Biotic games
bio tournament
Biohacking
workshops
hacking evolution
workshop
prototype nature
biodesign studies
Microfluidics
DIY automation
workshop
sci-tech
concepts
education format
bootstrapping
biotech startups
personalised
pharmaceutics
solve the
antibiotic crisis
advanced
Microbiology
Bio-Playstation
workshop
Copying DNA
workshop
one health
strategy
interplanetary
Bioteleportation
bacteria
cultivation
phage cultivation
We offer a large range of technology workshops, materials and tools
AI.Bio
Predict protein
structures f
Programming
DNA
patent strategy

6. Green Gold
Challenge ● Avocado is the holy grail of
healthy lifestyle and
nutrition.
● Avocado market grows at
5.72%.
● Global market size of
US$17.905 billion by 2025.
● High demand in developed
economies.

7. Why is it in
demand?
It‘s tasty
It has high nutritional value
It has high fiber content
It‘s health beneficial
It‘s trendy

8. Longevity
• rich sources of fiber
• potassium
• vitamins C, E, and K
• carotenoid antioxidants
• monounsaturated fats
(heart-healthy)

9. Problems
Propagation
● Suppling
nurceries
● Cloning
● Grafting
Sustainability
● Mass production
● Monocultures
● Deforastation
● Water footprint
● CO2 footprint
Fairtrade
● Ethical production
● High demand
● ‘Bloody Avocados’

10. Persea americana
Belongs to the Magnoliids
which are more than 150
million years old.
Avocados originated in
Mexico.
Domesticated and
cultivated since 5,000 BCE.
Avocado or as Aztec liked to call
them: “testicles.”

11. From giant sloth
to hungry human
The avocado developed in
Cenozoic Era when
megafauna was roaming
on the planet Earth
ghosts of evolution

12. Subtropical Tree
Varieties: ~500 cultivars, were 90% of cultivated avocado corresponds to
the cultivar ‘Hass’
Growth temperature: 10ºC-35ºC, cold hardy varieties can stand up to -9ºC
Soil requirement : well drained, 1M depth, pH 5-6
Water requirement: 600L per KG
Other environmental conditions: no wind, mild sun
Tree parameters: 6-10 years to bear fruit, average yield per tree 150 fruits

13. Growth range
● Endemic to Sauth-Central
America
● Mexican, Guatemalan and
West Indian types
● Cultivated in tropical,
subptropical and
mediterenian climats
● In competiton with natural
tropical forests

15. Pollination is needed to get the fruit grow
but
‘Protogynous Dichogamy’
Flowers opens as female one day and male another day
● Female ● Male
‘Ideal’ flowering temperatures (maximum 25°C and minimum 20°C)

16. Maximising cross-pollination for more and bigger fruits
● (type A flowering sequence).
trees will open first in the morning as
female, close and then reopen the next
afternoon as male
● (type B flowering sequence).
trees open first in the afternoon as
female, close and then reopen the next
morning as functionally male
Flower type
Day 1
Morning Afternoon
Day 2
Morning Afternoon
A female closed closed male
B male closed closed female
Increasing the period of potential pollination by inter-planting with complementary varieties
Decreases monoculture orchards

17. Cold hardy
varieties
• Can stand up to -9ºC (15ºF)
frost but not for long
• Varieties of Mexican breed
• Needs further optimisation

19. Market
compatibility
Fruits are selected on the bases of : • Size
• Skin properties
• Durability
• Damage tolerance
• Taste
• Shape
• Easy peel
• Appeal

20. Reproduction
Not true to the seed=
Due to genetic
variation the seed will
not yield the same type
of plant as the original
plant
The tree needs to be cloned
To get a fruity tree

21. Industrial
propagation
• Happens in nurseries
• With grafting techniques
• 90% of cultivated avocado are
‘Hass’, which are grafted on
P. americana var. drymifolia
rootstock
Ressource and Time
consuming, labour extensive,
pricy with high plant death rate

22. Clonal rootstock grafting
Requires:
1. random nurse seedling
2. clonal rootstock for rooting
3. scion from desired fruiting variety

23. Cloning 1. Piece of desired variety is cut off
(meristematic cells)
2. Followed by tissue surface sterilisation
3. Next put in the growth medium with
right hormones (sterile conditions)
4. Incubated for initiation at 25±1°C with
white fluorescent light
5. Shoot indication should occur in ~3
weeks
6. Root induction should occur in ~4
weeks
7. Refresh growth media and adjust
hormones every other week

24. Preparation
Experiment

25. Genome sequencing
• long vegetative period
complicates directed cross
breeding.
• Genome sequence
information is highly
valuable to breeding
programs, to speed up the
selection of cultivars and
rootstocks with desirable
traits.
Rendón-Anaya, Martha, et al. "The avocado genome informs deep angiosperm phylogeny, highlights introgressive hybridization, and reveals pathogen-
influenced gene space adaptation." Proceedings of the National Academy of Sciences 116.34 (2019): 17081-17089.
Nuclear genomes of the Mexican (P.
americana var. drymifolia),
Guatemalan(guatemalensis) and West
Indian (P. americana var. costaricensis),
hybrid cultivar: ‘Hass’ and disease resistant
rootstock ‘Velvick’ have been sequenced.

26. Breeding with genetic
modification
• Frost tolerance
• Anti herbicidal properties
• Disease and abiotic
stresses tolerance
• Plant defensing gene
1. Genome programing
2. Obtaining protoplasts
3. Agrobacterium tumefaciens or gene
gun + CRISPR cas-9 editing
4. Callus culture growth into plant

27. Ecologically sustainable avocado
supply for Europe

28. Solutions?
● Lab cultivation
● Cold-hardy breed selection
● Genome-assisted breeding
● Development of marketing
strategies
● Business model for European
avocado farming (e.g. in
greenhouses)
● Solutions for sustainability
● Precision agriculture
● Waste management
(bioplastics)
● Fairtrade solutions

29. The resulting avocado plants
will be given to the Munich
Urban Colab and can serve as
decorative tropical office
plants

30. Useful links:
● Export, market potential and entry EU (CBI): https://bit.ly/2Q9jpmp
● Avocado association: https://avocadofruitoflife.com/en/
● Lynwood Avocado rootstock cloning: https://www.youtube.com/watch?v=VyMogjYoldY
● Avocado Genome Consortium: https://bit.ly/3v4UvD1
● Bioplastic from seed: https://biofase.com.mx/
● Info on some varieties: https://bit.ly/3dzOEzH
● Climate database: https://weatherspark.com/
● One of many sidling sellers: https://www.fast-growing-trees.com/
● Forest garden concepts: https://bit.ly/3x65JJz
● Netflix – Rotten: The Avocado War