Zerihun Tadele Institute of Plant Sciences University of Bern 30 - 31 August 2018. Gent-Zwijnaarde, Belgium. IPBO conference 2018: “Scientific innovation for a sustainable development of African agriculture”

1. Lodging and Drought
Resistant Tef
Zerihun Tadele
Institute of Plant Sciences
University of Bern
IPBO, Ghent, August 31, 2018

2. Yield potential & gap
Goals:
• To increase
productivity
per unit area
• To narrow the
yield gap
2
Tadele. Agronomy 2017, 7:22

3. Challenges for boosting productivity
1. Population increase vs food production
2. Environmental stresses: biotic & abiotic
3. Inherent properties of the plant
4. Biofuel production
5. Land and investment policies
6. Climate change: impact on crop cultivation
Brazil (2030) Central America (2030)
Maize
-10%
Rice
-14%
Wheat
-14%
Wheat
-9%
Rice
-10%
Bean
-4%
http://www.elsevier.com/connect/how-will-climate-change-affect-food-security

4. Why research on tef?
• Staple food for > 60 mio.
People
• Cultivated > 3 mio. ha in
Ethiopia
• Adapts to extreme climatic
& soil conditions
• Life-style crop (super grain):
absence of gluten
• Low productivity
• Little research by global
community
% cereal area
0
10
20
30
40
0,0
1,0
2,0
3,0
4,0
Yield (t/ha)
4

5. Tef is closely related to finger millet
5
BMC Genomics (2014) 15: 581

6. 6
Source: IFPRI 2006
Tef map: environmental constraints
Lodging: universal
Drought
Soil acidity
Soil salinity

7. Tef Improvement Project: Goals
1. Tolerance to Lodging
• Tef has tall and weak stem
• Lodged plants produce low
yield (quantity & quality)
2. Tolerance to drought (climate
smart)
3. Genome sequencing
7

8. Technology Generation
Technology Transfer
Technology Delivery
Univ.Bern
NationalResearch
Seedprod.
Mutmap
Genotypic
screening
Trainings,
workshops
Introgression
On-station testing
On-farm testing
Variety release
Seed production & dissemination
SNP discovery
Mutagenized pop.
Phenotypic
screening
Natural accessions RNAseq
Genome
sequencing
TILLING
RAD/GBS
Technology
Genotypic
screening
Eco-
TILLING
Tef Improvement Project: strategy
Genomis

9. Exploiting diversity in tef
• 5 000 landraces: EBI,
Ethiopia
• Huge phenotypic
diversity
9
Fold-change

10. Genotypic difference: very low
10
No lodging tolerant
genotype

11. Mutation breeding
Varieties released: 3222; crop types: >210; countries: >70;
Mutagens: Physical (x-ray, gamma-ray) & chemicals (EMS)
Crop Variety Trait Country Value
Barley
Golden
promise
Semi-dwarf, salt
tolerant
UK 417 mio $
Durum
wheat
Creso
Grain quantity &
quality
Italy 1.8 bio $
Rice RD16 Aromatic Thailand 16.9 bio $
Chickpea
CM-
88/98
High yield, blast
resistant
Pakistan 9.6 mio $
Sunflower NuSun Quality oil USA 149 mio $
Cotton NIAb-78
Early muturity,
determinate
Pakistan 3 .0 bio $
[
Euphytica (2004) 135:187-204; http://www-naweb.iaea.org/nafa/pbg/mutation-breeding.html
11

12. Mutation
induction
M1 population
Mutation
detection
Mutation
breeding
Candidate line(s)
with improved
trait(s)
LiCOR DNA
analyzer
Agarose
Electrophoresis
Next Generation
Sequencing
(NGS)
Introgression to elite
cultivar(s)
TILLING population
Improved cultivar
with desirable
trait(s)
High Resolution
Melt (HRM)
Non-denaturing
Polyacrylamide
gel
Pollen
Seed
Callus M2 population
Mutation breeding: value-chain

13. Increase diversity through mutagenesis
Used for screening for lodging, drought & soil acidity tolerance
EMS mutagenesis
(four genotypes)
M1 population M2 population
~ 12 000~ 18 000
13

14. TILLING
(Targeting Induced Local Lesions IN Genomes)
LiCor
M2M1
Illumina
sequencer
Pollen
Seeds
14

15. Mutagenesis
M1 pop. (15 000) M2 pop. (7 000)
DNA extraction (6 000)
Screening
D11 (BR)
KO2 (GA3)
Candidates
D11 (26)
KO2 (25)
Confirmation
D11 (16)
KO2 (16)
Useful mutations
D11 (5)
KO2 (7)
Sequence Evaluation,
introgression
TILLING for lodging tolerance

16. Lodging tolerance
16Expt Botany (2015) 66: 933-944

17. Candidate gene approach
1717
Expt Botany (2015) 66: 933-944
2 µM Propyzymide
(microtublue depolymerization)
• Diverse hormones and inhibitors:
no obvious effect
• Mutations in microtubule
• twisted dwarf 1 (tid1) in rice
• lefty, spiral (spr) in Arabidopsis
• Taxol: micro-tubule stabilizing drug
(insignificant effect)
• Oryzalin: disrupts microtubule by
binding to α-tubulin
(mutant insensitive)

18. Mutated gene
18
• Mutation in Tua1
• CAPS marker: cla 1 restriction
• Three α-tubulin genes: Tua1, Tua2 and Tua3
Expt Botany (2015) 66: 933-944

19. Mechanical properties of tef plant
Pushing – root anchorage Bending – flexural rigidity
To provide information to breeders 19

20. Materials: ecotypes & cross sections
10
cm
10 cm
bottom
internode
(No.1) (hollow ellipse-
shaped culm section)
top internode
(No. 5)
(filled circular-shaped culm
section)

21. Safety factor for lodging
The safety factor is the
ratio of the weight of the
plant, its angle and center
of gravity to the breaking
force of the plant. It is
measure of how many
times its own weight the
plant can withstand
before breaking.
21

22. 22
Drought tolerant tef (dtt)
Stomata length Stomata number
• Adaxial (upper) side:
o High number of
stomata
o Smaller stomata
• dtt:
o less number of
stomata
o smaller stomata
tdt: terminal drought
tolerant
F7 progenies
being tested at
drought prone
areas in Ethiopia

23. Proline content
23

24. Tef Transcriptome: Novel Targets
qPCR
RNASeq
FoldchangeFoldchange
No.ofgenes
24

25. 25
J Plant Phys (2018) 224:163-172
microRNA response to drought

26. 26J Plant Phys (2018) 224:163-172
microRNA and targets

27. Embryo-rescue
Tef x E. nindensis
Super tef
High yield
Extreme drought tolerant
Wild Eragrostis species
• 80 Eragrostis species at IPS
• E. curvula: lodging tolerant
• E. nindensis: drought tolerant
Introduce key traits from wild species
Flow-
cytometry
Diploid
Tef
(Tsedey)
E. rigidior
E. lehimania

28. In vitro regeneration method for tef

29. Variety development
(through hybridization)
Improved Cv. Desirable property Undesirable property
• Tsedey wide adaptation lodging susceptible
• Magna consumer preference lodging susceptible
• Quncho high yield &
consumer preference lodging susceptible
Our candidates
• Kegne lodging tolerant low yield
• GA-10 lodging tolerant, tillers
• Dtt2 drought tolerant low yield
• Dtt13 drought tolerant low yield
>50% of the total of 95 crosses made by the National Tef
Research Program used our candidate lines as a parent
29

30. Muti-location testing
• 16 experiments at 24 traditional
tef growing areas.
– Distance from AA: 655km W, 955 N,
770 km NW
– Altitude: 1400 m We, 2500 m Bi/Sham
– Climate: Arid (Mehoni), semi-arid (Dh,
AT), Cool (Ho)
• 10 Non-traditional tef growing
areas (irrigated & rainfed).
– Distance: 750 km W, 900 km NW, 600
km E
30

31. Tef Improvement Project: Achivements
Lodging Tolerance: semi-dwarf cultivars
31

32. 32
abc
ab
bcd
abc
a
de
abcd
cd
de
e
bc
a
de cd
b
de
b
de
ef
f
Tesfa: new variety with desirable traits
Tesfa or RIL-181
• Compact panicle
• Non-shattering type
• Thick culm
• Suitable for irrigation
Euphytica (2018) 214:31

33. Tef Technology: Value-chain
Breeder
Agronomist
Soil scientist
Bioinformatician
Entomologist
Pathologist
Weed scientist
Food scientist
Socio-economist
Farmers (FPR)
Seed
Producers
Farmers
Market
Research
Policy
Inputs
Credit
Insurance
Consumers
Development
Improved
variety
Policy
&
Fund
33

34. 800
4500
3000
Import
Production
Area
+400
5700
3000
Import
Production
Area
Current yield:
1.5 ton/ha
Yield increase:
0.5 ton/ha
thousand ha
thousand ton
thousand ton
thousand ha
thousand ton
thousand ton
Impact of tef breeding
34

35. 35
Multiply
seeds
New
cultivar
Funding
(SFSA, UniBe,
SystemsX)
Seed agencies
(Private/Public)
NARS
(EIAR, RARIs)
Farmers
Implementation
(NARS, Syngenta-
Maribo, SFSA)
Tef Improvement
Project
(IPS, Bern)
Facilities
(Syngenta AG,
UniBe)
Service
(SIB, FGCZ, Fasteris,
MWG, Macrogen)
Research
(CDE, ETH Zurich,
Capetown Univ.,
KwaZulu-Natal, AAU,
Edinburgh Univ.)
Partnerships - PPP

36. Summary
36
• Goals & strategy
• Traits of interest: lodging & drought tolerance
• Omics: genome, transcriptome, RNAseq, microRNA
• Wild crosses
• Variety development & dissemination
• Value-chain approach: from research to development
(and consumption)
• Research on both major and orphan crops
• Involve all relevant stakeholders