Genetic modification of highland bananas:
biofortification with PVA and iron

G. Arinaitwe, H. Khana, P. Namanya, . Kiggun...
Introduction
Uganda has a population of 34 M people & is projected to be
53 M by 2025
Problems:
Inadequate food quantities...
Banana is synonymous to food

A

B

C
Bananas in Uganda
Banana weevils
Nematode effect
Banana bacterial wilt
Black Sigatoka disease
Micronutrient deficiencies in Uganda
Micronutrient undernutrition
The major micronutrient deficiencies in Uganda include:
...
Sources of genes for biofortification
psyB73
psy

psy2

Psy2a

crt I
Constructs used in biofortification
1st Batch

2nd Batch
(Opt series)

3rd Batch
(GEN series)

E1-ZmPsy1

E1-APSy2a

E1-Zm...
Biofortified product for low and highlands

Cultivars: ‘Nakinyika’ (EA-AAA)
And ‘Sukali Ndizi’ (ABB)

Cultivars: Hybrid M0...
ECSs work at Kawanda, Uganda
1. Male
bud

Genetic transformation

2. Inoculation stage

3. Embryogenic callus
4. Cell in s...
Materials and methods

Male bud

Immature flower

Embryogenic
callus

ECSs

ECS initiation and
transformation

ECS infecti...
Transformation
and selection

Shoot regeneration improved

2 Weeks

1.5 Months

2.5 Months

Shoot regeneration

Control: 0...
Weaning and hardening
First CFT at NARO, Uganda
CFTs at NARO-Kawanda
Several CFTs of
transgenic lines
containing disease
resistance genes and
micronutrient traits
Phenotypes observed
Carotenoid content at FG during storage
BCE (µg/g DW)
Cultivar

%Water content

Day1

Day7

Day 14

Mbwazirume

76.3

9.8
...
High carotenoid content in transgenic S. Ndizi
1
3

2

Control line

4

ZmPsy1 (12-39) line
Cross section of the control a...
PVA retention in target cooking bananas

Steamed

Boiled
FOC symptoms of the disease
Microconidia entry to the corm
is through wounded /cut root
hairs

Yellow
leaves

The conidia ...
FOC wilt in cultivar Sukali Ndizi

ND control

ND control

ND 1

ND 1

ND 2

ND 2

ND 3

ND 3

ND 4

ND 4
12
12

55

72

83

NTC
Acknowledgements
(Program Leader)

Dr. Wilberforce
Tushemereirwe

QUT

NARO

Rob Harding
Doug Becker
Marion Bateson
Jason ...
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Biofortification of highland bananas in Uganda - November 2012

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Presentation by Geoffrey Arinaitwe
Delivered at the B4FA Media Dialogue Workshop, Kampala, Uganda - November 2012
www.b4fa.org

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Biofortification of highland bananas in Uganda - November 2012

  1. 1. Genetic modification of highland bananas: biofortification with PVA and iron G. Arinaitwe, H. Khana, P. Namanya, . Kiggundu, A W.K Tushemereirwe and J. Dale Research Framework National Research Laboratories - Kawanda
  2. 2. Introduction Uganda has a population of 34 M people & is projected to be 53 M by 2025 Problems: Inadequate food quantities Imbalanced nutrients, containing mainly carbohydrates
  3. 3. Banana is synonymous to food A B C
  4. 4. Bananas in Uganda
  5. 5. Banana weevils
  6. 6. Nematode effect
  7. 7. Banana bacterial wilt
  8. 8. Black Sigatoka disease
  9. 9. Micronutrient deficiencies in Uganda Micronutrient undernutrition The major micronutrient deficiencies in Uganda include: Vitamin A deficiency (VAD) Iron deficiency Anaemia (IDA) Iodine Deficiency Disorders (IDD) National Nutritional Surveys 1994-1999 31 out of 36 districts had VAD 50% of children have VAD 40% of children > 5 years have IDA 50% of pregnant women had IDA 30% of maternal deaths were attributable to IDA
  10. 10. Sources of genes for biofortification psyB73 psy psy2 Psy2a crt I
  11. 11. Constructs used in biofortification 1st Batch 2nd Batch (Opt series) 3rd Batch (GEN series) E1-ZmPsy1 E1-APSy2a E1-ZmPsy1Q60 E1-Crt1 Acs-ZmPsy1Q60+E1-S.Fer E1-APsy2a E1-APsy2a Acs-ZmPsy1Q60 BT4act-APsy2a E1-S.Fer E1-ZmPsy1Q60 Ubi-APsy2a 35S-Fro2 E1-ZmPsy1 Ubi-ced9 35S-IrtI Ubi-mced9 Ubi-AtBag4 Ubi-Gus E1-Gus
  12. 12. Biofortified product for low and highlands Cultivars: ‘Nakinyika’ (EA-AAA) And ‘Sukali Ndizi’ (ABB) Cultivars: Hybrid M09 Tolerant to: Black Sigatoka, Nematodes and weevils
  13. 13. ECSs work at Kawanda, Uganda 1. Male bud Genetic transformation 2. Inoculation stage 3. Embryogenic callus 4. Cell in suspension 5. Somatic embryos formed from suspensions 6. Embryo germination 7. Multiple shoot formation and rooting 8. Weaning and potting of plantlets
  14. 14. Materials and methods Male bud Immature flower Embryogenic callus ECSs ECS initiation and transformation ECS infection Shoots from EC clones Selection Future activities Morphological analyses PCR analyses Southern blot analyses NARO-KARI Biotechnology Laboratory
  15. 15. Transformation and selection Shoot regeneration improved 2 Weeks 1.5 Months 2.5 Months Shoot regeneration Control: 0mg GA3 0.5 mg/L GA3 1.0 mg/L GA3 1.5mg/L GA3
  16. 16. Weaning and hardening
  17. 17. First CFT at NARO, Uganda
  18. 18. CFTs at NARO-Kawanda Several CFTs of transgenic lines containing disease resistance genes and micronutrient traits
  19. 19. Phenotypes observed
  20. 20. Carotenoid content at FG during storage BCE (µg/g DW) Cultivar %Water content Day1 Day7 Day 14 Mbwazirume 76.3 9.8 17.5 23.5 Nakinyika 73.9 76.9 75.1 74.5 58.2 65.2 13.5 19.6 9.1 4.3 18.2 0.8 21.0 23.4 18.0 11.0 22.3 1.7 26.5 28.3 18.8 16.8 23.1 2.2 Nakitembe Kisansa M9 Butobe Kayinja NAKINYIKA NAKITEMBE HYBRID M9 KAYINJA
  21. 21. High carotenoid content in transgenic S. Ndizi 1 3 2 Control line 4 ZmPsy1 (12-39) line Cross section of the control and the ZmPsy1 (12-39) line; above: Chromatogram peaks for 12-39 fruit extract identified at λmax=450nm, peak 1, lutein at 6min; peak 2, α-carotene at 12.2; peak 3, trans-β-carotene 13.1min & peak4, cis β-carotene at 13.9min
  22. 22. PVA retention in target cooking bananas Steamed Boiled
  23. 23. FOC symptoms of the disease Microconidia entry to the corm is through wounded /cut root hairs Yellow leaves The conidia spreads up the xylem and blocks water and nutrient movement Discoloured pseudostem Death of plant releases the dormant chlamydospores back into the soil Reinfection is through running Discoloured water and propagating suckers corm
  24. 24. FOC wilt in cultivar Sukali Ndizi ND control ND control ND 1 ND 1 ND 2 ND 2 ND 3 ND 3 ND 4 ND 4
  25. 25. 12 12 55 72 83 NTC
  26. 26. Acknowledgements (Program Leader) Dr. Wilberforce Tushemereirwe QUT NARO Rob Harding Doug Becker Marion Bateson Jason Geijskes Harjeet Khanna Anthony James Jen Kleidon Bulu Malazi A. Kiggundu G. Arinaitwe P. Lamwaka B. Magambo R. Ecodu P.Namanya Prof James Dale

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