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

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. 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. Banana is synonymous to food
A
B
C

4. Bananas in Uganda

5. Banana weevils

6. Nematode effect

7. Banana bacterial wilt

8. Black Sigatoka disease

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. Sources of genes for biofortification
psyB73
psy
psy2
Psy2a
crt I

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. 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. 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. 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. 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. Weaning and hardening

17. First CFT at NARO, Uganda

18. CFTs at NARO-Kawanda
Several CFTs of
transgenic lines
containing disease
resistance genes and
micronutrient traits

19. Phenotypes observed

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. 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. PVA retention in target cooking bananas
Steamed
Boiled

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. 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. 12
12
55
72
83
NTC

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