Biotechnology

Research-for-Development

at IITA

I Ingelbrecht

IITA-Ibadan, 31 August 09

Biotech complementary tool to conventional approaches

Multidisciplinary approach (breeding, genetic resources, pathology, etc)

Product oriented

Partnerships: national research programs across SSA

and advanced labs overseas

Integrate capacity building with research

Guiding principles

~8 full time PhD scientists with additional scientists

(breeding, virology, pathology, etc)

Two main hubs: HQ in Nigeria + BECA, Kenya

Crop Genetic

TransformationIn vitro Tissue Culture

Biotechnology R-4-D

Molecular Marker

Applications

‘traditional’ ‘modern’

Capacity building

I. In vitro Tissue Culture

yam

cassava

Banana, plantain

Activities

In vitro micropropagation for distribution of elite

cassava, yam and banana/plantain materials

Disease clean up: production of certified, disease-free

planting materials (with PQS)

Training + technical backstopping of NARS in all

aspects of in vitro micropropagation and set up of

tissue culture facilities

High demand by national programs across SSA

Technology well established: applications for breeding &

genetic resource conservation (fairly) routine

Technology transferred to national programs, in some cases

(high value crops) applied for commercial purposes

In vitro tissue culture recently integrated with Genetic

Resources Center at HQ, Ibadan, Nigeria

Impact

II. Crop Genetic Transformation

• GM crops developed and commercialized in industrialized as well as

developing countries with demonstrated benefits for producers and

environment

• With proper identification of targets, GM technology can complement

conventional breeding for intractable traits in SSA agriculture

• For ‘orphan’ crops tools and products mostly in stage of development,

some already available

• Need to consider regulatory environment lacking in most countries

in SSA; capacity building

Ongoing projects

Genetic transformation of cassava for resistance to the Cassava Brown Streak Disease, starch modification

Genetic transformation of banana for bacterial blight resistance,

DNA recombinant technology for construction of genes and analysis of transgenic plants

Biosafety with application for CFT of GM crops, by national programs

Three steps in genetic transformation protocol:

1. In vitro shoot regeneration method

2. Gene transfer method: Agrobacterium

3. Selection and/or screening for transgenic shoots

Focus on African genotypes

Example: cassava, banana

Uganda

Kenya

Tanzania

Mozambique

Zambia

DRC

ROC

Equatorial

Guinea

Malawi

CBSD reportedCBSD damagingCBSD devastating

Transformation of farmer-preferred cassava

for CBSD resistance

A transgenics approach for resistance to Potyviruses previously used and

grown commercially: eg papaya resistant to Papaya ringspot virus

Current transformation protocols are for ‘model’ genotypes, not used

by farmers or breeders in Africa: excellent research tool but

limited application in the field

Bottleneck since current protocols are genotype-dependent

develop protocol for (African) farmer-preferred lines

Why target cassava landraces?

Multiple shoots

Shoot org

anogenesis

Plantlets

Adventitious shoots

Primary SE

Em

bry

ogenesis

Em

bry

ogenesis

Explants; e.g. immature leaf lobes

Cotyledonary

stage SE*

Embryos

Friable embryogenic

Callus*

Embryogenic

suspensions

Protoplasts

Secondary SE*

Summary cassava regeneration responses

Fig. modified from Zhang et al. 2006

+/-

++++

++++

++

+/-+/- 0 - 40%

++ 20 - 40%

++++ >90%

++++

cv Albert TME12

SE produced for 9 genotypes: 4 landraces from ESA

4 landraces from WCA

1 IITA elite line

Somatic embryogenesis

Transgenic African cassava with reporter gene

Transgenic African cassava with reporter gene

b

a

d

c

e

3 mm

4 mm

5 mm

3 mm

1 cm

Non transgenic Transgenic – 1st ratoon Transgenic – 2nd ratoon

petiole

Young stem

Old stem

Fibrous root

tuber

Status

- Resistance genes developed and tested in N benthamiana:

resistance is strain specific

- Transformation of African cassava with resistance genes

ongoing

Banana transformation - status

- Transgenic plants produced with resistance gene(s) for

bacterial blight

- Field trial planned for 2010 (Uganda)

III. Molecular marker applications

• For characterization and management of germplasm and pathogens

(diversity) and for diagnostics purposes

• In marker assisted selection/breeder to accelerate breeding

process; extensively used in cereals and other crops of developed world

• As with GM technology, for ‘orphan’ crops tools are mostly in stage

of development

Rationale

Ongoing projects

Characterization of germplasm collections of cassava, yam, banana

QTL mapping of virus (CMD & CBSD) resistance in cassava, drought tolerance in cowpea, maize and cassava

MAS for striga resistance in cowpea

Development of SSR and SNP markers for cassava & yam

Diagnostic markers for bacterial blight + other pathogens

Development of DNA microarray for gene discovery in cassava

Total number of EST sequences investigated: 18,166

Total number of unique SSR loci appropriate for primer modeling: 646 (3.3%)

Number of candidate SSR investigated : 346

PCR successful: ~ 90% Failed PCR: ~ 10%

Eliminate

PCR products with expected sizes Amplification of introns

Number of unigenes used for in silico identification of SSRs: 8,577

Example: EST-SSR marker development

Screen on diversity panel

> 500 bp

Example cassava

- 3078 cassava accessions with minimal passport data held in trust in GRC

DNA fingerprinting for effective

management and use by breeding

program

New Cassava SSR markers

M 1 2 3 4 5 6 7 8 9 10 1112 1314 15 16 1718 19 20 21 22 23 24

180 new SSR markers

Used for fingerprinting collection, mapping and variety identification

Semi-automated genotyping using fluorescent labeling

Different technologies can be used

for diversity studies, QTL mapping

Example: cassava microarray

~14,000 genes or 1/4 to

1/3 of all cassava genes

- Hybridization and scanning

- Identify differentially expressed genes for use in transgenic

program or complement QTL mapping

IV. Capacity Building & partnerships

- Degree (MSc, PhD) and non-degree training of students or VS;

mostly integrated with ongoing research activities; topical also possible if funded

- Workshops: genetic transformation, molecular breeding, biosafety

- Infrastructure: eg MARI, Tanzania for GM

Advanced labs:

DDPSC, USA; ETH, Switzerland; KU, Copenhagen, Univ Virginia, USA; others

National research institutions and universities

NRCRI, Nigeria; MARI, Tanzania, NABDA, Nigeria; many universities in Nigeria

and other African countries for joint degree research.

Development investors

USAID

RF

BMGF

Generation CP

AATF

Gatsby Charitable Foundation

Masterfoods