Transcript AARINENA Agricultural Biotechnology Network Expert Meeting,15

ICARDA
Developments on Biotech and Biosafety
West Asia and North Africa
M. Baum,
The Internartional Center for Agricultural Research in the Dry Areas (ICARDA)
P.O. Box 5466 Aleppo
Syria
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Introduction
ICARDA
Gradients of Biotechnology
ICARDA
5
5. Biosafety
Proteomics
4. Genomics
4
Bioinformatics
Genetic engineering of animals
3. Genetic engineering of plants
Genetic engineering of microbes
Rising
cost
Recombinant DNA technology
2. Molecular breeding
2
Monoclonal antibody production
Embryo transfer in animals
1. Plant tissue culture
1
Biological nitrogen fixation
Introduction
Microbial fermentation
Increasing efficiency
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
3
1. Plant Tissue Culture
ICARDA
Introduction
Plant tissue culture
Date palm production via somatic embryogenesis or organogenesis:
UAE, Morocco, Kuwait, Saudi Arabia, Egypt, Iraq, etc.
Other tissue culture techniques used: production of virus free planting
material such as seed potatoes, citrus, bananas, ornamental plants
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Doubled Haploids in Cereals
ICARDA
Aim: Development of homozygous (true-breeding
lines) to speed up development of breeding lines
Introduction
Plant tissue culture
Doubled haploids (Algeria, INRA-Tunisia, INRAMorocco, Sudan, ICARDA)
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Gradients of Biotechnology
ICARDA
5
5. Biosafety
Proteomics
4. Genomics
4
Bioinformatics
Genetic engineering of animals
3. Genetic engineering of plants
Genetic engineering of microbes
Introduction
Plant tissue culture
Rising
cost
Recombinant DNA technology
2. Molecular breeding
2
Monoclonal antibody production
Embryo transfer in animals
1. Plant tissue culture
1
Biological nitrogen fixation
Microbial fermentation
Increasing efficiency
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
3
National Genebanks e.g. Morocco, Egypt, Iran, Algeria etc
International Gene bank efforts: ICARDA (133,000 accessions)
Genetic characterisation of germplasm accessions with
molecular markers: AFLPs, SSRs, SNP etc.
Development of Core Collections
Introduction
Plant tissue culture
Molecular breeding
ICARDA
2a. Collection and Molecular Characterisation of
Plant Genetic Resources
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Molecular breeding
Plant tissue culture
Introduction
2b. Molecular Breeding
ICARDA
Genetic mapping with molecular markers: AFLPs, SSRs, SNP etc.
Evaluation of resistances to biotic and abiotic stresses
Correlation and creation of QTL maps
Marker-Assisted Selection with diagnostic markers
ICARDA
Scald (Rhynchosporium secalis) Resistance
Gene Rrs1 on Chromosome 3H in Barley
Introduction
Plant tissue culture
Molecular breeding
Genetic engineering
Examples of Marker-Assisted Selection
Molecular marker efforts in: Egypt, Morocco,
Jordan, Tunisia, Sudan, Iran, Turkey, etc.
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Genetic engineering
Molecular breeding
Plant tissue culture
Introduction
Gradients of Biotechnology
ICARDA
5
5. Biosafety
Proteomics
4. Genomics
4
Bioinformatics
Genetic engineering of animals
3. Genetic engineering of plants
Genetic engineering of microbes
Rising
cost
Recombinant DNA technology
2. Molecular breeding
2
Monoclonal antibody production
Embryo transfer in animals
1. Plant tissue culture
1
Biological nitrogen fixation
Microbial fermentation
Increasing efficiency
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
3
Introduction
Plant tissue culture
Molecular breeding
Genetic engineering
Introduction
AARINENA Biotechnology Network Meeting, Cairo 15-16 December, 2007
Plant tissue culture
Molecular breeding
Genetic engineering
Introduction
AARINENA Network Meeting, Cairo 15-16 December, 2007
Plant tissue culture
Molecular breeding
Genetic engineering
Genetic engineering
Molecular breeding
Plant tissue culture
Introduction
Status of plant genetic engineering research at AGERI/Egypt
Discipline
Potato
Tomato
Cotton
Maize
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Virus
resistance
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Stress
tolerance
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Genome
mapping
and finger
printing
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Fungal
resistance
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Insect
resistance
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Faba
bean
Cucurbits
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Wheat
Banana
Date
palm
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AARINENA Biotechnology Network Meeting, Cairo 15-16 December, 2007
Genetic engineering
Molecular breeding
Plant tissue culture
Introduction
Status of plant transformation at ABRII/ Iran
Crop
Gene/Trait
Current status
Field release
Laboratory
Green house
Field test
Rice variety 1
CrylA(b), insect resistance
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Rice variety 1
Chitinase, sheath blight
resistance
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–
Rice variety 1
Salinity/drought tolerance
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Rice variety 2
CrylA(b)
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■
.
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Rice variety 2
Glucanase
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.
Rice variety 2
Glucanase + Chitinase
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.
Rice variety 3
CrylA(b)
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.
Cotton
CrylA(b)
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.
wheat
Salinity/drought tolerance
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wheat
Glucanase + Chitinase
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Maize
CrylA(b)
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Maize
Glucanase + Chitinase
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Sugar beet
CrylA(b)
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Canola
AGL1, AGL5, AGL8
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Canola
Herbicide resistance gene
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Cumin
GOX
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Potato
CrylII
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Genetic engineering
ICARDA Legume Crops
Introduction
Plant tissue culture
Molecular breeding
ICARDA
AARINENA Biotechnology Network Meeting, Cairo 15-16 December, 2007
Genetic engineering
Molecular breeding
Plant tissue culture
Introduction
Lentil transformation systems
ICARDA
1
2
Surface
sterilization
Wounding
8
7
3
Co cultivation
4
Selection
(Kanamycin, Basta)
6
5
5
confirmation
Transferring
into pots
Putative
transgenic
explants
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Elongation
Wheat plants growing in
the field with spikes
Immature embryos (10-12
days post-anthesis)
Induced callus and somatic
embryos on scutellum of
immature embryos
Genetic engineering
Introduction
Plant tissue culture
Molecular breeding
Wheat transformation procedure ollaboration with CBS, Tunisia and
AGERI, Egypt
Selection and regeneration of
transgenic wheat plants
Gene gun bombardment
Genetic engineering
Molecular breeding
Plant tissue culture
Introduction
Genes/Phenotype Category for Legume and Cereal
ICARDA
Transformation at ICARDA
Enhancing
drought/salinity
stress tolerance
Reporters
Anti-fungal
/Herbicide
resistant
BI-GST(Kanr & PPTr)
Dreb1A (Kanr & PPTr)
35S:Bar / GUS (PPTr)
vst-1 / bar (PPTr)
LeGST-T2 (Kanr)
rd29A / LeREBP (PPT r)
rd29A/GUS (Kanr)
Afp/bar(PPT)
LeGST-T3 (PPTr)
35S / LeREBP (PPTr)
Ubi1: 35S:Bar/GUS
LeGST-T5 (Kanr)
TVP1
LeTpx1 (Kanr & PPTr)
TNHx1
Enhancing
oxidative stress
tolerance
ICARDA
ICARDA
Establishment of a Containment Facility at
ICARDA, July 2007-July 2009 (AFESDbiotech Phase 2)
Duration: July 2007- June 2009
Budget: US$ 850.000
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Testing
of plant quarantine diseases e.g. rust pathogens
Rust Pathway
East Africa-onward
Occurrence and Movement of Air borne Pathogens
Molecular breeding Genetic engineering Genomics
Plant tissue culture
Gradients of Biotechnology
ICARDA
5
5. Biosafety
Proteomics
4. Genomics
4
Bioinformatics
Genetic engineering of animals
3. Genetic engineering of plants
Genetic engineering of microbes
Rising
cost
Recombinant DNA technology
2. Molecular breeding
2
Monoclonal antibody production
Embryo transfer in animals
1. Plant tissue culture
1
Biological nitrogen fixation
Microbial fermentation
Increasing efficiency
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
3
Molecular breeding Genetic engineering Genomics
Plant tissue culture
ICARDA
Identification of Candidate Genes with
Microarrays: Genomics
1.
DNA-microarrays is a modern method,
that allows to analyse the genes of cells
during different growth stages
2.
For the transcription analysis mRNA is
extracted from cells that were grown
under different environmental conditions,
and labelled with two different fluorescent
dyes
3.
The labelled nucleic acids are then
hybridised with the microarrays and the
fluorescence intensity of the individual
spots quantified through laser light. Upand down-regulated genes are identified
X
barley 1 Gene Chip: 22.000 Genes
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
2a. Relativer Chlorophyll Contents
2b. Ratio of Fv/Fm
ICARDA
tolerant
tolerant
susceptible
1.2
1.2
1.2
1
1
1
0.8
0.8
0.8
0.6
0.6
Genotype 1
Martin
Control
Control
Control
0.2
Drought-stress
0.2
Drought-stress
1
3
5
7
9
11
0
13
Tage nach Behandung
1
3
5
7
9
11
0
13
Tage nach Behandung
1
1
0.8
0.8
0.8
0.6
Genotyp1
0.4
0.6
H.spontaneum
HS41-1
Martin
0
1
3
5
7
9
11
Days after Treatment
13
7
9
11
13
Control
0.4
Drought stress
0.2
0.2
0.2
5
Genotyp 2
Drought stress
Drought stress
3
Morocco9-75
Control
0.4
Control
1
Tage nach Behandung
1
0.6
Drought-stress
0
0
0
0
Genotype 2
Morocco9-75
0.4
0.4
0.4
0.2
0.6
H.spontaneum
HS41-1
0
1
3
5
7
9
11
13
Days after Treatment
0
1
3
5
7
9
11
13
Days after Treatment
Figure 2 Chlorophyll contents and maximum quantum yield of PSII (Fv/Fm) of three
genotypes (Martin, HS41-1 and Morocco9-75) under well-watered conditions (70%
available water in the soil) and drought stress (10% available water in the soil). Results
are presented as mean ± SE of six individual measurements.
Genomics
Plant tissue culture Molecular breeding Genetic engineering
Use of Microarrays and qPCR
ICARDA
77 genes significantly regulated at severe drought stress during
heading stage could putatively be only responsive genes to
drought stress. e.g. dehydration-resp. protein RD22, heat
shock protein-like, dehydrin 3, protein kinase SPK-3, etc
•372 genes differentially are the specific/ responsible genes for
drought tolerance in Tadmor during heading stage. e.g. serine/
threonine kinase-like protein, pathogenesis related protein-1
and 4, PRB1-2, PRB1-4, ABC transporter family protein,
organic anion transporter, glutathione S-transferase, etc.
Biosafety
Genomics
Tissue C. Molecular breeding Gen.engineering
ICARDA
Current Status of Biotechnology for Crop production
in the dry areas
Biotech application
Countries
Tissue culture applications
Micro-propagation
Algeria, Egypt, Iran, Iraq, Jordan, Kuwait,
Lebanon, Morocco, Syria, Tunisia, Turkey,
United Arab Emirates
Doubled haploids
Algeria, Iran, Morocco, Sudan, Syria, Tunisia
Cell cultures
Egypt, Morocco, Tunisia, United Arab Emirates
Molecular breeding
Algeria, Egypt, Iran, Iraq, Jordan, Kuwait,
Lebanon, Morocco, Syria, Tunisia, Turkey
Genetic Engineering
Egypt, Iran, Morocco, Syria, Tunisia, Turkey
Genomics
Egypt, Iran, Syria, Tunisia, Turkey,
Proteomics
Egypt, Iran,
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Gradients of Biotechnology
ICARDA
5
5. Biosafety
Proteomics
4. Genomics
4
Bioinformatics
Genetic engineering of animals
3. Genetic engineering of plants
Genetic engineering of microbes
Rising
cost
Recombinant DNA technology
2. Molecular breeding
2
Monoclonal antibody production
Embryo transfer in animals
1. Plant tissue culture
1
Biological nitrogen fixation
Introduction
Microbial fermentation
Increasing efficiency
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
3
Biosafety
Genomics
Tissue C. Molecular breeding Gen.engineering
International Regulatory Developments
Cartagena Protocol on Biosafety
ICARDA
2003, September: Biosafety Protocol comes into force
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Legally binding framework for international movement of living GMOs
– Does not dictate national regulations;
– Provides strong incentive for harmonisation
Distinguishes between import for planting or for food
Establishes Biosafety Clearing House (BSC)
Does not oblige countries to create full regulatory system
Risk assessment and risk management
Unintentional /illegal transboundary movement
Capacity Building
Public Awareness and participation
Socio-economic considerations
Liability and redress
Financial mechanism
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Biosafety
Genomics
Tissue C. Molecular breeding Gen.engineering
National Biosafety Frameworks (NBFs)
Article 8g of the CBD and Article 2 of the CPB:
“…Each Party shall take necessary and appropriate legal, administrative
and other measures to implement its obligations ….”
NBFs vary from country to country, but usually have a number of
common components:
•
A policy on biotechnology and biosafety
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Regulatory framework for biosafety
•
System to handle notifications/requests for permits
•
Systems for enforcement and monitoring
•
Public information and public participation
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
ICARDA
Development of National Biosafety Framework
• Duration of the project:
– 18 months (September 2006)
• Finance:
– UNEP – GEF
• Responsibility:
– Ministry of Environment in cooperation with
other ministries and organizations. Source: B. Safadi
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Biosafety regulations have been enacted in Syria as of 2001
Relevant Laws
Syrian Agriculture Quarantine Law 237 dated 1960
Decree 91 of the Syrian Ministry of Agriculture dated 1991
Syrian Law 158 (prevention of fraud and cheating) dated
1960 and its modifications
Source: B. Al-Safadi
Biosafety
Genomics
Tissue C. Molecular breeding Gen.engineering
ICARDA
Current status of National Biosafety Frameworks
in West Asia and North Africa
11 Countries have completed their NBF under
UNEP-GEF projects:
Algeria, Egypt*, Iran, Jordan, Lebanon, Mauritania*,
Syria, Tunisia*, Yemen
Countries developing NBFs without GEF support:
Oman, Saudi Arabia
*These countries completed their NBFs under the
UNEP-GEF pilot projects 1997-99
Source: N. Mohamed
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Tissue C. Molecular breeding Gen.engineering
Genomics
Biosafety
Regional Project on Biotechnology and
ICARDABiosafety for Agriculture and Environment in
the WANA sub-region.
• Which Countries will participate in the Project?
• West Asia:
• NBF Implementation (5) – Iran*, Jordan*, Lebanon, Syria*,
and Yemen*;
• NBF Development (2) - Iraq, Palestinian Authority.
• Other partner countries (4) – Kuwait*, Oman*, Qatar*, Saudi
Arabia*.
• North Africa:
• NBF Implementation (8) – Algeria*, Egypt*, Ethiopia*, Libya*,
Mauritania*, Morocco, Sudan*, Tunisia*.
• Note:
• *These countries are Parties to the Cartagena Protocol on
Biosafety (CPB)
• This project is a joint initiative between FAO, ICARDA, and
UNEP, and will build on each agency’s comparative
advantage
Biosafety
Genomics
Tissue C. Molecular breeding Gen.engineering
ICARDA
What is the overall Project Objective?
To develop and strengthen biosafety capacity in WANA sub
region to promote agricultural and environmental sustainability
through the implementation of national biosafety frameworks
(NBF), capacity building for the safe development and
application of biotechnology, information sharing, regional
cooperation, and strengthening of regional institutions and
cooperation.
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Biosafety
Genomics
Tissue C. Molecular breeding Gen.engineering
What will the Project aim to achieve?
• Implementation of NBFs in the countries of the WANA subregion, resulting in fully operational and effective NBFs in place in
all CPB Parties. By the end of the project in 2012, each Party will
have:
– Integrated biosafety & biotechnology into national development
plans & policies;
– Regulatory systems (laws and regulations) in place for handling
GMOs;
– Systems in place for handling requests for GMOs including risk
assessment, decision-making, risk management and risk
communication;
– Systems in place for monitoring; enforcement, and emergency
responses for GMOs;
– Systems in place for public awareness, education, participation
and access to information;
– Increased capacity for Biosafety management including: trained
human resources; strengthened institutions; and the necessary
decision tools, technical and laboratory facilities for risk
assessment and management.
ICARDA
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Biosafety
Genomics
Tissue C. Molecular breeding Gen.engineering
Capacity building and establishment of mechanisms for regional
cooperation on biosafety and biotechnology in WANA
• resulting in a fully functional regional framework for cooperation on
biosafety and biotechnology. By the end of the project, the sub-region will
have in place:
– An Inventory of national and regional biosafety, biotechnology and
biodiversity resources and capacity building needs;
– National and sub-regional strategies for biosafety and biotechnology;
– Common approaches, including decision aiding tools, for risk
assessment, risk management, risk communication, and monitoring of
GMOs;
– Common approaches, including decision aiding tools, for consideration
of religious, ethical and socio-economic issues in decision-making on
GMOs;
– Strengthened national and regional centres of excellence and
ICARDA
reference laboratories;
– Regional networks, including a regional BCH, for sharing information,
reference data and on-line technical and training resources on
biosafety and biotechnology in Arabic and other regional languages.
Biosafety
Genomics
Tissue C. Molecular breeding Gen.engineering
Capacity building for safe development
and application of Biotechnology
•
resulting in improved food security and
sustainable management of agro-biodiversity
in the WANA sub-region. By the end of the
project, the sub-region will have in place:
–
–
–
–
An inventory of strategic crops and important biotic
and abiotic stresses;
Strategic application of GM and non-GM
biotechnology to resolve major constraints to
environmental and agricultural sustainability;
Characterization of national and regional
germplasm base for key agronomic crops;
Agreed mechanisms for sharing of genetic
resources within the sub-region.
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt
Thank you for your attention
AARINENA Agricultural Biotechnology Network
Expert Meeting,15-16 December 2007, Cairo-Egypt