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Plant Biotechnology

Access Course Presentation
by

Rechelle Reid

on 8 May 2015

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Transcript of Plant Biotechnology

Ideology
vs.
Reality
A. Tumefaciens
Global
Health
v
Global
Wealth
The
Science

Global health vs. global wealth
Ti Plasmid of
A. Tumefaciens
Bibliography
"The
protection
of human health and the environment are our overriding priorities."

GM
crops enter Britain mainly as
animal
feed.

GM
potatoes
and GM
wheat
UK
Position
Public
Debate
Disadvantages
Advantages
Supporting Study
Positioning
GM crops as ‘good
or
bad

We must focus
rationally
on which technological approach is most effective, including
GM
.

Economic
, political and
technical
challenges remain.

Global
consensus
on how to
regulate
GM crops.
Global
Monopoly
Pricing and Price
Discrimination

Monsanto
controls 95 percent of the market for insect and herbicide resistant cotton traits. They invented a type of
GM
seed that would be resistant to the
chemicals
. They were called "
Round-Up Ready
" seeds

If a
company
wants to produce or sell these
seeds
, they must pay a licensing fee to
Monsanto

Because
Biotech
companies gain so much revenue from these royalties, they make sure to put
15%
of their income from sales back into
R&D
. It is their primary business strategy.
Department for Environment, Food & Rural Affairs (2013 online) 'Genetic Modification (GM)' Available on http://www.defra.gov.uk/environment/quality/gm/ accessed on: 19-2-2013

Langreth R and Herper M (2009 online) 'The Planet Versus Monsantao' Available on http://www.forbes.com/forbes/2010/0118/americas-best-company-10-gmos-dupont-planet-versus-monsanto.html accessed on 25-02-2013
Nuffied Council on Biothics (2013 online) 'The use of genetically modified crops in developing countries' Available on http://www.nuffieldbioethics.org/sites/default/files/GM%20Crops%20short%20version%20FINAL.pdf accessed on 19-2-2013

Robin Mckie (2009 online) 'Why Britain faces a bleak future of food shortages' Availabe on http://www.guardian.co.uk/science/2009/dec/13/britain-faces-food-shortage accessed on 23-02-2013

World Health Organization (2013 online) 'Micronutrient deficiency' Available on http://www.who.int/nutrition/topics/vad/en/ accessed on: 18-2-2013

World Health Organization (2013 online) '20 Questions on Genetically Modified (GM) Foods Available on http://www.who.int/foodsafety/publications/biotech/20questions/en/ accessed on: 18-2-2013
Plant Biotechnology Applications
Agrobacterium Tumefaciens and Ti Plasmid
Direct Nuclear Transformation
Protoplast Transformation
Biolistic
Viral Vectors
Chloroplast Transformatio
Transgenic Technologies
Gene and Genome Analysis
Comparative Genomics
Functional Genomics
Developmental Studies
A. Tumefaciens
- Gram-negative soil bacterium belongs to the class alpha-proteobacterium.
Contains a 200- kb tumour inducing plasmid (Ti) that genetically transform plant cells leading to the formation of crown gall tumours in dicotyledonous plants.
Crown gall formation is the result of the transfer, integration and expression of T-DNA (Transferred DNA) into plant genome.
T-DNA contains 2 types of genes:
Oncogenic genes - auxins and cytokinins synthesis
Opine synthesis genes.
T-DNA is flanked by two 25-bp direct repeats border sequences :
Left and right border - mediates the integration of the T-DNA into the plant genome.
Opine catabolism and virulence (
vir
) genes.
Isolation of the genes of interest from the source organism.
Development of a functional transgenic constructs:
Gene of interest
Promoters to drive expression
Codon modification
Marker genes
Insertion of the transgene into the Ti-plasmid.
Introduction of the T-DNA-containing-plasmid into Agrobacterium.
Regeneration of the transformed cells into genetically modified plants.
Transformation of transgenic plants using Ti Plasmid
Scientific Debate
• Less labour intensive- does not require sophisticated equipment.
• More cost effective.
• Agrobacterium-mediated transformation typically results in a low copy number of the introduced gene as opposed to particle bombardment leading to introduction of multiple copies.
• Experiences with rice, potato and wheat suggest that higher transgene copy numbers correspond to higher expression levels, indicating that the transgenes are expressed efficiently.

Advantages
of Ti Plasmid
Technique
• Limited success has been reported with Agrobacterium-mediated transformation of:
− Seeds
− Somatic embryos
− colyledonary node
− Soybeans
− Immature zygotic cotyledons

Disadvantages of
Ti
Plasmid
Technique
The bacterium transfers part of its DNA to the plant, and this DNA integrates into the plant’s genome, causing the production of tumours and associated changes in plant metabolism.
Plant tissue and
Agrobacterium
must be compatible for process of T-DNA transfer and integration to occur.
DNA must be introduced into cells which are both susceptible to
Agrobacterium
and responsive to plant regeneration.
Agrobacterium
-mediated transformation relies on sonication-induced tissue wounding to provide an entry point for the bacterium.
Crown Gall Disease
Plant Biotechnology:
Agrobacterium tumefaciens

By: R.A.P
Plant Biotechnology
“Plant biotechnology describes a precise process in which scientific techniques are used to develop useful and beneficial plants’’
Council for Biotechnology
• Investigated the effect of two
A. tumefaciens
strains:
LBA4404
and
AGL0
• 3 different binary plasmid ORI on transformation frequency
• Vector backbone insertion
• Single copy event frequency
• Quality event frequency a
• Usable event quality frequency

Effect of
Agrobacterium
strain and plasmid copy number on transformation frequency, event quality and usable even quality in an elite maize cultivar.
Agrobacterium transformation significantly affects maize transformation frequency, vector backbone insertions and event quality.
Replication of origins on the binary vectors and plasmid copy number correlated with transformation frequency.
Higher copies of the super-binary vector improved overall transformation frequency.
Conclusion/Results
Methodology
T-DNA vectors were introduced into A. tumefaciens strains AGL0 and LBA4404 carrying the acceptor plasmids pSB1, PHP527 and PHP528 by electrophoration.
Plasmid DNA was extracted from the Agrobacterium strains were transformed into E. coli and analysed by restriction digestion.
Relative plasmid copy number was determined by quantitative real-time PCR
References
Full transcript