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Презентация на тему Plant biotechnology

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Plant Tissue CulturePlant cells differ from animals cells in that they are totipotentA totipotent cell is one that can develop into specialized cell types & regenerate an entire organismTissue culture of plants and the regeneration of
Plant Biotechnology Plant Tissue CulturePlant cells differ from animals cells in that they are http://www.webschoolsolutions.com/biotech/transgen.htmPlant Cloning http://catf.bcresearch.com/biotechnology/tissueculture_research.htmMicropropagationCallus, undifferentiated mass of plant cellsSeedlings, each from an individual cell How Do They Engineer Plants? Methods of producing transgenic plants http://www.colostate.edu/programs/lifesciences/TransgenicCrops/how.htmlPlant Genetic EngineeringTi plasmid of Agrobacterium tumefaciensA. tumefaciens is a soil microbe T-DNA portion of the Ti plasmid contains genes responsible for the diseaseT-DNA http://www.webschoolsolutions.com/biotech/transgen.htm http://faculty.abe.ufl.edu/~chyn/age2062/lect/lect_09/10_19A.GIFBiolistics (Biological Ballistics)Useful for engineering corn, rice, wheat, barley, & other crops Helios Gene Gunhttp://www.bio-rad.comUses an adjustable burst low-pressure helium to sweep DNA- or Genetically Engineered PlantsHerbicide toleranceInsect resistanceCrop improvementsFunctional foodsPlants as bioreactorsBiofuelsTimber improvementsBioremediation Currently 215 million acres of GM crops grown worldwideGenetically Modified (GM) Crops >60% of the foods we purchase have GM ingredients 95% of canola Who Produces GM Food?BASF Inc.Aventis CropscienceBayer CropscienceSyngenta Seed Inc.Pioneer Hi-Breed International Inc.Dow Agroscience LLCMonsanto Company FLAVR SAVR, The First GM Food http://resources.emb.gov.hk/envir-ed/globalissue/images/ModifiedTomato.jpgThe first GM food was the FLAVR SAVR tomatoIntroduced in 1994 it had delayed ripening characteristics Fruit softens because polygalacturonase degrades pectinAntisense technology was used to turn off PG genetranscriptionmRNAtranslationPG genetranscriptionmRNAAntisense mRNAtranslationAntisense Technology Polygalacturonase (PG) is an enzyme that breaks down pectin in ripening http://www.wachstumshormon.info/kontrovers/gentechnik/flavrsavr.html?gfx=2Most GM tomatoes were used only in canned pureeBy 1997 FlavrSavr was no longer marketed Current technologies aid the farmer not the consumerHerbicide ResistanceVirus ResistanceInsect Resistance Herbicide ToleranceMethods used to promote crop growth also promote weedsWeeds often outgrow Soybean with no herbicidesSoybean after herbicidesHerbicides are used for weed controlHerbicide Tolerance Non-selective herbicides  (Roundup Ultra and) Roundup® (chemical name: glyphosate) Breaks down EPSPS Transgene Introduced into PlantsCodon usage modifiedfor efficient expression in plantspromoterRegulatory sequences Move to greener herbicide Benefits of Glyphosate Tolerance in Crops Can Roundup® Ready CropsCornAlfalfaSoybeansCanolaSorghumCottonTomatoPotatoWheat 1996 Roundup® Ready Gene AgreementTerms: The farmer must pay a $5 per The farmer must give up his/her right to save and replant Concerns of Roundup® Ready CropsSpread of resistance genes to weedsProblems with quality Benefits of Roundup® Ready CropsFields no longer need tillingReduction in weed management Roundup® patent recently expiredResearchers have designed new method of resistanceSorted thru 100s There are also varieties of various crops resistant to:GlufosinateBromoxyilSulfonylurea Virus ResistanceYellow Squash resistant to three different viruses were developed by Asgrow Transgenic PRV-resistant papaya has been grown 	commercially in Hawaii since 1996Increased virus promoterRegulatory sequences recognised by plant(either from plant gene or plant virus gene).In Various Cry genes (CryIA(b), CryIA(c), & Cry9C) have been inserted crops such What is BTdoplnit z prednasky roslinna biotechnologie Molecular basis of the Bt action Bt Corn & Monarch ButterfliesCry toxin is expressed in all of the Biodiversity / NTO StudiesMonarch Butterfly, symbol of nature and “wildness” in North Bt Corn & AllergiesBt corn is approved for human consumptionHowever Starlink® brand Benefits of Bt CornCrop yield increases by up to 33%39% less insecticide Concerns associated with GM cropsPossible production of allergenic or toxic proteins not How to prevent development of Bt resistance in insects? at least 20% Crop ImprovementsCurrent research into crop improvements include: Increased growth rate Increased salt Drought / Salinity ResistanceTrehalose is a protectant against many environmental stresses; freezing, Modification of Seed Oil Content Plant Seed OilsOils either for cooking or industrial uses are usually extracted http://biology.clc.uc.edu/courses/bio104/lipids.htmFatty AcidsTriglyceride http://food.oregonstate.edu/images/fat/lard1.jpghttp://www.plattsalat.de/Gawang.htmlhttp://www.aces.edu/dept/extcomm/newspaper/feb23b01.htmlCanola Oil -10°C  Coconut Oil 20-25°C		Lard 40.5°C5% Saturated	  85% Saturated		100% Saturated http://biology.clc.uc.edu/courses/bio104/lipids.htmCis-fatty acids, naturally occuring unsaturated acidsTrans-fatty acids, artifically generated to keep unsaturated Soybeans with Modified Oil ContentSoybeans normally have high amounts of the unsaturated Over 60 million tons of seed oil are used for edible purposesAbout Rape field full of brillantly yellow flowershttp://www.tiscali.co.uk/reference/encyclopaedia/hutchinson/m0011652.html The first transgenic crop with modified oil content was a high lauric Other varieties of GM rapeseed could provide:	Steric acid as a substitute for http://www.teedrogen.info/systematik/7_bilder/ara-th-1.jpgArabidopsis thaliana (Cress)Arabidopsis is a model organism used by scientists to investigate http://www.thegutsygourmet.net/post-brassica.jpgBrassica (Mustard) Family Nutritional Enhancement Improving Protein Quality Nutritional value of seed storage proteins is often ‘Increased nutritive value of transgenic potato by expressing a nonallergenic seed albumin Changes in protein quality in Amaranthus albumin potatoes 5-8 fold higher essential http://www.princeton.edu/~fecelik/GMFoods/impactshumanconsumptionpros.htmlGolden RiceInserted genes from other plants & bacteria to produce –caroteneVitamin A Vitamin A Vitamin A (retinol) is essential to human growthOur bodies cannot 400 million people are at risk of vitamin A deficiency (VAD), particularly Vaccine FoodsIn the early 1990’s tomatoes, bananas, & potatoes were proposed as There are concerns about dosing when these crops are directly consumed. 	Would Plants as BioreactorsPlants (crops or cell culture) can be used to produce Large Scale Biology Corp. (LSBC) uses tobacco plants for drug manufacturingLSBC uses USDA just approved the use of rice to produce lactoferrin and lysozymeSigma-Aldrich Plant BioFuels Cellulosic EthanolHas higher yield due to the fermentation of sugar released from BioDiesel1900 Rudolph Diesel runs his engine on peanut oilBiodiesel is defined as http://www.biodiesel.org/pdf_files/fuelfactsheets/Production_Graph_Slide.pdfU.S. consumes 40 billion gallons of diesel/yr http://www.wired.com/wired/archive/11.04/genetics.html?pg=1&topic=&topic_set=Timber Biotechnology Reduction in generation timeTrees can take years to flowerThose overexpressing the LEAFY Alteration in tree size or formAltered expression of a gene involved in From left to right:antisense-GA 20-oxidase, wild type, & GA 20-oxidase overexpressingwww.upsc.se/tmoritz.htmSix Weeks Old Hybrid Aspen Leaves from GM poplarhttp://stacks.msnbc.com/news/947076.asp?0cl=cr&cp1=1 http://www.enn.com/news/enn-stories/1999/08/080999/trees_4724.aspTrees engineered to produce lower amounts of ligninThese trees grow faster and Bioremediation Bioremediation using bacteria has limitationsThe engineered or specialized bacteria used are often
Слайды презентации

Слайд 2 Plant Tissue Culture
Plant cells differ from animals cells

Plant Tissue CulturePlant cells differ from animals cells in that they

in that they are totipotent
A totipotent cell is one

that can develop into specialized cell types & regenerate an entire organism
Tissue culture of plants and the regeneration of complete plants from cells has been done since 1930s
This allows large-scale clonal propagation of plants

Слайд 3 http://www.webschoolsolutions.com/biotech/transgen.htm
Plant Cloning

http://www.webschoolsolutions.com/biotech/transgen.htmPlant Cloning

Слайд 4 http://catf.bcresearch.com/biotechnology/tissueculture_research.htm
Micropropagation
Callus, undifferentiated mass of plant cells
Seedlings, each from

http://catf.bcresearch.com/biotechnology/tissueculture_research.htmMicropropagationCallus, undifferentiated mass of plant cellsSeedlings, each from an individual cell

an individual cell


Слайд 5 How Do They Engineer Plants?

How Do They Engineer Plants?

Слайд 6 Methods of producing transgenic plants

Methods of producing transgenic plants

Слайд 7 http://www.colostate.edu/programs/lifesciences/TransgenicCrops/how.html
Plant Genetic Engineering
Ti plasmid of Agrobacterium tumefaciens
A. tumefaciens

http://www.colostate.edu/programs/lifesciences/TransgenicCrops/how.htmlPlant Genetic EngineeringTi plasmid of Agrobacterium tumefaciensA. tumefaciens is a soil

is a soil microbe that induces crown gall
Crown gall

is a ‘cancerous’ mass which forms at the site of infection in plants

Слайд 8 T-DNA portion of the Ti plasmid contains genes

T-DNA portion of the Ti plasmid contains genes responsible for the

responsible for the disease
T-DNA becomes incorporated into the genome

of the plant
Part of the T-DNA may be replaced with a foreign gene and used to incorporate this gene into the plant’s genome
A marker is also added to determine which cells have the recombinant gene

TIBS 1998, 19:500-506.


Слайд 9 http://www.webschoolsolutions.com/biotech/transgen.htm

http://www.webschoolsolutions.com/biotech/transgen.htm

Слайд 10 http://faculty.abe.ufl.edu/~chyn/age2062/lect/lect_09/10_19A.GIF
Biolistics (Biological Ballistics)
Useful for engineering corn, rice, wheat,

http://faculty.abe.ufl.edu/~chyn/age2062/lect/lect_09/10_19A.GIFBiolistics (Biological Ballistics)Useful for engineering corn, rice, wheat, barley, & other crops

barley, & other crops


Слайд 11 Helios Gene Gun
http://www.bio-rad.com
Uses an adjustable burst low-pressure helium

Helios Gene Gunhttp://www.bio-rad.comUses an adjustable burst low-pressure helium to sweep DNA-

to sweep DNA- or RNA-coated gold pellets from the

inner wall of a small plastic cartridge directly onto a target

Слайд 12 Genetically Engineered Plants
Herbicide tolerance
Insect resistance
Crop improvements
Functional foods
Plants as

Genetically Engineered PlantsHerbicide toleranceInsect resistanceCrop improvementsFunctional foodsPlants as bioreactorsBiofuelsTimber improvementsBioremediation

bioreactors
Biofuels
Timber improvements
Bioremediation


Слайд 13 Currently 215 million acres of GM crops grown

Currently 215 million acres of GM crops grown worldwideGenetically Modified (GM) Crops

worldwide
Genetically Modified (GM) Crops


Слайд 14 >60% of the foods we purchase have GM

>60% of the foods we purchase have GM ingredients 95% of

ingredients
95% of canola is biotech herbicide-tolerant
50% of corn

is biotech herbicide-tolerant
35% of corn is biotech insect-resistant
61% of cotton is biotech herbicide-tolerant
52% of cotton is biotech insect-resistant
93% of soybean is biotech herbicide-tolerant
(2005 Data, Source :GM Crops: The First 10 Years -- Global Socio-economic and Environmental Impacts; PG Economics Limited )

How Much of What We Eat Is GM?


Слайд 15 Who Produces GM Food?
BASF Inc.
Aventis Cropscience
Bayer Cropscience
Syngenta Seed

Who Produces GM Food?BASF Inc.Aventis CropscienceBayer CropscienceSyngenta Seed Inc.Pioneer Hi-Breed International Inc.Dow Agroscience LLCMonsanto Company

Inc.
Pioneer Hi-Breed International Inc.
Dow Agroscience LLC
Monsanto Company


Слайд 16 FLAVR SAVR, The First GM Food

FLAVR SAVR, The First GM Food

Слайд 17 http://resources.emb.gov.hk/envir-ed/globalissue/images/ModifiedTomato.jpg
The first GM food was the FLAVR SAVR

http://resources.emb.gov.hk/envir-ed/globalissue/images/ModifiedTomato.jpgThe first GM food was the FLAVR SAVR tomatoIntroduced in 1994 it had delayed ripening characteristics

tomato
Introduced in 1994 it had delayed ripening characteristics


Слайд 18 Fruit softens because polygalacturonase degrades pectin
Antisense technology was

Fruit softens because polygalacturonase degrades pectinAntisense technology was used to turn

used to turn off (silence) the polygalacturonase (PG) gene
Gene

encoding antisense RNA was inserted into tomato cells
The antisense RNA finds the normal RNA and hybridizes
The cell then degrades this complex, preventing the normal RNA from being translated

Слайд 19 PG gene
transcription
mRNA
translation
PG gene
transcription
mRNA
Antisense mRNA
translation
Antisense Technology

PG genetranscriptionmRNAtranslationPG genetranscriptionmRNAAntisense mRNAtranslationAntisense Technology

Слайд 20 Polygalacturonase (PG) is an enzyme that breaks

Polygalacturonase (PG) is an enzyme that breaks down pectin in

down pectin in ripening fruit walls
Plants with an

antisense PG
transgene produce less PG.
Walls soften more slowly

Many genes manipulated in the
same way to answer basic
questions:

- what is the role of hormones
in ripening?
- what do particular enzymes do in
fruit walls?

Wild-type
fruit

Antisense PG
fruit

PG activity

Days from 1st colour change

0

10

2

4

6

8

Altering Fruit Ripening with Antisense RNA


Слайд 21 http://www.wachstumshormon.info/kontrovers/gentechnik/flavrsavr.html?gfx=2
Most GM tomatoes were used only in canned

http://www.wachstumshormon.info/kontrovers/gentechnik/flavrsavr.html?gfx=2Most GM tomatoes were used only in canned pureeBy 1997 FlavrSavr was no longer marketed

puree
By 1997 FlavrSavr was no longer marketed


Слайд 22 Current technologies aid the farmer not the consumer
Herbicide

Current technologies aid the farmer not the consumerHerbicide ResistanceVirus ResistanceInsect Resistance

Resistance
Virus Resistance
Insect Resistance


Слайд 23 Herbicide Tolerance
Methods used to promote crop growth also

Herbicide ToleranceMethods used to promote crop growth also promote weedsWeeds often

promote weeds
Weeds often outgrow crops and reduce farm output
Even

though there are about 100 chemical herbicides, weeds still reduce crop productivity by ~12%
Problem is that many herbicides kill both crops & weeds
This has led to the creation of herbicide tolerant crops

Слайд 24 Soybean with no herbicides
Soybean after herbicides
Herbicides are used

Soybean with no herbicidesSoybean after herbicidesHerbicides are used for weed controlHerbicide

for weed control
Herbicide Tolerance
Weeds
drastically
reduce
crop
yield


and quality

Слайд 26 Non-selective herbicides (Roundup Ultra and)
Roundup® (chemical name:

Non-selective herbicides (Roundup Ultra and) Roundup® (chemical name: glyphosate) Breaks down

glyphosate)
Breaks down quickly in the soil,
eliminating
residual

carry-over problems

and
reducing environmental impact.

Roundup Ready®


transgenic varieties
of common crops
completely resistant
to those herbicides


Слайд 28 EPSPS Transgene Introduced into Plants
Codon usage modified
for efficient

EPSPS Transgene Introduced into PlantsCodon usage modifiedfor efficient expression in plantspromoterRegulatory

expression
in plants
promoter
Regulatory sequences recognised by plant (either from

plant gene or plant virus gene). In this case 35S CaMV promoter

Agro. EPSPS

Transit peptide from plant gene added to allow chloroplast import


Слайд 30 Move to greener herbicide
Benefits of Glyphosate

Move to greener herbicide Benefits of Glyphosate Tolerance in Crops

Tolerance in Crops
Can use at any time

- can wait until there is a problem

Reduced herbicide use

Very effective
- Weeds very sensitive
- GM crop very resistant

GM canola surrounded by weeds

- glyphosate

+ glyphosate


Слайд 31 Roundup® Ready Crops
Corn
Alfalfa
Soybeans
Canola
Sorghum
Cotton
Tomato
Potato
Wheat

Roundup® Ready CropsCornAlfalfaSoybeansCanolaSorghumCottonTomatoPotatoWheat

Слайд 32 1996 Roundup® Ready Gene Agreement

Terms:

The farmer

1996 Roundup® Ready Gene AgreementTerms: The farmer must pay a $5

must pay a $5 per bag "technology fee“

The

farmer must give Monsanto the right to inspect, monitor and test his/her fields for up to 3 years

The farmer must use only Monsanto's brand of the glyphosate herbicide it calls Roundup®

Слайд 33 The farmer must give up his/her right

The farmer must give up his/her right to save and

to save and replant the patented seed (replanting seed

is a practice as old as agriculture)

The farmer must agree not to sell or otherwise supply the seed to "any other person or entity."

The farmer must also agree, in writing, to pay Monsanto "...100 times the then applicable fee for the Roundup® Ready gene, times the number of units of transferred seed, plus reasonable attorney's fees and expenses..." should he violate any portion of the agreement.

Слайд 34 Concerns of Roundup® Ready Crops
Spread of resistance genes

Concerns of Roundup® Ready CropsSpread of resistance genes to weedsProblems with

to weeds
Problems with quality of crops
e.g.. Cotton bolls falling

off prior to harvest
Farmers required to purchase seed annually
The herbicide is still toxic at high doses

Слайд 35 Benefits of Roundup® Ready Crops
Fields no longer need

Benefits of Roundup® Ready CropsFields no longer need tillingReduction in weed

tilling
Reduction in weed management costs of up to 37%
Decrease

in herbicide use by >1lb/acre
Overall 74% increase in farmer profits1

1U.S. Corn Crop 2003


Слайд 36 Roundup® patent recently expired
Researchers have designed new method

Roundup® patent recently expiredResearchers have designed new method of resistanceSorted thru

of resistance
Sorted thru 100s of microbes to find a

detoxifying enzyme
Found 3 genes in Bacillus licheniformis which encode glyphosate N-acetyltransferase (GAT)
Using directed evolution generated an enzyme 10000x more efficient
~5yrs to market

Слайд 37 There are also varieties of various crops resistant

There are also varieties of various crops resistant to:GlufosinateBromoxyilSulfonylurea

to:
Glufosinate
Bromoxyil
Sulfonylurea


Слайд 38 Virus Resistance
Yellow Squash resistant to three different viruses

Virus ResistanceYellow Squash resistant to three different viruses were developed by

were developed by Asgrow Seed
Resistance was then transferred to

zucchini
Virus resistant papaya were developed in the mid ’90s
This was after a outbreak of papaya ring spot virus destroyed 40% of the Hawaiian crop
The varieties called Rainbow® & SunUp® are provided free to farmers

Слайд 39 Transgenic PRV-resistant papaya has been grown commercially in

Transgenic PRV-resistant papaya has been grown 	commercially in Hawaii since 1996Increased

Hawaii since 1996
Increased virus resistance: Papaya ringspot virus (PRV)
Virus

has had huge impact on papaya industry in Hawaii - reduction of fresh fruit production directly related to spread of PRV

No naturally occurring resistance genes - without GM, papaya industry in Hawaii would be destroyed


Слайд 40 promoter
Regulatory sequences recognised by plant
(either from plant gene

promoterRegulatory sequences recognised by plant(either from plant gene or plant virus

or plant virus gene).
In this case 35S CaMV promoter

and terminator

polyA+

PRV coat protein gene expressed from 35S CaMV promoter.
Papaya transformed by particle bombardment

PRV coat protein gene

Confers partial resistance to PRV in one variety (‘Rainbow’) and complete resistance in another (‘SunUp’)

Growers have to sign up to careful crop management - minimize virus pressure on transgenics to maintain resistance

Papaya Resistant to PRV


Слайд 41 Various Cry genes (CryIA(b), CryIA(c), & Cry9C) have

Various Cry genes (CryIA(b), CryIA(c), & Cry9C) have been inserted crops

been inserted crops such as corn, cotton, potatoes, &

rice
Pest must ingest a portion of the plant for the toxin to be effective
Within hours the gut breaks down and the pest dies

Insect Resistance

http://www.agbios.com/docroot/articles/03-314-001.pdf

Wt Corn

Bt Corn


Слайд 42 What is BT
doplnit z prednasky roslinna biotechnologie

What is BTdoplnit z prednasky roslinna biotechnologie

Слайд 43 Molecular basis of the Bt action

Molecular basis of the Bt action

Слайд 44 Bt Corn & Monarch Butterflies
Cry toxin is expressed

Bt Corn & Monarch ButterfliesCry toxin is expressed in all of

in all of the plant as well as pollen
Corn

pollen can blow onto milkweed growing near corn fields
Monarch caterpillars feed exclusively on milkweed
An early study showed a possible toxic effect of Bt pollen on monarch caterpillars

http://homepages.ihug.co.nz/~mostert/land%20photography/Insects/insects/monarch%20butterfly.jpg


Слайд 45 Biodiversity / NTO Studies
Monarch Butterfly, symbol of nature

Biodiversity / NTO StudiesMonarch Butterfly, symbol of nature and “wildness” in

and “wildness” in North America.
The reports of Bt effects

on Monarch butterflies have fueled much emotional debate on the use of biotech crops.

Слайд 51 Bt Corn & Allergies
Bt corn is approved for

Bt Corn & AllergiesBt corn is approved for human consumptionHowever Starlink®

human consumption
However Starlink® brand corn is approved only for

animal feed
Contains Cry9C which may be a potential allergen
In 2000 Starlink® was found in Taco Bell-brand taco shells
EPA determined that no one who ate the food was allergic, but they found that it had a moderate potential allergenicity
All the products were recalled and Starlink® corn is not approved for human consumption

Starlink® trademark of Aventis Corp.


Слайд 52 Benefits of Bt Corn
Crop yield increases by up

Benefits of Bt CornCrop yield increases by up to 33%39% less

to 33%
39% less insecticide used
Increase in monetary gains by

18%1

1U.S. Corn Crop 2003


Слайд 53 Concerns associated with GM crops
Possible production of allergenic

Concerns associated with GM cropsPossible production of allergenic or toxic proteins

or toxic proteins
not native to the crop

2.

Adverse effects on non-target organisms,
especially pollinators and biological control organisms

3. Loss of biodiversity

4. Genetic pollution (unwanted transfer of genes to other species)

5. Development of pest resistance

6. Global concentration of economic power and food production

7. Lack of "right-to-know"
(i.e., a desire for labeling transgenic foods)

Слайд 54 How to prevent development of Bt resistance in

How to prevent development of Bt resistance in insects? at least

insects?
at least 20%
of a farm's corn acreage


must be planted
to non-BT corn.
R = resistant European borer;
S = susceptible borer.

few Bt-resistant insects
surviving in the Bt field
would likely mate
with susceptible individuals
that have matured
in the non-Bt refuge.
Thus, the resistance alleles
would be swamped
by the susceptible alleles.

< 80%

> 20%

Strategy will not work if resistance is dominant !!!


Слайд 56 Crop Improvements
Current research into crop improvements include:
Increased

Crop ImprovementsCurrent research into crop improvements include: Increased growth rate Increased

growth rate
Increased salt tolerance
Increased drought resistance
Modification

of seed oil content

Слайд 57 Drought / Salinity Resistance

Trehalose is a protectant against

Drought / Salinity ResistanceTrehalose is a protectant against many environmental stresses;

many environmental stresses; freezing, osmotic pressure (salinity), heat and

dessication.

Trehalose (1--D-glucopyranosyl-glucopyranoside)
is synthesised in a two-step process in yeast.

Zygosaccharomyces rouxii is one of the most highly
osmo-tolerant yeasts – especially to salt

Kwon, S.J., Hwang, E.W. & Kwon, H.B. (2004). Genetic
engineering of drought resistant potato plants by
co-introduction of genes encoding trehalose-6-phosphate
synthase and trehalose-6-phosphate phosphatase of
Zygosaccharomyces rouxii. Korean J. Genet. 26, 199-206.

Transgenic potatoes morphologically identical to parents.

trehalose-6-P synthase

trehalose-6-P phosphotase

2A

RB

LB

CaMV
35S

nptII

NS ter

NS prom


Слайд 58 Modification of Seed Oil Content

Modification of Seed Oil Content

Слайд 59 Plant Seed Oils
Oils either for cooking or industrial

Plant Seed OilsOils either for cooking or industrial uses are usually

uses are usually extracted from seeds such as,
Corn, safflower,

sunflower, canola, coconut, flax
Seed rely on their stored oil as an energy & carbon sources for germination
Most seed oils are made up of unsaturated fatty acids
Some tropical oils such as palm & coconut have significant levels of saturated fatty acids

Слайд 60 http://biology.clc.uc.edu/courses/bio104/lipids.htm
Fatty Acids
Triglyceride

http://biology.clc.uc.edu/courses/bio104/lipids.htmFatty AcidsTriglyceride

Слайд 61 http://food.oregonstate.edu/images/fat/lard1.jpg
http://www.plattsalat.de/Gawang.html
http://www.aces.edu/dept/extcomm/newspaper/feb23b01.html
Canola Oil -10°C Coconut Oil 20-25°C Lard 40.5°C
5%

http://food.oregonstate.edu/images/fat/lard1.jpghttp://www.plattsalat.de/Gawang.htmlhttp://www.aces.edu/dept/extcomm/newspaper/feb23b01.htmlCanola Oil -10°C Coconut Oil 20-25°C		Lard 40.5°C5% Saturated	 85% Saturated		100% Saturated

Saturated 85% Saturated 100% Saturated


Слайд 62 http://biology.clc.uc.edu/courses/bio104/lipids.htm
Cis-fatty acids, naturally occuring unsaturated acids
Trans-fatty acids, artifically

http://biology.clc.uc.edu/courses/bio104/lipids.htmCis-fatty acids, naturally occuring unsaturated acidsTrans-fatty acids, artifically generated to keep

generated to keep unsaturated fatty acids from going rancid
Trans-fatty

acids increase LDL (bad cholesterol) increasing the risk of heart disease

Слайд 63 Soybeans with Modified Oil Content
Soybeans normally have high

Soybeans with Modified Oil ContentSoybeans normally have high amounts of the

amounts of the unsaturated fatty acid linolenic acid
Vistive by

Monsanto is Roundup Ready soybeans bred to have low amounts of linolenic acid
This eliminates the need to hydrogenate the oil

http://www.siberiantigernaturals.com/omega3.htm


Слайд 66 Over 60 million tons of seed oil are

Over 60 million tons of seed oil are used for edible

used for edible purposes
About 15 million tons is employed

in industrial usage
One of the first plants modified was Rape (Brassica napus oleifera)
A member of the mustard family, Rape has been grown for centuries as animal feed and natural lubricant
In the 1970s selective breeding led to strains of Rape lacking erucic acid
The removal of erucic acid made the oil fit for human consumption, LEAR (low-erucic acid rapeseed)

Слайд 67 Rape field full of brillantly yellow flowers
http://www.tiscali.co.uk/reference/encyclopaedia/hutchinson/m0011652.html

Rape field full of brillantly yellow flowershttp://www.tiscali.co.uk/reference/encyclopaedia/hutchinson/m0011652.html

Слайд 68 The first transgenic crop with modified oil content

The first transgenic crop with modified oil content was a high

was a high lauric oil rapeseed.
Rapeseed oil normally contains

about 0.1% lauric acid
Modified rapeseed contains ~40% lauric acid
Lauric acid from GM rapeseed would be used in place of oils from palm or coconut
The primary use of lauric acid is in detergents

CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-SO4- Na+

Sodium Lauryl Sulfate a.k.a SDS


Слайд 69 Other varieties of GM rapeseed could provide:
Steric acid

Other varieties of GM rapeseed could provide:	Steric acid as a substitute

as a substitute for hydrogenated oils
Jojoba waxes for use

in cosmetics and lubricants
Various acids for use in biodiesel
Phytases for animal feed
Novel peptides for pharmaceuticals

Why is rapeseed so versatile?
One reason is because it is related to…

Слайд 70 http://www.teedrogen.info/systematik/7_bilder/ara-th-1.jpg
Arabidopsis thaliana (Cress)
Arabidopsis is a model organism used

http://www.teedrogen.info/systematik/7_bilder/ara-th-1.jpgArabidopsis thaliana (Cress)Arabidopsis is a model organism used by scientists to

by scientists to investigate plant development and genomics
The Arabidopsis

genome was recently completed

Слайд 71 http://www.thegutsygourmet.net/post-brassica.jpg
Brassica (Mustard) Family

http://www.thegutsygourmet.net/post-brassica.jpgBrassica (Mustard) Family

Слайд 72 Nutritional Enhancement

Nutritional Enhancement

Слайд 73 Improving Protein Quality
Nutritional value of

Improving Protein Quality Nutritional value of seed storage proteins is

seed storage proteins is often limited
- may lack one

or more amino acid essential to human health
e.g. legume seeds lack cysteine and methionine; other seeds can lack lysine

Animals and humans are incapable of making 10 ‘essential’ amino acids
- must obtain in diet

Amino acid balance in seeds has been manipulated in laboratory experiments using a number of strategies:

- introduce seed storage protein from another species

- alter sequence of seed storage protein gene in vitro

- manipulate amino acid biosynthetic pathway to increase abundance of particular amino acids

Similar strategies have been used to improve protein content and composition in non-seed food crops...


Слайд 74 ‘Increased nutritive value of transgenic potato by expressing

‘Increased nutritive value of transgenic potato by expressing a nonallergenic seed

a nonallergenic seed albumin gene from Amaranthus hypochondriacus’
Chakraborty et

al., PNAS 97, 3724-3729 (2000)

Potato is the fourth most abundant global crop and used for food, animal feed and production of starch and alcohol

Limited in lysine, tyrosine, methionine and cysteine

Transformed potato with seed albumin from Amaranthus hypochondriacus which has good amino acid balance


p35S CaMV

Nos 3’

AmA1

pGBSS

Nos 3’

AmA1

2 alternative constructs. Promoters constitutive or tuber-specific

Expression in tuber 5-10 fold higher with GBSS promoter than with 35S promoter

pSB8

pSB8G


Слайд 75 Changes in protein quality in Amaranthus albumin potatoes

Changes in protein quality in Amaranthus albumin potatoes 5-8 fold higher


5-8 fold higher essential amino acids in pSB8G

transgenics

Total protein content also increased (35-45%)

Fold
increase

1-

8-

4-

D E S G H R T A P Y V M C I L F K

Amino acid


Слайд 76 http://www.princeton.edu/~fecelik/GMFoods/impactshumanconsumptionpros.html
Golden Rice
Inserted genes from other plants & bacteria

http://www.princeton.edu/~fecelik/GMFoods/impactshumanconsumptionpros.htmlGolden RiceInserted genes from other plants & bacteria to produce –caroteneVitamin

to produce –carotene
Vitamin A deficiencies affect >124 million children

worldwide

Слайд 77 Vitamin A
Vitamin A (retinol) is essential to

Vitamin A Vitamin A (retinol) is essential to human growthOur bodies

human growth
Our bodies cannot make vitamin A,
All carotenoids

that contain a -ring can be converted into retinol, and one of the most important carotenoid pro-vitamins is -carotene

-carotene is a pigment required for photosynthesis

- produced in all plant green tissues


Слайд 78 400 million people are at risk of vitamin

400 million people are at risk of vitamin A deficiency (VAD),

A deficiency (VAD), particularly in Asia and Africa
implicated in

up to 2.5 million deaths annually in children under 5

0.5 million children go blind each year
because of VAD

Supplementation programmes have reduced child mortality by up to 50% in target areas

supplementation not universal; expensive; misses remote areas

VAD makes children especially vulnerable to infections

Vitamin A deficiency

VAD is most serious in regions where rice is the staple food ; up to 70% children under 5 affected


Слайд 80 Vaccine Foods
In the early 1990’s tomatoes, bananas, &

Vaccine FoodsIn the early 1990’s tomatoes, bananas, & potatoes were proposed

potatoes were proposed as delivery vehicles for vaccines
Touted as

a simple method of delivering vaccines especially to developing countries
Studies have shown plant-produced oral vaccines to increase immunity in mice
Potatoes containing Hepatitis B vaccine have been shown to boost immunity in humans

Слайд 81 There are concerns about dosing when these crops

There are concerns about dosing when these crops are directly consumed.

are directly consumed.
Would a dose be? 2 bananas

and a tomato
What if a person eats too many vaccine potatoes?
Also there is concern if the vaccine foods enter the food supply of people who are vaccinated the traditional way

Слайд 82 Plants as Bioreactors
Plants (crops or cell culture) can

Plants as BioreactorsPlants (crops or cell culture) can be used to

be used to produce proteins currently produced by microbes

or animal cells
The advantage over microbes:
The proteins are more like human proteins
The advantage over animal cells:
Plants cannot become contaminated with mammalian pathogens

Слайд 83 Large Scale Biology Corp. (LSBC) uses tobacco plants

Large Scale Biology Corp. (LSBC) uses tobacco plants for drug manufacturingLSBC

for drug manufacturing
LSBC uses an engineered tobacco mosaic virus

(TMV)
Recombinant gene is inserted into TMV which infects & replicates in the plants
During replication large amounts of the drug are generated
The drug accumulates in the leaves which are harvested
-galactosidase A purified from tobacco is as effective in treating Fabry’s disease as the animal cell derived drug

Слайд 84 USDA just approved the use of rice to

USDA just approved the use of rice to produce lactoferrin and

produce lactoferrin and lysozyme
Sigma-Aldrich now sells aprotinin and typsin

made in tobacco
Duckweed is being used to produce interferon-

More and More Plants Are Being Used to Produce Proteins


Слайд 85 Plant BioFuels

Plant BioFuels

Слайд 86 Cellulosic Ethanol
Has higher yield due to the fermentation

Cellulosic EthanolHas higher yield due to the fermentation of sugar released

of sugar released from cellulose
Requires the addition of cellulase

or acid
Agricultural plant wastes (corn stover, cereal straws)
Plant wastes from industrial processes (sawdust, paper pulp)
Crops grown specifically for fuel production (switchgrass)

Слайд 87 BioDiesel
1900 Rudolph Diesel runs his engine on peanut

BioDiesel1900 Rudolph Diesel runs his engine on peanut oilBiodiesel is defined

oil
Biodiesel is defined as “a fuel comprised of mono-alkyl

esters of long chain fatty acids derived from vegetable oils or animal fats”
Transesterification converts triglycerides into methyl esters of fatty acids

http://www.campa-biodiesel.de/caengnof/caenkra2.htm


Слайд 88 http://www.biodiesel.org/pdf_files/fuelfactsheets/Production_Graph_Slide.pdf
U.S. consumes 40 billion gallons of diesel/yr

http://www.biodiesel.org/pdf_files/fuelfactsheets/Production_Graph_Slide.pdfU.S. consumes 40 billion gallons of diesel/yr

Слайд 89 http://www.wired.com/wired/archive/11.04/genetics.html?pg=1&topic=&topic_set=
Timber Biotechnology

http://www.wired.com/wired/archive/11.04/genetics.html?pg=1&topic=&topic_set=Timber Biotechnology

Слайд 90 Reduction in generation time
Trees can take years to

Reduction in generation timeTrees can take years to flowerThose overexpressing the

flower
Those overexpressing the LEAFY (LFY) gene can flower in

as little as 7 months.
This is of particular value in fruit bearing trees
Also allows for rapid analysis mature traits

Слайд 91 Alteration in tree size or form
Altered expression of

Alteration in tree size or formAltered expression of a gene involved

a gene involved in hormone synthesis can give wildly

differing results
Overexpression of GA 20-oxidase gives faster growing trees both in height and diameter and longer wood fibers
Inhibition of GA 20-oxidase gives dwarf trees

Слайд 92 From left to right:
antisense-GA 20-oxidase, wild type, &

From left to right:antisense-GA 20-oxidase, wild type, & GA 20-oxidase overexpressingwww.upsc.se/tmoritz.htmSix Weeks Old Hybrid Aspen

GA 20-oxidase overexpressing
www.upsc.se/tmoritz.htm
Six Weeks Old Hybrid Aspen


Слайд 93 Leaves from GM poplar
http://stacks.msnbc.com/news/947076.asp?0cl=cr&cp1=1

Leaves from GM poplarhttp://stacks.msnbc.com/news/947076.asp?0cl=cr&cp1=1

Слайд 94 http://www.enn.com/news/enn-stories/1999/08/080999/trees_4724.asp
Trees engineered to produce lower amounts of lignin
These

http://www.enn.com/news/enn-stories/1999/08/080999/trees_4724.aspTrees engineered to produce lower amounts of ligninThese trees grow faster

trees grow faster and have greater cellulose content
Lignin is

a glue-like compound that must be chemically removed from pulp prior to papermaking

Слайд 95 Bioremediation

Bioremediation

Слайд 96 Bioremediation using bacteria has limitations
The engineered or specialized

Bioremediation using bacteria has limitationsThe engineered or specialized bacteria used are

bacteria used are often unable to compete with indigenous

soil bacteria
An alternative is phytoremediation, the use of plants to mop up toxic waste
A standard technique for treating soils contaminated with heavy metals (lead or cadmium), or organic pollutants (pesticides) in a cost-effective way

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