better health trhough nutricionally enhanced transgenic crops
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Better health through nutritionally enhanced transgenic crops
Multivitamin maize through genetic engineering
CYTALIA XIV April 22, 2009
747.psd -
The Drivers
Why we do what we do?
How we do what we do?
Applied Plant Biotechnology Laboratory
1311.psd -
Structure of the talk
Food insecurity and nutritionRole of Biotechnology and its contribution to poverty alleviation and nutritional improvement in the developing worldThe biofortified corn (maize) paradigmThe politics and regulation of GE cropsWhat does the future hold? -
Millennium Development Goals 2008
Eradicate extreme poverty and hunger
Achieve universal primary education
Promote gender equality and empower women
Reduce child mortality
Improve maternal health
Combat HIV/AIDS, malaria and other diseases
Ensure environmental sustainability
Develop a global partnership for development
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Copenhagen Consensus 2008 ranking, based on the costs and benefits of the solutions-Ranked in descending order of desirability
ChallengeSolution1Malnutrition Micronutrient supplements for children (vitamin A and zinc) 2Trade The Doha development agenda 3Malnutrition Micronutrient fortification (iron and salt iodization) 4Diseases Expanded immunization coverage for children 5Malnutrition Biofortification 6Malnutrition/EducationDeworming and other nutrition programs at school 7Education Lowering the price of schooling 8WomenIncrease and improve girls schooling 9Malnutrition Community-based nutrition promotion 10WomenProvide support for womens reproductive role -
Interventions to alleviate malnutrition
Food fortification (dairy products and salt)Supplementation (pills or mineral solutions)Biofortification-Agronomic interventionsBiofortification-Plant breedingBiofortification-Genetic engineering -
Predicted changes in population
Year 1995 2025
Population 5702 8122
(millions)
Nature 2008
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The 15 hungriest countries (% of population undernourished)
1979-81
1996-98
80%
60%
40%
20%
10%
0%
Som Afg Bur Erit Hait Cong Moz NKor Ethi Lib Nig Mong Zam SLeo Ang
Somalia
Afganistan
Burundi
Eritrea
Haiti
Congo DR
Mozambique
North Korea
Ethiopia
Liberia
Niger
Mongolia
Zambia
Sierra Leone
Angola
1305.psd -
In the developing world 840 million people are chronically undernourished
Many more people, perhaps half of the worlds population, suffer from diseases
caused by dietary deficiencies and inadequate supplies of vitamins and minerals
Despite the prevalence of hunger and malnutrition, global food production has
outpaced population growth over the last 40 years thanks mainly to the successes
of the Green Revolution
Today's food insecurity is caused not by insufficient food production, but by poverty,
with nearly 1.3 billion people living on less than $US1 per day and another
2 billion only marginally better off
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Projected world cereal demand
Production1999, Mt)Needed 2025, MtYield t/ha1999Yield t/ha2025 NeededWheat5859002.73.8Rice6079003.14.3Maize60510004.15.9All cereals(includingminor crops)207431002.94.1
N. Borlaug, April 2001 -
80% of all nutritional calories come from 20 crops.
70% from five.
40% from two
1304.unknown -
If all Chinese ate five more
Big-Macs a year, this would
wipe out the entire US feed
corn productionJerry Coldwell, CEO Mycogen
The Corn Question
1303.unknown -
Evolution in Food & Feed Production
domestication of plants and animals
animal power
mechanical power
genetics (traditional breeding)
chemical assistance
biotechnology
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Traditional Breeding and Genetic Engineering (GE not GM!)
humanity has been shaping its environment for milleniawheat, rice and corn are all the product of breedingGE permits introduction of desirable traitsall our staple crops are GE through plant breedingtraditional breeding is at its limitGE gives us new opportunities1302.unknown -
Increased
yields
Manipulation of
plant architecture
Diversion of biomass
to edible organs
Protection against
pests and pathogens
Reduced losses due to
disease and infestation
Tolerance towards
abiotic stresses
Increased use
of marginal soils
Increased
planting density
Increase in levels
of utilizable
carbohydrate,
protein or fat
Fortification with
vitamins and
minerals
Manipulation of
plant development
Shorter generation intervals
Multiple production cycles
Eliminate
anti-nutritional
factors
Enhanced
photosynthesis
and nutrient uptake
Increased
accumulation
of biomass
Higher
nutritional
value
Remove constraints
Increase potential
HUNGER/MALNUTRITION
Modification of
carbohydrate,
protein or fat content
Bioremediation
Tolerance of
extreme weather
Impact of Transgenic Plants on Food Security
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Plant Biotechnology-product development timeline
time
product launch
resistances to
viruses, insects
herbicides
male
sterilitydisease
resistance
nematode
resistance
improved quality
health food,
nutraceuticals
improved
yields1996 1998 2000 20052007 2010
therapeutics
vaccines
diagnostics
1306.psd1307.psd1308.psd1309.psd1310.psd -
Plant Genetic Engineering
Nature at work
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Creation of the first transgenic plant
Agrobacterium tumefaciens
Nature, 303: 209 1983
NOS-OCS i NOS-CAT
1294.psd1295.psd -
Two alternative methods to create transgenic plants
Argobacterium tumefaciensDirect DNA transfer through particle bombardment1293.psd -
Plant transformation vector
GENE OF INTERST
targeting
signal
tag, anchor,
retention
ATG
start
TAA
stop
5UTR
3UTR
PROMOTOR
Term.
Seed-specific expression
Constitutive
marker genes
1296.unknown1297.unknown
gusA, GFP, Luciferase, DsRED -
Donor plant
Transgenic plant
Corn transformation using
direct DNA transfer
Corn seeds = 14 dap
Immature embryo
(2,4 D)
Immature embryo derived callus
Callus ready
for bombardment
3 weeks
Callus tissue under phosphinothricin selection
Callus regenerating shoots
3-4 weeks darkness
To the light
In the dark
3-4 weeks
3-4 cm tall
Plant on rooting media
1292.bin -
Agronomic properties focusing on insect and herbicide resistance
Durability, sustainability and environmental friendliness
Bacillus thuringensis (Bt)First generation of transgenic plants
1300.psd -
Antama,
Spain
Antama,
Spain
Lleida January 2007
1513.psd1514.psd -
Effect on chemical use
30-40% reduction in herbicide use with herbicide-resistant plants, because farmer can evaluate weed pressure before applicationMarket share shifting from narrow to broad-spectrum herbicidesUp to 80% reduction in insecticide use with insect-resistant plantsConclusion: transgenic plants lead to
ecology friendly agriculture!
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Multi-gene engineering is a significant hurdle in complex pathway analysis due to the diminishing rate of return as more transgenes are introduced simultaneously into target plants
791.psd -
RECONSTRUCTION AND EXTENSION OF THE CAROTENOID PATHWAY IN MAIZE THROUGH COMBINATORIAL NUCLEAR TRANSFORMATION*
* Relies on the mechanism of co-integration of multiple
independent transgenes via direct DNA transfer into
one genetic locus
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Carotenoids are naturally occurring biologically active compounds with exquisite health promoting properties
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Experimental system: South African elite white maize inbred M37W, which lacks carotenoids in the endosperm due to the absence of the enzyme phytoene synthase (PSY1)
743.psd -
WHITE vs YELLOW MAIZE
M37W
White endosperm Total carotenoids: 1.1g/g DW Lutein : Zeaxanthin 0.5 : 0.27 g/g DWA632
Yellow endosperm Total carotenoids: 28 g/g DW Lutein : Zeaxanthin 15.61 : 7.77 g/g DW -
WHITE vs YELLOW MAIZE: A Comparison
RT-PCR
psy1
psy2
pds
zds
crtISO
lyc
lyc
bch 1
Actin
M37W
A632
bch 2
RNA extracted from endosperm tissues psy1 is not expressed in M37W psy2, pds, zds, crtISO and bch exhibit similar levels ofexpression in both genotypes
Phytoene synthase 2 is the only enzyme responsible forphytoene synthesis in white maize (leaves)
Lyc and lyc expression is lower in M37W as compared to A632mRNA blot analysis
M37W
A632
M37W
A632
psy1
lyc
M37W
A632
bch
Expression of endogenous genes in the carotenoid biosynthetic pathway
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psy1 (Zea mays - yellow) U32636
crtI (Erwinia uredovora) D90087
lyc (Gentiana lutea) D017367
bch (Gentiana lutea) AB027187
crtW (Paracoccus sp.) D58420
For ketocarotenoids (Astaxanthin)
EXPERIMENTAL STRATEGY
M37W (white endosperm)
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The resulting combinatorial population can be mined for phenotypes corresponding to the production of specific carotenoids, which in turn correlate with specific transgene expression and metabolic profiles
941.psd -
psy1+crtI
psy1+crtI+lycb+bch+crtW
Endosperm specific expression
337.psd331.psd332.psd333.psd334.psd335.psd336.psd338.psd -
mRNA ANALYSIS
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HPLC PROFILES (carotenoid phenotypes)
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CAROTENOID COMPOSITION
M37W-WT
A632-WT
Phenotype 1
Phenotype 2
Phenotype 3
Phenotype 4
Phenotype 5
Phenotype 6
Phenotype 7
Psy1 + crtI
Astaxanthin
Adonixanthin
Epoxy-Lutein
Lutein
Zeaxanthin
OH-Echinenone
-cryptoxanthin
-cryptoxanthin
Echinenone
Lycopene
-carotene
-zeaxanthin
-carotene
-carotene
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INCREASE IN TOTAL CAROTENOIDS
43-fold
69-fold
60-fold
93-fold
132-fold
2.4-fold
150-fold
PhenotypesTransgenes1psy12crtI3psy1+crtI4psy1+crtI+lycb5psy1+crtI+bch+crtW6psy1+crtI+lycb+bch+crtWpsy1+crtI+lycb+bch+crtW7Chart1M37WA6321234567g/g DWTotal Carotenoids (g/g DW)1.12847.42.69165.476.1966.66103.5146.1Sheet1g/g DWM37W1.1A63228147.422.693165.4476.19566.666103.57146.12.15S3.992.16S4.962.3S102.13S202.7S44.0811.9117.394.692.95000.710.764.5947.052.28S33.39.9918.253.682.49000.90.936.1747.41.20S300.530.8600000001.41.25S301.561.1200000002.698.8915.64.274.412.261.0801.8415.9668.491.6S2.10006.6916.836.255.156.341.5503.6426.3976.191.10S33.768.7214.719.4904.741.7501.321.120.771.324.0858.052.14S108.9515.2723.1304.32.8502.251.290.71.415.3166.662.4-S21.7M37W012.0913.2919.1236.763.146.931.991.440.83.3631.57103.5A6321.1816.5210.6115.922.3613.281.635.9129.031.1604.7340.98146.120.0212.2712.664.9443.664.641.146.4159.61165.412.7921.774.675.8623.771.910.852.834.71157.40.50.270.070.090.141.1estimatedtotal 0.7815.617.771.991.090.3328Sheet1000000000g/g DWTotal Carotenoids (g/g DW)Sheet2Sheet3 -
The approach provides a unique and surprisingly straightforward
strategy for pathway analysis and multi-gene engineering in plants
It involves the introduction and coordinated expression of multiple
transgenes followed by the selection of stable lines expressing the
specific combination of transgenes required for particular metabolic
outputs
Individual lines, producing specific metabolites, can be goals in
themselves if the aim is to engineer particular molecules. However,
by examining the entire diverse population of plants, it becomes
possible to dissect the pathway and subsequently reconstruct it
either in its original form or with modifications
This provides a basis for understanding and subsequently
engineering the synthesis of novel metabolites
- Transgene combinations Independent transformantsexhibiting the same phenotype1psy12 (Ph-1)2psy1+crtI3 (Ph-2)3crtI4 (Ph-3)4lycb35crtI+lycb56psy1+crtI+lycb4 (Ph-4)7bch28crtW39psy1+crtI+bch+crtW2 (Ph-5)10psy1+crtI+lycb+crtW3 (Ph-6)11psy1+crtI+lycb+bch+crtW5 (Ph-7)12crtI+lycb+bch+crtW2
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High Astaxanthin corn
psy1+crtI+lycb+bch+crtW
Powerful biological antioxidant (100 times the strength of Vitamin E)
801.psd - PSY1 is the rate limiting enzyme in M37W (white maize)
endosperm
Over-expression of PSY1 in transgenic M37W plants increased total carotenoids
Conversion of lycopene to -carotene (lyc) is a second limiting step in white maize expressing PSY1 Combinatorial Genetic Transformation: A novel technique to produce mutants that provide useful data to study a complex biosynthetic pathway The present study provides a platform to understand the carotenoid biosynthesis in maizePh 3
Ph 4
Ph 7
Enhanced levels of carotenoids in maize endosperm (lycopene, -carotene, zeaxanthin, ketocarotenoids) New strategies for carotenoid production in maize can be optimized on the basis of the mutant profiles -
The approach is much simpler than traditional methods for the modification of the carotenoid or other complex pathways
797.psd
it relies on probability and random sampling to generate a library of metabolic variants and a rapid visual selection to identify lines of interest -
The approach is analogous to standard mutagenesis screens although the mutants are generated not by random mutagenesis to create loss-of-function phenotypes, but by random multiplex transgene insertion to create partially reconstructed pathways
1000.psd -
High provitamin A corn
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Golden Rice 1 beta-carotene 1.6g/g DW
Golden Rice 2 beta-carotene 37g/g DW
57.35
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Multi vitamin corn
-
D-Man-P
GDP-D-Man
GDP-L-Gal
L-Gal -P
L-Gal
L-GalL
L-Ascorbate
GDP-L-Gul
L-Gul-P
L-Gul
L-GulL
GDP-L-Gul
UDP-D-galacturonate
D-galacturonate
L-Galactonate
L-GulLoxidase
D-Glucuronate
L-Gulonate
myo-Inositol
myo-Inositol
oxidase
Monodehydroascorbate
Dehydroascorbate
Dehydroascorbate
reductase
Ascorbate
oxidase
Dehydroascorbate reductase DHAR
Vitamin C
1001.psd -
Folate biosynthesis in plants
ADC: aminodeoxychorismate
DHN: dihydroneopterin
DHNTP: dihydroneopterin triphosphate
GCHI: GTP cyclohydrolase IHMDHP: hydroxymethyldihydropterin
glu: glutamate
-PP: pyrophosphatepABA: p-aminobenzoate
folE
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Multi-vitamin corn
-carotene 60 g/g DW (PSY1+CrtI)
200 g/g DW folate (folE)
110 g/g FW ascorbate (DHAR)
WT-M37W
L-1 (PSY1+CrtI)
Plus folE and dhar
838.psd -
100-200 g of grain provides full RDI of -carotene (as a sole source of vitamin A), more than enough folate, and about 20% of the RDI of ascorbate
Harvest Plus
7-10 years to create high vitA corn in locally adapted varieties
1002.psd -
Multi vitamin x yellow corn
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Food security vs politics
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The Politics of Plant Biotechnology
Some personal thoughts
and reflections
-
Key words
Poverty
Food Security
Developing countries
Agriculture
World trait
Protectionism
Big business
Politics
Biotechnology
Genetic modification
Public perception
Hostile press
Safety
Environmental impact
Social impact
Intellectual Property
Regulation
EU vs US trade
wars
Economics
Research funding
National policies
Globalisation
GATT
Cartagena protocol
Urbanisation
Migration
Regional conflicts
Government corruption
Overpopulation
Birth control
Clean water
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Has it occurred to you how astonishing the culture of Western society really is?
Industrialized nations provide their citizens with unprecedented SAFETY, HEALTH and COMFORTAverage life spans increased 50% in the last centuryYet modern people live in abject fearThey are afraid of strangers, of disease, of crime, of the environmentThey are afraid of the homes they live in, the food they eat, the technology that surrounds them -
28 agosto 2003
Zaragoza Espaa
Accin de Greenpeace en un campo de maz transgnico en Zaragoza
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Mad Potato Disease
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Aspects of the political dimension of transgenic plants
GREENPEACE
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The precautionary approach as applied to Geneticaly Enhanced products in Europe
Regulators do not need to show scientifically that a biotech crop or product is unsafe but rather that it has not been proven harmless
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Applying the precautionary principle (approach?) and its implications on developing countries
Research is being slowed down without any scientific valid reason and if this situation continues, the worlds poor will be the ones to suffer
1126.unknown -
In the 35 years since the environmental movement came into existence, science has undergone a major revolution
This revolution has brought new understanding of nonlinear dynamics, complex systems, chaos theory, catastrophy theoryIt has transformed the way one should be thinking about evolution and ecologyYet these no-longer-new ideas have hardly penetrated the thinking of environmental activists, which seems oddly fixed in the concepts and rhetoric of the 1970s -
We need a new environmental movement, with new goals and new organisations
We need more people working in the field, in the actual environment, and fewer people behind computer screensWe need more scientists, many few politicians and even fewer lawyers -
And that is why the intermixing of science and politics is a bad combination, with a bad history
We must remember the history, and be certain that what we present to the world as knowledge is disinterested and honest
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Alston Chase
When the search for truth is confused with political correctness and advocacy, the pursuit of knowledge is reduced to the quest for power
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Chemical X
An abundunt chemical found in the environement in most parts of the worldIt is found in lakes and riversIt remains on fruits and vegetables after they are washedIt makes you sweatIt is responsible for the deaths of thousands of people every year in developing and industrialised countries alike -
Even if the precautionary principle were to be applied mildly, this chemical should be banned
In fact banning the chemical through legislation at the EU level is what 59% of Europeans said in a survey in the UK, Germany, France, Austria and Scandinavia
83% of the same people expressed very strong views about the need to
control global warming, objected totally to the use of any animals for
medical research and of course they were overwhelmingly anti-biotech!
-
Dihydrogen monoxide
Otherwise known as
Water
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Jettisoning scientific risk-benefit assessment and replacing it with a precautionary approach will result in arbitrary and politically motivated decisions that will decide the fate of Genetically Enhanced crops and products
The Cartagena biodiversity protocol is the single most
important threat to transgenic crop development
This is because the original focus of the protocol has
been highjacked for politial and economic reasons
aiming solely towards protectionism and restoration of
trait barriers under the pretext of biosafety!
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Plant Biotechnology has a huge potential to contribute substantially to food security and poverty alleviation, in addition to creating a better and healthier environment
The major issues we now have to address are political and economic in nature
and this is where scientists have a role to play in putting the record straight!
1127.psd1128.psd1129.psd1130.psd1131.psd -
European objections to transgenic crops and food derived from them are aiming to protect the CAP and NOTHING ELSE!
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Transgenic plant releases and commercialization are governed by Draconian rules
unparalleled elsewhere in any other sector. The European Union in a report following
a 15 year study (1985-2000) involving 400 public research institutions, to the cost of
70 million Euros stated genetically modified plants and products derived from them
present no risk to human health or the environmentthese crops and products are
even safer than plants and products generated through conventional processes
(EC Research, 2001).
The claim that antibiotic resistance genes in transgenic plants
will escape into natural bacterial populations that will subsequently become resistant
to them, thus creating super-bacteria is at best odd, as these genes are already
present in bacterial population in nature. It is worth remembering that the selectable
marker genes were isolated from these very naturally occurring bacteria in the first
place, for use in the laboratory!
EC Research (2001) EC-Sponsored Research on Safety of Genetically
Modified Organisms: A review of results:
http://europa.eu.int/comm/research/quality-of-life/gmo/
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A degree of realism on the part of regulatory agencies is necessary
to assure that over-regulation stops being undully cumbersome and
prohibitively expensive.
Regulatory agencies need to divorce themselves from environmental
and political activists that threaten to put a stop transgenic plants.
Safety and efficacy should be paramount but they need to be
realistic and proportionate, and need to consider risk-benefit ratios
Image by permission of Fotosearch.com
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Biotech Crop Countries and Mega-Countries, 2007
2008/9
-
Create super-nutritious biofortified rice and maize seeds which will
represent a quantum leap in current efforts to address issues
of poverty alleviation and food insecurity, through biotechnology
in the developing world with durability and sustainability in mind,
mostly by limiting key agronomic inputs imposed by the multitude of
insects and noxious/parasitic weeds in marginal environments,
focusing on Sub-Saharan Africa and the Indian subcontinent
Simultaneous multi-pathway engineering in crop plants through
combinatorial genetic transformation: creating complete biofortified
cereal grains for food security (BIOFORCE)
-
BIOFORCE targets
Vitamines A, C, folate (B9), EMinerals Fe, Zn, Se, CaInsect resistance and eliminate StrigaEuropean
Research
Council
-
Vitamin A deficiency is prevalent in the developing world, and is probably responsible
for the death of 2 million children every year.
In surviving children, vitamin A deficiency is a leading, but avoidable cause of blindness. Effects of vitamin A deficiency are manifested as xerophthalmia
(visual impairment), blindness, and increased mortality due to increased severity
of children diseases such as measles, diarrhoea, and increased maternal
transmission of virus such as HIV.
Humans can synthesize vitamin A if provided with the precursor molecule
beta-carotene (also known as provitamin A). Endosperms of food crops, such as maize
and wheat, are low in provitamin A (1-10%) as compared with non-provitamin A
carotenoid.
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Teresa Capell
SPAIN
Changfu Zhu
CHINA
Koreen Ramessar
(PhD student-South Africa)
Sonia Gomez
(PhD student-Spain)
Ariadna Peremarti (PhD student-Spain)
Svetlana Dashevskaya
(PhD student-Israel)
Gemma Farr
(PhD student-Spain)
Maite Sabalza
(PhD student-Spain)
Dawie Yuan
(PhD student-China)
Ludovic Bassie
FRANCE
Bruna Miralpeix
(Final year project student
Spain)
Shaista Naqvi
(PhD studen-Pakistan)
MICINN
Generalitat de Catalunya
UdL
ICREA
EU FP 6 & 7
ERA NET
Bill & Melinda Gates Fd
G. Sandmann, Uni Frankfurt
G. Ross, Univ Murcia
745.psd746.psd -
If you desire peace, cultivate justice, but.
Norman E. Borlaug, Oslo, Norway, December 11, 1970
Nobel lecture
.at the same time cultivate the fields
to produce more bread.
otherwise there will be no peace
0
20
40
60
80
100
120
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180
M37W
A632
1234567
Total Carotenoids (g/g DW)