New Trends in Biotechnology

Martina Newell-McGloughlin

Director, University of California Systemwide

Biotechnology Research and Education Program

(UCBREP)

Paper: Nutritional and Safety Assessments of Foods and Feeds

Nutritionally Improved through Biotechnology:

http://www3.interscience.wiley.com/journal/119423779/issue

Newell McGloughlin, Plant Physiology, July 2008

http://www.plantphysiol.org/cgi/content/full/147/3/939

• High yielding affordable high quality food feed and fuel with minimum inputs

• 17% of land under cultivation degraded by human activity 1945 to 1990. Ag land shrinks by 20,000 ha yearly. (World Bank)

• Without yield increase land use will 2X by 2050.

• Latin America: greatest yield increase had lower land use (less deforestation)

• High yield “land sparing” better than “wildlife”-friendly inefficient land use farming

(Green, Royal Soc. Bird Protection 2005)

• EU pursuing 19th C technology, young scientists will flee. If the EU engages rational harmonized regulatory framework it will encourage a more rapid international diffusion of the technology.

• EU Commission "need to take urgent action to avoid negative implications for EU livestock production and agriculture overall".

1997 acreage

Reality check

Agriculture: A history of

Technology

8,000 BC

19thC

Ea 20th C

Md 20th C

1930s

1940s

1950s

1970s

1980

1990s

2000s

21st C

Cultivation

Selective Cross breeding

Cell culture

Somaclonal variation

Embryo rescue

Mutagenesis and selection

Anther culture

Recombinant DNA

Marker assisted selection

---omics - Bioinformatics

Epigenetics/RNAi/Paramutation

Adaptive technology/transgenomics

Systems Biology

Quality Traits - ($210B by 2015)

Improved post harvest characteristics

Shelf life, processing, taste

Improved Nutrition –Improved Functionality

Macro: protein, oils, carbs, fibre

Micro: Vitamins, minerals,

Phytochemicals – Antioxidants

Remove Antinutrients/allergens/ Toxins

CO2

Opportunities/Challenges for Biotech Crops

Agronomic Traits – $30B Biotic-

pest/disease/weeds/ Abiotic Stress:

Drought salinity marginal soils,

Yield

Value

Renewable Resources

Biomass conversion,

feedstocks, biofuels,

Phytoremediation

Concerns land/ water

use Perennials: Trees

Plants as Factories

Pharmaceuticals/ Industrial products

(Ventria – Rice Lactoferin Lysozyme

30% Diarrhea, recovery 3/6 days,

Concerns gene flow co-mingling

Source: ISAAA

• 2009, 14 M farmers 134 M hac (330 M acres) of biotech crops in 25 countries,

• 13.3 M farmers/125 M hacs (7%) in 2008. 13 /14 M (90 %) resource-poor LDC.

• 46% global age of biotech crops LDC- Total 57 countries have reg approvals.

• 6 EU planted 94,750 hacs in 2009, down from 7 and 107,719 hacs 2008, Germany

discontinued. Spain planted 80 % Bt maize maintained record adoption 22%.

• $44 billion 1996 to 2007, 44% yield gains, 56% reduction costs (including 359,000 tonne a.i. in pesticides); gains of 141 million tons, would require 43 M additional hectares

• Environmental pesticide footprint down by 15.4 %. GM reduction in 286 million kg of CO2 emissions equivalent to removing 6 M cars from the roads (Barfoot and Brookes 2008)

• Additional soil carbon sequestered since 1996 has been equivalent to 63,859 million tonnes of CO2 that has not been released into the global atmosphere.

• In 1994-95 farmers spent $78 Ha in herbicides; today they spend $37/Ha and insecticide use has decrease 90%.

• HT- increase in no- till: reduction in erosion, soils much healthier, organic matter, less soil compaction, better H2O usage, fuel use down by 20 gals/acre (Fawcett & Towery 2005 )

Environmental Impact

• China: Bt rice has the potential to increase yields up to 8 percent, decrease pesticide use by 80 percent (17 kg/ha) and generate US$4 billion in benefits annually (James, 2010). Significant decrease in adverse health effects – Lives saved

• Organisms in “Bt crops” fields fared better in trials than those with insecticides Monarch butterflies increase (Marvier, 2007)

• BT corn 90% reduction in mycotoxin fungal fumonisins -total US benefit estimated at $23m annually. (Wu, 2006)

• Origin Agritech China also approved Phytase maize (2009)

• CP papaya saved Hawaii papaya industry (and helped organic farmers!) may be the outcome for plum pox –C5 PTGS insurance against typhoid Mary in nurseries

• Blight-resistant potato (BASF -Rpi-blb1 and Rpi-blb2 NBS-LRR) -UI study concluded for the major potato-producing regions of the world would be $4.3 billion.

• ISAAA expects the number of biotech farmers globally to reach 20 million or more in 40 countries on 200 million hectares in just more than five years in 2015.

Two Down One doing Well

Spain: farmer Jose Victor Nogues” Most people can appreciate the huge benefits and lack of negative effects -Introducing GM maize was definitely the way forward

• France: Thierry de l'Escaille, European Landowners' Organization, - “wide-scale adoption of these three biotech crops in Europe could significantly increase annual production, improve farmer income by more than 1 billion Euros and reduce spraying. With results like these, it's easy to understand why farmers want access to this technology," said l'Escaille

• Sarkozy dit “mais non”!

Bt corn farmers earn about $85

more per acre, ISAAA (2007), while at

the same time producing a healthier

feed that is better for the environment.

Romania: Buzdugan farmers reported

price premiums of up to 10 percent for

biotech soybeans due to fewer weed

impurities. Average price gain 2%.

Production gain 29-33.5K M tonnes

(16-19% ) Earnings increase 35

million and 62.4 million euros (2004).

I can tell you that soybean farmers in

Romania are very interested in biotech

seeds," “Although the seeds are 10 to

15 percent more expensive, the income

gains make the extra cost more than

worthwhile” Now EU so no GM Soy!

• Other key highlights approval of SmartStax, a novel biotech

maize 8 different genes for insect and herbicide resistance

• SmartStax protects crops from major corn pests, including

European and southwestern corn borer, northern and

western corn rootworm, western bean cutworm, black

cutworm, corn earworm, and fall armyworm. SmartStax

will also provide resistance to glyphosate and glufosinate

herbicides. Dow AgroSciences field research trials

conducted in 2007-2008 confirmed SmartStax provides a

broader spectrum of insect protection in corn hybrids.

.

Genuity™ SmartStax™ Corn technology

-- Above-ground insect control: Dow's Herculex® I

technology with Monsanto's second-generation, two-gene

lepidopteran control technology called YieldGard® VT

PRO, SmartStax will provide protection against European

corn borer, southwestern corn borer, corn earworm, fall

armyworm, western bean cutworm, and black cutworm;

-- Below-ground insect control: Combining Dow

AgroSciences' Herculex® RW technology and Monsanto's

YieldGard VT Rootworm/RR2 technology, SmartStax will

provide protection against Western, Northern and Mexican

corn rootworms; and,

-- Weed control: Monsanto's Roundup Ready® 2 technology

with Liberty Link® herbicide tolerance, SmartStax will

offer broad spectrum weed and grass control with the

option of two different modes-of-action.

- asked the EPA to consider reduced refuge requirement due

to different modes of action - broader spectrum of control

and lessened likelihood of target pests developing resistance

Genuity™ SmartStax™ Corn technology

• Cross licensing Monsanto Syngenta

• Four years of field trials across six U.S. states showed 7 to

11% higher yields, compared to the first generation of

Roundup Ready soybeans.

• “Roundup Ready 2 Yield did very well for me last season,”

Jeff Barth, a farmer from Illinois, said. “They were planted

late and still performed five to six bushels better than the

first-generation Roundup Ready soybeans that were planted

earlier. I’m anticipating similar results this year, and that’s

why I will dedicate all of my 1,100 soybean acres to the

product when they become available.” This is also Barth’s

second year growing Roundup Ready 2 Yield soybeans.

Roundup Ready 2 Yield

Roundup Ready 2 Yield soybeans – the first product

of a new class of technology that allows more

efficient, precise gene insertion to directly impact

yields.

“Resistance” Genes natural and otherwise

• Rootknot nematodes R in tomato (Mi) and

(aphids). Alternate to fumigation (Williamson

UC Davis)

• Xa21 rice R gene confers resistance to several

Xoo. Defense response triggered by Xo

molecule, AvrXa21. Transgenic more resistant

due to copy number (Ronald)

• Use of apoptosis inhibition to protect plants

from mycotoxin damage (Gilchrist, UC Davis)

• Sclerotinia-resist sunflower oxalate oxidase

Pioneer wheat

• Zinc Fingers Dow/Sangamo

Abiotic Stress:

Drought, Cold, Heat, Salinity

Abiotic stress limiting factor for

crops reaching genetic potential

Improved water

conservation –

Fewer crop losses –

Higher yields on all

acres through

improved water

utilization –

Expand in drylands

BASF Drought-

tolerant corn 12

bushels more an acre

+ Gene EControl+ Gene DControl

Abiotic Stress:

Drought, ColdHeat, Salinity

Drought Stressed Rice

Abiotic stress limiting factor for crops

reaching genetic potential

Improved water conservation -Fewer crop

losses -Higher yields on all acres through

improved water utilization -Expand in

drylands - Nuclear Factor Y B subunit

Crops limited by salinity on 40 % world's

irrigated land (25 % US)

Cold: Engineering with COR15a Tf, role

in freezing tolerance.

Plants engineered with Choline oxidase

(codA) soil tolerated saline and cold

Homeodomain-leucine zipper (HD-Zip)

transcription factors respond to H2O &

osmotic stress, exongenous abscisic acid

Transport protein. Grow and fruit even in

irrigation water that is > 50X saltier than

normal. > 1/3 salty as seawater.

Blumwald and Zhang)

Abiotic stress limiting factor for crops reaching genetic

potential

Drought Tolerant Oilseed Rape The plant is engineered to

reduce the levels PARP [poly(ADP-ribose) polymerase], a key

stress-related protein in plants. This results in a crop which

was far better able to survive drought than reference plants.

Show relative yield increases of up to +44% compared to non-

drought tolerant varieties.

Bayer CropScience is currently conducting research work on

maize, cotton, oilseed rape and rice, with the objective of

developing a new generation of stress-tolerant, high-

performance crop varieties. The drought tolerant varieties have

a mutation that changes the activity of farnesyltransferase.

Pioneer Hi-Bred International is developing hybrids and

varieties that use water sources more efficiently and therefore

perform better during water deficits.

Maintaining yields during water stress will help preserve

grower incomes and yield more grain for the food and energy

value chain as well as reducing the need for irrigation.

- Improve Nitrogen Assimilation- Increase Sucrose hydrolysis,

Starch biosynthesis- Increase O2 availability - Modify photosynthesis

Yield Gene Control

Increased Yields

Forage Crops: This short-day

sorghum plant was used to map

the Ma-1 gene (genes which

modify photoperiodic behavior

and thus maturity). This gene

which works in other cereals

would offer particular benefits to

biomass and forage crops in

which flowering is undesirable

Nitrogen Use

Efficiency (NUE)

• Nitrogen fertilizer accounts for one-third of

the GHGs produced by agriculture

• Nitrous oxide 300 times more global

warming potential than carbon dioxide.

• Farmers spend $60 billion annually for 150

million tons of fertilizer (Stern Review 2006).

• Arcadia's Nitrogen Use Efficiency (NUE)

plants with equivalent yields that to

conventional varieties but which require

significantly less nitrogen fertilizer because

they use it more efficiently

• This technology has the potential to reduce

the amount of nitrogen fertilizer that is lost

by farmers every year due to leaching into

the air, soil and waterways. In addition to

environmental pressures, nitrogen costs can

represent a significant portion of a farmer's

input costs and can significantly impact

farmer profitability.

• licensed its technology to DuPont for use in

corn and to Monsanto for canola.

30% less applied N fertiliser

BioFuels

• LDC 30 % global energy. Growth driven by population and economic . Of

the world's 47 poorest countries, 38 are net oil importers, and 25 import all

of their oil, consuming much of their national income to pay for it.

• The challenge: 5-10 times more efficient - 2001 $5/gal -2005 18c/gal

• Biomass Conversion: Organic polymeric material, lignin, starches celluloses

bioconverted ethanol; hemicellulose hydrolyzed to sugars, xylose , glucose .

• Modify Plants and algae to improve enzymatic conversion.

• Modify enzymes to improve conversion and fermentation

• Maize other cereals, Switch grass, elephant grass poplars

• Biodiesel is biodegradable and non-toxic - alkyl esters made from the

transesterification of vegetable oils or animal fats., (60% less CO2)

• Rapeseed, Botryococcus braunii (Bb) colloidal microalgae

• Concerns: Food trade off – Efficiency of production – ecological impact

Many common food crops not perfect for nutritional

requirements of humans or animals.

Proteins: Maize, wheat, Sweet potato and cassava

WHO: 800 million people suffer from malnutrition, Protein-

energy malnutrition (PEM), the most lethal form, affects

1 in 4 children:

70% live in Asia, 26% Africa, 4% Latin America, Caribbean

• Grains low in Lysine – LDCs food - Feed Rations/pollution

• High Lysine maize: Use non feedback- enzyme (5X ppm)

• N assimilation modified pathway GDH 12% increase protein

• SRP Nonallergenic Amaranthus Albumin for potato

• High Protein: Cytokinin rescue flower pair kernels fused

single kernel two embryos - high protein/oil low CHO

• NAC Tfs (NAM) senescence and nutrient remobilization

leaves to grains, RNAi delay senes 30% protein, Zn, Fe

• Artificial Proteins:

• ASP-1-sweet potato 67% increase protein (EAA 80%)

Improved Nutritional Content

Improved Nutritional ContentCarbohydrates

• Starch High Amylose (resistant starch) inhibit 2 SBE

• Sorbitol role in fruit carbon metabolism and affects

quality attributes sugar-acid balance and starch accum

• Wheat puroindoline genes in rice better starch/flour

Fibre – Humans increase

• Polymers, Inulins, Fructo-oligosaccharides (FOS)

• SC Fructans sucrose taste: GI Tract health- fermented

colonic – bifidobacteria (compete pathogenic bacteria)

SC Fatty acid – anticancer/ inhibit HMG-CoAR less LDL

• SC fructans 1-SST Jerusalem artichoke. 90% sucrose

converted "fructan beets“ (Koops, 2000)

• Potato synthesize the full spectrum of inulins from

globe artichoke roots

• Lignans: enterodiol/lactone estrogen-dependent cancer

Forage Crops Fibre – Animals Decrease

• Brown midrib (COMT)–Decreased lignin increase

digestibility better feed conversion, livestock prefer

(Sorghum)

Improved Nutritional Content

Oils and Fatty acids

• Altering chain length and saturation level•

• Novel genes to produce unusual fatty acids in oilseed

• MUFA: High Oleic Acid: more stable than PUFA heat/

oxidation resistant, little or no postrefining

(hydrogenation): AS oleate desaturase soybean gave

>80% oleic acid (23%), Less SF milk/meat of animals

• MCT: medical foods, ergogenic aids. Acyl-ACPT

canola, increase in capric (C10) and caprylic (C8)

• High-CLA: Antiox- free radicals heart disease/cancer

• Omega -3 DHA-EPA “Fish Oil” CV/thrombosis/

Cancer/ Arthritis/ Cognitive/Mental/ premies -

D6 Desaturase: Canola/soybean precursor SDA 3.6X

ALA in generating EPA

• GLA safflower oil (C18:3n-6) anti-inflammatory effect,

improved skin health and maintaining weight loss

• Sitostanol: phytosterol phospholipid Block

cholesterol

Improved Nutritional Content

Micro nutrients Vitamins/ minerals:

• Vit A Golden rice II b-carotene-Rice 25X (CBP)

• Biofortified cassva flour- Field trial Nigeria (Sayre)

• Vit B Folate increase in rice (pregnancy deficinicies)

• Vit E a-tocopherol g-TMT; Vit C increase corn DHAR

Minerals: Ferritin (bean S protein), Metallothionein (Rice,

wheat). Ca/proton antiporter (sCAX) Ca transport into

vacuoles. Ca-fortified carrots enhanced absorption.

Multi vitamin Corn• Combinatorial direct DNA transformation rapid

production of multi-complex metabolic pathways

• transferred 5 constructs controlled by different

endosperm-specific promoters into white maize.

Different enzyme combinations show distinct metabolic

phenotypes – resulting in

• 169X beta carotene (60 mg/g v. 14 by breeding)

• 6X vitamin C, and

• 2X folate (Christou, 2009)

(bacteria)

(daffodil)

(daffodil)

Introduced

enzyme

(maize)

Improved Nutritional Content

Functional components - effects greater than nutrient value

alone

Phytochemicals:

• Carotenoids: Golden Rice, Sweet Potato - (sight,

development)

• Lycopene: polyamine Tomato – (reduce LDL, cancer)

• Isoflavones: genistein and daidzein, tyrosine kinase inhibitor

• Phenolics: resveratrol antioxidant Sirtuins (anti-aging)

• Flavanols: Catechins, Flavones: quercetin (less adjuncts)

Anti-nutrients: Trypsin Inhibitors; oxalic acid; furans;

Phytate, Bioavailability Phosphate, divalent ions: Phytase

(Rice, alfalfa)

Allergens: soy P34 removal; peanut; gluten digestion

Toxins: Glycoalkaloid (potato) AS solanine

• Cyanogenic glucoside (cassava) hydroxynitril lyase

• Concerns:

• High levels, Novel proteins, metabolites (pleiotropic effects)

Increased susceptibility to pathogens

Risk Management

Antibiotic Resistance (perceived not scientific)Transposon taggingPositive selection – exclusive energy source

Gene Flow-SpaceMale sterility“Terminator” technologyChloroplast transformation

Lack of specificity – pleiotropic effects (expected, unexpected)Site-specific recombinationCre-Lox

Effect on non-target speciesTissue specific expressionChloroplast transformation

Loss of effectiveness – resistance managementRefugiaGene Stacking, Gene Pyramiding, Gene shuffling

Reduced diversityMore sources of genetic diversity – rescue heritage varieties and

landraces Novel animals – Food safety, feral potential

Physical and Biological containment

Cooperation works

No

yellow

kernels

Organic Blue Cornfield near yellow non-organic field Fred Yoder Ohio

No cross

pollination

(no blue

kernels)

Biotech Corn Organic Corn

• Historically, worldwide the market adequately addressed

economic liability issues relating to trace presence of

unwanted material in any agricultural crop.

• US organics cannot be (legally) downgraded or growers

decertified by unintentional presence when all required

measures and best practices are adhered to and no

producer has been so impacted to date

Primum non nocere• Commercialization: 7 to 10 years -at least 9 review stages

• Biotech crops and foods more thoroughly tested than

conventional varieties ( “assumed” to be safe)- One

biotech soybean subjected to 1,800 separate analyses

• 23 feeding studies - dairy, beef, poultry, soy/corn

equivalent in composition, digestibility and feeding value

to non-GM. Clarke et al 2000

• Substantial equivalence with parent - Molecular

characterization (17) Toxicity studies (5) - marker genes

(4) - Nutritional content (7+)- Allergenicity potential -

Anti-nutritional effects - Protein digestibility

• Environmental aspects (5 items)- Ecological impact (5

items) OECD, CBD, CODEX

Omic studies

Wheat ( Baker 2006), Potato (Catchpole 2005)

Transcriptomic and Metabolomic studies show greater variation between

conventional bred cultivars and even growth locations than between GM and

parental variety (except of course for the intended modification!) - differences

between sites were generally greater than differences between lines

Greatest Challenges going forward

• Technical

• Intellectual Property: PIPRA - Specialty crops – FTO

• Liability

• Biosafety: so–called – LDCs – Specialty crops

• Acceptance: - countering fear and misinformation

(ethical) - moral imperative real need v. hypothetical risk

I hope that there is nothing

genetically modified in this