United Nations Pavilion, EXPO 2015 Milano

photo: E. Pedrazzini

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sugar cane maize rice wheat milk (cow) potato vegetables cassava soybean eggplant

The great cropsGlobal production (Gtons) 2013

FAOSTAT

Barley yield through millennia

Stanca et al.

• Natural genetic variability of a given species may be wide, but it is neverunlimited

• Beyond a certain limit, you have two options:• search for alternative crops• artificially introduce mutations, using whatever method is available

The hidden hunger

Phytic acid is the principal storage form of phosphorus in seeds

• Phytic acid has strong affinity to metals, making them far less absorbable in the intestines

• It is estimated that only 5% of the Fe and 25% of the Zn in legumes and cerealseeds is bioavaliable

cytosol

vacuole

The bean lpa1 mutant has a non functional PHYTIC ACIDtransporter (Pvmrp1) at the vacuolar membrane

glutamic acid 1155 → lysine

90% reduction of phytic acid in the seed

Panzeri et al (2011)Sparvoli & Cominelli (2014)Sparvoli & Cominelli (2015)

Biscuits made with 1/3 lpa1 bean flour

From molecules, through the field, to our table

PROTEIN 37,5 %

ESSENTIALAMINO ACIDS

PHYTIC ACID 51,5 %

Lys 244,0 %Trp 134,0 %

Sparvoli et al. in preparation

Average global protein supply quantity(g/capita/day)

cereals (excluding beer) 31.90 grams

animal products 31.70 grams

pulses 4.0 grams

FAOSTAT

Wheat gluten structure: still an open question 10.000 years after domestication

Gluten

After Salamini et al., 2002

Gliadins Glutenins

Monomeric Polymeric

The fertile crescent Tecnological properties

Technological quality

Ceriotti & Grasso

food lysine tryptophan methionine + cysteinerice 36 13 36wheat 26 12 35maize 29 7 40soybean 64 13 26common bean 72 10 19beef 87 12 37milk 79 14 34fish 100 11 45egg 70 16 56

estimated human requirements (FAO 2011)0 – 6 months 69 17 336 months – 3 years 57 8.5 27> 3 years 48 6.6 23

Some essential amino acids (mg/g protein)

Nutritional quality

THE FIRST….

• Plant mRNAs and genes to be isolated and sequenced• Plant genes introduced into a transgenic plant• Plant proteins modified through genetic engineering

BUT….

The paradox of seed storage proteins

“…….despite the investment of millions of dollars bythe major seed/plant biotechnology companies, theconsumer is yet to see a transgenic cereal or legumecrop expressing higher levels of essential amino acids”

Rao (2008) Plant Physiol. 147: 6–12

Possible explanations

• Major seed biotech companies have rapidly shifted their interest toproducers’ rather than consumers’ expectations: yield has beenconsidered more important than quality

• Seed storage proteins belong to allergenic protein families

• Altering protein structure without side effects on stability is not easy,we need to understand more

alternative crops

Chenopodium quinoaAmaranthus

food lysine tryptophan methionine + cysteinerice 36 13 36wheat 26 12 35maize 32 6 40Quality Protein Maize 45 10 39soybean 64 13 26common bean 72 10 19quinoa 61 11 48amaranth 51 – 61 9 - 18 41 - 49camelina1 52 15 51flax2 38 19 43cannabis2 42 16 41beef 87 12 37milk 79 14 34fish 100 11 45egg 70 16 56

estimated human requirements (FAO 2011)0 – 6 months 69 17 336 months – 3 years 57 8.5 27> 3 years 48 6.6 23

2Russo et al (2015)

1Colombini et al (2012)

crop Yield (tons/hectar)

year 2000 2013

quinoa 0,8 0,9

wheat 2,7 3,3

rice 3,9 4,5

maize 4,3 5,5

soybean 2,2 2,5

common bean 0,7 0,8

faostat.fao.org

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QuinoaGlobal production (tons)

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PerùBoliviaPrice (USD/ton)

food lysine tryptophan methionine + cysteinerice 36 13 36wheat 26 12 35maize 32 6 40Quality Protein Maize 45 10 39soybean 64 13 26common bean 72 10 19quinoa 61 11 48amaranth 51 – 61 9 - 18 41 - 49camelina1 52 15 51flax2 38 19 43cannabis2 42 16 41beef 87 12 37milk 79 14 34fish 100 11 45egg 70 16 56

estimated human requirements (FAO 2011)0 – 6 months 69 17 336 months – 3 years 57 8.5 27> 3 years 48 6.6 23

2Russo et al (2015)

1Colombini et al (2012)

name % lysine and tryptophan

seed phenotype Storage proteins

opaque2 nearly doubled opaque, fragile less α-zeins

opaque2 QPM nearly doubled normal (vitreous) less α-zeins

Maize quality improvement

Holding (2014)

name % lysine and tryptophan

seed phenotype Storage proteins

opaque2 nearly doubled opaque, fragile less α-zeins

opaque2 QPM nearly doubled normal (vitreous) less α-zeins

Maize quality improvement

Holding (2014)

Parent-of-origin effects on α-zein expression

o2-Parent 1c

o2-Parent 2

♂P2 x ♀P1

♀P1 x ♂P2

zeins

Hybrid production for better quality may be influenced by epigenetic phenomenathat we still do not understand in detail

Lauria et al, 2004, 2014Ciceri et al, 2000

PNAS 2010 107 (29) 12810

Elimination of 27 kD and 16 kD γ-zeins abolishes the vitreousrestored phenotype in QPM maize

What is peculiar in these proteins?

αγ 16 kDβ

δ

γ 27 kD

γ 50 kD

zeinsMainieri et al

Clarissa LancilliMassimiliano Lauria

Davide MainieriClaudia MarranoIride Mascheretti

Emanuela PedrazziniRaul Pirona

Angelo Viotti

Eleonora CominelliMaria Gloria Daminati

Roberto BolliniDario Panzeri

Francesca Sparvoli

Incoronata GalassoRemo ReggianiRoberto Russo

Aldo CeriottiAldo Grazzo

• Università degli Studi, Milano• Università. Di Pavia• Università di Modena• University of East Anglia• CREA, Bologna, Bergamo,

Montanaso Lombardo, Lodi• CINVESTAV, Irapuato• USDA, Madison• BOKU, Vienna

Framework agreement CNR – RegioneLombardia

maize

common bean

camelinaflaxcannabis

wheat

• The boundaries between established genetic modification (GM) and non-GM techniques will become increasingly blurred as techniques develop.

• A regulatory system based on the characteristics of a novel crop, by whatever method it has been produced, would provide more effective and robust regulation than current EU processes, which consider new crop varieties differently depending on the method used to generate them.

• We ask that EU regulators confirm that the products of new breeding techniques, when they do not contain foreign DNA, do not fall within the scope of GMO legislation.

• The aim in the EU should be to regulate the specific agricultural trait and/or product, not the technology.• At the same time, there is also continuing need for wide-ranging engagement on critical issues and this should

include re-examination of the appropriate use of the precautionary principle.

NEW TECHNIQUES FOR GENETIC CROP IMPROVEMENT Position statement

NEW BREEDING TECHNIQUES Statement

The European Commission’s delays in clarifying the legal status of the NPBTs weaken the competitiveness of the EU plant breeding sector. EU farmers already suffer unfair competition from primary producers in other regions of the world regarding access to all appropriate tools including genetic engineering and NPBTs.

CROP GENETIC IMPROVEMENT TECHNIQUES Statement