Benefici e rischi di OGM dalla ricercapubblica Italiana
Prof. Bruno MezzettiDepartimento di Scienze Agrarie, Alimentari
ed AmbientaliUniversità Politecnica delle Marche
Mola di Bari, 08/07/2015
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BIOTECHNOLOGY IN THE ANCIENT GREEK
Philosopher and botanist of the ancient greek, disciple of Aristotle.371 b.C., : 287 b.C.
Teofrasto
A grafted plant it is already a combination of 2
genomes
Agrobacterium tumefaciens: the natural
biotechnologist
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6
27
9
2
70
2
25
15
3
38
6
50
0 10 20 30 40 50 60 70
Insect resistance
Fungi resistance
Virus resistance
Plant yield
Plant Habitus
Fruit development-quality
Nutritional Value
Herbicide resistance
Male sterility
Male Ster.+ Herbicide Res.
Insect + Herbicide Res.
Other
Type of Genes
GM Research Trend in Italy 1992-2003
Source CIV
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64
2
4
8
1
14
14
17
72
10
31
8
69
17
1
15
0 10 20 30 40 50 60 70 80
Veneto
Umbria
Toscana
Sicilia
Sardegna
Puglia
Piemonte
Marche
Lombardia
Liguria
Lazio
Friuli Ven.Giulia
Emilia Romagna
Campania
Calabria
Basilicata
REGIONS where GM trials were carried out
GM Research Trend in Italy 1992-2003
Source CIV
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100
2
1
2
1
4
7
45
7
2
1
7
95
1
1
3
4
1
1
5
1
3
12
1
8
38
0 10 20 30 40 50 60 70 80 90
Zucchina
Grape
Tobacco
Statice
Soia
Rice
Tomato
Potato
Oliv
Melon
Eggplant
Mais
Lettuce
Raspberry
Kiwi
Weat
Ginestrino
Geranio
Strawberry
W.Melon
S.cherry
Cicory
Canola
Calendula
Sugarbeet
24 different GM crops
The most studied were maize (95
notifications) and tomato (45
notifications), but as a peculiarity of
our country, there were also research
activities carried out on many
horticultural crops, such as fruit,
vegetables and flowers.
Fruit without Fertilization
1.Novelty - especially for fruits such as cherry and
raisin tomatoes, squash, eggplant, pepper, strawberry,
melon and watermelon. 2. Improved taste - since
seedless fruits often have higher Total Soluble Sugar
(TSS) levels than seeded fruit. 3.Greater reliability of
crop yield under varying climatic conditions - due to
elimination of pollination as a factor in fruit set.
4.Cost reductions of greenhouse grown fruits, since
pollination-related expensed are avoided. 5.Increased
profitability for the fruit processing industry: a)
because the longer harvesting period of parthenogenic
crops enables processing equipment to be more
efficiently used, b) the elimination of the seed
removal stage reduces expenses (for instance in
tomato paste production or in melon canning), or
increases the value of the product (as in oleoresin
extraction from paprika).
Eggplant has been produced
using Auxin gene (iaaM)
under the control of
Antirrhinum majus
ovule-specific promoter
Antirrhinum majusP. savastanoi
Ba
mH
I
DefH9-IaaM Gene (Spena – UniVR)
It confers expression within the placenta and the ovules and it has been used
to confer parthenocarpic fruit development to
several plant species and varieties
(Rotino et al., 1997) From regeneration and selection
to the open field (2001)
Eco
RI E
coR
I
Kp
nI
Eco
RI
Hin
dII
I
DefH9 PromoterIaaM codingTerminator
LB RB
The experimental trial: ‘Thompson Seedless’ Control and GM line: 32 plants each ‘Silcora’ Control and GM lines (line A and line B): 16 plants each Vines were spaced at 2.5 x 1.5 m, trained with the ‘double guyot’ system
Mean+SE Duncan’s statystical test, P < 0.05
Production cycles 2004-05-06
DefH9-iaaM gene effects on plant production, fruit and berry size cv. ‘Thompson seedless’
0,80
0,82
0,84
0,86
0,88
0,90
0,92
0,94
0,96
0,98
Th CT Th DefH9-iaaM
Berr
y w
eig
ht
(g)
0
50
100
150
200
250
300
N°b
err
ies/
clus
ter
Berry weight
N°berries/clustera
b
a
b
0
1
2
3
4
5
6
7
8
Th CT Th DefH9-iaaM
N°c
lust
ers
/pla
nt
0
25
50
75
100
125
150
175
200
225
Clu
ster
weig
ht
(g)
N°clusters/plant
Cluster weight (g)
a
a
b
b
1. Thomphson seedless yield
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DefH9-iaaM - Table grape
Effect on shoot fruitfulness (Duncan’s Multiple Range Test; p < 0.01)
Thompson seedless Control Thompson seedless DefH9-iaaM
Laboratory Field fruitfulness
2003 Bud Fertility and Position - Laboratory Field fruitfulness
Total bud
with shoots
% Buds with
fertile shoots
% Buds
without
fertile shoot
Buds
fertility
position
Total number of
bunches per shoot
TH - CT 141.00 14.90 85.10 12.62 0.08
TH - GM 108.00 25.00 75.00 10.81* 0.26*
SI - CT 176.00 50,00 50.00 11.89 0.33
SI - GM 166.00 59.64 40.36 10.51* 0.46*
CLONES
Inflorescence on a shoot
arose from a basal bud of
DefH9-iaaM Thompson
seedless, unusual for this
variety.
Fruit developmental phases: PB= pre-blooming; A= antesys; FS= fruit-set; SH= small hard fruit; LH= large hard fruit. Values are referred to PB, arbitrary set to 1.
DefH9-iaaM gene expression during berry development
Elisa Costantini - Ph.D. discussion - Marche Polytecnic University 26/01/07
DefH9-iaaM thomp
0
10
20
30
40
50
60
70
PB* A FS SH LH V H
rela
tive
expre
ssio
n
DefH9-iaaM silcora
0
10
20
30
40
50
60
70
PB* A FS SH LH V H
rela
tive
expre
ssio
n
cv. ‘Thompson seedless’ cv. ‘Silcora’
2. DefH9-iaaM gene expression and effects on IAA content and on VvHT1 gene
IAA content during berry development
Fruit developmental phases: IS=inflorescences separated; PB=pre-blooming; A= antesys; FS=fruit-set; SH=small hard fruit
Elisa Costantini - Ph.D. discussion - Marche Polytecnic University 26/01/07
GC-MS analysis on tissues samples Cv ‘Thompson seedless’ and ‘Silcora’ (control and GM line)
0
5
10
15
20
25
30
35
40
45
IS PB A FS SH
IA
A p
mol
/g
Thompson CTThompson DefH9-iaaM line
0
20
40
60
80
100
120
140
IS PB A FS SHIA
A p
mol
/g
Silcora CT
Silcora DefH9-iaaM line A
2. DefH9-iaaM gene expression and effects on IAA content and on VvHT1 gene
DefH9-iaaM - Table grape
Effect on yield capacity (0ne plot production)
TH – Control SI – ControlTH – GM SI – GM
(a) F. vesca - Control, Line 1, Line 2
(b) F. x ananassa - Control and Line 1
(c) R.idaeus – Control and Line 1
GMO at Marche Polytechnic University
Strawberry and raspberry fruits from control and DefH9-iaaM plants
DefH9-iaaM raspberry differs for Increased No.
of Inflorescence and of No. Fruits per laterals.
Ruby – Control Ruby – Line 1
GMO at Marche Polytechnic University
Raspberry fruits from control and DefH9-iaaM plants
Raspberry - Plant Fecundity
Cisgenic plants
(Solanum tuberosum)
DNA from
- the species itself
- a cross-compatible
species
Wild Solanum
Agrobacterium tumefaciens
Cell and Tissue culture
T-DNA
Biolistic techniques
CISGENESIS/INTRAGENESIS
NEW PLANT BREEDING TECHNIQUES
• difference among gm clones.
Same symptoms
(PNAS, 20 gennaio 2004, vol. 101, n.3, fig.3)
Papayas are grown in many tropical countries. But papaya cultivation is being threatened by Papaya Ringspot-Virus,
a disease that is sharply lowering yields.
Since 1998, genetically modified papayas have
been cultivated in Hawaii, USA. They are resistant to a domestic viral disease.
In the EU, genetically modified papayas are not
approved.
Transgenic VS Non transgenic Papaya
SEQUENCE
HOMOLOGOUS TO THE
PROMOTER
SMALL DOUBLE
STRANDED RNA
Plant cell
Plant
genome
RdDM plants
x
SILENCED
GENE
RdDM: RNA-DEPENDENT
DNA METHYLATION
NEW PLANT BREEDING TECHNIQUES
VIRUS Resistance – an emergency in Prunus cultivation
Agrobacterium
tumefaciens
Transgenic
plantTransgenic
rootstock
Non-
Transgenic
scion
Non-
Transgenic
fruits
GRAFTING ON GM ROOTSTOCK
NEW PLANT BREEDING TECHNIQUES
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Assessment of risks and benefits
Assessment of benefits and risk
Why we need the open field trials with GM plants:
Which risks: environment?, Human Health?, the agricultural economy????
Which benefits: increased production efficiency and quality
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‘SU BASE SCIENTIFICA’
•Rischio Salute Uomo •Rischio Ambiente
•Rischio sistemi agricoli
•‘CASO PER CASO’(Gene-Specie-Pianta)
The European Commission adopted Directive 2001/18/EC (repealing Directive 90/220/EEC) to govern the deliberate release of GMOs into the
environment.
APPLICARE LE DIRETTIVE COMUNITARIEE LE L.G. EFSA
‘DA METODO A PRODOTTO’
‘DA METODO A PRODOTTO’
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TRANSGENIC TRIAL
PLANNING A GMO FIELD EXPERIMENTAL TRIAL
10 m border with trap plant
10 m non cultivated Border
Protection nets:
One with holes of 5 x 5 cm max, 180 cm high; and
one with holes 1,5 x 1,5 cm and 90 cm high,
but half under soil (40 cm).
A Screen House net type of
protection can be requested depending to
the type of crop.
TO CREATE A NET OF RESEARCH ON OPEN FIELD GMO RISK AND BENEFITS EVALUATION
Notification Costs for Public research:Italy 1500 Euro
The Netherlands, Poland, Spain = 0
Strawberry Grape