PROJECT LIFE12 ENV/IT/000336
“Anti-infective environmental friendly
molecules against plant pathogenic bacteria
for reducing Cu”
“AFTER Cu”
Start: 01/01/2014
End: 31/12/2015
Worshop Progetti LIFE
Dal confronto di esperienze diverse
lo sviluppo di tecniche sostenibili
Sala Centuria - RIT, via Granarolo, 62 – Faenza
2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
Copper is an essential microelement
for any living organisms
BUT…
Some nasty side effects of copper treatments
used in plant protection…
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
Here a list of the main nasty side effects of
copper treatments in plant protection:
 accumulation into the topsoil
 negative impact on soil biology
 negative impact on transformation/dynamic of nutrients
 secondary pollution in watercourses
 serious negative ecotoxicological effects
… last but not least
increase of copper- and antibiotic-resistant
bacteria in agroecosystems
following copper treatments
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
Antibiotics are allowed for the control of
bacterial diseases of plants in the USA!!!
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
Accordingly, recent restrictions have been established by EU
concerning the use of copper compounds in plant protection and
their maximum residue levels in food and feed






Directive 2009/37/EC
Regulation 396/2005/EC
Council Directive 91/414/EEC
Commission Regulation 149/2008/EC
Regulation 473/2002/EC
….
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
Other EU funded projects focused on reduction/replacement
of copper in plant disease control
PROJECT LIFE12 ENV/IT/000336
Other EU funded projects focused on reduction/replacement
of copper in plant disease control…
BUT
Just some alternatives
to copper fungicides
were designed or developed…
No substitutes for copper bactericides!
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
AFTER-CU project will aim at demonstrating
the use of innovative
anti-infective peptide-based molecules
for the environmentally friendly and sustainable
control of bacterial diseases of plants
Worshop Progetti LIFE
Faenza - 2015, January 22nd
BACTERIA
PLANTS
disease
DI
Courtesy of E.Colombi, 2012
«The AFTER Cu approach»
A
Courtesy of E.Colombi, 2012
PROJECT LIFE12 ENV/IT/000336
Host-Pathogen Interaction Model
Pseudomonas savastanoi
Pseudomonas savastanoi is the causal
agent of olive and oleander knot disease,
characterized by hyperplastic knots on
stems, branches, and occasionally on
leaves and fruits.
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
Type Three Secretion System
HrpA subunits
Buttner & Yang He (2009)
The EMBO Journal (2002) Vol.21 N.8
= HrpA
© 2014 Simone Giuntini
PROJECT LIFE12 ENV/IT/000336
Prove di Patogenicità
Piante in vitro di Oleandro
Knot21Formation
Giorni
on Nerium oleander
21 days
WT
Psn23
ΔhrpA
PF5aa
PF3aa
Worshop Progetti LIFE
Faenza - 2015, January 22nd
= HrpA
= Peptide
© 2014 Simone Giuntini
PROJECT LIFE12 ENV/IT/000336
PROJECT LIFE12 ENV/IT/000336
Peptide efficacy evaluation
Co-inoculation
with synthetic peptides
Knot Formation
on Nerium oleander
inoculated with synthetic
peptides 30µM
14 days
WT
Psn23
∆hrpA
Psn23+
Li27
Psn23 +
Ap17
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/00036
“Disarming” the plant pathogenic bacteria without killing them
using as a target conserved mechanisms for their pathogenicity
Advantages of AFTER Cu strategy
 Low risk of drug resistance emergence
 Active against copper- and antibiotic- resistant bacteria
 Broad spectrum strategy against Gram- plant pathogenic bacteria
 Easily transferable for the control of all Gram- pathogenic bacteria
 Low cost of biotechnological production
 Low environmental impact both in production and application
 ……
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/00036
Advantages of AFTER Cu strategy
To be demonstrated by


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The design and synthesis of anti-infective peptides
Evaluation of their in vitro and in vivo performances
Evaluation of their ecotoxicological profiles
Model plants used: olive, kiwi and citrus
Challenged with P. savastanoi, P. syringae pv. actinidiae, P.
syringae pv. syringae, respectively
in comparison with effects of traditional copper treatments
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/00036
AFTER Cu:
skills, expertise, technological advanced instruments
• Chemistry platform
• Molecular biology platform
• In planta screening platform
• Microbiological platform
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
AFTER Cu kick off meeting – 2014, January 10th
Laboratorio Patologia Vegetale Molecolare, DISPAA, Università degli Studi di Firenze,
Polo Scientifico e Tecnologico, Sesto Fiorentino (Firenze)
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/00036
AFTER Cu Associated beneficiary
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
AFTER Cu Associated beneficiary
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
AFTER Cu Associated beneficiaries
Worshop Progetti LIFE
Faenza - 2015, January 22nd
PROJECT LIFE12 ENV/IT/000336
AFTER Cu Coordinating beneficiary
Dept. of Chemistry
Dept. of Biology
Maria Rosa Moncelli
Patrizia Bogani
Dept. of Agrifood Production and Environmental Sciences
Carlo Viti
Stefano Biricolti
Stefania Tegli
Thank for your attention
from all
AFTER Cu people!
Courtesy of E.Colombi, 2012
La Resistenza agli Antibiotici
è il risultato di una classica
selezione darwiniana!
...la sopravvivenza e la moltiplicazione
del più “adatto”...
perchè l’intera popolazione
possa sopravvivere!
applicazione
antibiotico X o Cu
gene per la resistenza
all’antibiotico X o per Cu