25 mM NaCl +EM - Test sulla sicurezza

University of Padova, Department of Agronomy, Food, Natural Resources,
Animal and Environment
Biostimolanti, corroboranti, induttori di resistenza e micronutrienti nelle
colture ortoflorofrutticole
29-30 maggio 2014, Ancona
Giornate tecniche SOI
Verso l’EXPO 2015
" Biostimolanti:
nuovo scenario per il futuro dell'agricoltura" .
Prof.ssa Serenella Nardi
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
The European Biostimulants industry Council
ha definito i biostimolanti come:
“Plant biostimulants contain substance(s) and/or micro-organisms whose function when applied to
plants or the rhizosphere is to stimulate natural processes to enhance/benefit nutrient uptake, nutrient
efficiency, tolerance to abiotic stress, and crop quality. Biostimulants have no direct action against
pests, and therefore do not fall within the regulatory framework of pesticides” .
Finally, the EBIC concept of biostimulants includes products with some nutrients, provided that the
effect on plant growth is not through direct fertilization: “Biostimulants operate through different
mechanisms than fertilisers, regardless of the presence of nutrients in the products” (EBIC 2012).
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
in Europe >6.2 million hectars were treated with biostimulants in 2012
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
The First World Congress on Biostimulants in Agriculture
26-29 November, 2012, Strasbourg Congress Center, France
The First World Congress on the use of Biostimulants in Agriculture was an international
scientific and technical conference to review the latest knowledge on these products, which are
increasingly used in crop production around the world. A panel of prestigious international
speakers discussed the impact of Biostimulants on Plant Nutrition, Abiotic Stresses, Plant
Disease Responses, Plant Growth and Development, as well as the various aspects of legislation
on these products in the main markets, with a special focus on Europe.
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
I biostimolanti si suddividono:
1. Inoculanti microbici,
2. Sostanze umiche,
3. Idrolizzati proteici ed aminoacidi,
4. Prodotti a base di alghe.
produzione Ha
peso secco
germinazione
peso fresco
2. SOSTANZE UMICHE
crescita radicale
crescita fogliare
Attività BIOLOGICA
Effetti diretti ed indiretti
 Crescita ( dimensione e sviluppo)
Effetti sulla morfologia
Canellas et al., 2008
Piante di mais cresciute per 6 giorni in presenza o assenza di 5 mg C org/l.
Schmidt et al, Plant Soil (2007)
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
Control
Treated plant
Concheri, G., Nardi, S., Reniero, F., Dell'Agnola, G., 1996. The effects of humic
substances within the Ah horizon of a Calcic Luvisol on morphological changes related
to invertase and peroxidase activities in wheat roots. Plant and Soil 179, 65-72.
r
ELISA immuno-enzimatic test by
monoclonal antibodies
Radio-immunoassay (RIA) in fluid phase
by polyclonal antibodies
HS from earthworm faeces 0.3 – 0.5%
HS from forest soils
Antibodies
Muscolo, A., Cutrupi, S., Nardi, S., 1998. IAA detection in humic
substances. Soil Biology & Biochemistry 30, 1199-1201.
D., Nicolini, G., Nardi, S., 2001. Hormone-like activity of humic
0.0045 – 0.0363% Pizzeghello,
substances in Fagus sylvaticae forests. New Phytologist 151, 647-657.
GC-MS
GAS
CHROMATOGRAM of the
standard methylated IAA (A) and
the gas chromatogram of the
methylated HA isolated from
earthworm compost
Canellas, L.P., Olivares, F.L., Okorokova-Facanha, A.L., Facanha, A.R., 2002. Humic acids isolated from earthworm compost enhance root
elongation, lateral root emergence, and plasma membrane H+-ATPase activity in maize roots. Plant Physiol. 130, 1951-1957.
INTRODUCTION
Nardi et al., 1996. Humic substances in terrestrial ecosystems
Photographs of leaf explants of Nicotiana plumbaginifolia treated with HS and hormones
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
IAA1 auxin-responsive protein
Response of Arabidopsis marker genes IAA-responsives
H2 O
IAA1
IAA11
Act2
TQ
IAA
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
HS Biological Activity
Fisiologia e metabolismo
Nutrizione / Trasporto dei nutrienti (macro e microelementi)
Induzione del trasporto del nitrato / attività H+-ATPasi
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
Effetti di stimolazione……
Trasporto dei nutrienti
• sintesi proteica
•Dell’Agnola and Ferrari, 1971
•Vaughan et al., 1985
• Nardi et al., 2000
• Cinetica di trasporto
• Cacco et al., 2000
• Maggioni et al., 1987;
•
attività-ATPasica
• Nardi et al., 1991;
• Pinton et al., 1992
• Varanini et al., 1993
• attività H+-ATPasica del plasmalemma e
del tonoplasto
• Pinton et al., 1997
• Pinton et al., 1999
• Canellas et al., 2002
HS Biological Activity
(Ion uptake : NO3 – )
influx
andH+-ATPase
mRNA
NO33-– uptake
Westernblotand
analysis
(H+synthesis
of maize
H+-ATPase
activity
measured
in PM
ATPase
PMA2
antibodies)
Mha2 and
NO3- transporters
vescles
isolated
from roots
(Zea mays)
Pinton et al., 1999
Canellas et al., 2002
Quaggiotti et al., 2004
DAFNAE -Department of Agronomy, Food,
Natural Resources, Animals and Environment
Università di Padova - Italy
Humic Substances biological
activity:
a proteomic approach
Buchanan B.B., Gruissem W., Jones R.L. (2001).
Biochemistry & Molecular Biology of Plants. American
Society of Plant Physiologists
The aim of this study was to establish which
proteins were differentially expressed after
exposure to HS in PM-enriched root extracts.
The plasma membrane represents the site for the exchange of information and substances
between the cell and its environment.
Carletti, P., Masi, A., Spolaore, B., De Laureto, P.P., De Zorzi, M., Turetta, L., Ferretti, M., Nardi, S., 2008. Protein expression changes in
maize roots in response to humic substances. Journal of Chemical Ecology 34, 804-818.
7-8 November 2012
NUDIBA ANNUAL MEETING
16/Slides
cDNA-AFLP
T0
TRANSCRIPTOMICS
Sequencing
C30 HS30 C2h HS2h
Isolation and
purification
Bioinformatic analyses
Selection of the
differentially expressed
transcripts
Identification of the HS
biological targets
Gene Ontology
Analisys
Cellular Components
Biological processes
Molecular Functions
Mora V., Bacaicoa E., Baigorri R., Zamarreno
A., Garcia-Mina J.M. 2013. NO and IAA key
regulators in the shoot growth promoting action
of humic acid in cucumis sativus. J. Plant
Growth Regul.
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
I biostimolanti si suddividono:
1. Inoculanti microbici,
2. Sostanze umiche,
3. Idrolizzati proteici ed aminoacidi,
4. Prodotti a base di alghe.
•
•
•
•
•
•
•
•
•
•
Corte et al., 2014 J. Sci Food. Agric.
Kauffman et al., 2007. Crop Sci.
Sharma et al., 2006. J. Ex. Bot.
Zhang and Ervin, 2004. Crop Sci.
Zhang and Ervin, 2008. Crop Sci.
Quartieri et al., 2002. Acta Hort
Mladenova et al., 1998. Agro Food Industry Hi-Tech
Cavani et al., 2006. J. Agric. Foof Chem.
Jannin et al., 2013. J. Plant Regul.
Calvo P. et al., 2014. Agricultural uses of plant biostimulants. Plant Soil
Marschner Review
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
Piante di erba medica =EM=
ILSACON
L’idrolizzato proteico è stato prodotto
con
un
una
idrolisi
enzimatica
(FCEH®) usando materiale vegetale.
Schiavon et al., (2008) J. Agric. Food Chem. 56 ,
11800-11808.
CONTROLLO
a
b
c
Ertani et al., 2013 J. Agr. Food Chem.
a
b
c
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
Schiavon et al., (2008) J. Agric. Food Chem. 56 , 11800-11808.
Effetto del trattamento con EM sul peso secco di radici e foglie di plantule di mais
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
Schiavon et al., (2008) J. Agric. Food Chem. 56 , 11800-11808.
Effetto del trattamento con EM sul contenuto di nitrato ed azoto in radici e foglie di plantule
di mais.
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
NH4+
*FROM
KREBS CYCLE
NR
NO2-
NiR
NO3-
University of Padova, Department of Agronomy, Food, Natural Resources, Animal and Environment
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
ROOT
+27%
+39%
Schiavon et al., (2008) J. Agric. Food Chem. 56 , 11800-11808.
+10%
+19%
Root enzyme activity of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT), malate dehydrogenase
(MDH), isocitrate dehydrogenase (IDH), citrate synthase (CS), aspartate aminotransferase (AspAT) in Z. mays plants grown for 12 d in Hoagland
modified complete nutrient solution and treated for 2 d with EM at 0.01 or 0.1 mg L-1. Data are the means of 5 values each from three independent
experiments (±SE). Different letters on bars indicate significant differences between treatments (P<0.05). C = control
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
LEAF
+78%
+24%
Schiavon et al., (2008) J. Agric. Food Chem. 56 , 11800-11808.
+24%
+43%
Leaf enzyme activity of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT), malate
dehydrogenase (MDH), isocitrate dehydrogenase (IDH), citrate synthase (CS), aspartate aminotransferase (AspAT) in Z. mays plants grown for 12 d in
Hoagland modified complete nutrient solution and treated for 2 d with EM at 0.01 or 0.1 mg L-1.
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
Schiavon et al., (2008) J. Agric. Food Chem. 56 , 11800-11808.
Gene expression (mRNA level) and relative transcript accumulation of nitrate reductase (NR), malate dehydrogenase (MDH), isocitrate
dehydrogenase (IDH), citrate synthase (CS), asparagine synthetase (AS) in roots of Z. mays plants grown for 12 d in Hoagland modified
complete nutrient solution and treated for 2 d with EM at 0.01 or 0.1 mg L-1.
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
Schiavon et al., (2008) J. Agric. Food Chem. 56 , 11800-11808.
Gene expression (mRNA level) and relative transcript accumulation of nitrate reductase (NR), malate dehydrogenase (MDH),
isocitrate dehydrogenase (IDH), citrate synthase (CS), asparagine synthetase (AS) transcript accumulation in leaves of Z. mays plants
grown for 12 d in Hoagland modified complete nutrient solution and treated for 2 d with EM at 0.01 or 0.1 mg L-1.
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
EM= ILSACON
Control
EM
25 mM NaCl
25 mM NaCl +EM
75 mM NaCl
75 mM NaCl +EM
150 mM NaCl
150 mM NaCl +EM
ERTANI A., Schiavon M., Muscolo A., Nardi S. (2013). Alfalfa plant-derived biostimulant stimulate short-term growth of salt
stressed Zea mays L. plants. PLANT AND SOIL. Vol. 364, pp. 145-158
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
Treatment
GS
GOGAT
(µmol -1 g -1 fw)
Control
4.27±0.12b 14.46±0.32c
EM
6.11±0.32a 16.82±0.18a
25 mM NaCl
3.13±0.10c 12.50±0.11d
25 mM NaCl +EM
5.50±0.17a 14.90±0.23c
75 mM NaCl
3.81±0.11c 11.90±0.20e
75 mM NaCl +EM
5.00±0.28b 15.52±0.17b
150 mM NaCl
3.72±0.11c 12.57±0.35d
150 mM NaCl +EM 4.75±0.30b 14.78±0.33c
ERTANI A., Schiavon M., Muscolo A., Nardi S. (2013). Alfalfa plant-derived biostimulant stimulate short-term growth of salt
stressed Zea mays L. plants. PLANT AND SOIL. Vol. 364, pp. 145-158
PAL activity
Treatment
PAL
(nmol cinn acid mg-1 prot min-1)
Control
3.94± 0.31 f
EM
3.04± 0.29 f
25 mM NaCl
8.03± 0.20 e
25 mM NaCl +EM
12.92± 0.15 c
75 mM NaCl
12.34± 0.09 d
75 mM NaCl +EM
15.27±0.22 b
150 mM NaCl
12.69±0.18 d
150 mM NaCl +EM
17.47± 0.33a
ERTANI A., Schiavon M., Muscolo A., Nardi S. (2013). Alfalfa plant-derived biostimulant stimulate short-term growth of salt
stressed Zea mays L. plants. PLANT AND SOIL. Vol. 364, pp. 145-158
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
ERTANI A., Schiavon M., Muscolo A., Nardi S. (2013). Alfalfa plant-derived biostimulant stimulate short-term growth of salt
stressed Zea mays L. plants. PLANT AND SOIL. Vol. 364, pp. 145-158
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
Landlab Studio Associato, Quinto Vicentino, Vicenza
University of Padova, Department of Agronomy, Food, Natural Resources, Animal
and Environment
Grazie dell’attenzione!

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