NeMo Looking at Root of Nematode Molecular Interactions - Cra-Abp

NeMo
Looking at Root of Nematode Molecular Interactions
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20° NeMo of Activities
2014- Plant-Nematode Interaction
2004- Development of nematode cryostorage
1994 Molecular Diagnosis
1994-
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NeMo: surveys and collection
FAIR UE-Program 1996-2000 «Pest Risk Analysis of Pine Wood
N
Nematode-related
t d l t dB
Bursaphelenchus
h l h species
i iin view
i off South
S th E
European
pine wilting and wood imports from Asia»
Fungi and nematode culture
NeMo: Molecular identification by ITS-PCR & ITS-RFLP
- Ambrogioni, Irdani T. and S. Caroppo
Redia, LXXXVI 2003:139-146.
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NeMo: PREVENTO e CRYO-UP project 2012-2014
CRYOPRESERVATION OF BIOLOGICAL COLLECTION
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CRYOPRESERVATION UNIT
A COMFORTABLE
C MF R BLE METHOD
ME H D FOR
F R THE
HE OPERATORS
PER
R
MORE HANDY EQUIPMENTS
TEMPERATURE UNIFORMITY COMPARED TO LN2
LESSENED RISK OF LEAKING VIALS
NeMo: CRYOPRESERVATION SLOW-FREEZING vs. VITRIFICATION
The technology is based on the induction of “cell vitrification” during a decrease
of temperature.
temperature
«VITRIFICATION»
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VITRIFICATION
A technique for freezing: microorganisms and
cells
to preserve for use at a later date.
A wide range of cells of biological
biological, medical and agricultural importance can be
cryopreserved at ultra-low temperatures below -196°C, in liquid nitrogen
DRAWBACKS
Cp toxicity
Varied and specialized skills required
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NeMo: CRYO-UP project 2012-2014
SOFT VITRIFICATION TECHNIQUE: Low CP & UltraRapidCooling
Simple and unique-procedure
unicellular
multicellular
A UNIQUE CRYOPRESERVATION METHOD FOR SEVERAL GENERA: FUNCTIONALITY AND PROSPECTS
Irdani T. et al., Low-Temperature Society London 2014, personal communication.
CRYOPRESERVATION of PLANT-PARASITIC NEMATODES
SEVERAL GENERA HAVE BEEN
Bursaphelenchus eremus
ALREADY CRYOPRESERVED
WITH THIS UNIQUE
PROCEDURE
Meloidogyne incognita
Globodera tabacum
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STEROPLANTS
Studi molecolari riguardanti la difesa delle piante dall’attacco di
nematodi su piante modello,
modello «non
non alimentari
alimentari» come il tabacco e la specie selvatica
S. torvum
NEMAMEL
NeMo: plant defence programs
Le piante non possono muoversi, perciò tutto quello che
possono fare per difendersi è sintetizzare
PROTEINE
Proteine che aiutano la pianta a reagire contro
«l’invasore»: proteine di rinforzo della parete cellulare e
fitoalessine
- una risposta di ipersensibilità (HR), che porta al
«suicidio» della cellula invasa, evitando la progressione
dell infezione.
dell’infezione
Quando però l’invasore elude le barriere e penetra…
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NeMo: nematode-infection
The infective-stage juveniles (J2), hatch from eggs, penetrate the root tip and move
te ce u a y, caus
causing
g minimum
u da
damage,
age, to reach
eac tthe
e vascular
ascu a e
element
e e t where
e e tthey
ey
intercellularly,
establish elaborate feeding sites known as giant cells
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NeMo lab: plant-parasitic development
5 dpi (days after infection)
Specialized cells, multinucleate that provide a
source of nutrients for the nematode
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NeMo:STEROPLANTS 2008-2011
plant & nematodes in vitro
gall
STEROPLANTS 2008-2011
Steroid receptor, plant-development & nematode
infection.
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NEMAMEL 2008-2011 coordinato da CRA-ORL
Root-knot nematodes (RKN, Meloidogyne spp.) were employed in infection experiments and global
transcript profiling by microarrays analysis were performed.
S. torvum
Resistant plants
NemR genes:
Chiti
Chitinases
sesquiterpenoids
Solanum torvum responses to the root-knot nematode Meloidogyne incognita
Bagnaresi P,
P Sala T,
T Irdani T,
T Scotto C,
C Lamontanara A,
A Beretta M,
M Rotino GL,
GL Sestili S,
S Cattivelli L and Sabatini E.
E BMC genomics
14, 540 (2013)
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NeMo: proteins & plant-response
Dagli studi molecolari informazioni utili su:
-Quali le proteine essenziali?
Proteine di difesa: R-genes come Mi-1 (NB-LRR protein);
Expansine: EXP5
Enzimi: chitinase
-Quali procedure alternative di difesa mettere in campo?
Vegetali «vaccinati» ? con siRNA
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Ringrazio tutti voi per l’attenzione
e
Tutti i partecipanti ai diversi progetti
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