L'attività di ricerca attuale si concentra sullo studio e
ottimizzazione di metodi alternativi e non invasivi per
l'identificazione dei microrganismi associate con le piante
e per la diagnosi precoce delle fitopatie. Tra queste metodologie,
la ricerca si è focalizzata sulle tecniche basate sul vis-NIR e sul
monitoraggio dei composti volatili
(VOC) tramite l'utilizzo del naso elettronico (e-nose). In
particolare, lo studio dei VOC ha permesso di indagare anche il
loro possibile ruolo biologico nelle
interazioni tra pianta e microrganismi. Tra i VOC maggiormente
studiati ci sono stati alcune molecole segnale come l'etilene, il
metil-salicilato e l'acido giasmonico
che possono influenzare i meccanismi di difesa della pianta. Dal
2009-2010, la ricerca si è anche estesa allo studio delle
interazioni tra le specie di Actinidia (A.
deliciosa and A. chinensis) il batterio patogeno Pseudomonas
syringae pv actinidiae. Lo studio di tali interazioni è stato
condotto principalmente in planta utilizzando
batteri marcati con proteine auto fluorescenti e monitorando le
interazioni tra pianta e patogeno in tempo reale tramite
microscopia confocale e stero microscopia
fluorescenza.
Development of novel e-nose based diagnostic system
Summary: the research deals with the development of
alternative methods (electronic nose, near infrared spectroscopy
NIRs) for the diagnosis of plant disease and the identification of
plant-associated microorganisms. The most recent researches
investigate also the possible biological role of VOCs and the
airborne plant-to-pant or plant-microbe interactions.
Possible Thesis Duration: 6 months
Expertises/competences achieved during the research
thesis:electronic nose use and data-processing, basic
bacteriology, micropropagation, VOCs analysis and identification
ability, nucleic acid extraction as well as PCR detection of
bacterial species.
External collaborations:
Prof. S. Cristescu - Radboud University, Life Science
Trace Gas Facility, Heyendaalseweg 135, 6525 AJ, Nijmegen, The
Netherlands
Dr. J.L. Vanneste – Plant and Food, Ruakura Research Centre,
East Street Private Bag 3123, Hamilton, New Zealand
Develop of new methods for the real-time monitoring of
Pseuomona syringae pv. actinidiae directly in
plant.
Summary: The research aims to develop a method allowing
to monitor in vivo the plant-bacteria interactions during the
infection process. These non-destructive methods will allow the
direct visualization of the spatio-temporal interactions of P.
syringae pv actnidia (PSA) in intact Actinidia chinensis and A.
deliciosa tissues.
To reach this objective, a stable and broad host- range plasmid
vector, pDSK-GFPuv, which expresses GFPuv (driven by a constitutive
chloroplast promoter, psbA) at high levels will be inserted in
competent PSA cells.
The GFPuv-labeled plant pathogenic bacteria not only can be
easily visualized at the cellular level under a fluorescence
microscope but also are clearly visible, as bacterial colonies, to
the naked eye at the whole-plant level under longwavelength UV
light.
The plant-pathogen interaction will be successively studied be
means of fluorescent steromicroscope and by confocal laser scanning
microscopy.
Once the protocols have been optimized for this pathosystem,
they will allow to clarify crucial aspects of the infection process
such as:
-
how the bacteria invade the plant tissues
-
how far the bacteria spread inside the plant tissues
-
how the plant defenses can contain the bacteria spread
The knowledge of this aspects will help in developing the most
appropriate cultural strategies to contain the disease and to
evaluate the most effective treatments to reduce the infection
incidence and severity.
Finally, these methods may also be applied for Determination of
temperature thresholds for PSA infection.
Possible Thesis Duration: according with the specific
subject at least 3 days per week for 6 months.
Expertises/competences achieved during the research
thesis:nucleic acid extraction, bacterial transformation,
microscopy techniques, diagnostic methods
External collaborations: University of Siena, Department
of BioPhysics-Univeresity of Bologna, Plant&Food Research (New
Zealand).