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Abstract
The focus of this research was the development and evaluation of different complex
liquid and solid media for the isolation and growth of phytoplasma strains infecting
grapevine plants. Previously reported media supporting phytoplasma isolation are commercial
and not easy to modify in order to improve performance and selectivity towards obtaining
pure cultures of 'Candidatus Phytoplasma' species. Three media (Piv®, CB and MB) were
therefore evaluated for phytoplasma isolation and colony formation under microaerophilic
growing conditions, using grapevine canes from plants showing yellows symptoms, and
infected by "flavescence dorée", "bois noir" and aster yellows phytoplasmas as sources.
The newly developed methodology was applied for two years at three sample collection
times. Broad applicability and a good repeatability in supporting phytoplasma colony
formation were obtained in Pivs® and CBs media. While the MB medium did not support
phytoplasma isolation and growth, the CB media support a phytoplasma growth comparable
to the one obtained in the previously reported media. This medium has the advantage
of a formulation that allow its modification to implement specificity towards selective
phytoplasma growth. Moreover preliminary trials on serial dilutions and tetracycline
addition confirmed some phytoplasma growth behaviours.