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67 results on '"Lo Piccolo, S."'

54. Mango pathogens in Sicily

Author

LO PICCOLO, Sandra, MONDELLO, Vincenzo, TORTA, Livio, BURRUANO, Santa, LO PICCOLO S, MONDELLO V, TORTA L, and BURRUANO S

Published
2005

62. Evoluzione morfologica di Albugo candida in Eruca sativa

Author

BURRUANO, Santa, CONIGLIARO, Gaetano, LO PICCOLO, Sandra, TORTA, Livio, Burruano, S, Conigliaro, G, Lo Piccolo, S, and Torta, L

Subjects
Albugo candida, patogeno, eruca sativa, Albugo candida, Eruca sativa, morfologia
Published
2002

66. An Optimized and Rapid DNA Extraction Method From Leaves of Grapevine Suitable for PCR-DGGE Based Analysis

Author

Nicola Francesca, Gaetano Conigliaro, S. Burruano, Luca Settanni, Giancarlo Moschetti, Sandra Lo Piccolo, Lo Piccolo, S, Conigliaro, G, Francesca, N, Settanni, L, Burruano, S, and Moschetti, G

Subjects
Bioengineering, Biology, Applied Microbiology and Biotechnology, High-performance liquid chromatography, Anisodamine, law.invention, chemistry.chemical_compound, Laboratory flask, Erlenmeyer flask, law, Bioreactor, DNA extraction, PCR-DGGE, Chromatography, Inoculation, Settore AGR/12 - Patologia Vegetale, CTAB, General Medicine, polyphenol, chemistry, Polyphenol, polysaccharide, Vitis vinifera, Aeration, Settore AGR/16 - Microbiologia Agraria, Biotechnology
Abstract

Molecular techniques and genetic studies require a fair amount of DNA of high quality in order to produce reliable and clear results (Li et al., 2007). In grapevine, the yield and quality of DNA can be significantly affected by secondary metabolites such as polyphenols, polysaccharides and tannins, that may be consistently abundant during the different stages of leaf development (Iandolino et al., 2004). The development of a simple, rapid and reliable method for the extraction of genomic DNA from grape leaves collected at several stages during development, as well as from healthy grape leaves and leaves infected by pathogenic microorganisms, was the main objective of this study. The protocol reported, based on a modified cetyltrimethylammonium bromide (CTAB) extraction procedure (Doyle and Doyle, 1990), allowed the rapid DNA extraction from little amounts of leaf material without employment of liquid nitrogen for initial tissue grinding. The protocol included polyvinylpyrrolidone to bind phenolic compounds, β-mercaptoethanol to inhibit the oxidation of polyphenols, and a high concentration of NaCl (2.5 M) to increase the solubility of polysaccharides, thus, reducing their co-precipitation with DNA. Final DNA solution did not contain polysaccharides, polyphenols and other major contaminants. The quality of the DNA extracted with the above protocol was first evaluated for its accessibility for polymerase chain reaction (PCR); genomic DNA from healthy and Plasmopara viticola infected leaves of Vitis vinifera was successfully amplified by traditional PCR. Subsequently, the same DNA solution was used for amplification with a primer pair specific for PCR-denaturing gradient gel electrophoresis (DGGE), containing a GC-clamp at the 5′-end of the forward oligonucleotide. PCR produced amplicons with the expected molecular size (200 bp) suitable to be analyzed by means of a denaturing containing polyacrilamide gel for the bacterial ecology composition.

Published
2010
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67. Lasiolactols A and B Produced by the Grapevine Fungal Pathogen Lasiodiplodia mediterranea.

Author

Andolfi A, Basso S, Giambra S, Conigliaro G, Lo Piccolo S, Alves A, and Burruano S

Subjects
Ascomycota pathogenicity, Methanol chemistry, Methanol metabolism, Spectrophotometry methods, Ascomycota metabolism, Methanol analogs & derivatives, Vitis microbiology
Abstract

A strain of Lasiodiplodia mediterranea, a fungus associated with grapevine decline in Sicily, produced several metabolites in liquid medium. Two new dimeric γ-lactols, lasiolactols A and B (1 and 2), were characterized as (2S*,3S*,4R*,5R*,2'S*,3'S*,4'R*,5'R*)- and (2R*,3S*,4R*,5R*,2'R*,3'S*,4'R*,5'R*)-(5-(4-hydroxymethyl-3,5-dimethyl-tetrahydro-furan-2-yloxy)-2,4-dimethyl-tetrahydro-furan-3-yl]-methanols by IR, 1D- and 2D-NMR, and HR-ESI-MS. Other four metabolites were identified as botryosphaeriodiplodin, (5R)-5-hydroxylasiodiplodin, (-)-(1R,2R)-jasmonic acid, and (-)-(3S,4R,5R)-4-hydroxymethyl-3,5-dimethyldihydro-2-furanone (3 - 6, resp.). The absolute configuration (R) at hydroxylated secondary C-atom C(7) was also established for compound 3. The compounds 1 - 3, 5, and 6, tested for their phytotoxic activities to grapevine cv. Inzolia leaves at different concentrations (0.125, 0.25, 0.5, and 1 mg/ml) were phytotoxic and compound 5 showed the highest toxicity. All metabolites did not show in vitro antifungal activity against four plant pathogens., (© 2016 Verlag Helvetica Chimica Acta AG, Zürich.)

Published
2016
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