Poly(lactic-co-glycolic) acid (PLGA)-based NPs are currently considered among the most promising drug carriers, nevertheless their use in plants has never been investigated. Our study shows the ability of PLGA NPs to cross the plant cell wall and membrane both of in vitro Vitis vinifera cells and some grapevine-pathogenic fungi. Viability tests demonstrated that PLGA NPs were not cytotoxic for V. vinifera-cultured cells. By means of fluorescence microscopy and TEM analysis, we established that PLGA NPs ≤ 50 nm can enter in grapevine cells, while bigger ones remained attached to the cell wall. Fluorescence analysis suggested that PLGA NPs penetrate through endocytic vesicles but endocytosis in plants has been poorly studied. To date, the research has been mainly focused on clathrin-mediated endocytosis and only a limited amount of evidence on clathrin-independent pathways is available, partly because there are no specific markers for non-clathrin vesicles. Through the combined use of TEM and confocal analysis, fluorescent PLGA NPs loaded with coumarin-6, probe FM4-64 specific for clathrin vesicles, and clathrin endocytosis inhibitors, we demonstrated the involvement of clathrin-indipendent endocytosis in the internalization of PLGA NPs. Consequently, PLGA NPs are accumulated in the cytoplasm and not directed to the vacuole in which should be digested. Moreover, experiments on protoplasts highlighted the different selectivity of the cell wall and membrane on nanoparticle size. Moreover, we performed experiments in planta. PLGA NPs can enter into leaf tissues through stomata, in the roots through root hairs and transported to the shoot through vascular tissues. Uptake of PLGA NPs by Aspergillus spp and Botrytis cinerea cells has been also observed. The obtained results suggest that PLGA NPs could represent a novel strategy to deliver gradually agrochemicals in the prevention of damages caused by pathogenic fungi in agriculture.
Bio-compatible and environmental-friendly nanoparticles as a possible strategy to deliver agrochemicals in planta and in phytopathogenic fungi / Donati, Livia; Valletta, Alessio; Bramosanti, Marco; Chronopoulou, Laura; Palocci, Cleofe; Barbara, Baldan; Pasqua, Gabriella. - ELETTRONICO. - 1:(2016), pp. 52-52. (Intervento presentato al convegno Nanoinnovation 2016 Conference and Exhibition tenutosi a Roma nel 20-23 settembre 2016).
Bio-compatible and environmental-friendly nanoparticles as a possible strategy to deliver agrochemicals in planta and in phytopathogenic fungi
DONATI, LIVIA;VALLETTA, ALESSIO;BRAMOSANTI, MARCO;CHRONOPOULOU, LAURA;PALOCCI, Cleofe;PASQUA, Gabriella
2016
Abstract
Poly(lactic-co-glycolic) acid (PLGA)-based NPs are currently considered among the most promising drug carriers, nevertheless their use in plants has never been investigated. Our study shows the ability of PLGA NPs to cross the plant cell wall and membrane both of in vitro Vitis vinifera cells and some grapevine-pathogenic fungi. Viability tests demonstrated that PLGA NPs were not cytotoxic for V. vinifera-cultured cells. By means of fluorescence microscopy and TEM analysis, we established that PLGA NPs ≤ 50 nm can enter in grapevine cells, while bigger ones remained attached to the cell wall. Fluorescence analysis suggested that PLGA NPs penetrate through endocytic vesicles but endocytosis in plants has been poorly studied. To date, the research has been mainly focused on clathrin-mediated endocytosis and only a limited amount of evidence on clathrin-independent pathways is available, partly because there are no specific markers for non-clathrin vesicles. Through the combined use of TEM and confocal analysis, fluorescent PLGA NPs loaded with coumarin-6, probe FM4-64 specific for clathrin vesicles, and clathrin endocytosis inhibitors, we demonstrated the involvement of clathrin-indipendent endocytosis in the internalization of PLGA NPs. Consequently, PLGA NPs are accumulated in the cytoplasm and not directed to the vacuole in which should be digested. Moreover, experiments on protoplasts highlighted the different selectivity of the cell wall and membrane on nanoparticle size. Moreover, we performed experiments in planta. PLGA NPs can enter into leaf tissues through stomata, in the roots through root hairs and transported to the shoot through vascular tissues. Uptake of PLGA NPs by Aspergillus spp and Botrytis cinerea cells has been also observed. The obtained results suggest that PLGA NPs could represent a novel strategy to deliver gradually agrochemicals in the prevention of damages caused by pathogenic fungi in agriculture.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
