Microfluidic synthesis of methyl jasmonate-loaded PLGA nanocarriers as a new strategy to improve natural defenses in Vitis vinifera

The objective of the present work was to synthesize biopolymeric nanoparticles (NPs) entrapping the resistance-inductor methyl jasmonate (MeJA) to be employed as a novel and alternative strategy in integrated pest management. NPs were prepared by using a continuous flow microfluidic reactor that allows to precisely control some features that are crucial for applications such as size, polydispersion, morphology and reproducibility. Poly(lactic-co-glycolic acid) (PLGA), a biopolymer largely studied for its use in biological applications, was chosen for the production of NPs entrapping MeJA, a biotic endogenous elicitor able to trigger plant’s defense responses. The effect of different fluid-dynamic conditions, PLGA molecular weight and concentration on NP properties (dimensions, polydispersion, morphology, stability) was evaluated. DLS and SEM were employed to characterize the obtained NPs. MeJA-loaded PLGA NPs ranging from 40 to 70 nm were administered to Vitis vinifera cell cultures, in order to evaluate the biological response in terms of stilbene biosynthesis. HPLC investigations showed a faster response when the elicitor was administered by PLGA NPs in comparison with free MeJA. This result demonstrates that the encapsulation in PLGA NPs significantly promotes MeJA cell uptake and the activation of MeJA-induced responses.

The role of endogenous elicitor peptides in plant resistance to biotic stress

Plant elicitor peptides (Peps) are one class of elicitor substances, which are formed in plant cells in response to various biotic stressors and induced of nonspecific plant resistance. They are present and active in angiosperms, including many important agricultural crops, and can be considered as a promising class of compounds for creating environmentally safe drugs that induce phytoimmunity and increase the resistance of plants to stress. In this paper, an analysis of current literature data on the functional activity of endogenous plant elicitor peptides, the mechanisms of Pep-signaling and their role in plant resistance to biotic stresses is carried out.

Danger-associated peptide signaling in Arabidopsis requires clathrin

The Arabidopsis thaliana endogenous elicitor peptides (AtPeps) are released into the apoplast after cellular damage caused by pathogens or wounding to induce innate immunity by direct binding to the membrane-localized leucine-rich repeat receptor kinases, PEP RECEPTOR1 (PEPR1) and PEPR2. Although the PEPR-mediated signaling components and responses have been studied extensively, the contributions of the subcellular localization and dynamics of the active PEPRs remain largely unknown. We used live-cell imaging of the fluorescently labeled and bioactive pep1 to visualize the intracellular behavior of the PEPRs in the Arabidopsis root meristem. We found that AtPep1 decorated the plasma membrane (PM) in a receptor-dependent manner and cointernalized with PEPRs. Trafficking of the AtPep1-PEPR1 complexes to the vacuole required neither the trans-Golgi network/early endosome (TGN/EE)-localized vacuolar H+-ATPase activity nor the function of the brefeldin A-sensitive ADP-ribosylation factor-guanine exchange factors (ARF-GEFs). In addition, AtPep1 and different TGN/EE markers colocalized only rarely, implying that the intracellular route of this receptor–ligand pair is largely independent of the TGN/EE. Inducible overexpression of the Arabidopsis clathrin coat disassembly factor, Auxilin2, which inhibits clathrin-mediated endocytosis (CME), impaired the AtPep1-PEPR1 internalization and compromised AtPep1-mediated responses. Our results show that clathrin function at the PM is required to induce plant defense responses, likely through CME of cell surface-located signaling components.