GOAL 1: Enhancing drought tolerance through nanoparticle priming.

BACKGROUND Given the prevailing challenges, our principal aim is to enhance the drought resilience of crops in accordance with the principles of sustainable agriculture. A promising green technology for achieving this is priming, which fortifies plants against stress by inducing defense processes. Nanoparticles possess advantageous physical and chemical properties that can be harnessed to enhance the efficiency of priming. The administration of nitric oxide (NO) and its derived reactive forms as chemical donors or in the form of plasma-activated liquids (PAL), along with smoke-derived karrikins (KAR), has been shown to have a stress-tolerance enhancing effect.

QUESTION The main question in this practical, interdisciplinary work is whether the newly developed nano NO donors, nano KAR, and PAL stabilized with nanoparticles are more effective agents in enhancing drought tolerance during priming compared to their non-nano counterparts.

EXPERIMENTAL SYSTEM To answer this question, we conduct experiments in greenhouse model systems using a multi-level approach. We examine responses to drought at the cellular, organ, and organism levels and employ transcriptomics, proteomics, and metabolomics to characterize the stress-tolerance enhancing effects of nano materials.

Outline of the workflow
OUTLINE OF THE WORKFLOW (seed priming, PANC= plasma-activated nanocolloid, PAW=plasma-activated water, KAR2-CHT= chitosan-encapsulated karrikin2, KAR2=karrikin2, DMON-SNO= dendritic, mesoporous,organic silicon dioxide nanoparticle with S-nitrosothiol modification, GSNO=S-nitrosoglutathione, PAW-M= plasma-activated water supplemented with metal, pre-germination, control, drought/osmotic treatment, PEG-8000=polyethylene glycol-8000, water deprivation, decapping, plant analyses, drought tolerance parameters, root hair development, root system architecture, stomatal movements, transcriptomics, metabolomics, proteomics, root structure).

GOAL 2: Identification of Novel Nitrogen Monoxide-Linked Genes Associated with Osmotic Stress Response through Genetic Screening

BACKGROUND Previously, a collection of 40,000 transgenic Arabidopsis thaliana lines was generated, consisting of a cDNA library from a halophytic plant species, Lepidium crassifolium, known for its tolerance to salt, osmotic stress, and oxidative stress. These lines serve as the genetic screening material.

QUESTION Can we screen for transgenic lines that exhibit altered responses to NO compared to the wild type, and thereby, can we identify new NO-linked genes playing a role in the osmotic stress response?

EXPERIMENTAL SYSTEM We cultivate transgenic Arabidopsis thaliana plants in the presence of NO and observe their growth and developmental responses compared to the wild type. Promising candidates undergo detailed genetic and molecular biological investigations.

GOAL 3: Developing a new scientific website containing information related to plant NO.

BACKGROUND The project aims to support the research community by compiling scientific knowledge about plant NO on a dedicated website. Our hope is to present the information in a user-friendly and easily accessible manner on a single platform.

GOAL 4: Inducing plant fungal resistance by reactive nitrogen species liberating nanomaterials

BACKGROUND Fungal pathogens are among the dominant causal agents of plant diseases that threaten food safety across the world. Plant defence against pathogens can be triggered by the eco-friendly priming approach during which an external stimulus is perceived by the plant as a warning signal that leads to a moderate activation of induced defence mechanisms resulting in a faster and/or stronger defence response upon subsequent pathogen attack. The gaseous signal molecule, nitric oxide (NO) and its reaction products (collectively reactive nitrogen species, RNS) have been proven to act as a broad defence initiator to potentiate plants' defence mechanisms.

EXPERIMENTAL SYSTEM The priming effect of these novel RNS liberating nanoagents will be studied in model plant-fungus pathosystems such as Arabidopsis thaliana-Alternaria brassicicola and Solanum lycopersicum-Botrytis cinerea.