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Anionic phospholipids connect barley ROP signaling with resistance and susceptibility to fungal cell entry

ABSTRACT: Rho-of-plant small GTPases (ROPs) are regulators of cell polar growth processes and of plant-pathogen interactions. The barley ROP protein RACB is involved in susceptibility towards infection by the barley powdery mildew fungus Blumeria hordei, but little is known about the cellular pathways that connect RACB-signaling to disease susceptibility. We now used an untargeted co immunoprecipitation-liquid chromatography-coupled mass spectrometry screening from Bh-infected epidermal layers of barley plants, which expressed green fluorescing protein-tagged constitutively activated (CA) RACB. This uncovered novel RACB interaction partners of plant and fungal origin. Three of those proteins, a plant phosphoinositide phosphatase, a plant phosphoinositide phospholipase and one putative Bh-effector protein appear to be involved in the barley-Bh-pathosystem and function in disease resistance or susceptibility, respectively. RACB and its plant interactors bind to overlapping anionic phospholipid species in vitro, and in case of RACB, this lipid-interaction is mediated by its carboxy-terminal polybasic region (PBR). Four diverse fluorescent plant anionic phospholipid-markers show altered subcellular distribution during Bh-attack. Phosphatidyl-4-phosphate and phosphatidylserine show a distinct enrichment at the haustorial neck region, suggesting a connection to subcellular targeting of RACB at this site. We suggest that the interplay of ROPs with anionic phospholipids and phospholipid–metabolizing enzymes may steer subcellular enrichment of components that are pivotal for fungal penetration success or failure.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Hordeum Vulgare (barley) Blumeria Hordei A6

TISSUE(S): Leaf Epidermis

SUBMITTER: Julia Mergner

LAB HEAD: Bernhard Kuster

PROVIDER: PXD056811 | Pride | 2024-12-12

REPOSITORIES: pride

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			Action	DRS
	02806_BA1_P031351_B00_A00_R1.raw	Raw
	02806_BA2_P031359_B00_A00_R1.raw	Raw
	02806_BA3_P031367_B00_A00_R1.raw	Raw
	02806_BA4_P031375_B00_A00_R1.raw	Raw
	02806_BA5_P031383_B00_A00_R1.raw	Raw

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Project description:The urgent need to address water scarcity underscores the importance of enhancing plant drought resistance. This study investigates whether pretreatment with abscisic acid (ABA) activates early stress signaling, thereby improving barley drought response when subsequently exposed to drought conditions. Although the individual responses to drought and ABA are well-documented, their synergistic effects in barley warrant further investigation. This study examines the impact of ABA on barley drought resilience through an experimental design that incorporates four distinct treatments: optimal watering, ABA application at 60 days post-sowing, and two drought stress treatments - one with and the other without prior ABA application. Key physiological parameters, such as photosynthesis, stomatal conductance and chlorophyll content, were analyzed in conjunction with transcriptomics. The results suggest that ABA pretreatment initiates early stomatal closure and elevates the expression of essential genes like NCED1, BG8, and HvA22, priming barley for improved drought resistance. During the drought, ABA-pre-treated barley maintained high chlorophyll levels, indicating sustained photosynthetic activity, a trend that persisted across treatments during the post-drought recovery phase. Furthermore, ABA pre-treatment was found to preserve photosystem II efficiency during drought conditions. Transcriptomic analyses revealed distinct gene expression profiles, alternative splicing profile and isoform switching, highlighting the molecular complexities of ABA role in drought response. These alterations span stress response, metabolic pathways, and DNA modification processes, providing a comprehensive view of ABA treatment's regulatory and metabolic impacts. In conclusion, ABA pretreatment strengthens barley drought defense by fostering stomatal closure and gene activation, guiding research strategies grounded in ABA and suggesting that genotypes with elevated ABA levels could have enhanced resilience and recovery capabilities.

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Anionic phospholipids connect barley ROP signaling with resistance and susceptibility to fungal cell entry

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