Subcellular changes caused by late leaf rust in raspberries and the effect of water deficit on disease severity: confocal and transmission electron microscopy analyses

• Grant number: 24/02834-0
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: April 01, 2024
• End date: June 29, 2024
• Field of knowledge: Biological Sciences - Botany - Applied Botany
• Principal Investigator: Beatriz Appezzato da Glória
• Grantee: Lucas Henrique Santos Barbosa
• Supervisor: Tonni Grube Andersen
• Host Institution: Escola Superior de Agricultura Luiz de Queiroz (ESALQ). Universidade de São Paulo (USP). Piracicaba , SP, Brazil
• Institution abroad: Max Planck Society, Munich, Germany
• Associated to the scholarship: 22/00888-0 - "Can abiotic stresses alter plant-pathogen relationships in the Thekopsora americana - Rubus idaeus pathosystem? Epidemiological, physiological and histopathological analyses", BP.DR

Abstract

The BEPE project is intended to improve ongoing experiments, by incorporating microscopy techniques. The aim is to clarify plastid changes in raspberry leaves colonized by Aculeastrum americanum and investigate whether abiotic stress (water limitation) alters the colonization of the pathogen. To achieve this, Raspberry plants (Rubus idaeus L.) cv. Heritage with and without water limitation will be inoculated with a suspension of A. americanum urediniospores, adjusted to a concentration of 5x104 urediniospores mL-1. Leaf samples, collected at 7 and 14 days after inoculation, will be processed for two analyses. For transmission electron microscopy, the samples will be fixed in 3% glutaraldehyde and cacodylate buffer (0.2 M pH 7.2) for 48 h, processed and the blocks containing samples will be sent to Max Planck Institute for Plant Breeding Research (MPIPZ) in advance via postal service. For confocal laser scanning microscopy, samples will be fixed in 0.15% trichloroacetic acid in 4:1 ethanol/chloroform. The samples will be sent to MPIPZ, where the WGA-AF488 and Propidium Iodide co-staining protocol will be applied and confocal laser scanning microscopy performed. These analyses will be important to complement my results obtained so far. The BEPE will be carried out at MPIPZ, Cologne-Germany under the supervision of Dr. Tonni Grube Andersen, a specialist in histological and molecular studies of plant-pathogen and plant-environment interactions, over a period of 4 months.