Effectors as a new technology for the development of innovative bioinputs for crop protection

• Grant number: 24/15766-3
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: July 01, 2025
• End date: March 31, 2026
• Field of knowledge: Agronomical Sciences - Agronomy - Plant Health
• Principal Investigator: Diogo Maciel de Magalhães
• Grantee: Diogo Maciel de Magalhães
• Company: Ideelab Biotecnologia Ltda
• CNAE: Pesquisa e desenvolvimento experimental em ciências físicas e naturais
• City: Piracicaba
• Associated researchers: Sérgio Florentino Pascholati

Abstract

Effectors play essential roles in determining the outcome of interactions between plants and microorganisms. Considered the main virulence proteins of pathogenic microorganisms, they are indispensable for the colonization of host cells. During the co-evolution process with microorganisms, plants have developed receptors capable of recognizing these molecules and activating an immune system pathway known as Effector-Triggered Immunity (ETI), resulting in long-lasting and efficient disease control responses. Effector molecules from different microorganisms have been explored to induce resistance (IR) against phytopathogens and increase tolerance to environmental stresses. Using bioinformatics tools and a pipeline for effector prediction, we have identified numerous candidate effector proteins to be explored. In addition to these proteins, another class of molecules that has garnered interest for stimulating defense responses in plants are rhamnolipids (RLs), biodegradable surfactant glycolipids primarily produced by Pseudomonas and Burkholderia species. Mainly employed in soil bioremediation and the pharmaceutical and cosmetics industries, they have recently been explored in agriculture due to their bioprotective effect and antimicrobial properties, acting as activators of the ETI immune system. Considering the high IR potential of effector molecules and the agricultural market demand for natural, effective, and innovative products, the overall objective of this proposal is to develop a platform for the development of products based on microbial effectors, aiming to increase plant resistance against biotic and abiotic stresses. In this first stage, the goal is to obtain proteins and RLs with elicitor activity in plant defense. To achieve this, from the available list of effector candidates, a more in-depth characterization and selection will be carried out to select three leads to be used in a heterologous expression system. The recombinant proteins obtained will have their elicitor activity evaluated in vitro through ROS detection in leaf discs in the presence of the actives. RL molecules will also be evaluated for elicitor activity, both individually and in combination with recombinant proteins. In phase 2, recombinant proteins validated for in vitro elicitor activity will be produced from the fermentation of recombinant strains in bench and pilot bioreactors, with capacities of 10 L and 100 L, respectively. After adjusting the best production conditions, the potential effectors based on recombinant proteins and RLs, in optimized formulations, will be applied to plants in the presence of pathogens to evaluate IR under controlled greenhouse conditions and, subsequently, in the field. This is a technology that is still underexplored, and its development could have a significant impact, not only on the company's entry into this market but also on the overall market for biological agricultural products, representing a disruptive innovation in the sector and contributing to the reduction of agrochemical use, which causes significant environmental and human health impacts. (AU)