Validation of reference genes and differential gene expression by RT-qPCR in extremophile plants subjected to drought stress

• Grant number: 24/15249-9
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: November 29, 2024
• End date: May 28, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Helaine Carrer
• Grantee: Pedro Boscariol Ferreira
• Supervisor: Leon Aloys Bravo Ramirez
• Host Institution: Escola Superior de Agricultura Luiz de Queiroz (ESALQ). Universidade de São Paulo (USP). Piracicaba , SP, Brazil
• Institution abroad: Universidad de La Frontera (UFRO), Chile
• Associated to the scholarship: 22/09094-7 - Transcriptome analyses of desiccation-tolerant extremophytes to uncover targets of crop yield improvement, BP.PD

Abstract

Drought stress is one of the greatest challenges facing modern agriculture, particularly in regions where precipitation can determine crop productivity. Extremophile plants, which thrive in hostile environments, may serve as valuable sources of genetic information for stress tolerance. Our research has identified potential stress-related genes by comparing desiccation-tolerant species such as Strombocarpa tamarugo, Deschampsia antarctica, and Colobanthus quitensis (Antarctic ecotype) with phylogenetically related but less tolerant species. Biochemical validation of the expression differences in these genes is crucial to confirm results obtained from RNA-seq expression analyses, especially in non-model species. Thus, the aim of this proposal is to validate reference genes using RT-qPCR and analyze differential gene expression related to stress under control and drought conditions. The drought experiments have already been conducted at the Universidad de La Frontera (UFRO, Chile), and plant RNA has been extracted. We will validate the reference genes by evaluating their stability across species under both conditions, and then quantify the expression of at least four target genes involved in drought response for each species pair. The expected outcomes include the identification of stable reference genes and the characterization of gene expression patterns, contributing to the broader project by improving the quality of differential expression analysis in these non-model species. This will deepen our understanding of stress tolerance mechanisms in extremophile plants and potentially assist in the development of crops with enhanced resilience to climate change.