# Functional analysis of StWRKY75 gene in response to heat stress in potato (Solanum tuberosum L.)

## TL;DR

This study identifies the StWRKY75 gene in potatoes as a key player in improving heat tolerance through enhanced growth and stress responses.

## Contribution

The novel contribution is the functional characterization of StWRKY75 as a positive regulator of thermotolerance in potato under heat stress.

## Key findings

- Overexpression of StWRKY75 improves thermotolerance in potato plants under heat stress.
- StWRKY75 upregulates antioxidant enzymes, proline metabolism, and heat shock proteins.
- RNAi knockdown of StWRKY75 compromises heat stress responses in potato.

## Abstract

Heat stress severely restricts potato tuber growth and development, yet the roles of WRKY transcription factors (TFs) in mediating heat responses remain poorly understood. Using quantitative reverse transcription PCR (qRT-PCR), we identified StWRKY75 in potato cultivars ‘Atlantic’ and ‘Desiree’ as a heat-inducible gene, showing significant upregulation under 30°C and 35°C treatments. Phylogenetic analysis classified StWRKY75 into the WRKY Group II family, with close evolutionary homology to tomato SlWRKY75, and subcellular localization confirmed its nuclear targeting. To characterize its function, we generated transgenic lines overexpressing StWRKY75 (OE) or knockdown by RNA interference (RNAi). Under heat stress conditions, OE plants demonstrated superior thermotolerance compared to non-transgenic (NT) controls, manifested by improved growth parameters (plant height, tuber weight per plant, fresh weight, dry weight, root fresh weight, and root dry weight), enhanced activities of antioxidant enzymes (ascorbate peroxidase [APX], catalase [CAT], peroxidase [POD], and superoxide dismutase [SOD]), increased proline accumulation, elevated photosynthetic efficiency, and reduced malondialdehyde (MDA) content. Conversely, RNAi lines exhibited compromised thermotolerance with reversed growth parameters and biochemical characteristics. qRT-PCR analysis revealed that StWRKY75 positively regulated key heat-stress responsive genes, including those encoding antioxidant enzymes (StAPX, StCAT, StSOD, StPOD), proline biosynthesis Pyrroline-5-carboxylate synthase (StP5CS), and heat shock proteins (StHSP20, StHSP70, StHSP90). These findings demonstrate that StWRKY75 acts as a positive regulator of potato thermotolerance by enhancing growth, antioxidant capacity, proline metabolism, photosynthesis, and heat-responsive gene expression, providing a valuable target for improving crop heat resilience in breeding programs.