The pathogenicity and regulatory function of temperature-sensitive proteins PscTSP in Pseudofabraea citricarpa under high temperature stress.

Citrus fruit rich in beneficial health-promoting nutrients used for functional foods or dietary supplements production. However, its quality and yield were damaged by citrus target spot. Citrus target spot is a low-temperature fungal disease caused by Pseudofabraea citricarpa, resulting in citrus production reductions and economic losses. In this study, transcriptome and gene knockout mutant analyses were performed on the growth and pathogenicity of P. citricarpa under different temperature conditions to quantify the functions of temperature-sensitive proteins (PscTSP). The optimum growth temperature for P. citricarpa strain WZ1 was 20 °C, while it inhibited or stopped growth above 30 °C and stopped growth below 4 °C or above 30 °C. Certain PscTSP-key genes of P. citricarpa were identified under high temperature stress. qRT-PCR analysis confirmed the expression levels of PscTSPs under high temperature stress. PscTSPs were limited by temperature and deletion of the PscTSP-X gene leads to changes in the integrity of citrus cell walls, osmotic regulation, oxidative stress response, calcium regulation, chitin synthesis, and the pathogenicity of P. citricarpa. These results provide insight into the underlying mechanisms of temperature sensitivity and pathogenicity in P. citricarpa, providing a foundation for developing resistance strategies against citrus target spot disease.

Functional Evolution of Pseudofabraea citricarpa as an Adaptation to Temperature Change.

Citrus target spot, caused by Pseudofabraea citricarpa, was formerly considered a cold-tolerant fungal disease. However, it has now spread from high-latitude regions to warmer low-latitude regions. Here, we conducted physiological observations on two different strains of the fungus collected from distinct regions, and evaluated their pathogenicity. Interestingly, the CQWZ collected from a low-latitude orchard, exhibited higher temperature tolerance and pathogenicity when compared to the SXCG collected from a high-latitude orchard. To further understand the evolution of temperature tolerance and virulence in these pathogens during the spread process, as well as the mechanisms underlying these differences, we performed genomic comparative analysis. The genome size of CQWZ was determined to be 44,004,669 bp, while the genome size of SXCG was determined to be 45,377,339 bp. Through genomic collinearity analysis, we identified two breakpoints and rearrangements during the evolutionary process of these two strains. Moreover, gene annotation results revealed that the CQWZ possessed 376 annotated genes in the "Xenobiotics biodegradation and metabolism" pathway, which is 79 genes more than the SXCG. The main factor contributing to this difference was the presence of salicylate hydroxylase. We also observed variations in the oxidative stress pathways and core pathogenic genes. The CQWZ exhibited the presence of a heat shock protein (HSP SSB), a catalase (CAT2), and 13 core pathogenic genes, including a LysM effector, in comparison to the SXCG. Furthermore, there were significant disparities in the gene clusters responsible for the production of seven metabolites, such as Fumonisin and Brefeldin. Finally, we identified the regulatory relationship, with the HOG pathway at its core, that potentially contributes to the differences in thermotolerance and virulence. As the global climate continues to warm, crop pathogens are increasingly expanding to new territories. Our findings will enhance understanding of the evolution mechanisms of pathogens under climate change.