Comparative transcriptomic and metabolic profiling provides insight into the mechanism by which the autophagy inhibitor 3-MA enhances salt stress sensitivity in wheat seedlings

Abstract

Background Salt stress hinders plant growth and production around the world. Autophagy induced by salt stress helps plants improve their adaptability to salt stress. However, the underlying mechanism behind this adaptability remains unclear. To obtain deeper insight into this phenomenon, combined metabolomics and transcriptomics analyses were used to explore the coexpression of differentially expressed-metabolite (DEM) and gene (DEG) between control and salt-stressed wheat roots and leaves in the presence or absence of the added autophagy inhibitor 3-methyladenine (3-MA). Results The results indicated that 3-MA addition inhibited autophagy, increased ROS accumulation, damaged photosynthesis apparatus and impaired the tolerance of wheat seedlings to NaCl stress. A total of 14,759 DEGs and 554 DEMs in roots and leaves of wheat seedlings were induced by salt stress. DEGs were predominantly enriched in cellular amino acid catabolic process, response to external biotic stimulus, regulation of the response to salt stress, reactive oxygen species (ROS) biosynthetic process, regulation of response to osmotic stress, ect. The DEMs were mostly associated with amino acid metabolism, carbohydrate metabolism, phenylalanine metabolism, carbapenem biosynthesis, and pantothenate and CoA biosynthesis. Further analysis identified some critical genes (gene involved in the oxidative stress response, gene encoding transcription factor (TF) and gene involved in the synthesis of metabolite such as alanine, asparagine, aspartate, glutamate, glutamine, 4-aminobutyric acid, abscisic acid, jasmonic acid, ect.) that potentially participated in a complex regulatory network in the wheat response to NaCl stress. The expression of the upregulated DEGs and DEMs were higher, and the expression of the down-regulated DEGs and DEMs was lower in 3-MA-treated plants under NaCl treatment. Conclusion 3-MA enhanced the salt stress sensitivity of wheat seedlings by inhibiting the activity of the roots and leaves, inhibiting autophagy in the roots and leaves, increasing the content of both H2O2 and O2•—, damaged photosynthesis apparatus and changing the transcriptome and metabolome of salt-stressed wheat seedlings.

The Metacaspase TaMCA-Id Negatively Regulates Salt-Induced Programmed Cell Death and Functionally Links With Autophagy in Wheat

“…A unigene (accession no. TraesCS4D02G358100 ) released from EnsemblPlants was chosen and cloned, which was conferred to wheat seedlings' tolerance to salt stress in an RNA-Seq assay (Yue et al, 2021b ). Its complete coding sequence (CDS) was amplified from the first-strand cDNA using TransStart Fast Pfu DNA polymerase (TransGen, China), with a specific pair of primers presented in Supplementary Table 1 .…”

“…The staining procedures were carried out as previously described (Yue et al, 2021a ). The quantitative determination of H 2 O 2 and was performed following a previous study (Yue et al, 2021b ). Evans blue staining was carried out to visualize the degree of cell damage in the wheat leaves under NaCl treatment.…”

The Calcium-Dependent Protein Kinase TaCDPK27 Positively Regulates Salt Tolerance in Wheat

“…From the salt stress-induced differentially expressed genes (DEGs) of wheat roots and leaves identified via an RNA-Seq assay, a unigene (accession no. TraesCS5B02G109300) released from EnsemblPlants was chosen and cloned [46]. Based on its CDS, the specific primers were designed to amplify the full length of ORF with reverse transcription-polymerase chain reaction (RT-PCR) (Table 3).…”

“…The staining procedures were carried out as previously described [45]. The quantitative determination of H 2 O 2 and O 2− was performed following a previous study [46]. Evans blue staining was carried out to visualize the degree of cell damage in the wheat leaves under NaCl treatment.…”

De novo transcriptome analysis of high-salinity stress-induced antioxidant activity and plant phytohormone alterations in Sesuvium portulacastrum

“…Therefore, a highsalinity response mechanism was studied using the RNA-seq technology to screen for salt-tolerance genes. Differentially expressed genes (DEGs) are predominantly enriched in cellular amino acid catabolic processes and responses to ROS biosynthetic processes (Yue et al, 2021). Plant hormone signal transduction genes have been discovered in salt-treated Solenostemma argel (Ahmad et al, 2021).…”