Drought is the major limiting factor that directly or indirectly inhibits the growth and reduces the productivity of sorghum (Sorghum bicolor (L.) Moench). As the main vegetative organ of sorghum, the response mechanism of the leaf to drought stress at the proteomic level has not been clarified. In the present study, nano-scale liquid chromatography mass spectrometry (nano-LC-MS/MS) technology was used to compare the changes in the protein expression profile of the leaves of drought-sensitive (S4 and S4-1) and drought-resistant (T33 and T14) sorghum varieties at the seedling stage under 25% PEG-6000 treatment for 24 h. A total of 3927 proteins were accurately quantitated and 46, 36, 35, and 102 differentially abundant proteins (DAPs) were obtained in the S4, S4-1, T14, and T33 varieties, respectively. Four proteins were randomly selected for parallel reaction monitoring (PRM) assays, and the results verified the reliability of the mass spectrometry (MS) results. The response mechanism of the drought-sensitive sorghum leaves to drought was attributed to the upregulation of proteins involved in the tyrosine metabolism pathway with defense functions. Drought-resistant sorghum leaves respond to drought by promoting the TCA cycle, enhancing sphingolipid biosynthesis, interfering with triterpenoid metabolite synthesis, and influencing aminoacyl-tRNA biosynthesis. The 17 screened important candidate proteins related to drought stress were verified by quantitative real-time PCR (qRT-PCR), the results of which were consistent with the results of the proteomic analysis. This study lays the foundation for revealing the drought-resistance mechanism of sorghum at the protein level. These findings will help us cultivate and improve new drought-resistant sorghum varieties.

Wheat is a staple crop in China’s arid and semi-arid regions. Drought and low nitrogen (LN) are two major constraints to wheat growth and production. However, the molecular mechanism underlying wheat response to both drought and LN stress remains unknown. Accordingly, we conducted a proteomic study on the roots of two wheat varieties, Chang6878 (drought tolerant) and Shi4185 (drought sensitive) and compared the differences between drought and combined drought and LN stress treatments. In total, 5143 proteins were identified, of which 163 differentially abundant proteins (DAPs) were uniquely upregulated under drought and LN stress in Chang6878. Enrichment analysis showed that DAPs were mainly involved in mitogen-activated protein kinase signaling, phenylpropanoid biosynthesis, glutathione metabolism, ethylene biosynthesis, ethylene signal transduction, and oxidation–reduction reactions. These DAPs were verified via parallel reaction monitoring and quantitative real-time polymerase chain reaction. Chang6878 was treated with the ethylene synthesis precursor 1-aminocyclopropanecarboxylic acid, and its resistance to drought and LN stress improved. After treatment with the ethylene synthesis inhibitor silver nitrate and ethylene signal transduction inhibitor 1-methylcyclopropene, drought and LN stress resistance reduced. These results provide novel insights into the tolerance mechanisms of Chang6878 to drought and LN stress by altering ethylene synthesis and signal transduction. This study provides a reference for breeding drought- and low-nitrogen-tolerant wheat germplasm resources and a theoretical basis for maintaining food security in arid, barren areas.

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