Quantitative phosphoproteomic analysis provides insights into the aluminum-responsiveness of Tamba black soybean

Abstract

Aluminum (Al 3+) toxicity is one of the most important limitations to agricultural production worldwide. The overall response of plants to Al 3+ stress has been documented, but the contribution of protein phosphorylation to Al 3+ detoxicity and tolerance in plants is unclear. Using a combination of tandem mass tag (TMT) labeling, immobilized metal affinity chromatography (IMAC) enrichment and liquid chromatography-tandem mass spectrometry (LC-MS/MS), Al 3+-induced phosphoproteomic changes in roots of Tamba black soybean (TBS) were investigated in this study. The Data collected in this study are available via ProteomeXchange with the identifier PXD019807. After the Al 3+ treatment, 189 proteins harboring 278 phosphosites were significantly changed (fold change > 1.2 or < 0.83, p < 0.05), with 88 upregulated, 96 downregulated and 5 up-/downregulated. Enrichment and protein interaction analyses revealed that differentially phosphorylated proteins (DPPs) under the Al 3+ treatment were mainly related to G-protein-mediated signaling, transcription and translation, transporters and carbohydrate metabolism. Particularly, DPPs associated with root growth inhibition or citric acid synthesis were identified. The results of this study provide novel insights into the molecular mechanisms of TBS post-translational modifications in response to Al 3+ stress.

GmMYB183, a R2R3-MYB Transcription Factor in Tamba Black Soybean (Glycine max. cv. Tamba), Conferred Aluminum Tolerance in Arabidopsis and Soybean

“…Based on data from quantitative phosphoproteomics of TBS, a protein with significantly up-regulated phosphorylation induced by Al stress was screened [ 44 ]. The full-length coding sequences (CDSs) of GmMYB183 were amplified using cDNA from TBS.…”

“…Nonetheless, research exploring the post-translational modification of transcription factors in response to Al stress is scarce. Remarkable shifts in phosphorylation have been observed in GmMYB183 in response to Al stress, without any alterations in the transcript levels of GmMYB183 [ 44 ]. Interestingly, the phosphorylation of ALR1, a recently discovered Al receptor, is also regulated by Al stress [ 59 ].…”

“…TBS seedlings were cultured as previously described [ 44 ]. Briefly, pre-treated roots were transferred to a 50 µmol/L AlCl 3 solution (containing 0.5 mmol/L CaCl 2 , pH 4.5) for 3 h, 6 h, 9 h, 12 h, 24 h, 48 h or 96 h. Arabidopsis seeds (ecotype Columbia, Col-0) were directly germinated on mixed soil (substrate: vermiculite = 1:1) and grown in a controlled environment at day/night temperatures of 25/22 °C, with 16 h of light (120 µmol photons m −2 ·s −1 ).…”

“…First-strand cDNA was synthesized using Prime Script TM reagent Kit with a gDNA Eraser kit (Takara, Shiga, Japan). Quantitative real-time PCR (RT-qPCR) was performed according to our previous study with 40SrRNA gene (GenBank: XM_003549836.4) as an internal control [ 44 ]. Primers used in this study are presented in Table S1 .…”

“…Previously, we studied the effect of GmMYB183 on the Al stress response in TBS. We observed significant phosphorylation shifts, including upregulated phosphorylation of a serine residue (Ser36) [ 44 ]. In this study, our objective was to further investigate the function of GmMYB183 in the Al stress response.…”