Nhlakanipho Msomi Mhlonipheni, Prinsloo Gerhard, Nogemane Noluyolo
Department of Agriculture and Animal Health, Florida Science Campus, University of South Africa, Johannesburg, Gauteng Province, South Africa.
Heliyon. 2024 Sep 7;10(18):e37466. doi: 10.1016/j.heliyon.2024.e37466. eCollection 2024 Sep 30.
Soybean ( L.) serves not only as food for humans, animals, and industrial purposes, but is also a plant that can be used to comprehend molecular mechanisms occurring in stress response to various development techniques. To reveal the effect of applying dicarboxylic acids as stress priming agents on a metabolic level in soybean leaf extracts, the chemical profile of methanolic extracts were collected at different time points (1 h, 2 h, 12 h, 24 h, 1 week, 2 weeks and 3 weeks) after spraying were analyzed using H-NMR based metabolomics by way of PCA and OPLS-DA. The OPLS-DA revealed several metabolites, including organic acids (fumarate, citrate and malate) and amino acids (asparagine, alanine and GABA), which accumulated in higher amounts, with fumarate accumulating the highest in L. leaf extracts compared to untreated leaves. Denaturing 1DE gels were prepared for MS-based protein analysis and the presence of fatty acids (linolenic, oleic and α-linolenic acid) were confirmed by gas chromatography coupled with mass spectrometry (GC-MS), which served as jasmonic acid precursors. The MS-based profiling of proteins on the denaturing 1DE gels revealed several proteins that were differentiated between the treated and untreated leaf extracts. These proteins included ferritins, CaM, ferredoxin-thioredoxin reductase and chalcone-flavanone isomerase 1A. Following the treatment, fumarate was significantly elevated at 12 h to 3 weeks, compared to other compounds. It is, therefore, proposed that elevated quantities of fumarate could be related to the KEAP1-NRF2 metabolic pathway. This study represents the initial investigation of the effect of dicarboxylic acid application as a stress priming agent on L. using H-NMR metabolomic analysis, GC-MS and proteomic analysis.
大豆（L.）不仅是人类、动物的食物和工业原料，也是一种可用于理解各种发育技术应激反应中发生的分子机制的植物。为了揭示将二羧酸作为应激引发剂应用于大豆叶片提取物代谢水平的影响，在喷洒后不同时间点（1小时、2小时、12小时、24小时、1周、2周和3周）收集甲醇提取物的化学图谱，通过主成分分析（PCA）和正交偏最小二乘法判别分析（OPLS-DA），采用基于氢核磁共振（H-NMR）的代谢组学进行分析。OPLS-DA揭示了几种代谢物，包括有机酸（富马酸、柠檬酸和苹果酸）和氨基酸（天冬酰胺、丙氨酸和γ-氨基丁酸），它们的积累量更高，与未处理的叶片相比，富马酸在大豆叶片提取物中积累量最高。制备变性一维凝胶用于基于质谱的蛋白质分析，并通过气相色谱-质谱联用（GC-MS）确认了脂肪酸（亚麻酸、油酸和α-亚麻酸）的存在，这些脂肪酸是茉莉酸的前体。基于质谱的变性一维凝胶蛋白质谱分析揭示了几种在处理和未处理的叶片提取物之间有差异的蛋白质。这些蛋白质包括铁蛋白、钙调蛋白、铁氧还蛋白-硫氧还蛋白还原酶和查尔酮-黄烷酮异构酶1A。处理后，与其他化合物相比，富马酸在12小时至3周时显著升高。因此，有人提出富马酸含量的升高可能与KEAP1-NRF2代谢途径有关。本研究代表了使用H-NMR代谢组学分析、GC-MS和蛋白质组学分析对二羧酸作为应激引发剂应用于大豆的影响进行的初步调查。
