转录组和代谢组联合分析外源纳米硅对盐胁迫下水稻耐盐性的影响

    Transcriptomic and Metabolomic Integrated Analysis of the Effect of Exogenous Nano-silicon on Rice Salt Tolerance Under Salt Stress

    • 摘要:
      目的 探究外源纳米硅浸种对水稻耐盐性潜在生理和分子机制,重点聚焦协同转录和代谢反应,为盐渍地水稻抗逆栽培提供理论依据与技术指导。
      方法 以水稻‘9311’为材料,试验设置4个处理:蒸馏水浸种无NaCl处理(对照,CK),2 mmol/L外源纳米硅浸种无NaCl处理(外源纳米硅浸种,N),蒸馏水浸种+ 60 mmol/L NaCl处理(盐胁迫,S),2 mmol/L外源纳米硅浸种+60 mmol/L NaCl处理(外源纳米硅浸种+盐胁迫,N+S),三叶一心期取样,测定幼苗叶片和根系形态、活性氧水平与膜损伤、抗氧化酶活性、离子含量、渗透调节物质及内源激素含量等指标,利用转录组和代谢组分析差异表达基因(DEGs)和差异积累代谢物(DAMs)。
      结果 盐胁迫显著抑制水稻幼苗生长,与CK相比,S处理水稻幼苗地上部干质量和根系干质量分别显著下降35.53%和50.69%,叶片和根系POD活性分别显著提升32.02%和38.14%,MDA含量分别显著提升55.57%和18.23%,Na+含量分别显著提升1 382.48%和951.52%。与S处理相比,N+S处理水稻幼苗地上部干质量和根系干质量分别显著提升26.28%和49.53%,根系SOD、POD、CAT活性分别显著提升57.14%、20.53%和80.59%,叶片SP和Pro含量分别提升14.37%和21.86%,叶片和根系MDA含量分别下降26.37%和10.20%,叶片和根系Na+含量分别下降38.20%和25.75%。转录组分析可得,与S处理相比,N+S处理水稻幼苗叶片和根系分别鉴定出61和276个DEGs。N+S处理水稻幼苗叶片DEGs显著富集在赖氨酸生物合成和ABC转运蛋白等通路,根系DEGs显著富集在植物激素信号传导和亚油酸代谢等通路。代谢组分析可得,与S处理相比,N+S处理水稻幼苗叶片和根系分别鉴定出104和64种DAMs。进一步分析发现,N+S处理水稻幼苗叶片的烟酸盐和烟酰胺代谢显著富集,根系的亚油酸、半胱氨酸和甲硫氨酸代谢显著富集。转录组和代谢组联合分析可得,与S处理相比,N+S处理水稻幼苗叶片DEGs和DAMs富集在嘧啶代谢和丙酮酸代谢,根系DEGs和DAMs富集在玉米素生物合成。
      结论 外源纳米硅浸种处理通过调控玉米素生物合成的转录和代谢途径,激活水稻幼苗抗氧化系统、强化渗透调节、平衡离子稳态和内源激素,从而缓解盐胁迫对水稻幼苗的伤害。

       

      Abstract:
      Objective Exploring the potential physiological and molecular mechanisms of nano-silicon soaking on salt tolerance in rice, especially the synergistic transcriptional and metabolic responses, to provide theoretical basis and technical guidance for stress resistant cultivation of rice in saline soil.
      Method Rice cultivar '9311' was used as the experimental material, with four treatments: Seed soaking with distilled water without NaCl treatment (control, CK), seed soaking with 2.00 mmol/L exogenous nano-silicon without NaCl treatment (exogenous nano-silicon, N), seed soaking with distilled water + 60 mmol/L NaCl treatment (salt stress, S), and seed soaking with 2.00 mmol/L exogenous nano-silicon + 60.00 mmol/L NaCl treatment (nano-silicon + salt stress, N+S). Samples were collected at three-leaf-one-heart stage to measure morphological indicaes, antioxidant enzyme activities, osmotic regulatory substances, and endogenous hormones in leaves and roots. Transcriptomic and metabolomic analyses were integrated to identify differentially expressed genes(DEGs) and differentially accumulated metabolites (DAMs).
      Result Salt stress markedly inhibited the growth of rice seedlings. In comparison to the CK, the aboveground dry weight and root dry weight of seedlings subjected to S decreased significantly by 35.53% and 50.69%, respectively. Additionally, the activities of POD in leaves and roots increased significantly by 32.02% and 38.14%, respectively, while MDA content rose significantly by 55.57% in leaves and 18.23% in roots. The Na+ content increased significantly by 1 382.48% and 951.52%, respectively. Compared with the S treatment, the N+S treatment significantly increased the aboveground dry weight and root dry weight of seedlings by 26.28% and 49.53%, respectively. The activities of SOD, POD, and CAT in the root system increased significantly by 57.14%, 20.53%, and 80.59%, respectively. The contents of SP and Pro in the leaves increased by 14.37% and 21.86%, respectively. Furthermore, the MDA content in leaves and roots decreased by 26.37% and 10.20%, respectively, and the Na+ content decreased by 38.20% and 25.75%, respectively. Transcriptomic analysis indicated that, compared to S treatment, N+S treatment identified 61 and 276 DEGs in leaves and roots, respectively. The DEGs in leaves treated with N+S were significantly enriched in pathways such as lysine biosynthesis and ABC transporters. In contrast, the DEGs in roots were significantly enriched in pathways related to plant hormone signal transduction and linoleic acid metabolism. Metabolomics analysis identified 104 and 64 DAMs in leaves and roots treated with N+S, respectively, compared to S treatment. Further analysis revealed that the metabolism of nicotinate and nicotinamide was significantly enriched in leaves treated with N+S, while the metabolism of linoleic acid, cysteine, and methionine in roots was significantly enriched. Through the integrated analysis of transcriptomics and metabolomics, compared to S treatment, the DEGs and DAMs in the leaves treated with N+S were enriched in pyrimidine metabolism and pyruvate metabolism, whereas those in the root system were enriched in zeatin biosynthesis.
      Conclusion Exogenous nano-silicon seed soaking treatment activates the antioxidant system, enhances osmotic regulation, and balances ion homeostasis and endogenous hormone levels by modulating the transcriptional and metabolic pathways involved in zeatin biosynthesis, thereby mitigating salt stress damage in rice seedlings..