College of Life Sciences, Northwest A&F University, Yangling, Shaan Xi, China.
Plant Biol (Stuttg). 2024 Jun;26(4):544-559. doi: 10.1111/plb.13645. Epub 2024 Apr 8.
CircRNAs (circular RNAs) reduce the abundance of miRNAs through ceRNA (competing endogenous RNA), to regulate many physiological processes and stress responses in plants. However, the role of circRNA in drought stress is poorly understood. Through ring identification and sequencing verification of ath-circ032768, bioinformatics analysis predicted the interaction of ath-circ032768-miR472-RPS5, and further obtained transgenic plants overexpressing ath-circ032768 and silencing STTM-miR472. The change in drought stress was analysed using biochemical and molecular biological methods. Sequencing and biological analysis confirmed that ath-circ032768, miR472 and RPS5 were responsive to drought stress, and changes in gene expression were consistent with the prediction of ceRNA. The silencing vectors ath-circ032768 and STTM-miR472 were constructed using molecular biology techniques, and stable transgenic plants with drought tolerance obtained. Further physiological and biochemical studies showed that ath-circ032768 could bind to miR472, and that miR472 could bind to the RPS5 gene, resulting in decreased expression of RPS5. Hence, ath-circ032768 can competitively inhibit degradation of RPS5 by miR472 through ceRNA. This process is accompanied by increased expression of DREB2A, RD29A and RD29B genes. Through the ath-circ032768-miR472-RPS5 pathway, the RPS5 stress resistance protein interacts with DREB2A protein to enhance expression of downstream drought resistance genes, RD29A and RD29B, and participate in the regulation mechanism of plant drought resistance, thereby improving drought tolerance of plants.
circRNAs(环状 RNA)通过 ceRNA(竞争内源性 RNA)减少 miRNA 的丰度,从而调节植物中的许多生理过程和应激反应。然而,circRNA 在干旱胁迫中的作用还知之甚少。通过 ath-circ032768 的环状鉴定和测序验证,生物信息学分析预测了 ath-circ032768-miR472-RPS5 的相互作用,进一步获得了过表达 ath-circ032768 和沉默 STTM-miR472 的转基因植物。使用生化和分子生物学方法分析干旱胁迫的变化。测序和生物学分析证实 ath-circ032768、miR472 和 RPS5 对干旱胁迫有反应,基因表达的变化与 ceRNA 的预测一致。使用分子生物学技术构建了 ath-circ032768 和 STTM-miR472 的沉默载体,并获得了具有耐旱性的稳定转基因植物。进一步的生理生化研究表明,ath-circ032768 可以与 miR472 结合,miR472 可以与 RPS5 基因结合,导致 RPS5 表达减少。因此,ath-circ032768 可以通过 ceRNA 竞争性抑制 miR472 对 RPS5 的降解。这一过程伴随着 DREB2A、RD29A 和 RD29B 基因表达的增加。通过 ath-circ032768-miR472-RPS5 途径,RPS5 应激抗性蛋白与 DREB2A 蛋白相互作用,增强下游干旱抗性基因 RD29A 和 RD29B 的表达,参与植物抗旱调节机制,从而提高植物的抗旱性。