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<p><i>Elymus sibiricus</i>, a perennial grass of the Poaceae family, plays a vital role in ecological protection and animal husbandry in temperate regions. In this study, an efficient regeneration system was developed using young spikes and mature embryos as explants. Young spikes (0.7–2.6&#xa0;cm) were cultured on Murashige and Skoog (MS) medium supplemented with 5&#xa0;mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D) and 0.15&#xa0;mg/L 6-Benzylaminopurine (6-BA) for callus induction. Callus was then transferred to MS medium containing 1.5&#xa0;mg/L 2,4-D and 0.15&#xa0;mg/L 6-BA to promote shoot differentiation. A regeneration system based on young spikes was successfully developed for the variety Gaomu No.1, achieving a regeneration efficiency of 27.55%. For mature embryos, embryogenic callus was successfully induced under light conditions. Among 44 accessions screened, embryogenic callus characterized by cream-yellow color, granular texture, and compact structure appearance was induced exclusively in genotype C1, with a differentiation rate of 21.11%. To explore the molecular mechanisms underlying somatic embryogenesis, the transcriptomic profiles of embryogenic and non-embryogenic calli derived from mature embryos were compared. The differentially expressed genes (DEGs) in embryonic callus were primarily enriched in chromosomes, chromatin remodeling and abiotic stress response pathways. Nitroblue tetrazolium (NBT) staining further confirmed substantial reactive oxygen species (ROS) accumulation in embryogenic callus, in contrast to the absence of ROS in non-embryogenic callus. The efficient regeneration system established in this study provides a valuable platform for propagation and future genetic transformation in <i>E. sibiricus</i>, while transcriptomic analysis reveals key molecular pathways involved in somatic embryogenesis.</p>

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Establishment of young spike and mature embryo-based regeneration system and transcriptomic analysis of different callus types in Elymus sibiricus

  • 期刊论文

  • 发表日期：

    2025年8月6日

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  • Jialei Guo,
  • Bo Liu,
  • Wenjie Shen,
  • Ying Yang,
  • Jie Chen,
  • Yixin Meng,
  • Quanwen Dou

摘要

Elymus sibiricus, a perennial grass of the Poaceae family, plays a vital role in ecological protection and animal husbandry in temperate regions. In this study, an efficient regeneration system was developed using young spikes and mature embryos as explants. Young spikes (0.7–2.6 cm) were cultured on Murashige and Skoog (MS) medium supplemented with 5 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D) and 0.15 mg/L 6-Benzylaminopurine (6-BA) for callus induction. Callus was then transferred to MS medium containing 1.5 mg/L 2,4-D and 0.15 mg/L 6-BA to promote shoot differentiation. A regeneration system based on young spikes was successfully developed for the variety Gaomu No.1, achieving a regeneration efficiency of 27.55%. For mature embryos, embryogenic callus was successfully induced under light conditions. Among 44 accessions screened, embryogenic callus characterized by cream-yellow color, granular texture, and compact structure appearance was induced exclusively in genotype C1, with a differentiation rate of 21.11%. To explore the molecular mechanisms underlying somatic embryogenesis, the transcriptomic profiles of embryogenic and non-embryogenic calli derived from mature embryos were compared. The differentially expressed genes (DEGs) in embryonic callus were primarily enriched in chromosomes, chromatin remodeling and abiotic stress response pathways. Nitroblue tetrazolium (NBT) staining further confirmed substantial reactive oxygen species (ROS) accumulation in embryogenic callus, in contrast to the absence of ROS in non-embryogenic callus. The efficient regeneration system established in this study provides a valuable platform for propagation and future genetic transformation in E. sibiricus, while transcriptomic analysis reveals key molecular pathways involved in somatic embryogenesis.