Seed aging is a complex biological process and its fundamentals and mechanisms have not yet been fully recognized. This is a key issue faced by research teams involved in the collection and storage of plant genetic resources in gene banks every day. Transcriptomic changes associated with seed aging in the dry state have barely been studied. The aim of the study was to develop an efficient protocol for construction of RNA-Seq libraries from long-term stored seeds with very low viability and low RNA integrity number (RIN). Here, barley seeds that have almost completely lost their viability as a result of long-term storage were used. As a control, fully viable seeds obtained in the course of field regeneration were used. The effectiveness of protocols dedicated to RNA samples with high and low RIN values was compared. The experiment concluded that library construction from low viable or long-term stored seeds with degraded RNA (RIN < 3) should be carried out with extraordinary attention due to the possibility of uneven degradation of different RNA fractions.

中文翻译：

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从长期保存的种子构建 RNA-Seq 文库的 RIN 值较低：大麦种子的案例研究


种子老化是一个复杂的生物过程，其基本原理和机制尚未得到充分认识。这是每天参与基因库植物遗传资源收集和储存的研究团队面临的一个关键问题。与干燥状态下种子老化相关的转录组变化几乎没有被研究过。该研究的目的是开发一种有效的方案，用于从长期储存的活力极低和 RNA 完整性数 (RIN) 较低的种子构建 RNA-Seq 文库。这里使用的大麦种子由于长期储存而几乎完全失去了活力。作为对照，使用在田间再生过程中获得的完全有活力的种子。比较了专门用于具有高 RIN 值和低 RIN 值的 RNA 样品的方案的有效性。该实验得出的结论是，由于不同 RNA 组分降解不均匀的可能性，应特别注意从低活力或长期保存的 RNA 降解种子 (RIN < 3) 进行文库构建。