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Abstract：
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.
摘要：
种子衰老是一个复杂的生物学过程，其基本原理和机制尚未得到充分认识。这是每天在基因库中收集和存储植物遗传资源的研究团队面临的关键问题。几乎没有研究与干燥状态下种子老化相关的转录组变化。该研究的目的是开发一种有效的方案，用于从具有非常低的活力和低RNA完整性数（RIN）的长期储存种子构建RNA-Seq文库。这里,使用了由于长期储存而几乎完全失去生存能力的大麦种子。作为一种控制，使用在田间再生过程中获得的完全可行的种子。比较了专用于具有高和低RIN值的RNA样品的方案的有效性。实验得出结论，由于不同RNA部分不均匀降解的可能性，从具有降解RNA（RIN<3）的低活力或长期储存种子的文库构建应格外注意。
