Saccharina latissima, known as sugar kelp, is a brown macroalga with huge ecological and economic values. In marine intertidal environment, S. latissima has to cope with both biotic and abiotic stress, which can cause the reduction of the yield during cultivation. To better understand the physiological responses of S. latissima under different stress conditions, large-scale transcriptomic analyses are useful to explore global metabolic pathway regulations. In addition, real-time quantitative polymerase chain reaction (RT-qPCR) is a powerful and rapid method for further quantifying changes in gene expression, and for targeting specific defense-related gene pathways. However, its level of accuracy is highly related to the expression stability of reference genes used for normalization and those still need to be evaluated in S. latissima. In this study, we therefore experimentally tested eight candidate reference genes identified from in silico screening of public transcriptomic datasets of S. latissima from different abiotic and biotic stress treatments. The stability analysis using complementary statistical approaches showed that EIF5B and ATPase are the most stable reference genes under biotic stress, whereas, under temperature and light stress, their combination with NDH gene is the best choice for RT-qPCR normalization. The validated reference genes were used to monitor the expression of target genes, related to oxidative responses, such as those involved in oxylipin pathways, in S. latissima plantlets submitted to different stress in laboratory-controlled conditions.

Saccharina latissima，被称为糖海带，是一种棕色的大型藻类，具有巨大的生态和经济价值。在海洋潮间环境中，S。latissima必须应对生物和非生物胁迫，这可能会导致种植期间产量下降。为了更好地了解S. latissima的生理反应在不同的胁迫条件下，大规模的转录组分析可用于探索全球代谢途径的调控。此外，实时定量聚合酶链反应（RT-qPCR）是一种功能强大且快速的方法，可用于进一步量化基因表达的变化并靶向特定的防御相关基因途径。但是，其准确性水平与用于标准化的参考基因的表达稳定性高度相关，尚需在lat。S. latissima中进行评估。因此，在这项研究中，我们通过实验测试了从latissima的公共转录组数据集的计算机筛选中鉴定出的八个候选参考基因。来自不同的非生物和生物压力疗法。使用互补统计方法进行的稳定性分析表明，EIF5B和ATPase在生物胁迫下是最稳定的参考基因，而在温度和光照胁迫下，它们与NDH基因的结合是RT-qPCR标准化的最佳选择。经验证的参考基因用于监测在实验室控制条件下经受不同胁迫的拉丁美沙门氏菌幼苗中与氧化反应有关的目标基因的表达，例如与脂蛋白途径有关的基因。