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Identification of reference genes for RT-qPCR in the Antarctic moss Sanionia uncinata under abiotic stress conditions
PLOS ONE ( IF 3.7 ) Pub Date : 2018-06-19 , DOI: 10.1371/journal.pone.0199356
Mira Park , Soon Gyu Hong , Hyun Park , Byeong-ha Lee , Hyoungseok Lee

Sanionia uncinata is a dominant moss species in the maritime Antarctic. Due to its high adaptability to harsh environments, this extremophile plant has been considered a good target for studying the molecular adaptation mechanisms of plants to a variety of environmental stresses. Despite the importance of S. uncinata as a representative Antarctic plant species for the identification and characterization of genes associated with abiotic stress tolerance, suitable reference genes, which are critical for RT-qPCR analyses, have not yet been identified. In this report, 11 traditionally used and 6 novel candidate reference genes were selected from transcriptome data of S. uncinata and the expression stability of these genes was evaluated under various abiotic stress conditions using three statistical algorithms; geNorm, NormFinder, and BestKeeper. The stability ranking analysis selected the best reference genes depending on the stress conditions. Among the 17 candidates, the most stable references were POB1 and UFD2 for cold stress, POB1 and AKB for drought treatment, and UFD2 and AKB for the field samples from a different water contents in Antarctica. Overall, novel genes POB1 and AKB were the most reliable references across all samples, irrespective of experimental conditions. In addition, 6 novel candidate genes including AKB, POB1 and UFD2, were more stable than the housekeeping genes traditionally used for internal controls, indicating that transcriptome data can be useful for identifying novel robust normalizers. The reference genes validated in this study will be useful for improving the accuracy of RT-qPCR analysis for gene expression studies of S. uncinata in Antarctica and for further functional genomic analysis of bryophytes.



中文翻译:

非生物胁迫条件下南极苔藓桑翁草中RT-qPCR参考基因的鉴定

Sanionia uncinata是海洋南极的主要苔藓种。由于其对恶劣环境的高度适应性,这种极端嗜热植物被认为是研究植物对各种环境胁迫的分子适应机制的良好靶标。尽管S的重要性。作为用于鉴定和表征与非生物胁迫耐受性相关的基因的代表性南极植物,uncinata尚未确定对RT-qPCR分析至关重要的合适参考基因。在本报告中,从S的转录组数据中选择了11个传统使用的基因和6个新的候选参考基因。Uncinata并使用三种统计算法在各种非生物胁迫条件下评估了这些基因的表达稳定性;geNorm,NormFinder和BestKeeper。稳定性排名分析根据压力条件选择了最佳参考基因。在这17个候选对象中,最稳定的参考标准是POB1UFD2用于冷胁迫,POB1AKB用于干旱处理,UFD2AKB用于南极洲不同含水量的田间样品。总体而言,新基因POB1AKB不论实验条件如何,都是所有样品中最可靠的参考。此外,包括AKBPOB1UFD2在内的6个新候选基因比传统上用于内部对照的管家基因更稳定,这表明转录组数据可用于鉴定新型稳健的标准化基因。在这项研究中验证的参考基因将有助于提高S基因表达研究的RT-qPCR分析的准确性。uncinata在南极洲和苔藓植物的进一步的功能基因组分析。

更新日期:2018-06-20
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