Singh Vivek Vikram, Naseer Aisha, Sellamuthu Gothandapani, Jakuš Rastislav
Faculty of Forestry and Wood Sciences, Czech University of Life Sciences Prague, Kamýcká 129, Praha-Suchdol, 165 00 Prague, Czech Republic.
Institute of Forest Ecology, Slovak Academy of Sciences, Štúrova 2, 960 53 Zvolen, Slovakia.
Plants (Basel). 2024 Jan 28;13(3):389. doi: 10.3390/plants13030389.
Since the development of next-generation sequencing techniques and with the growing interest in transcriptomic studies, there is a demand for high-throughput RNA extraction techniques. General RNA extraction protocols are unreliable when it comes to the quality and quantity of isolated RNA obtained from different tissue types of different plant species. Despite Norway spruce () being one of the most significant and commercially valuable tree species in European forests, only limited genetic research is available. In this study, we developed a cetyltrimethylammonium bromide (CTAB) protocol by modifying the original method. We compared this CTAB protocol with other widely used methods for extracting RNA from different tissues (needle, phloem, and root) of Norway spruce, known for its richness in polyphenols, polysaccharides, and secondary metabolites. The modified CTAB method proves to be superior to the kit-based and TRIzol-based methods for extracting RNA from the metabolite-rich tissues of Norway spruce, resulting in high RNA quality and integrity values (RIN~7-9). The modified CTAB RNA extraction method is rapid, cost-effective, and relatively simple in yielding the desired RNA quality from Norway spruce tissues. It is optimal for RNA sequencing and other downstream molecular applications.
自从下一代测序技术发展以来，随着对转录组学研究兴趣的不断增加，对高通量RNA提取技术的需求也日益增长。当涉及从不同植物物种的不同组织类型中分离得到的RNA的质量和数量时，一般的RNA提取方案并不可靠。尽管挪威云杉（Picea abies）是欧洲森林中最重要且具有商业价值的树种之一，但现有的遗传研究却很有限。在本研究中，我们通过对原始方法进行修改，开发了一种十六烷基三甲基溴化铵（CTAB）方案。我们将此CTAB方案与其他广泛使用的从挪威云杉不同组织（针叶、韧皮部和根）中提取RNA的方法进行了比较，挪威云杉以富含多酚、多糖和次生代谢产物而闻名。改良的CTAB方法在从富含代谢产物的挪威云杉组织中提取RNA方面被证明优于基于试剂盒和基于TRIzol的方法，能产生高质量的RNA以及完整性值（RIN~7 - 9）。改良的CTAB RNA提取方法快速、经济高效，并且相对简单地就能从挪威云杉组织中获得所需的RNA质量。它对于RNA测序和其他下游分子应用而言是最佳的。
