Hainan Key Laboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China
Department of Agriculture, Abdul Wali Khan University Mardan, 23200 Pakistan.
Biosci Rep. 2019 Jan 3;39(1). doi: 10.1042/BSR20181163. Print 2019 Jan 31.
Complex plant tissues vary in hardness, i.e. some are succulent, while others are complex to break. Besides, plant metabolites, such as polysaccharides, proteins, polyphenols and lipids, can greatly interfere with the RNA extraction. So, in order to obtain a high-quality RNA from the complex tissues (like coconut endosperm, coconut apple and coconut leaf bud) rich in secondary metabolites, a robust method is demanded. Several methods (MRIP, CTAB and TRIZOL) have been used previously for the isolation of quality RNA from the coconut tissues, but without any success. The present study will provide with the details of a new method (Quick and Reliable RNA Extraction Method or QRREM), which have efficiently isolated the intact RNA form the complex tissues of coconut compared with CTAB, Trizol and RNA plant. The method has been validated for the isolation of high-quality intact RNA from the other available plant species (Areca/betel nut, mint and spring onion). The method has various advantages over the other methods in terms of time and cost effectiveness. Furthermore, the resulted RNA from various tissues of coconut performed well in the downstream experiments, i.e. reverse transcription and PCR for the production and amplification of cDNA.
复杂的植物组织在硬度上有所不同，例如有些多汁，而有些则难以破碎。此外，植物代谢物，如多糖、蛋白质、多酚和脂质，会极大地干扰 RNA 的提取。因此，为了从富含次生代谢物的复杂组织（如椰子胚乳、椰子果肉和椰子叶芽）中获得高质量的 RNA，需要一种稳健的方法。先前已经使用了几种方法（MRIP、CTAB 和 TRIZOL）从椰子组织中分离高质量的 RNA，但均未成功。本研究将提供一种新方法（快速可靠的 RNA 提取方法或 QRREM）的详细信息，该方法与 CTAB、Trizol 和 RNA plant 相比，能够有效地从椰子的复杂组织中分离完整的 RNA。该方法已在其他可利用的植物物种（槟榔、薄荷和葱）中验证了其从高质量完整 RNA 中分离的有效性。与其他方法相比，该方法在时间和成本效益方面具有多种优势。此外，从椰子的各种组织中提取的 RNA 在下游实验中表现良好，例如反转录和 PCR 用于 cDNA 的生成和扩增。
