Together with their undeniable role in the ecology of arid and semiarid ecosystems, Agave species are emerging as a model to dissect the relationships between crassulacean acid metabolism and high efficiency of light and water use, and as an energy crop for bioethanol production. Transcriptome resources from economically valuable Agaves species, such as Agave tequilana and A. salmiana, as well as hybrids for fibers, are now available, and multiple gene expression landscape analyses have been reported. Key components in molecular mechanisms underlying drought tolerance could be uncovered by analyzing gene expression patterns of roots. This study describes an efficient protocol for high-quality total RNA isolation from phenolic compounds-rich Agave roots. Our methodology involves suitable root handling and collecting in the field and using saving-time commercial kits available. RNA isolated from roots free of lignified out-layers and clean cortex showed high values of quality and integrity according to electrophoresis and microfluidics-based platform. Synthesis of long full-length cDNAs and PCR amplification tested the suitability for downstream applications of extracted RNA. The protocol was applied successfully to A. tequilana roots but can be used for other Agave species that also develop lignified epidermis/exodermis in roots.

连同它们在干旱和半干旱生态系统生态中不可否认的作用，龙舌兰物种正在成为剖析景天酸代谢与光和水的高效利用之间关系的模型，以及作为生物乙醇生产的能源作物。来自具有经济价值的龙舌兰物种的转录组资源，例如龙舌兰龙舌兰和A. salmiana，以及纤维的杂种，现在都可以使用，并且已经报道了多种基因表达景观分析。通过分析根的基因表达模式，可以揭示耐旱分子机制中的关键成分。本研究描述了一种从富含酚类化合物的龙舌兰根中分离高质量总 RNA 的有效方案。我们的方法包括在田间进行合适的根处理和收集，并使用可节省时间的商业套件。根据电泳和基于微流体的平台，从没有木质化外层和清洁皮层的根中分离的 RNA 显示出高质量和完整性的高价值。长全长 cDNA 的合成和 PCR 扩增测试了提取 RNA 下游应用的适用性。该协议已成功应用于A. tequilana根，但可用于其他龙舌兰品种，这些龙舌兰品种也在根中形成木质化的表皮/外皮。