Improved Protocol of RNA Isolation for Transcriptome Analysis of Poaceae Plants

The existence of metabolites that interfere with isolation procedures and downstream applications makes plant RNA extraction difficult. The current research used a standardized RNA extraction protocol from maize (Zea mays L.). We developed a protocol for extracting pure RNA from plant tissues using both extraction buffer and the TRIzol reagent, and show that this RNA extraction method works not only at Low temperatures but also at room temperatures, making it the easiest and most efficient method for extracting pure and undegraded RNA directly from tropical plants in the field. RNA isolation methods based on our modified protocol yielded good results in maize leaf, seed, flowers and other grass species. The isolated RNA was found to be suitable for both PCR and RT-PCR amplification and transcriptome analysis. The method is repeatable and can be used to isolate high-quality RNA and conduct gene expression studies. RNA extraction paves the way for deciphering the complex regulatory network involved in multiple stress responses by studying gene-environment interactions at the transcriptome level.