Purification of Total RNAs and Small RNAs from Fruit Tree Leaf Tissues

Abstract

Perennial fruit crops are susceptible to many viral pathogens, which often lead to declines in quality and yield. For the production of good quality and virus-free propagation materials, conventional molecular detection methods combining high throughput sequencing technology have been widely applied to virus detection and discovery in fruit trees. Recovery of high-quality RNAs from fruit tree leaf tissues, the critical step for the subsequent molecular analysis, is often complicated by the presence of high levels of RNases and problematic biomolecules. Therefore, the universal extraction methods often require modification according to different properties of various tissues. In this chapter, we provide a set of methods that have been used successfully to isolate total RNAs and small RNAs from various fruit tree leaf tissues and as examples, presented in detail of a modified TRIzol method for total RNAs purification from mulberry (Morus alba L.) leaf tissues and an alternative small RNAs purification protocol using mirVana™ miRNA isolation kit (Ambion/Life Technologies) for some fruit tree leaf tissues. The protocols described here aim to provide examples of what have worked successfully for a range of fruit trees and may be successful for a given sample in the future.

Keywords: Fruit tree; Leaf tissue; Small RNA purification; Total RNA purification.