RNA-Sequencing of asparagus samples during fall

Asparagus (Asparagus officinalis L.) is an important vegetable crop in southern Ontario, Canada, where winter air and soil temperatures below zero degrees are common. Consequently, cultivars growing in this area must be winterhardy. Research to date in asparagus freezing tolerance has determined the importance of cryoprotectants such as fructans, glucose, protein, and proline providing a narrow perspective on cellular metabolism and signaling. This study aimed to conduct transcriptomic analysis (RNA-seq) during fall acclimation and spring deacclimation in two cultivars (Guelph Millennium, GM and UC157, UC) with varying adaptation to southern Ontario. RNA was extracted from bud and rhizome tissues, sampled on three dates during early spring and late fall. High-quality RNA (260/280 ratio > 2, RIN > 7, concentration > 1000 ng/ul) was used for sequencing. NEB (New England Biolabs Ltd.) Next Ultra II Directional polyA mRNA library prep kit (Cat #E7760L, New England Biolabs, Whitby, Ontario, Canada) was used to construct libraries from the mRNA. The cDNA libraries were sequenced on two lanes of an Illumina NovaSeq 6000 SP flowcell (RRID:SCR_016387) with a read length of 2x 100 bases (paired-end). The findings of this project will help researchers understand the molecular mechanisms of cold acclimation and freezing tolerance in asparagus, leading to breeding and genetic strategies to improve the trait.