RNA sequencing of leaves and roots of transgenic potato (Solanum tuberosum) plants overexpressing the StCLE4 and StCLE4G6T genes
Nitrogen is an essential macronutrient for plant growth and development, and it is often a limiting factor in crop production. We found that StCLE4 gene was positively regulated by nitrogen supply and suggest that StCLE4 could be negative regulator of tuberization. Overexpression of StCLE4 affected shoot apex and root growth and also promoted stolon conversion into branches. We performed of RNA-Seq analysis of leaves and roots of transgenic (StCLE4 and StCLE4G6T overexpression) and control plants. The plants were propagated by two-node stem cuttings and were cultivated in vitro on a Murashige and Skoog (MS) medium containing 0.8% (w/v) agar and 1% sucrose under long-day conditions (16 h light:8 h dark) at 22C. N-depleted or N-rich medium was prepared as described in (Ohkubo et al., 2017). For RNA-Seq analyses, total RNA was extracted from samples using the RNeasy Plant Mini Kit (Qiagen, USA). Enrichment of mRNA was performed using NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, USA). The libraries were prepared using NEBNext Ultra II Directional (New England Biolabs, USA). For sequencing HiSeq PE Rapid Cluster Kit v2 and HiSeq Rapid SBS Kit v2 was used. The libraries were sequenced on the Illumina HiSeq 2500 sequencing platform.