Aminoacyl-tRNA synthetase gene alignments from multiple Sileneae species generated from full-length transcripts using Iso-Seq and raw microscopy image files

Trimmed and untrimmed alignments for the final aminoacyl-tRNA synthetases in Sileneae species and Arabidopsis thaliana. We investigated the evolution of subcellular localization of aaRS enzymes in five different species from the plant lineage Sileneae that has experienced extensive and rapid mitochondrial tRNA loss. By analyzing full-length mRNA transcripts with single-molecule sequencing technology (PacBio Iso-Seq) and searching genome sequences, we found instances of predicted retargeting of an ancestrally cytosolic aaRS to the mitochondrion as well as scenarios where enzyme localization does not appear to change despite functional tRNA replacement.
Nikon .nd2 raw microscopy files for the transient expression and imaging of predicted transit peptides and colocalization assays in N. benthamiana epithelial cells. The amino acid sequence plus 10 upstream amino acids of the protein body were fused to GFP and co-transfected with an eqFP611-tagged transit peptide from a known mitochondriall..., For trimmed and untrimmed alignments:
RNA was extracted from A. githago (hermaphrodite), S. conica (hermaphrodite), S. latifolia (male), and S. vulgaris (male-fertile hermaphrodite) with a Qiagen RNeasy Plant Mini Kit, using RLT buffer with 10 µl beta-mercaptoethanol. RNA was DNase treated with a Qiagen RNase-Free DNase Set. Separate RNA extractions were performed on leaf tissue and an immature flower sample (~5 days post flower development) for A. githago, S. vulgaris, and S. latifolia. Two different tissues were used to increase detection of diverse transcripts, but the two RNA samples were pooled equally by mass for each species prior to library construction, so individual reads cannot be assigned to leaf or floral tissues. Only leaf tissue was used for S. conica as the individual had not yet begun flowering at the time of RNA extraction. Both tissue types were harvested at 4 weeks post-germination, and RNA integrity and purity were checked on a TapeStation 2200 and a Nanodrop 2000.
..., These files can be viewed using Nikon's Element software, can also be viewed using ImageJ for PC (but not Mac).