RNA-Seq of Allium sativum cloves slices
Healthy garlic cloves with no visible symptoms were selected and a 2 cm2 x 3 mm thick slice of garlic was used for each treatment. As negative-infection control we used garlic slices without nematodes at 0, 24 and 72 hours after the initiation of the experiment (A_0, A_24, and A_72, respectively), and the treatment samples include garlic slices with nematodes during its first contact, and 24 and 72 hours post infection (NA_0, NA_24, NA_72) (Fig. 1). The nematode-garlic (NA) samples were inoculated with 100 L of a suspension containing approximately 10,000 nematodes, mostly in the juvenile stages. The inoculated and non-inoculated garlic slices were placed in a porcelain mortar at room temperature for the time of their respective incubation times (24 or 72 h). Afterwards, liquid nitrogen was added to macerate the samples, which were stored at -80 C until RNA extraction was performed. Three biological replicates of each control and treatment were analyzed. The macerated samples were sent to Langebio Genomic Services Laboratory (LABSERGEN-CINVESTAV, Guanajuato Mexico), for RNA extraction, and the resulting RNA was sent to Macrogen's sequencing RNA-Seq service for library preparation and Illumina sequencing. Paired-end (101bp x 2) library preparation was performed using the TruSeq Stranded mRNA LT Sample Prep Kit