transcriptome of wild-type and clv1-101 clv2-101 rpk2-5 upon beet cyst nematode infection

The goal of this project is to identify Arabidopsis CLE signaling downstream genes involved in syncytium formation during beet cyst nematode (Heterodera schachtii) infection.Wild-type (Col-0) and clv1-101 clv2-101 rpk2-5 seedlings were grown on modified Knops media in square plates (100 x 100 x 15 mm) vertically for 7 days. The growth chamber was set at 24 degree Celsius with 12 h light/12 h dark light cycle. About 10-15 sterilized BCN (H.schachtii) pre-parasitic 2nd stage juveniles were inoculated to each root. Developing syncytium (about 8 mm root segments) were collected from primary roots at 4 dpi (days-post-inoculation) under stereoscope and immediately frozen in liquid nitrogen.Samples treated with HsCLE2 peptide were also included to filter out genes specifically response to the HsCLE2 peptide effector. 11-day-old seedlings grown in square plates were treated by covering all roots by adding 3 ml of 5 uM HsCLE2 solution to the plate. Plates were then kept in the growth chamber for 3 hours. Root segments corresponding to BCN infection sites (about 8 mm) were collected under stereoscope and immediately frozen in liquid nitrogen for RNA isolation.