Amplification primers

PCR products were sequenced using the amplification primers (Eurofins Genomics, Germany). Sequence data quality was checked and edited using Geneious Prime software (version 2020.0.3, Biomatters Ltd.) [51 (link)]. The resulting sequences were aligned and compared with the Ae. aegypti reference sequence from VectorBase (AaegL5.0 Transcript: AAEL023266; Liverpool strain). Following convention, kdr mutations are numbered with reference to the homologous house fly Musca domestica vgsc. Due to the 233-base-pair (bp) intron in domain IIS6, between the S989P and V1016G mutations, some PCR products were cloned when sequences could not be read from direct sequencing of the PCR product (see the method in Additional file 1: Text S1).

Total RNA extraction and reverse transcription were performed using a RNeasy Mini Kit (Qiagen, Hilden, Germany) and a PrimeScript RT MasterMix (Perfect Real Time; Takara Bio, Shiga, Japan). Transcript levels of TSLP, IL-25, IL-33, CCL4, CCL5, CCL11, CCL26, TSLPR, TLR3, CCR5, ICAM-1, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were quantified by RT-PCR using a Rotor-Gene SYBR Green PCR kit (Qiagen, Hilden, Germany) on a Rotor-Gene Q HRM (Corbett Research, Cambridge, UK). Amplification primers (Eurofins Genomics, Tokyo, Japan) are shown (Supplementary Table S1).

To amplify SSPbP53, genomic DNAs (gDNA) of Plasmodiophora brassicae pathotype 3 H (Pb3H) obtained from Dr. Gary Peng (AAFC-Saskatoon Research Center) and single spore isolates SACAN-SS₃ (Pb2), ORCA-SS₃ (Pb5), AbotJE-SS₃ (Pb6), and CDCN-SS₁ (Pb8) [31 (link)], were used. PCR amplification used Phusion High-Fidelity DNA Polymerase (Thermo Fisher Scientific, CAD) in a 50 μL final volume containing 300 nM of each primer (gSSPbP53F/gSSPbP53R, Supplementary Table S1) and 2 to 4 ng of gDNA. The 530 bp amplicon generated for each pathotype was purified using GeneJET PCR Purification Kit (Fermentas Life Science, CAD) and directly sequenced with the amplification primers (Eurofins Scientific, CAD).