## 7 Building a Molecular Toolbox for Engineering of Targeted Disease Resistance
### Materials and Methods for the Wheat Orthologue of Arabidopsis DORN1  
The action of extracellular ATP in Arabidopsis, acting through its receptor DORN1, has led us to consider characterizing the wheat orthologue of DORN1 in interactions with nematodes and fungal necrotrophic root pathogens. A collection of wheat and barley orthologues predicted to contain lectin-binding, kinase, and transmembrane domains were obtained from published literature [113; 114]. Candidate wheat or barley orthologues were identified using HMMER 3.0 [115] and TMHMM [116] and databases IWGSC URGI and International Barley Sequencing Consortium genome coding sequences [110], for wheat and barley, respectively. Another wheat orthologue search based on amino acid sequence identity was conducted in 2019 using Blast and the IWGSC RefSeq v1.0 coding sequence database. For this search, GenBank accessions of \(DORN1\) nucleotide sequences from the brassicas _Arabidopsis thaliana_, _A. Iyrata_, and _Camelina sativa_ were obtained. The coding regions of the brassica sequences were used to find IWGSC (wheat) accessions with the highest sequence identity. Deduced \begin{table}
\begin{tabular}{c c c c c c c c} \hline \hline  & & & \multicolumn{3}{c}{**RPKM 1**} & \multicolumn{3}{c}{**Fold-induction 2**} \\ \hline
**Annotation/locus ID 3** & **Cont** & **Pn** & **Pt** & **Pn+Pt** & **Pn** & **Pt** & **Pn+Pt** \\ \hline
**Dehydrin DHN2** & & & & & & & \\
**Tases\_5 DL\_134F29727.1** & 26.6 & 18.52 & 161.92 & 133.76 & ns & 6.09 & 5.03 \\
**German-like protein 2** & & & & & & & \\  
ate embryogenesis abundant protein, group 3** & & & & & & & \\
**Tases\_1 BL\_558 CE4 FE2F1.1** & 10 & 6 & 263.51 & 249.8 & ns & 26.35 & 24.98 \\
**Tases\_1 AL\_34404D5D8.2** & 7.34 & 3.79 & 149.17 & 140.26 & ns & 20.31 & 19.1 \\
**Tases\_1 BL\_5 CD8 BF94C.2** & 4.09 & 2.62 & 152.77 & 145.16 & ns & 37.36 & 35.5 \\
**Tases\_1 AL\_34404D5D8.4** & 3.91 & 1.83 & 80.85 & 74.72 & ns & 20.68 & 19.12 \\
**Tases\_1 BL\_55 DR94C.3** & 3.12 & 2.58 & 132.86 & 128.74 & ns & 42.58 & 41.26 \\
**Tases\_1 BL\_58 CE7 FE1.2** & 2.61 & 2.46 & 69.06 & 64.1 & ns & 26.46 & 24.56 \\
**Tases\_1 BL\_SEAAF6001.1** & 2.42 & 1.2 & 61.75 & 64.26 & ns & 25.49 & 26.52 \\
**Tases\_1 BL\_SDSR94C.3** & 2.31 & 1.21 & 51.63 & 47.21 & ns & 22.35 & 20.44 \\
**Tases\_1 BL\_SAF6001.1 & 2.09 & 1.94 & 63.53 & 69.99 & ns & 30.35 & 33.44 \\
**Tases\_1 BL\_SCDBF894C.1** & 1.89 & 0.88 & 55.33 & 49.77 & ns & 29.31 & 26.36 \\
**Major pollen allergen Set v1-D/H** & & & & & & & \\
**Tases\_4S\_CA\_SE1 BBBDD1** & 21.2 & 60.77 & 150.59 & 169.3 & 2.87 & 7.1 & 7.98 \\
**Major pollen allergen Set v1-F/H** & & & & & & & \\
**Tases\_2 DL\_B766A2857.1 & 45.11 & 94.39 & 201.8 & 215.2 & 2.09 & 4.47 & 4.77 \\
**Tases\_2 DL\_B76A2857.2 & 41.7 & 87.33 & 179.9 & 191.89 & 2.09 & 4.31 & 4.6 \\
**Tases\_4 DL\_SC868874E.1** & 36.53 & 56.19 & 145.46 & 154.27 & ns & 3.98 & 4.22 \\
**Tases\_2 DL\_194C61 EBA.1 & 33.63 & 53.16 & 137.44 & 145.51 & ns & 4.09 & 4.33 \\
**Metascalesser** & & & & & & & & \\
**Tases\_5 AL\_SD58A83 BF8.1 & 98.48 & 82.64 & 122.31 & 216.42 & ns & ns & 4.2 \\
**Pathogenesis-related protein 1** & & & & & & & \\
**Tases\_2 DL\_90 ZD78D4C1.1** & 39.84 & 65.82 & 169.57 & 167.29 & ns & 4.26 & 4.2 \\
**Unknown protein** & & & & & & & \\
**Tases\_1 DS\_57B13D45E6.3 & 171.05 & 368.87 & 177.81 & 365.82 & 2.16 & ns & 2.14 \\
**Tases\_1 DS\_57B13D45E6.1** & 48.48 & 119.79 & 52.23 & 122.46 & 2.47 & ns & 2.53 \\ \hline \hline \end{tabular} \({}^{1}\) Cont, control; Pr, P. neglectus; Pt, P. thornei; PnPt, P. neglectus and P. thornei. 2 Fold-induction in nematode-treated roots compared to the control. ns = fold-change <2.0. 3 Transcript annotation was assigned using Blast2 GO (_BioBam Bioinformatics S.L., Valencia, Spain)_; transcript locus identifiers (ID) were determined by mapping transcript group sequences to the International Wheat Genome Sequencing Consortium (IWGSC) RefSeq v1.0 database [110].  
\end{table}