Figure 3: Ectopic expression and heat induction of Mds-1 in resistant wheat.

(a) Ectopic expression of Mds-1 correlates with a phenotype switch of Molly plants from resistance to susceptibility to Hessian fly biotype GP infestation. Northern blot analysis of Mds-1 transcript in samples from transgenic Molly lines T640, T1166 and T1417 (Supplementary Table S2, Supplementary Fig. S9); non-transgenic, resistant Molly; and the susceptible wheat Newton with (I) and without (U) Hessian fly (biotype GP) infestation. 28S, 18S and Mds-1 (900 bp) along with arrows on the left of the northern blot represent the locations of 28S rRNA, 18S rRNA and Mds-1 mRNA. An 18S rRNA image is given as loading control. Interaction phenotypes are given under the 18S rRNA images with: R, resistant; S, susceptible. (b) Heat stress induces Mds-1. Northern blot and phenotypic analyses of resistant genotype Molly seedlings that were either unstressed (20 °C) or stressed at 30 or 35 °C. (c) The decay of heat-induced Mds-1 transcript and protein correlates with restoration of Molly resistance to Hessian fly. Northern (upper panel), western (lower panel) and phenotypic analyses on Molly plants were conducted during recovery after stress. Seedlings were stressed at 37 °C for 12 or 24 h, and some 24 h-stressed plants were allowed to recover at 20 °C for 3–48 h. Denotations for northern blot and phenotypic analysis are the same as in a. The positions of protein size markers and the location of the Mds-1 protein are given on the left of the western blot. A coomassie-blue stained gel image is give under the blot as loading control. Plants were infested with Hessian fly biotype GP.