The wheat cultivar Henong 6425 was chosen as the experimental material and was acquired from Tianjin Academy of Agricultural Sciences. Henong 6425 is susceptible to powdery mildew. The wheat seedlings were grown in hydroponics as described previously [38 (link)]. At the two-leaf stage, uniformly growing seedlings were chosen for subsequent PW inoculation. The fresh conidia of Bgt, whose virulence type is E09, on heavily diseased seedlings were shaken off onto the second leaves, which were flattened. Leaves were at 0, 3, 6, 12, 24, and 48 h after PW infection, immediately frozen in liquid nitrogen and then stored at -80 °C for subsequent RNA extraction.
A cDNA fragment of 131 bp (+ 1612 bp to + 1742 bp) was employed to acquire the TaCDPK27-silenced vector. TaCDPK27-silenced wheat seedlings were acquired with barley stripe mosaic virus (BSMV)-based virus-induced gene silencing (VIGS), as described by Yue et al. (2022) [14 (link)]. Control (containing wild-type (CK) seedlings and fourth leaf-stage BSMV-VIGS-inoculated seedlings that displayed chlorosis (the materials of which included BSMV-VIGS-GFP-inoculated seedlings (γG) and TaCDPK27-silenced seedlings) were inoculated with PW causal agent conidia. Fungal structures were subsequently stained withCoomassie brilliant blue and visualized via microscopy.
A cDNA fragment of 131 bp (+ 1612 bp to + 1742 bp) was employed to acquire the TaCDPK27-silenced vector. TaCDPK27-silenced wheat seedlings were acquired with barley stripe mosaic virus (BSMV)-based virus-induced gene silencing (VIGS), as described by Yue et al. (2022) [14 (link)]. Control (containing wild-type (CK) seedlings and fourth leaf-stage BSMV-VIGS-inoculated seedlings that displayed chlorosis (the materials of which included BSMV-VIGS-GFP-inoculated seedlings (γG) and TaCDPK27-silenced seedlings) were inoculated with PW causal agent conidia. Fungal structures were subsequently stained withCoomassie brilliant blue and visualized via microscopy.
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