Table 1.
Description of the seven selected CSEPs.
Fig 1.
HIGS-mediated silencing of CSEP0081 and CSEP0254 decreased Bgh haustorial formation success.
Barley leaves, bombarded with RNAi and GUS reporter constructs, were infected with Bgh and scored for fungal haustoria formation. The haustorial formation rate was calculated as the ratio of haustoria-containing transformed cells (GUS expressing cells) divided by the total number of transformed cells. Relative haustorial formation rate was computed relative to the empty vector control of each experiment, which was set to 100%. Data shown are mean values of five independent experiments, but CSEP0145, CSEP0216, CSEP0222, CSEP0398 and Mlo were excluded from two of those. All values are presented ± SE. ** refers to significant differences compared to the empty vector control (P < 0.01). Arrow indicates haustoria. Scale bar, 20 μm.
Fig 2.
Relative expression levels of CSEP0081 and CSEP0254 during Bgh infection.
The CSEP transcripts were monitored using RT-qPCR. Total RNA was isolated from Bgh-infected barley leaves. Haustorial and epiphytic expression was analysed separately at 24 and 48 hpi (H and E, respectively). Expression of Bgh glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used to normalize the CSEPs expression in each sample. Expression was determined relative to 0 hpi, arbitrarily set to one. Data represent means of three independent biological repeats, each with two technical repeats, ± SE. Statistical analysis of biological repeats: t test relative to 0 hpi; **, P<0.01; ***, P<0.001.
Fig 3.
CSEP0081 fused to YFP localizes to the cytosol and the nucleus of barley leaf epidermal cells.
Construct encoding CSEP0081, lacking signal peptide, with YFP fused to its C-terminus was co-expressed with a free mCherry marker construct in barley leaf epidermal cells using particle bombardment. Localization was monitored 1 to 3 days later. The CSEP0081-YFP fusion proteins co-localized with mCherry in the cytosol and the nucleus, shown by a complete merge of yellow and red signals. C, cytosol. N, nucleus. Scale bar, 20 μm.