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. 2023 Feb 11;4(4):100561. doi: 10.1016/j.xplc.2023.100561

Figure 6.

Figure 6

MoLrp1 is involved in the cAMP signaling pathway and is critical for accumulation of MoMsn2 from the cytosol to the nucleus during the early infectious stage.

(A) HPLC analysis of intracellular cAMP levels of Guy11 and ΔMolrp1 cultured in CM with or without Ole treatment. Error bars are SDs of three biological replicates, and asterisks indicate significant differences by two-tailed Student’s t-test at ∗∗p < 0.01. CK, no treatment.

(B) Pathogenicity of the indicated strains on detached barley leaves with or without exogenous cAMP treatment.

(C) Statistical analysis of disease area caused by the indicated strains. Error bars are SDs of three biological replicates, and asterisks indicate significant differences by two-tailed Student’s t-test at ∗∗p < 0.01.

(D) Hyphae of Guy11 and ΔMolrp1 co-expressing MoMsn2-GFP and H1-RFP proteins were observed under a confocal microscope after treatment with cAMP, Ole, or MDL-12330A (a type of adenylate cyclase inhibitor) for 10 min. Insets highlight areas analyzed by line scan. The green line represents MoMsn2-GFP; the red line represents H1-RFP. Scale bar, 10 μm.

(E and F) Conidial suspensions of Guy11 and ΔMolrp1 co-expressing MoMsn2-GFP and H1-RFP proteins were inoculated into rice sheaths, and we observed MoMsn2-GFP accumulation in IHs at 24 hpi. Insets highlight areas analyzed by line scan. The green line represents MoMsn2-GFP; the red line represents H1-RFP. Scale bar, 20 μm.

(G) Expression profiles of MoLRP1 and MoDCI1 at different infectious stages. RNA was extracted from hyphae cultured for 36 h (HY) and rice leaves inoculated with wild-type conidia for 24, 48, and 72 h, respectively. Error bars are SDs of three biological replicates, and statistical analysis was performed by two-tailed Student’s t-test.

(H) qRT–PCR analyses of the expression levels of MoLRP1 in Guy11 and MoDCI1 in Guy11 and ΔMolrp1 with or without Ole treatment. Error bars are SDs of three biological replicates, and asterisks represent significant differences by two-tailed Student’s t-test at ∗∗p < 0.01.