Figure Legends
Figure 1. (a) Relative expression of Os10g39300 after BPH infestation. Asterisks represent significant differences based on one-way ANOVA and Tukey test (*, P < 0.05; **, P< 0.01). (b) Phylogenetic analysis of the conserved ASP domain from rice OsAPs and RBPH16. The phylogenetic tree was constructed by the neighbor-joining method in MEGA 4.0 software (p-distance model; 1000 bootstraps). Scale bar represents 0.2 amino acid substitution per site.
Figure 2. BPH resistance evaluation of lines overexpression (OE) and knockout (CR) of OsAP79 . BPH-resistance assay of lines OE, CR and Nipponbare (NIP) at seedling (a) or adult stage (c). Scale bar, 5 cm. Survival rates of seedling (b) and adult (d) detected with the homozygous T2 lines of OE and CR of OsAP79 . BPH weight gain (e) and Honeydew excretion (f) by newly emerged female adult BPHs fed on the lines of OE, CR and NIP for 48 h. Asterisks represent significant difference based on one-way ANOVA and Tukey test (*, P < 0.05; **, P < 0.01).
Figure 3. Molecular characteristics of OsAP79 . (a) Relative expression of gene OsAP79 in Nipponbare (NIP) after BPH infestation. (b) Relative expression of OsAP79 in different organs of NIP. Asterisks in (a) and (b) represent significant differences based on one-way ANOVA and Tukey test (*, P< 0.05; **, P < 0.01. (c) Rice protoplasts were co-transfected with endoplasmic reticulum-red fluorescent protein (ER-RFP) and OsAP79-GFP. The transfected cells were examined under a confocal microscope. The images were then processed using ZEISS ZEN confocal microscope software. Overlapping fluorescence signals show that OsAP79 localizes to the ER. Bright-field images are shown on the right. Scale bars, 10 μm.
Figure 4. Root length detection of lines overexpression (OE), knockout (CR) and Nipponbare (NIP). (a) Five days after germination, plants were grown hydroponically for 30 days under normal nutrient solution conditions. Scale bars: 1 cm. (b) The root growth phenotype of plants growing in the soil. Adults grown on were separated carefully from the soil transferred. Scale bars: 10 cm. Main root length was measured at seedlings (c) and adults (d). (e) Relative root length rate in 5d and 10d (1μM IAA treated versus untreated in each genotype, respectively). IAA (f) and SA (g) content of 20-day-old normal grown seedlings. Asterisks in (c, d, e, f and g) represent significant differences based on one-way ANOVA and Tukey test (*, P< 0.05; **, P < 0.01).
Figure 5. Rice sclerenchyma layer thickness detection. Representative images of making free hand sections to detect the sclerenchyma layer thickness in stem cross-sections of lines overexpression (OE), knockout (CR) and Nipponbare (NIP). (a) seedling; (b) adult. Scale bars, 1000μm. The thickness of sclerenchyma layer in stem transverse sections at seedling (c) and adult stage (d) was measured. Asterisks in (c,d) represent significant differences based on one-way ANOVA and Tukey test (*, P < 0.05; **, P< 0.01).
Figure 6. Metabolites were mapped onto network of metabolic pathways. The contents of metabolites after BPH feeding at different time were represented by Graphics with different Position. Each graph represents the fold change considering the time 0h as a control; columns represent the two evaluated times for each metabolite. The first row corresponds to the Nipponbare (NIP), whereas the second row corresponds to the overexpression (OE) (a) or knockout (CR) (b). Red symbols denote significant increases (P < 0.05); black symbols denote no significant changes (P > 0.05); green symbols denote significant decreases (P < 0.05). The upstream and downstream relationship of these metabolites was indicated by direction of arrow. Solid lines denote direct correlations among each of metabolites. Dotted lines denote that relative long pathway transformation occurred among each of metabolites.
Table 1 Agronomic trait detection of Nipponbare, overexpression (OE) and knockout (CR) lines.