Fig. 8

Analysis of immune effects induced by transient overexpression of three candidate sugarcane miRNAs in Nicotiana benthamiana. A PCR products of the three candidate sugarcane pre-miRNAs detected by electrophoresis in the same gel. M, DNA marker 15,000 + 2,000 bp; 1, pre-nov_3741; 2, pre-nov_22650; and 3, pre_nov_40875. B The transcript of three candidate miRNAs in N. benthamiana leaves at 2 d after agroinfiltration by qRT-PCR analysis. C DAB staining in N. benthamiana leaves infiltrated by Agrobacterium tumefaciens for 2 d. D The transcript of the immune-related marker genes at 2 d after agroinfiltration by qRT-PCR analysis, including the hypersensitive response marker genes HSR201, HSR203 and HSR515, the salicylic acid-responsive genes NPR1, PR-1a/c, PR2 and PR3, and the ethylene synthesis-depended genes EFE26 and Accdeaminase. EF1-α was used to normalize the transcript level. All data points are the means ± SE (n = 3). Different lowercase letters indicate a significant difference (p-value < 0.05). 35S::00 represents the A. tumefaciens strains carrying the empty vector pCAMBIA1301. 35S::3741, 35S::22650 and 35S::40875 represent the A. tumefaciens strains carrying the recombinant vector pCAMBIA1301-nov_3741, pCAMBIA1301-nov_22650 and pCAMBIA1301-nov_40875, respectively