Figure 4.
The Function of miR172c in Regulating Nodule Development Is Dependent on NFRs.
(A) qRT-PCR analysis of miR172c in response to NF treatment. Four-day-old seedlings were treated with distilled, deionized water (−NF) or 10−8 M B. japonicum NF (+NF), and the roots were collected at 3 DAI (n = 5).
(B) to (D) qRT-PCR analysis of miR172c expression in wild-type cv Bragg (B) and its isogenic nonnodulation mutants nod49 (C) and nod139 (D), which harbor mutations in NFR1α and NFR5α, respectively (n = 5). Seven-day-old seedlings were inoculated with B. japonicum strain USDA110. The roots were harvested at 0, 1, 3, 5, and 10 DAI.
(E) Image of hairy roots from the NFR1α mutant nod49 expressing empty vector (EV) or 35S:miR172c at 28 DAI. Bar = 700 μm.
(F) qRT-PCR analysis of miR172c expression in transgenic roots of the NFR1α mutant nod49 expressing 35S:miR172c (n = 5).
For all gene expression results shown, miR1520d was used as an internal control. Expression levels are shown as means ± se from three replicates. Asterisks represent statistically significant differences (Student’s t test, ***P < 0.001).
[See online article for color version of this figure.]