Figure 3.

Rescue Assays in M. truncatula dmi1 Mutant with Lj-CASTOR, Lj-POLLUX, DMI1, and Modified Alleles.

(A) to (C) Rescue of the M. truncatula dmi1 mutation by Lj-CASTOR and Lj-POLLUX.

(A) Lj-CASTOR expressed alone does not rescue nodulation phenotype in dmi1 when inoculated with S. meliloti (pXLGD4).

(B) Lj-POLLUX expressed alone does not rescue nodulation phenotype in dmi1 mutant as only immature nodules or nodule bumps were observed in few instances.

(C) Lj-CASTOR and Lj-POLLUX together rescue dmi1 as mature functional nodules were formed.

(D) and (E) Rescue of M. truncatula dmi1 mutation by DMI1, Lj-POLLUX^S329A, and DMI1^A294S.

(D) DMI1 rescues nodulation phenotype in dmi1 mutant.

(E) Lj-POLLUX^S329A gains function and rescues dmi1 without Lj-CASTOR.

(F) DMI1^A294S does not rescue dmi1 when paired with Mt-CASTOR-RNAi.

Mature nodules are shown by white arrows; nodule bumps are shown by white arrowheads. In (C), the double transgenic roots of dmi1 line expressing Lj-CASTOR and Lj-POLLUX were colocalized for GFP (for Lj-CASTOR) and DsRED1 (for Lj-POLLUX), yielding yellow to orange fluorescent roots. In (F), the double transgenic roots of dmi1 line expressing DMI1^A294S and Mt-CASTOR-RNAi were colocalized for GFP (for DMI1^A294S) and DsRED1 (for Mt-CASTOR-RNAi), yielding yellow to orange fluorescent roots. In the figures, lowercase, italicized words correspond to the mutant line and uppercase, italicized words correspond to wild-type gene used for rescue assay. Bars + 5 mm.