Transgenic Complementation of nena-1.
(A) to (D) A. rhizogenes–mediated transformation of nena-1 mutants with genomic NENA including 2 kb of 5′ regulatory sequence fused to RFP (NENA:RFP) led to restoration of RNS ([A] to [C]) and AM establishment (D).
(E) to (H) A. rhizogenes–mediated transformation of nena-1 with the 2-kb 5′ regulatory sequence fused to RFP (NENApro:RFP) did not restore RNS ([E] to [G]) and AM (H). Epifluorescence microscopy images show GFP expression in transgenic roots ([A] and [E]) and DsRed expression by M. loti in root nodules ([B] and [F]) using a GFP and a RFP filter, respectively. Corresponding white light illumination images are shown in (C) and (G). Root segments containing AM fungal structures (green) stained with WGA-Alexa Fluor 488 were visualized by DIC/epifluorescence microscopy using a GFP filter ([D] and [H]).
Bars = 2 mm in (A) to (C) and (E) to (G) and 40 μm in (D) and (H).
[See online article for color version of this figure.]