Seedlings of the sweet fern, Comptonia peregrina (L.) Coult., grown aeroponically, were inoculated with a nodule suspension to allow infection by the actinomycete-like organism which causes nodule formation. Roots with early stages of infection and nodule initiation were fixed, embedded in resin, sectioned, and examined. Infection is infrequent in Comptonia with only a few nodules per seedling root system. Infection via root hair invasion causes the retention of the curled and deformed root hair in an intensely cytoplasmic state with ramification of multiple filamentous strands of the endophyte. A limited cortical proliferation occurs in response to the infection forming the prenodule; endophyte filaments grow radially inward from the base of the infected epidermal root hair and invade a portion of the prenodular cells resulting in their hypertrophy. Distal and proximal to the prenodule site, a number of primary nodule primordia are initiated, varying from a few up to a dozen or more. These primordia appear to develop more or less simultaneously under the stimulus of the invading endophyte; they are like lateral roots in their site of origin, occurring largely opposite the protoxylem poles and involving pericyclic and endodermal cell proliferation. They differ in that the cortical cells external to each primordium are stimulated to undergo divisions and these cortical cell derivatives are incorporated into the developing primordium. The endophyte enters the cortical tissues of the lateral root on which the prenodule has formed and then invades proximal and distal to the infection site, progressing into the cortical tissues of each of the developing nodule primordia. A cork-like layer develops on the original lateral root in areas not occupied by primordia by initiation of subepidermal cell divisions and wall thickening. Normal lateral root primordium formation occurs in the pericycle opposite the protoxylem poles and involves cellular derivatives of the pericycle and endodermis but no cortical cells, which instead are crushed and displaced by the lateral root primordium as it develops. Nodule formation clearly involves complex chemical interactions, which remain for further study, between the host cells and the invading endophyte.
The Auxin-Regulated AP2/EREBP Gene PUCHI Is Required for Morphogenesis in the Early Lateral Root Primordium of Arabidopsis