Anatomical and histochemical characterization of in vitro haustorium from roots of Castilleja tenuiflora

In vitro induction of haustoria from Castilleja tenuiflora Benth. was achieved by applying 25 μM catechin, 25 μM vanillin, or 25 μM H₂O₂. Of the treatments tested, 25 μM vanillin was the strongest inducer of haustoria in C. tenuiflora roots in vitro (up to 3 haustoria per root). Haustorium development occurred laterally and was observable 14 d after inducer application. It was characterized by elongation of the epidermal cells and division of the inner cortical cells which also possessed abundant nuclei. Histochemical analysis using 3,3-diaminobenzidine (DAB) and diphenylboric acid 2-aminoethyl ester (DBPA) indicated that the formation of haustoria was associated with the accumulation of H₂O₂ and flavonoids.