Abstract
Among four full-length cDNA clones encoding cell-wall purple acid phosphatase(NtPAP) from tobacco cells, the mRNA level of NtPAP12 increased in proportion to the wall regeneration. In order to study the role of NtPAP, NtPAP12 was constitutively over expressed in cultured tobacco XD-6 cells. At the early stage of wall regeneration of protoplasts isolated from the 7day-old cells over-expression of NtPAP accelerated the deposition of beta-glucan, which was observed using Calcofluor stain. The enhancement of the deposition reached maximum at 60 min on the surface of plasma membrane. In order to evaluate the amount of the deposition of NtPAP on the surface of plasma membrane of the transformed protoplasts within 60 min during cell wall regeneration, the concentration of NaCl to elute NtPAP was first determined using batch method. The amount of the deposition of NtPAP increased about two times as much as that of the wild-type protoplasts.
