Changes in proteins and peroxidases induced by compatible pollination in the ovary of Nicotiana tabacum L. ahead of the advancing pollen tubes

Abstract

Proteins and peroxidases produced by the ovules and placenta of tobacco (Nicotiana tabacum L.) in response to compatible pollination were analyzed by two-dimensional polyacrylamide gel electrophoresis and by enzyme staining in flat-bed native isoelectric focusing gels. For two-dimensional gels, ovaries were sampled at 36 h after pollination, at which time pollen tubes have penetrated much of the length of the style but have not yet entered the ovary. At least 11 major proteins from pollinated ovaries had no detectable counterparts in unpollinated ovaries. These showed a range of molecular mass and pI. For peroxidase isozyme assays, ovaries were sampled at 0, 12, 24, 36 and 48 h after pollination. At 45–50 h, pollen tubes were beginning to enter the top of the ovary but could still be separated from the ovules and placenta during sampling. Ovules and placentae from unpollinated pistils showed only one form of peroxidase, whereas those from pollinated pistils showed additional isozymes at pH 5.4 and pH 10.0. Both new isozymes increased in staining intensity over the first 36 h after pollination. At 48 h, however, the acidic peroxidase had continued to increase, while the basic component had declined so as to be barely detectable. The observations are discussed in relation to accumulating evidence that some form of pollination-induced signal reaches the ovary ahead of the advancing pollen tubes. The nature of this signal and possible involvement of peroxidases are also briefly discussed.