Differential scanning calorimetry（ DSC） was used to measure the thermal hysteresis activity(THA) of plant antifreeze proteins(AFPs). The results reveal that DSC is a good method to screen and study AFPs. In the sixteen components extracted from Ammopipanthus mongolicus leaves, one(P3S1) was found to have apparent thermal hysteresis activity by DSC. As the amount of ice nuclei in the sample decreased, the THA of P3S1 increased from 0.01℃ to 0.65℃ . It is notable that the two- peak thermal hysteresis effect was observed. Two endothermic peaks appeared in the melting process of P3S1, while the freezing peak also consisted of two peaks. The peaks appeared antecedently showed larger thermal effect. This phenomenon shows P3S1 has two different kinds of interaction with water and ice crystal. It is probably an important property of a class of AFPs.
The conventional protein chromatography technique was adopted to purify the antifreeze proteins (AFPs) from the leaves of Ammopiptanthus mongolicus (Maxim.) Cheng f. Two bands on native PAGE gel showed thermal hysteresis activity, one was band B1, whose thermal hysteresis was 0.46 ℃ at 8 g/L, which showed two bands (67 kD, 21 kD) on SDS_PAGE gel; the other was B3, whose thermal hysteresis was 0.45 ℃ at 10 g/L, and it contained only a single protein (39.8 kD). Both B1 and B3 are not glycoproteins, because neither do they interact with Shiff_reagent, nor show ultraviolet characteristics of a typical glycoprotein.
Ultrastructure of sperm cells in pollen tube of Amaryllis vittata Ait. has been investigated in details by electron microscopy ,with particular emphasis on the organization and distribution of microtubules.The two newly formed sperm cells are arranged in tandem and sometimes in transverse at the right angle to the long axis of the pollen tube.Thevegetative nucleus is ahead and closely associated with the two sperm cells in all examined pollen tubes. The microtubules are distributed in the region between the common cell wall and the proximity of the sperm nucleus,they are singles and dispersed with mainly orientation of longitudinal and oblique,forming a loose bucket-like structure as a whole.In late stage of development,all of the microtubule array longitudinally and enclose the sperm nucleus.This configuration is very similar to the basket-like structure of microtubule in the generative cell. These results show that the organization and distribution of microtubules in the sperm cells are dynamic during cell development.
