Immunolocalization And The Roles Of Arabinogalactan Protein Epitopes During Zygotic Embryo Development Of Arabidopsis Thaliana

Morphologic character of pollen tube growth and embryo development were observed during sexual reproductive process of Arabidopsis thaliana (Columbia ecotype). Then distribution and the roles of Arabinogalactan proteins (AGPs) during embryo development of Arabidopsis thaliana were also investigated. The main results are as follows:1. With the aid of aniline blue staining, pollen germination and pollen tube growth in pistil tissue were observed in Arabidopsis, and the growth speed of pollen tubes in style and ovary were calculated. The results showed that after pollen germination, the pollen tube grew into the style and the ovary, the growth speed was different when at different stages, and the pollen tube grew faster as time goes on.2. Embryos at different developing stages were manually isolated and observed to understand their morphologic character change as time goes on, then developmental situation difference were compared between the Columbia ecotype and the Landsberg ecotype, the results showed that the developmental speed of the Columbia ecotype was slower than that of the Landsberg ecotype. Furthermore, the lengths of siliques atdifferent developing stages were got and relation between the silique length and time were well understood, the results showed that the siliques grew slower as time goes on.3. Monoclonal antibody (MAb) JIM13 was chosen out by Western blotting to localize AGPs on Arabidopsis embryos at different developing stages by immunolocalization, the results showed that from the octant embryo stage to the heart embryo stage, the green fluorescence distributed mainly on the surface of embryo proper, the top 1-2 cells and basal part of suspensors. And the fluorescence at shoot apex meristem was also extremely strong when at the triangular embryo stage, which was at the transition stage from the globular embryo to the heart embryo stage. However, the green fluorescence disappeared gradually after torpedo stage, only on the shortening suspensors. Red auto fluorescence appeared from the late globular embryo stage.4. BGlcY, a reagent specifically binds to AGPs, with different concentrations (10-lOOjuM) were added into the Arabidopsis in vitro ovule culture medium, the results showed that the development of ovules at zygotic stage, and the embryo differentiation of ovules at globular embryo stage were suppressed, indicating that the AGPs were involved in zygotic embryo development and differentiation in Arabidopsis.In this present paper, isolation methods of embryos at different developing stages in Arabidopsis were established, and the distribution character of AGPs on embryos at different developing stages were detected, furthermore, BGlcY were added in an in vitro ovule culture system to investigate the roles of AGPs in Arabidopsis embryo development process.