桃果实生长素结合蛋白ABP1的组织定位及蛋白表达分析

doi:10.3864/j.issn.0578-1752.2015.05.10

Abstract

Abstract: 【Objective】Auxin is almost involved in all aspects of plant development regulation, and ABP1 (Auxin binding protein 1) is established as a crucial component of auxin signaling．ABP1 as a kind of auxin fast response protein participates in a series of physiological processes such as regulation of fruit development. It is distributed mainly in the developing embryo and surrounding tissue of embryo. The purpose of this paper is to study whether ABP1 plays a role in peach fruit development and explore its expression characteristics and hence to provide a theoretical foundation for the further research．【Method】The peach ‘No.24’ was used as experimental material. Three periods of development were divided through the assay of growth curve. The fruits from different stages were separated into mesocarp, endocarp and seeds, small tissue was cut from seed and mesocarp, then put them into the EDAC solution, fixed by glutaraldehyde and paraformaldehyde after vacuumizing. The fixed samples continued dehydrated, paraffin embeded, and then used in paraffin section. The other samples which was used for extracting protein immediately froze in liquid nitrogen, then stored at -80℃. ABP1 rabbit polyclonal antibody was made successfully. The immunohistochemical localization technology was used to analyze the ABP1 distribution in seed and mesocarp at different development stages of peach fruit．The protein expression in mesocarp, endocarp and seed was analyzed by Western blot．【Result】According to peach fruit growth curve, it could be divided into three periods that are the first fast growing period, the hardcore period, and the second fast growing period. The results of immunohistochemical localization showed that ABP1 was distributed in both seed and mesocarp at all stages. ABP1 was evenly distributed in different parts of the seeds of all stages, the signal had no difference among different parts of seed at the same development stage. The stronger signals were detected in cells of outer and inner integument and around vascular tissue of seed coat. In the whole growth period, ABP1 signal did not significantly change. Between the outer and inner integuments, there were sporadic, zonal signals. The expression of ABP1 was around vascular tissue in mesocarp during the hardcore period of fruit development. The level of ABP1 expression was higher at the first rapid growth period（46DAFB）and the start of second rapid growth period (76DAFB) according to Western blot result．Its expression level was the lowest at 39 DAFB. In different tissues, the expression of ABP1 in the mesocarp and endocarp were lower than that in seed．【Conclusion】The distribution and expression level of ABP1 have a difference in tissue and at developmental stage in peach fruit, ABP1 plays an important role in peach fruit development.

Key words: peach fruit, ABP1, immunohistochemistry, expression analysis

YU Jia, LI Yang, GONG Shuo, GUAN Wei, LIU Yue-ping. Tissue Location and Protein Expression Analysis of Auxin Binding Protein ABP1 in Peach Fruit (Prunus persica L.)[J].Scientia Agricultura Sinica, 2015, 48(5): 921-930.

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