Primary Differential Proteome Analysis Of Slef-And Cross-Pollinated Pistils In Tea Plant (Camellia Sinensis)

Tea plant (camellia sinensis L.), which belongs to the Theaceae family, is a cross-pollinated perennial plant. It has adopted self-incompatibility for its very lower seed setting rate after self-pollination. This study was conducted to explore the molecular mechanism underlying self-incompatibility (SI) in tea plant at the proteome level. This research includes the establishment of a proper two-dimensional gel electrophoresis protocol for proteomic study of tea plant pistils; differential protein expression of pistil proteomes at different stages after self-and cross-pollination; and differential protein expression of pisitl proteomes after self-pollination. And the results are as follows:1. This study first establish a proper two-dimensional gel electrophoresis (2-DE) protocol for proteomic study of tea plant pistils, and the optimized system includes:total proteins of sample extracted using TCA/acetone method followed with clean up kit, separating the proteins with 17 cm pH5-8 IPG strips,13% SDS-PAGE, and CBB R-250 stain method. With about 1200 spots can be detected, this system resulted in large quantities of protein with better 2-DE separation.2. Two-dimensional electrophoresis revealed 63 protein spots were differentially expressed in self-and cross-pollinated pistils following different stages.59 differentially expressed proteins were successfully identified by MALDI-TOF/TOF to match diverse protein species. These proteins involve in different cellular and metabolic processes with obvious functional skew toward metabolism, protein biosynthesis, protein fate, cell sturcture, storage protein, transport, stress response, energy metabolism, and nuclesome assembly. Self-pollination resulted in increased expression of antioxidative enzymes APX, Catalase isozyme 3, sHSP, TCTP after 24 h and 48 h pollination; Protein syntheses related protein expressed increasedly within 48 h after self-pollination but no significant difference at 72 h, while 26S proteasome regulate subunit A expression was only decreased after 72 h self-pollination; Structure related protein and storage protein expression increased in self-pollinated pistil; TIM and FBA, which involved in energy metabolism, increasedly expressed within 48 h after self-polliantion; Caffeine synthesis relate protein increased and then greatly declined.3. Proteome expression of tea plant pistils after self-pollinaiton at different stages was studied.17 differentiallly expressed proteins were successfully identifed. These protiens belong to eight functional groups, stress response, metabolism, signal transduction, cell structure, storage protein, protien fate, transport and unclassified proteins. Among them,5 stress response related protein takes the biggest part, and followed with metabolism and protien fate, PPIase FKBP12,40S ribosomal protein S12, GTP-binding protein Ran/TC4 increase the most. Profilin increased rapidly after 72 h pollination.