Studies On Different Proteomic Among The Phases Of Adventitious Rooting Of Xanthoceras Sorbifolia Cuttings

In our research, IAA, IB A, ABT were uded for the rooting of Xanthoceras sorbifolia’s cuttings. Baded on two-dimensional electrophoresis (2-DE) and MS, we detected the differentially expressed proteins during the phase of adventitious root formation of Xanthoceras sorbifolia. We have got these results as below:1. Establishing a2-DE system for the phloem of Xanthoceras sorbifolia. As the phloem had a lot of fiber but only a little protein, we also created a protocol for protein extraction. The established2-DE system was:Extracting proteins with Lysis buffer Ⅱ(7M urea,2M thiourea,4%(W/V) CHAPS,65mmol.L-1DTT,0.5%(V/V) carrier ampholytes), oscillation in27℃for1hour is good for extraction. The IPG strip with ph4-7and17cm length is good for show the separated proteins, and the appropriate content is180μg per strip which should Isoelectric focusing for80000Vh. In our study,12%was the suitable gel concentration, and the gel should stained with silver.2. We had separated about900proteins in one gel from the phloem of Xanthoceras sorbifolia, and90of them were identified as differentially expressed proteins by PDQuest7.4between control (fresh water treatment for4hours) and the best treatment (the IBA concentration of800mg.L-1for4hours). Fourteen of the significantly expressed proteins were identified by MALDI-TOF/TOF, and eleven of them were finally identified. These idenfified proteins were involved in stress response, calcium signaling pathway, cell division and proliferation and so on.3. A simple analysis of differentially expressed fragments during adventitious root formation of Xanthoceras sorbifolia was done based on cDNA-AFLP technique. With16pairs of primers, we had found at least30differentially expressed bands between different phase of adventitious rooting. We hope that through this experiment, to lay a certain foundation for the laboratory follow-up study.