Studies On Different Proteomic During Adventitious Rooting Of Tetraploid Robinia Pseudoacacia Cuttings

Tetraploid Robinia pseudoacacia is one of species that take root difficultly, mainlyproduces by grafting seedling.Restricted by factors such as grafting area effect and longseedling period,it is difficult to promote in large area.At present, the research on theregulation mechanism of rooting is hysteretic while the technology on that is still immature.There has been no breakthrough on rapid seedling.In order to establish effective system for rapid propagation of seedlings of Tetraploidrobinia pseudoacacia,this study takes Tetraploid robinia pseudoacacial hard graft asmaterical,studies the protein dynamic metabolism of cutting rooting during differentstages,and reveals the regulation mechanism of rooting from the aspects of proteomics.Thispaper obtained the following results:1. By optimizating some detail technology of hard branch cutting,one can overcome theproblem that hard branch cuttage has low rooting rate.Applying a series of measures such aschanging the substratum,using heat continuously on the soil, we can get more than80%rooting.2. Establishment of Tetraploid robinia pseudoacacial two-dimensional electrophoresis(2-DE) system. TCA/acetone method is used on the segments of robinia pseudoacacial toextracte all phloem tissue protein.We build up two-dimensional electrophoresis system forTetraploid robinia pseudoacacial phloem protein through searching and optimizing theinterpublic pH gradient of dry strip, the concentration of separation gel isoelectric focusing,the amount of protein sample and total hours of volts.The results show that using theTCA/acetone to extract all proteins of robinia pseudoacacial,we can select17cm drystrip with pH4~7, stain180μg silver nitrate on samples, increase IEF from60000vh to80000vh, combined with two-dimensional electrophoresis can get a clear background, moreprotein points, good separating degree and repeatability two-dimensional electrophoresis.3.Using the established two-dimensional electrophoresis system of Tetraploid robiniapseudoacacial cuttings to take root in three different stages (stage of callus and adventitiousroot primordium formation stage, the stage of adventitious root elongation) of cuttings basephloem tissue protein electrophoresis separation, all three phases have610,619and619 clear protein seperately.Take adventitious root primordia formation phase as a keyanalysis,take callus stage as contrast and select24disparate spots for MALDI-TOF/TOF MSanalysis, and successfully identified seven spots that has a direct or indirect effect on theformation process of adventitious root primordia.