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Chestnut (Castanea mollissima BL.) Is China's famous special dried fruit, nutritious fruit, is an important species of woody food. Chestnut seed breeding seedling grafting propagation coefficient. Plant tissue culture characteristics, consistent with seedlings obtained in the short term, but also the technical basis of the genetic improvement of chestnut varieties. Therefore, the academics to carry out the study of the chestnut plant tissue culture, and to explore its mechanism of adventitious root formation. Chestnut tissue culture persists rooting low, following long generation time, need further study. In this paper, on the basis of previous trials, still Yanshan Red Li 2-3 months seedlings stem segments as explants for tissue culture, explant sterilization method has been improved, and following the generation of proliferation culture exists problem analysis. By two-dimensional electrophoresis technology adventitious roots occur in different periods Chestnut in Plant Tissue protein changes. The results show that: the stem segments as explants, 5% NaClO (?) Inch explant surface sterilization 20min alternative to highly toxic mercuric chloride and sterile system quickly. Chestnut Subculture shoot tip necrosis second generation necrosis was 19.5%, 37.1% to the 7th generation. With the increase in the number of subculture, shoot tip necrosis increasingly serious phenomenon. Inferred by the results of previous studies Chestnut adventitious root formation is divided into three phases, namely 0-3d, root primordia induction stage; 3-7d root primordia formation stage; adventitious root elongation growth stage after the first 7d. Chestnut in Plant Tissue micro-branches of the three periods of this thesis were collected using two-dimensional electrophoresis (2-DE) technology, protein changes chestnut adventitious root formation in different periods. To determine the best method of Chestnut protein extract modified TCA / acetone precipitation method, to establish a suitable Chestnut Pearson rhizome section of two-dimensional electrophoresis method to obtain a two-dimensional electrophoresis map resolution and reproducibility was good. There are significant differences in protein spots with ImageMaster software analysis, analysis, results show that: (1) adventitious root formation process, the the protein total number essentially unchanged, but the expression level downward; (2) to the root primordia from root primordia induced 5 during the formation of the up-regulated protein spots, down 9; 6 new protein spots disappear 9; (3) from the root primordia formation adventitious roots during the differentiation of growth increased expression levels of four proteins, 7 reduced; 9 new protein spots disappear 8. Select chestnut adventitious roots occurred three stage-specific expression of six proteins for MALDI-TOF-MS analysis, the determination of the peptide mass fingerprinting, search the possible homology of protein in the NCBI database, the preliminary identification of these proteins: the composition PS Ⅱ the oxygen complex OEE protein 1, is closely associated with the Calvin cycle Rubisco and SBPase, etc., most of these proteins is closely related to plant photosynthesis, may indirectly affect the chestnut adventitious root formation and its mechanism needs further study .
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