Sifting Of Cold Resistance Protein In The Leaves Of Camellia Sinensis

Plants in low temperature induce new proteins due to the change of geneexpression. These new proteins are highly hydrophilic and thermal stability, canprotect plant cells from low-temperature damage. Currently, for most plants on thecold resistance induction experiment we found cryogenic stress can change solubleprotein content of plant cells and product specific protein of plant hardiness closelyrelated.The protein content of tea leaves accounted for about25percent of the drymatter weight, most of which exists in the cytoplasm or combines membrane notclosely, can be extracted with nonpolar solvent (such as water, dilute acids, dilutealkali or dilute salt solution), the protein collectively referred to as soluble protein,including thermal stability and thermal unstable protein, mainly for various enzymesand some physiological activity protein. The leaf tissue cells of tea accumulate a hugemass of soluble protein with decomposition histone or other material and transformunder low temperature stress to increase the concentration of the cell so as to reducethe freezing point to resist low temperature environment.The content of polyphenols, pigment in tea tree is particularly high, the teapolyphenols is extremely easily oxidized again in the grinding leaves to form massiveoxidation product which substances serious interference with electrophoresis process.In this experiment, we extract protein of tea leaf using acetone dry powder preparationmethod, improved acetone settlement method and TCA-ACE method, the SDS-PAGEelectrophoresis results display improved acetone settlement method not onlyextraction efficiency high, but also impurity less disturbance, the protein channel ofelectrophoresis results is clear and large quantity. Satisfied reults were achieved in theapplication of the optimized method to the cold resistance protein analysis of severaltea plants,23ku down-regulation protein band in yun kang43and51ku up-regulationprotein band in shu cha zao related to cold resistance were obviously showed out inthe analysis of SDS-PAGE.Further to extract proteins of annual shu cha zao branches under4℃low-temperature processing three days for two-way electrophoresis experiment,PDQuest software analysis of protein expression spectrum display the46sportsquantitative change multiples greater than1.5times, including low-temperatureprotein expression point31, abate of protein expression point15, speculate that these differences protein points may relate to cryogenic stress related.According to the information, the isoelectric point of tea plant is generallyconcentrated in the4-6.5, and considering the factors of the reproducibility, separation,the expression on the map of2-DE, we selected and indentified7protein spots usingpeptide mass fin-reprinting and matrix-assisted laser desorption ionizationtime-of-flight mass spectrometry. Seven identified proteins and their putativefunctions were presented. Six of them were identified as ribulose-1,5-bisphosphatecarboxylase protein family, and one was identified as transketolase.