Study On Cell Proliferation Capacity Of Chinese Fir After Synergistic Effect Of Calcium,phosphorus And Aluminum

Chinese fir is one of the most important forests in South of China, however, in the current,land degradation,productivity decline etc impact on the development of forestry of Chinaseriously. Research shows that one of the important reasons which affect the growth of Chinese firis aluminum toxicity which is induced by soil acidification. So, How to reduce the adverse effectsof aluminum toxic and remission aluminum toxicity is the key problem to be solved in Chinese firproduction.In view of this, in this paper, the author took aluminum toxicity as a major research field andtook Chinese fir as the study object. On the basis of previous research, the paper used three factorsand four levels orthogonal experiment design,set an acidic environment (pH=4.0) and artificialsimulation water stress on annual seedling of Chinese fir. Using flow cytometry to measure cellcycle of various material handling,relative cell number and then analysis cell proliferation abilityof each processing. Lastly, the paper analyzed the response of cells on Chinese fir seedlings whichwere treated differently and analyzed the capacity and mechanism of each factor in remission ofAl stress. In order to seek a method that can remit aluminum toxic of Chinese fir and improve theproductivity and economic benefits of Chinese fir. The main results are summarized as follows:1,This paper first researched the method of making single cell suspensions of Chinese firleaves, summed up a method of single nuclear dissociation which suitable for Chinese firleaves.The experiment results showed that the suitable nuclear dissociation for Chinese fir leavesis Galbraith's, and the experiment showed that among the16different treatments the sixteenthprocessing (dissociation liquid is Galbraith's, dissociation time is4hours, centrifugal speed is800r/min, centrifugal time is10min) is the most suitable method for Chinese fir leaves nucleardissociation which dissociated the most nucleus number and the least fragments.2,This paper studied the staining method and summed up the cell staining method ofoperation which is the most suitable for Chinese fir, in order to detect the cell proliferation abilityof Chinese fir after the stress of calcium, phosphorus and aluminum. This experiment set9different processing by regulating dye concentration and the time of dyeing, and the resultsshowed that among the9processings the9th processing (the dye concentration of PI is300vl, thedyeing time is60min) is the most suitable DNA labeled operation processing for Chinese fir.3,Using flow cytometry detect the cell division cycle during different stress periods, analysis the change of the percentage of S phase during16treatments and then reflects the ability of cellproliferation. The results as follows: S phase of each treatment ratio changed greatly after stress.The percentage of S phase value increased of the treatments7,5,3,4and16. While, the others'percentage of S phase decreased in different degree. It descript that the treatment7,5,3,4and6can promot cell proliferation ability effectively; While, the others have different degrees ofweakening in cell proliferation ability after stress. Among them the12,15and13treatments' cellproliferation ability weaken mostly.4,Using flow cytometry detect relative cell number in different periods of stress. Throughthe analysis of the detection time of16treatments we got the conclusion as follows: before stresseach treatment material is relatively uniform, while, after stress the time that detect30000cellsrequired have changed a lot during each treatment. Among them, the detect time of7,3,5and4treatments no change basically. While, the others detect time has increased in different degreesapparently. It showed that the7,3,5and4treatments can promote the proliferation of cellseffectively; While, the other treatments have inhibition of different degrees on cell proliferationability, and the12,13treatments have the greatest inhibition effect.In summary, when pH=4.0, under the solution culture, phosphorus and calcium has asignificant impact on the regulatory effects of aluminum toxicity in cell proliferation. Among the16treatments in the orthogonal test, the seventh treatment(Ca0.5mmol/L,P3.0mmol/L,AL6.0mmol/L)is the best aluminum,calcium,phosphorus concentration combination which is themost effective in alleviating aluminum toxicity on Chinese fir, followed by the fifth treatment(Ca0.5mmol/L,P0mmol/L,AL2.0mmol/L).While, the worst concentration combined are the twelfth(Ca1.0mmol/L,P4.5mmol/L,AL2.0mmol/L)and thirteenth(Ca1.5mmol/L,P0.0mmol/L,AL6.0mmol/L)treatments.