The Programmed Cell Death In Bright Yellow-2Tobacco Suspension Cells Induced By5-Azacytidine

Programmed cell death(programmed cell death,PCD),also known as apoptosis,is thenumber of cells in the organism’s own gene regulation through a proactive way to die.5-azacytidine is a DNA methylation inhibitor that incorporation of DNA,DNA replicationand DNA binding process, inhibition of DNA methyltransferase (DNA methyl transferase,DNMT),to achieve demethyl capabilities, and can be re-activated due to excessivemethylation and inactivation of genes,genomic DNA demethylation of genomic changesthat led to biological mutation.Thus,5-azacytidine(5-Azacytidine)in the induction ofnew species has an important significance. However,5-azacytidine inhibits DNA methyltransferase enzyme,it will affect programmed cell death.In the National Natural Science Foundation of China(30771746)under the auspices of paper with5-azacytidine(5-Azacytidine)Processing grown to logarithmic phase(3-4d)of tobacco BY-2(bright yellow-2)suspension cells,observation and measurement of tobacco BY-2cells programmed cell death morphological and physiological changes.The overall results have been summarized as follows:l.To observe the BY-2tobacco cells that induced by5-azacytidine,discovering the apoptosis of plant cells experienced the process of early phase,medium phase and later phase,with the prolong of the time.In the early phase cell membrane of apoptotic cells wrinkled, departed with cell wall becoming a gap,that just was similar with plasmolysis.A part of cell nucleus marginalized.In the medium phase cell membrane wrinkled deeper,cell nucleus agglutinated like a cap.Cytoplasm was turbid,nucleus became sone wads in the later phase.2.Dealt with5-azac,extracted the DNA of apoptosis tobacco cells on agarose gel electrophoresis,DNA Ladder was observed,and DNA degraded into2-4nucleosomes.TUNEL detection observed a strong positive signal.The above tow results are the important biochemical feature of PCD.These results suggest that the cell dead induced by5-azac is PCD,and possesses the typical feature of PCD.3.To observe the content of ROS of BY-2tobacco cells that treated with5-azac,discovering the phenomenon of ROS burst in cells.With the prolong of the time,the content of ROS was a curve.with some peaks,At5.5hour,the content of ROS was the highest,that was0.1123,declared that ROS burst was the biggest.We can guess that ROS participated the pathway of signal transduction.