| Apoptosis is controlled by genes,involves a series of gene activation,expression and regulation in order to better adapt to the living environment,and an initiative to fight the death process.Laser tweezers Raman spectroscopy(LTRS)is combined with the laser tweezers Raman spectroscopy and a new type of biological photon technology,the three-dimensional optical trap(optical tweezers)imprison the single living cell in the solution by highly convergent laser beam,then transient increase the laser power and at the same time excitat and collect the Raman spectra of the cells.LTRS has the advantages of the non-invasive,sensitive and rapid,more importantly,will real-time and non-invasive to observe the information of physiological and biochemical changes of single living cell.The topic on the mitochondrial level,LTRS as a single cell analysis tools combined with conventional biochemical analysis techniques,different concentration of aluminum ions as apoptosis-inducing solution,dynamic record the Raman spectral data of single mitochondrion induced by aluminum,help us to understand the change reasons of its internal biological macromolecule in the process of a single mitochondrion induced by different concentration of aluminum ions,reveal a deeper mechanism in plant cells apoptosis induced by aluminum.The main findings are as follows:(1)Research the stability and integrity of the mitochondrion of Vicia faba L.at 4 ? and room temperature conditions using LTRS technology,combined with conventional method of physiological and biochemical analysis to detect physiological changes of the mitochondrion under the same conditions for verification.The results show that the intensity of the nucleic acid peaks(825 and 1262 cm-1),the intensity of cytochrome C(Cyt C)peaks(750 and 1127 cm-1),the intensity of mitochondrial respiratory characteristic peak(1602 cm-1),the intensity of the main composition of phospholipid peaks(1065,1127 and 1457 cm-1),the intensity of protein peaks(750,930,1262,1330,1360,1457 and 1360 cm-1)are decrease significantly as a function of the time at the both storage conditions,but in 2 hour,compared with the control,the storage conditions at 4 ?,the decrease far less than the storage conditions at normal temperature,using conventional analytical methods to measure mitochondrial respiration rate(RCR),phosphorus oxygen ratio(P/O),Cyt c oxidase activity,NADH oxidase activity,succinate oxidase activity,in 2 hour,compared with the control,the storage conditions at 4 ?,the decrease far less than the storage conditions at normal temperature.Indicate the stability and integrity of mitochondrion under 4 ? is better than the room temperatur,so study the mitochondrion in vitro is best to ensure completion within 2 h at 4 ?,also shows the LTRS technology is a non-invasive,fast and effective tool for mitochondrion analysis at a single mitochondrial level.(2)LTRS is use to research the biological macromolecules change of the mitochondrion of root tip cells of Vicia faba L.after induced with aluminum,combined with traditional physiological and biochemical analysis tools to detect physiological changes of the mitochondrion under the same conditions for verification.The results show that the intensity of the nucleic acid peaks,the intensity of Cyt C peaks,the intensity of mitochondrial respiratory characteristic peak,the intensity of the main composition of phospholipid peaks,the intensity of protein peaks are decrease significantly as a function of the duration of the aluminum stress,which is similar with the physiological and biochemical indexes of elongation of root,RCR,P/O,Cyt c oxidase activity,cyt c content,cyt c/a and malondialdehyde(MDA)content.Indicate that membrane lipid peroxidation deepened,the inclusions in the mitochondrion such as proteins,nucleic acids,lipids,Cyt c are gradually released into the cytoplasm,the respiratory function is reduced,affect the normal growth of Vicia faba L,thereby further causing apoptosis,as a function of the duration of the aluminum stress.(3)LTRS is use to research the biological macromolecules change of the mitochondrion of root tip cells of Vicia faba L.in vitro after induced with aluminum,combined with traditional physiological and biochemical analysis tools to detect physiological changes of the mitochondrion under the same conditions for verification.The results show that the intensity of the nucleic acid peaks,the intensity of Cyt C peaks,the intensity of mitochondrial respiratory characteristic peak,the intensity of the main composition of phospholipid peaks,the intensity of protein peaks are decrease significantly as a function of the duration of the aluminum concentration and time,which is similar with the physiological and biochemical indexes of RCR and P/O.The Raman spectra peaks of the proteins,nucleic acids,lipids,Cyt c and respiratory characteristic of mitochondrion of root tip cells of Vicia faba L.in vitro and in vivo after induced with aluminum are decrease significantly as a function of the duration of the aluminum stress.Indicate that aluminum could penetrate into the cell interior and directly impact the mitochondria possibly,result in membrane lipid peroxidation deepened,the inclusions in the mitochondrion such as proteins,nucleic acids,lipids,Cyt c are gradually released into the cytoplasm,the respiratory function is reduced,cause the apoptosis of Vicia faba L.cells which relies on the mitochondria pathways. |
PDF Full Text Request