Effect Of Nitric Oxide And Ruthenium Red On MPTP And MtDNA Cope Number In Peaches

Mitochondria not only plays an important role in the aerobic respiration and energy production,but also takes part in the process of electron transport,cell apoptosis and cell metabolism,in which play an important role in the normal physiological activity of the body.Nitric oxide?NO?is a bioactive substance which is fat-soluble,participating in multiple physiological reactions of plants in life information delivery as the first messenger.NO with a suitable concentration can influence the growth of plant seedlings,the ripening and storage of the fruits by controlling the physiological functions of mitochondria.Mitochondrial permeability transition pore?MPTP?controls the ions which passed by mitochondria and participates in the cell apoptosis process,but its mechanism in plants is not clear.That mitochondrial DNA?mtDNA?is easy to suffer damage because of its self structure,resulting in the mutation and missing of mtDNA makes mtDNA an important control factor of the cell apoptosis.Calcium ion(Ca²⁺)is the second messager in organisms and is also an important composition of the living body.Figuring out the influence of the NO on the copy number of mt DNA and the opening of MPTP,can help us to explore the specific mechanism about the influence of NO on the storage of the Feicheng peaches after picking.It plays a significant role in extending the storing time of the peaches.Our experiment is based on the Feicheng peach.NAD1 gene-design primer was obtained from National Center for Biotechnology Information?NCBI?.We used all of the DNA at different temperature and management as the template to observe the change of the copy number of mtDNA by real-time fluorescence quantification PCR and fluorescence microscope.Peaches which suffered the worst cold at 5 ? were chosen and were treated with single 15?mol L^-1 NO,5 ?mol L^-1 c-PTIO and mixed 20 ?mol L^-1 RR,to measure the content of ROS,the activity of antioxidase,the potential of mitochondrial membrane and the permeability of mitochondrial so as to explore the influence of NO on mitochondrial MPTP.Target gene fragment with the length of 159 bp was successfully obtained.Compared the the real-time fluorescence quantification PCR and fluorescence microscope of,that under thecondition of 0 ? and 5 ?,15 ?mol L^-1 NO can promote the decrease of the mitochondrial copy number in peaches.On the contrary,15 ?mol L^-1 NO can restrain the decrease of the mitochondrial copy number in peaches stored under room temperature.After the treatment of ruthenium red,the copy number of mtDNA enhanced in peaches stored under 0 ? and 5 ?.In The results from MPTP in mitochondrial of Feicheng peach fruit showed: 15 ?mol L^-1 NO can decrease the content of ROS in peach mitochondrial and enhance the activity of antioxidase?CAT,SOD,POD?;15 ?mol L^-1 NO decrease mitochondrial oxygen consumption restrain the respiratory action of the cell and the decline of mitochondrial membrane potential,then maintained the membrane potential at high level;enhance the resistance of mitochondrial membrane potential,restrains the enhancement of the permeability of mitochondrial membrane.The content of ROS in mitochondrial decreases within a short time after handled with 15 ?mol L^-1 NO-20 ?mol L^-1 RR,and enhanced the activity of CAT,restrained the decline of the film potential and the opening of the permeability of mitochondrial membrane and can restrain the opening of the mitochondrial MPTP within a short time.This result demonstrates that exogenous NO can restrain the opening of the mitochondrial MPTP,maintain the stabilization of the environment in mitochondria,slow down the process of cell apoptosis to extend the storage time of Feicheng peach fruit.