Regulatory Mechanisms By Nitric Oxide On Adenine Nucleotide Translocator In Mitochondria Of Feicheng Peach Fruits

Adenine nucleotide translocator is an important part of the mitochondrial permeability transition pore, involved in the regulation of physiological functions MPTP can induce apoptosis, but its mechanism of action in the plant is still unclear. Nitric oxide as a gas molecule, it has a fat-soluble characteristics, it belongs to first messenger of the three messaging messenger in life body. It can mediate a variety of physiological functions in plants, For example, it can regulate a variety of physiological processes in the mitochondria, and promote the growth of plant seedlings, and regulate the ripening of fruits. And the appropriate concentrations of nitric oxide can extend the storage time of the fruit. Therefore, studying on the regulation of nitric oxide for adenine nucleotide translocator molecular mechanism is helpful to confirm the influence of adenine nucleotide translocator to the storage of Feicheng peach fruit.In this paper, the experimental materials were Feicheng peach fruit. Getting adenine nucleotide translocator's mRNA sequence of maize, rice and other plants were from the NCBI, and obtained degenerate primers. Through the rapid amplification of cDNA ends(RACE) won the cDNA sequence of adenine nucleotide translocator. And the full-length cDNA sequence of adenine nucleotide translocator was 1432 bp, coding region of 1155 bp, which encodes 384 amino acids. Through simulation analysis, the adenine nucleotide translocator glutathione reaction occurred more easily than nitroso chemical reaction. The interaction between nitric oxide and adenine nucleotide translocator protein is only one possible. The recombinant adenine nucleotide translocator protein was expessed in Transetta(DE3), purified through Nichelating affinity chromatography, and renaturation with dialysis.It was proved that adenine nucleotide translocator protein can interact with phosphate carrier protein by Yeast two hybrid. In fluorescence spectrum of nitric oxide with adenine nucleotide translocator protein reaction, we found that nitric oxide could reduce the fluorescence degree of adenine nucleotide translocator protein and cause fluorescence quenching, and this fluorescence quenching belongs to static quenching. From the result of Scatchard model, we found that nitric oxide could react with adenine nucleotide translocator protein to form a 2:1 compound. The result from realtime fluorescence quantitative PCR showed that 15 ?mol L-1 nitric oxide could reduce the expression of adenine nucleotide translocator. Real-time PCR from ANT study found, 15 ?mol L-1 NO treatment decreased expression of ANT. The results from biochemical indexes in mitochondrial of Feicheng peach fruit showed: 15 ?mol L-1 nitric oxide could reduce the content of ROS in mitochondria, while enhancing SOD, POD and CAT activity. It could reduce the content of MDA in mitochondria and decrease mitochondrial oxygen consumption also. Conversely, c-PTIO could increase the content of ROS and MDA in mitochondria, while decrease SOD, POD and CAT activity. In addition, it also increased mitochondrial oxygen consumption. With the increase of peach fruit storage time, the mitochondrial membrane potential showed downward trend, while 15 ?mol L-1 nitric oxide treatment effectively slowed down the rate of decrease in mitochondrial membrane potential. 80 mmol L-1 N-Acetyl-L-cysteine-15 ?mol L-1 nitric oxide co-processing could decrease the content of ROS and MDA in mitochondria and increase SOD activity. And it slowed down the rate of decrease in mitochondrial membrane potential also. The results showed that nitric oxide could mediate the mitochondrial function to extend the storage time of Feicheng peach fruit.