Mingliang Yu

Potential role of reactive oxygen species and antioxidant genes in the regulation of peach fruit development and ripening.

The roles of reactive oxygen species (ROS) as both toxic by-products and as signaling molecules have been reported in fruit development and ripening. Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) play important roles in balancing the induction and removal of ROS in plants, and are respectively encoded by families of closely homologous genes. In the present study, we investigated the roles of ROS and the above-mentioned antioxidant genes during the development and ripening of peach fruit. The experimental results indicated that O2(-) and H2O2 acted as potential signaling molecules in the middle stage of fruit development, and only H2O2 might function as a main toxic molecule to stimulate lipid peroxidation and oxidative stress in the late stage of fruit ripening. PpaCu/Zn-SODs were the most abundant members in the PpaSOD gene family and they expressed steadily in peach fruit development and ripening. Low temperature (4xa0°C) postponed and suppressed the climacteric peaks of respiration and ethylene, significantly enhanced the activities of CAT and GPX, and up-regulated the expression of PpaCAT1 and PpaGPX6 in the late stage of fruit ripening. PpaCAT1 and PpaGPX6 were two key genes in alleviating oxidative stress in the late stage of fruit ripening.

Proteomic analysis of changes in mitochondrial protein expression during peach fruit ripening and senescence.

UNLABELLEDnRipening and senescence define the last step of fruit development, which directly affects its commercial value, and mitochondria play a crucial role in these processes. To better understand mitochondrial roles in maintaining and regulating metabolism in storage tissues, highly purified mitochondria were isolated from peach tissues (Prunus persica. cv. Xiahui-8) stored at 4°C and 25°C, respectively, and their proteome was conducted using the method of 2-DE and MALDI-TOF/TOF. Twenty-four (24) differentially expressed proteins (2-fold, p≤0.01) were identified out of more than 300 spots and were divided into six categories by PIR and Uniprot, including oxidative stress (34%), carbon metabolism (29%), respiratory chain (17%), amino acid metabolism and protein biosynthesis (8%), heat shock protein (4%), ion channels (4%). Proteins involved in antioxidative systems, gluconeogenesis, glycolysis, ethanol fermentation were changed significantly in response to high temperature. Storage at 4°C dramatically delayed ripening and senescence processes by postponing the climacteric peak, slowing down carbon metabolism and degradation of cell structure. Besides, low temperature induced the expression of formate dehydrogenase and some amino acid metabolism proteins. Proteins classified in respiratory chain, ion channels showed high coherence with climacteric respiratory burst, and the antioxidative enzymes showed relatively important symptoms on ROS scavenging through orderly expressions.nnnSIGNIFICANCEnWith the advent of proteomics and mass spectrometry (MS), it becomes possible to identify the specific functions of differentially abundant proteins in peach mitochondria. In the present study, a procedure to isolate mitochondria from peach fruits was established, and the mitochondrial proteome was systematically analyzed by 2-D gel electrophoresis procedures in combination with protein identification by mass spectrometry. Differentially expressed proteins in peach mitochondria during different stages of peach fruit ripening and senescence were characterized. Our data provide a great deal of information likely to enhance the understanding of the mitochondrial function in peach ripening and senescent process during storage.

Quantitative proteomic analysis of pre- and post-harvest peach fruit ripening based on iTRAQ technique

To further elucidate the molecular mechanism involved in peach (Prunus persica cv. Xiacui) fruit ripening, a differential proteomic analysis was conducted using iTRAQ based strategy for studying peach fruit ripening from day 15 before commercial harvesting (D0) to 15-day storage period at 25xa0°C. 64 differentially expressed proteins (≥1.5-fold, pxa0≤xa00.05) were successfully identified. Lipoxygenase (LOX) family members exposed the status of causing disorder metabolism and senescence by increased expressions. Some well-known antioxidant enzymes did not show significant expressions especially in later storage time. Nevertheless, proteins involved in ABA metabolism were up-regulated, demonstrating the important role of ABA in adaptation of plants to various environmental stresses. Glutathione S-transferase, cysteine proteinase inhibitor, transcription factor bHLH135 all showed great potential in response to ROS or other stresses. Proteins associated with flavonoid biosynthesis and amino acid metabolism also revealed critical roles in metabolism during storage.

1-Methylcyclopropene Treatment on Phenolics and the Antioxidant System in Postharvest Peach Combined with the Liquid Chromatography/Mass Spectrometry Technique

In the present study, the potential effect of 1-methylcyclopropene (1-MCP) treatment on phenolics and antioxidant capacity in postharvest peach was assessed. Peach fruit (cv. Xiahui-8) treated with 1-MCP or without treatment was stored in 25 °C for 2, 4, 6, and 8 days. The phenolic composition and change trend were evaluated by liquid chromatography/mass spectrometry. The reactive oxygen species production and scavenging capacity against DPPH, O2•u202f-, and HO• were determined. Gene expression of enzymes in the flavonoid biosynthetic pathway was assayed by quantitative reverse transcription polymerase chain reaction analysis. 1-MCP application inhibited the ethylene and CO2 production and stimulated the total phenol and total flavonoid contents. Total anthocyanin formation may be influenced directly or indirectly by the level of ethylene. The scavenging capacities of DPPH, HO•, and O2•u202f- after 1-MCP treatment were enhanced. 1-MCP treatment affected the tissue color change, stimulated gene expression of PpaPAL, PpaCHS, PpaF3H, and PpaUFGT, and promoted the biosynthesis of flavonoids and stability of anthocyanin. PpaDFR and PpaUFGT played crucial roles in rapid color change stages. Kaempferol and kaempferol 3- O-galactoside increased distinctively during storage time.

Regulation of the protein and gene expressions of ethylene biosynthesis enzymes under different temperature during peach fruit ripening

Ethylene has profound effect on fruit development and ripening, and the role of ethylene biosynthesis enzymes involving 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS), ACC oxidase (ACO), and S-Adenosyl-l-methionine synthetase (SAMS) in peach fruit (cv. Xiahui-8) was characterized under 25 and 4xa0°C, respectively. All these enzymes in ethylene synthesis pathway were identified using 2-DE and real-time PCR. Both protein and gene expressions of ACO and SAMS were much higher at 25xa0°C than at 4xa0°C. Among five members of ACS family, PpaACS4 may belong to system II ethylene biosynthesis, while PpaACS3 involved in system I during development stage, and low temperature can induce PpaACS1 expression. The ethylene release and low expressions of proteins and genes of most enzymes indicated that low temperature can effectively postpone ripening stage by reducing ethylene evolution. High gene expression of PpaSAMS did not cause excessive expression of SAMS protein under low temperature, and over-expression of PpaACS1 at low temperature still did not induce increase of ethylene production. The mechanism underlying the phenomenon about how temperature affects ethylene release was also discussed.