Optimization Of Technology And Function Analysis Of H₂O₂ And NO In Fraxinus Mandshurica Somatic Embryogenesis

F. mandshurica, belong to oleaceae ash, is one of the three big hard broad-leaf species of China’s northeast. F. mandshurica mature zygotic embryos as explants, this paper analyzed its role in F. mandshurica embryogenesis and cell death of the explant, and probed into its place in the signal network through studying effects of exogenous H2O2 and NO on somatic embryogenesis and the total amount of cell death, intracellular H2O2 content and the intracellular NO content. This paper screened for materials of F. mandshurica embryo proliferation and germination training, and then embryonic callus as the subculture material, this paper studyed the subculture number influences on embryo proliferation and plant regeneration, analysis of northeast after successfully embryo induced, analyzed embryonic maintain and plantlet regeneration abilities in the process of F. mandshurica embryo subculture, providing cover for germplasm resources preservation of F. mandshurica.The main results were as follows:1. H2O2 and NO were plays an important role in somatic embryogenesis of F. mandshurica. Increasing appropriate explantation intracellular H2O2 content promoted somatic embryogenesis; eliminating explantation intracellular H2O2 content was bad for somatic embryogenesis and late development. Increasing explantation intracellular NO content was beneficial to somatic embryogenesis and late development, improving the quality of the somatic embryo; reducing explantation intracellular NO content was bad for somatic embryogenesis and late development, reducing the quality of the somatic embryo.2. During proliferation culture of Fraxinus mandshurica somatic embryo, direct proliferation effect of 4 mm not green somatic embryo was best. Compared with direct proliferation material, the best material for subculture multiplication culture was the callus.Proliferation coefficient of Fraxinus mandshurica somatic embryo increased with the increase of the number of proliferating subculture.3. During germination culture of Fraxinus mandshurica somatic embryo, germination effect of 4-8 mm not green somatic embryo was best. With the increase of of proliferation subculture number, the germination rate of Fraxinus mandshurica secondary somatic embryo decreased gradually. With the increase of germination culture time, somatic embryo had a large number of the phenomenon of browning and death. With the increase of of proliferation subculture time, the rooting rate of Fraxinus mandshurica secondary somatic embryo increased at first and decreased subsequently.4. Combination of H2O2 and NO could be used as the signal factor involved cell death process of in somatic embryogenesis induced by the high concentration of sucrose stress, and speculated that NO as upstream signal of H2O2 in the signal network. Amount of H2O2 treatment inhibited total cell death, and promoted the production of intracellular H2O2 and NO; CAT treatment inhibited total cell death and inhibited the production of intracellular H2O2 and NO.5. SNP treatment processed inhibition of explant cell death, and inhibition of intracellular H2O2 production, and promoted the generation of intracellular NO; PTIO inhibited cell death and the production of intracellular H2O2 and NO.