The Preliminary Research Of In Vitro Regeneration Optimization,infected Leaves Of Differentially Expressed Proteins For Cymbidium Hvbridum And Model Plant Antiviral Mechanism

Orchidaceae is a large family after the family Asteraceae, is the largest Division of the monocots, and truly be called the world's flowering plants, ornamental and medicine has significant economic benefits and application prospectsvalue in use. However, the orchids by serious violations of incurable viral diseases, is hindered the normal growth of the orchid plant germplasm conservation and orchid industry, orchid treasure.Orchidaceae perennial transgenic get outside the explant regeneration frequency is low, long life cycle in vitro culture, vulnerable to pollution problem is still the bottleneck of the perennial plant disease-resistant strains. N.benthamiana this s easy cultivation, rapid growth, reproduction and strong in vitro regeneration frequency characteristics, usually laboratory an ideal model plant and research materials. In this study, plant tissue culture techniques to optimize the Cymbidium hybridum in vitro regeneration and propagation system. Also differentially expressed proteins of the virus-infected mature leaves of Cymbidium hybridum. Cymbidium hybridum and the model plant's tobacco anti-virus genetically engineered plant-based RNA silencing mechanism, preliminary research, the results of their research on the orchid industry has an important value and economic benefits.In this study, the main findings are as follows:1,Using plant tissue culture techniques, the establishment of the regeneration of Cymbidium and N.benthamiana breeding system.Shoot-tips from Cymbidium hybridum were used as the explants to investigate the effects of6-BA and NAA in different concentration combinations on PLB (Protocorm-Like Body) induction, propagation and differentiation. Under the concentration combinations of1.0mg/L6-BA+0.5mg/L NAA,0.8mg/L6-BA+0.4mg/L NAA, and1.0mg/L6-BA+0.3mg/L NAA respectively,93.3%of shoot-tips were able to survive,85.8%of them were induced into PLBs and the propagation rate of PLBs was reached to4.59individually. The optimized concentration combination of0.8mg/L6-BA+0.5mg/L NAA resulted in8%higher ratio of PLBs induction (80.8%) and differentiation (87.5%) than that with currently reported72.7%and76.8%respectively.2,This experiment will contain CymMV and ORSV silencing vector selective marker vector pCAMBIA1301directly by particle bombardment into protocorms of Cymbidium, protocorm proliferation,sprouting, rooting,plant-based RNA silencing mechanism, foreignthe edge CymMV and ORSV disease juice infection Cymbidium, and observe the disease results. Through experimental testing, CymMV and the ORSV the silencing vector has been transferred to the Cymbidium hybridum.3. This study were healthy, infected with ORSV infection CymMV+ORSV virus mature Cymbidium hybridum Extraction of Total Protein, during the two-dimensional electrophoresis, and then get the map of the protein spots obtained by the analysis software, health, infection ORSV,CymMV+ORSV virus Cymbidium hybridum two-dimensional electrophoresis maps of protein spots, respectively,199,256,278, healthy two-dimensional electrophoresis map of ORSV virus infection, there are eight points up or down, health and infection CymMV+ORSV the virus of two-dimensional electrophoresis Figure4point upward or downward, while ORSV virus infection in the two-dimensional electrophoresis map with or without point9, whether the two-dimensional electrophoresis map of the infection CymMV+ORSV virus12expression analysis, mass spectrometry difference on the above points, respectively, the results from the protein level to reveal the host of the virus-mediated interaction mechanisms and possible virus interacting protein interaction of physiological and biochemical Cymbidium hybridum and virusesand to explore the molecular mechanism of the the Cymbidium virus infection response, provides a theoretical basis for the cultivation of new resistant varieties.4,In this study, plant-based RNA silencing mechanism, injection vaccination RNA enzyme water control plants, the PVX/GFP plants, PVX/NbRBRl-GFP plants, the PVX/NbPCNAGFP the plant to observe this's tobacco plant virus induced genethe life cycle of the silent phenotype,'s tobacco NbRBR1and the NbPCNA gene silencing on ACMV DNA replication. The results show that the virus induced gene silencing the endogenous NbRBR1, and NbPCNA gene expression occurred, and NbRBR1and NbPCNA the gene silencing caused by the abnormal phenotype observed, lay the foundation for the next step to depth the study NbRBR1and NbPCNA gene silencing mechanism.