Establiashment Of Populus Tomentosa Carrière Genetic Transformation System And Phenotypic Analysis Of Transgenic Plants With High Photosynthetic Efficiency

Poplar is the most useful model plants for the regeneration and genetic transformation of woody plants.Populus transgenosis technology can overcome the incompatibility of distant hybridization and break the chains on different species of plants.These characteristics led to its widespread use as a new technical approach in forestry genetic engineering.This study intends to introduce multi-gene vector GOC-PYL1305 into Populus to create a photospiration metabolism bypass,aiming to improving the photosynthesis meanwhile reducing photorespiration loss,and finally achieving engineering Populus plant with high photosynthetic and biomass e fficiency.The results obtained are summarized as follows:1.An efficient regeneration system was established.MS medium supplemented with0.4mg/L6-BA,0.1mg/LNAA and 0.01mg/LTDZ were used as leaf regeneration medium;MS medium supplemented with 0.4mg/L6-BA and 0.1mg/LNAA were used as shoot elongation medium;MS medium were used as root differentiation medium.The results show that the growth coefficient was up to 7.98,differentiation rate was up to 100%.Regeneration system can provide an effective transgenic acceptors fo r genetic transformation.2.Experimental result showed that 10 mg/L Hyg was suitable for differentiation selection,200 mg/L Cef for bacteriostatic was suitable.The experimental result showed that in process of genetic transformation bacteria conce ntration are O D₆₀₀=0.6,and the infection time 15-20 min were suitable for transformation,adding 200?mol AS in the cocultivation medium for 2 days can improve the transformation efficiency.3.Based on multi-gene transform technology,a multi-gene expression vector GOC-PYL1305 cloning form the rice was transformed into Poplar chloroplast.Aim to metabolize part of the glycolate produced by photorespiration to the oxalate and finally decomposed in into CO2 within the chloroplast.Thus eliminate to peroxide and improve the efficiency of plant photosynthesis.4.Populus tissue culture plants were used to genetic transformation materials.300explants inoculated by Agrobacterium-mediated genetic transformation combination,30Hyg-resistant plants apperared spec ific bands.The positive plants were tested by PC R,17plants Specific amplification bands,the positive rate was56.67%,then 20 plants were tested by RT-PCR and GUS dyeing,10 plants presented the same hybridization band as positive CK,transformation efficiency was 60%,the result showed that multi-gene vector was introduced into genome of Populus.5.Observed the phenotype of the transgenic plants and measured the photosynthetic indexes.Compared with wild type,the height,ground diameter,flag leaf length and width of GOC-PYL1305 transgenic plants were increased,the leaf color was greener,indicating that the photosymthetic and biomass accumulation of transgenic plants became more efficient.the GOC pathway was successfully diverted part of the photorespiration,similar to the CO2 concentrating mechanism in C4 plants.