Optimization Research On Open Tissue Culture Of Potato

Plant tissue culture has been increasingly widely used,but this technology requires long-term aseptic environment and demands for qualified technicians,long time of high-pressure sterilization.It is energy-consuming and costly.In contrast,open issue culture needs neither high-pressure sterilization nor ultra-clean workbench.It can be operated in a sterile natural environment,so it is simple,energy saving and cost-saving.In this study,tissue culture potato seedlings were used as materials to change the medium preparation and inoculation methods of tissue culture.The sterilization concentration of sodium hypochlorite soaking tissue culture container was preliminarily screened,and the concentration of bacteriostatic agent,agar and sucrose were added into the screening medium.Response surface optimization was performed for the concentration of sodium hypochlorite,concentration of bacteriostatic agent added in the medium,concentration of agar and concentration of sucrose.An ordinary worktable in the inoculation room was used to replace the ultra-clean worktable,and the effects of different inoculation methods and different inoculation sites on potato tissue culture seedlings were compared to find the appropriate inoculation methods and sites.The cleanliness of the inoculation room was compared with the ultra-clean table.The cost of open tissue culture is compared with that of traditional tissue culture in order to guide the practical application of open tissue culture.The results are as follows:1.When the concentration of sodium hypochlorite used for soaking and disinfection in tissue culture containers was 2% or above,no pollution occurred but albinism happens.When the concentration of sodium hypochlorite was 1.5%,the contamination rate of tissue culture seedlings reduced to 10% and the growth condition was good.In the open tissue culture medium,the contamination rate of tissue culture seedlings was 0 when the concentration of the bacteriostatic agent Iperon was 0.8ml/L.When sucrose concentration was 7.5g/L,the growth index of tissue culture seedlings under open inoculation condition was better,and the contamination rate was 10%.When the agar concentration in the open tissue culture medium was 8g/L,the contamination rate was 0.The tissue culture seedlings grew normally and were easy to operate.There was no difference in growth index and photosynthetic index between open inoculation and traditional inoculation.Openinoculation operation is simple and cost saving.Open inoculation was used in the study of response surface optimization design.2.The response surface design was optimized for the four factors.The optimal medium for open tissue culture of potatoes was obtained as follows: sodium hypochlorite 1%+Iperon0.5ml/L+ sucrose7.5g/L+ agar6.6g/L.The significant effects of four factors on the effective growth rate of the potato tissue culture seedlings were:sodium hypochlorite > sucrose > Iperon > agar.3.The cleanliness of the five worktables in the open inoculation room reached30,000 grade.In the actual inoculation process,the plants cultivated after open inoculation grew well.The pollution rate can be controlled under 5%.The total cost of traditional tissue culture preparation of 100 L culture medium for tissue culture seedlings for 28 days was 805.39 yuan.The cost of open tissue culture was about 2times of 426.06 yuan.The sterilization cost of traditional tissue culture was 312.73 yuan,which was 26 times of that of open tissue culture.The inoculation cost of traditional tissue culture was 83.11 yuan,which was 2.3 times of 36.59 yuan for open tissue culture.