Establishment Of Linum Usitatissimum L.callus Culture System And Determination Of Its Effective Component Content

Linum usitissimum L.belongs to Linaceae(Linaceae),which is an annual herb.People usually use its seeds as medicine.It is grown in a wide range in more than 20 countries,including the United States,Canada,Belgium,the Netherlands and India.It contains many chemical components such as alpha-linolenic acid and linoleic acid,which are significant in anti-cancer,cardiovascular and cerebrovascular diseases,blood pressure lowering and blood glucose lowering.Full and dry seeds of Linum usitissimum L.were used in this experiment.75% ethanol,mercuric chloride disinfectant time,types ofmedium were uesd as the independent variable,to investigate their effects on aseptic seedlings.Linum usitissimum L.callus will induce successful by using aseptic seedlings as experiment material,the explants tape,theage of explants,explants inoculation method,media tapy,light conditions and hormone combination will be set as the independent variables,to investigate their influence on Linum usitissimum L.callus induction.The condition of investigating Linum usitissimum L.callus were determined by using tweezers make the hypocotyl of Linum usitissimum L.Which cultivate 10 d into the MS medium with 3 mg/L IBA + 1.5 mg/L KT.Culture conditions: temperature 25 ?,humidity,light intensity for 2000 lx,light cycle is 12 h cultivation / 12 h dark light.Callus proliferation conditions of Linum usitissimum L.were optimized on the basis of the above,set the typeof medium,quantity of inoculation,cycle of light,pH,concentration of carbon source,subculture cycle and different hormone combinations as independent variables,through data analysis of their impact on Linum usitissimum L.callus growth.Finally,the best secondary conditions of Linum usitissimum L.were obtained: MS+1.5 mg/L 6-BA+0.2 mg/L IAA;The pH of medium was 6.5.The carbon source concentration is 30g/L;The cycle of light is in 12 h/d light culture;The subculture cycle is 28 d.After the establishment of a stable and genetically great Linum usitissimum L.callus culture system,change the light cycle,adjust the type and concentration of hormones,and establish the Linum usitissimum L.regeneration system.Suitable medium for Linum usitissimum L.'s regrowth germination was MS+1.00 mg/L KT+0.50 mg/LIAA;Suitable medium for the regrowth of Linum usitissimum L.to take root was MS+1.00 mg/L NAA.Finally,with the help of previous experiment,the chromatographic conditions should be optimized to determine and compare the alpha linolenic acid content and alpha linolenic acid content from Linum usitissimum L.seed,original plant of Linum usitissimum L.,Linum usitissimum L.regeneration plant,Linum usitissimum L.callus.The alpha linolenic acid content were 17.13%,1.17%,0.96% and 0.81% respectively,and the linoleic acid content were 4.01%,0.26%,0.23%,0.18% respectively.The establish of Linum usitissimum L.callus culture system and the Linum usitissimum L.regeneration system laid a foundation for further genetic operation of flax linen and expansion of medicinal resources.