Study On Key Technologies Of Somatic Embryo Suspension Culture Of Broussonetia Papyrifera

This paper is mainly study on Broussonetia papyrifera.In order to obtain somatic embryo by establishing the system.of callus.Establish stable suspension cell system of Broussonetia papyrifera by creating dynamic model of cell suspension culture. At the same time, the key influencing factors of somatic embryo induction were studied. The main results are as follows:(1) In the process of Broussonetia papyrifera callus induction,while the blade was induced material,the callus induction rate was the highest.And the induced callus cells grow best. The best medium was :MS+1.0 mg/L 6-BA+2.0 mg/L 2,4-D, the induction rate reached96.55%.(2) After 20 days of subculture on solid MS medium,the phytohormone combinations of1.0 mg/L 6-BA+0.5 mg/L 2,4-D showed the best subculture effect.(3) The.growth curve of Broussonetia papyrifera callus suspension cell showed typical "s" type.After thirteen days cultured,the proliferation rate of callus was up to 4.32,The speed of growth was becoming slowly.From the 19 th day,callus started the stagnant period of growth.(4) In the phytohormone combination optimization test of Broussonetia papyrifera suspension cell, the mass concentration of 6-BA was very significant.The concentration of2,4-D and KT had significant influence on the suspension cultured results.The optimization results showed that the optimal hormone combinations for suspension culture was: 1 mg/L6-BA+0.5 mg/L 2,4-D+0.4 mg/L KT+0.4 mg/L NAA.(5) Under the combination of 4 g fresh weight and 50 ml liquid medium, the proliferation effect of callus was the best.(6) In the test of creating dynamic model of cell suspension culture The analysis of response surface test showed that the fresh weight ratio of suspension cultured cell Y=+5.72-0.48A+0.067B-0.079C-0.85D-0.1AB-0.27AC-0.21AD-1.09BC+0.05BD+0.038CD-1.05A2-1.02B2-0.50C2-0.24D2. The factor A(shaking speed) and D factors(the concentrationof carbon source) had significant effect in the equation.B factor(temperature) and C factors(PH) had obvious synergistic effect in the equation.(7) After the orthogonal test of optimal conditions of somatic embryo induction, the result showed that the best combination of conditions for inducting somatic embryo genesis of Broussonetia papyrifera was:cultured at the temperature of 25 and under the light illumination of 1800 lx, MS + 2.0 mg/L 6-BA+ 1.0 mg/L NAA,pH 6.4.