Induced Calli And Establishment Of Regeneration System Of Mature Maize Embryo

The maize as forage,grain,economical crop has great potential in our country. Studying the maize variety improvement mostly concentrate on quality and resistance, and there are less research based on the maize mature embryo as explant establishing regeneration systems. Because of less time restriction and the quantity guarantee, It is a good test system for biotechnology research and varity resources innovation. But mature embryo embryogenic calli has bad quality and most of them difficult to regenerate.so it severely interfer the research of gene-engineering.So studying the influencing factors in maize transgenic , the frequency of plant regeneration and the regeneration system has a very important effect on maize mature embryo tissue culture. In this research ,we used mature maize K12 as the material,through different cutting mode, carbons sources,mediums and hormones inducement for it .This test provide a theoretical basis for seeking a suitable reference on differentiation,subculture,induct calli,hormones and its concentration ratio and so on.1.By studying influence of inducement for calli in different cutting methods (Crosscut,Slitting Line),the result showed,inducement frequency of seeds embryo Calli with crosscut was higher than Sitting line. Some crosscut seeds can not induct calli.for this cuting method similar to whole mature seeds(restriction in tissue and not easly forme calli).The cuting in line not only helpful for forming calli because of the effect directly two half-embryo machine injury, but also helpful for inducting calli for inhibiting embryo occurrence directly.2. Through studying inducted effect of different explants(embryo,radicali,hypocotyl) to induct calli, The result showed,different explants require different hormones and its concentration, at the same time, the different explants have significant difference on the hormones and its Concentration.3. By studying effect of carbons sources (maltose,Sucrose)and mediums (LS,N6,MS)to differentiatio,subculture and induction calli. The result showed,maltose was more suitble to maize embryo calli induction,subculture and differentiation than Sucrose;LS medium helpful to calli induction,subculture and differentiation of mature maize embryo,the second was N6 medium.4. By studying effect of 2.4-D,NAA on calli induction and subculture, The result showed ,low and high concentration of 2.4-D all was not suitble to calli induction and subculture. The growth process of embryo,radicali,hypocotyl calli induction and subculture was slow when the concentration of 2.4-D low to 1.0mg/L. embryo,hypocotyl calli were injuried when the 2.4-D concentration of 2.4-D high to 1.0mg/L.when the concentration of 2.4-D high to 2.5mg/L go against growth of Radicali calli.5. By studying effect of NAA on calli induction and subculture, The result showed,it suitable embry and hypocotyl calli subculture when the concentration of NAA 0.5mg/L.It promote radical calli growth when the concentration of NAA 0.5-0.6mg/L;if the concentration of 2.4-D was 2.0-2.5mg/L, it radically promote calli growth;It was helpful to calli differentiation for different explants when concentration of NAA 0.5mg/L.6. By studying effect of 6-BA on calli induction and subculture, The result showed,it was suitable to embry, radical and hypocotyl calli subculture when the concentration of 6-BA 0.2mg/L.But the high concentraion was the important factors to variation of plantlet.So it unnecessary for calli differentiation with high concentration of 6-BA .It was helpful to calli differentiaton for different explants when the concentration of 6-BA 1.5mg/L .7. By studying effect of 2.4-Dand 6-BA on calli subculture, The result showed,the ratio of 2.5mg/L 2.4-D and 0.5mg/L NAA were helpful to calli subculture of embryo,radicali and hypocotyl .8. By studying effect of NAA and 6-BA on calli subculture.The result showed,if adding 0.5mg/L NAA and 1.5mg/L 6-BA in medium, it helpful for calli differentiation of mature maize seeds.In addition, the time of subculture affect the quality of calli severely.The low subculture frequency, the more serious of water socking.The more subculture frequency, the more degree of browning.