| Maize mature embryo is good acceptor system suitable for biological research techniques and an improved maize varieties, maize mature embryo as explants regeneration system is an important area in recent research. Because the mature embryos of maize are easily obtioned, not limited by time and quantity, But the mature embryos of maize are difficult to induce, the quality of the embryonic callus is poor, most varieties are difficult to regenerate, and resrtricted by genotype. So the study of the tissue conditions of maize mature embryos, the select of good kinds of maize varieties, establish efficient system of maize mature embryo regeneration have significance for study maize transgenic. This experiment through different varieties, different hormones, different explants research, to be looked for optimal system of the callus inducted by mature embryo, mainly in the following aspects:1、By stuying influence of inducement for callus in different maize varieties, The result showed, under the condition of4mg/L2,4-D, the rate of induction by18-599W was53.33%, HI-II was49.33%, CML295was47.33%, yidan947was9.33%. In the same culture condition, there are remarkable different between varieties of callus growth conditions and induction rate.18-599W was most suitable for callus induction, the second was CML295、HI-II, the worst was yidan947.2、By studying influence of inducement for callus in IAA、BL、4mg/L2,4-D mixed with BL, the result showed:under the condition of IAA culture,8mg/L was the best concentration, the rate of induction by18-599W was34.67%, HI-II was34.00%, CML295was32.00%; under the condition of BL culture, the best concentration was0.2mg/L, the rate of induction by18-599W was56.67%, HI-II was50.00%, CML295was52.00%; in4mg/L2,4-D and BL mixed culture conditions, the best concentration was4mgL2,4-D+0.2mg/L BL, the rate of induction by18-599W was93.33%, HI-II was90.00%,CML295was88.67%.3-. By studying influence of inducement for callus in different hormones, the result showed:the rate of induction by18-599W in the condition of4mg/L2,4-D+0.2mg/L BL.0.2mg/L BL、4mg/L2,4-D、8mg/L IAA respceted is93.33%,56.67%,53.33 %、34.67%.2,4-D+BL was most suitable for callus induction, the second was BL、2,4-D, the worst is the IAA.4、 Different explants(root、mature embryo、hypocotyl、spire) callus inducton different comparison, the result showed: different explants in the culture condition of4mg/L2,4-D and0.2mg/L BL had the same efficient in callus induction, also had a good effect,especially4mg/L2,4-D+0.2mg/L BL most sutiable callus induction of different explants. To18-599W for example, in the condition of4mg/L2,4-D+0.2mg/L BL, the root tip of induction rate was77.33%, the mature embryo of induction rate was93.33%, the hypocotyl of induction rate was33.33%, the spire of induction rate was16.00%. So, the best for callus induction was mature embryo, the second was the root, the next was the hypocotyls, the last was spire.5、The preliminary research of agrobacterium-mediated genetic transformation by the mature embryo and the callus induced by mature embryo, the result showed:By the mature embryo culture systemcan be used as acceptor material in Agrobacterium transformation. This experiment succeded by Agrobacterium transformation method put pEGAD plasmid into mature embryo, and transgenic plant obtained through this system. This experiment selected the variety and eplant callus to adapt to induce callus, and the methods by induction of callus from mature embryos was optimized. Through the result of induction and genetic transformation, proved that the callus induced by mature embryo of18-599W in the condition of4mg/L2,4-D+0.2mg/L BL suitable for the receptor of genetic transformation. |
PDF Full Text Request