The AFLLP Analysis Of Several NarcissusGermplasm Resources And The Hexaploid Induction Of Chongming Narcissus

We collected the narcissus varieties of Shanghai and Zhangzhou areas, using AFLP markers to test their genetic diversity. Which provide a molecular basis for protecting of the germplasm resource of narcissus. The variety of Chinese narcissus is single, and there is no new variety breeding in the long time. We through tissue culture with colchicine treatment to try to induce Chongming narcissus hexaploid, in order to seek the optimal condition of polyploid induction, we had three factors level of orthogonal test design, including colchicine concentration, processing temperature and time. The main conclusions are as follows:(1) The result discovered that Chongming narcissus and Zhangzhou narcissus genetic relationship is nearly. Zhuangyuan narcissus、Chongming narcissus and Zhangzhou narcissus genetic relationship is nearly. Chinese narcissus with daffodils genetic relationship is far away. The polymorphisms of the AFLP marker exceed that of the ISSR, found that can effectively reveals the specificity mark of daffodils.(2) This experiment has also established tissue culture system of Chongming narcissus. Tissue culture results show that:A short time sterilized was not effectively, butlong time HgCl2sterilization to explants cause harm, and the rate of pollution was still higher. The ability of differentiation of differentexophyte is different, is as follows:the intact scale stems> pedicel> ovary, the intact scale stems is the best exophyte to induce callus. Different hormone combination has a different effect of exophyte, KT and2,4-D are more good for callus induction than BA and NAA, and the MS+KT1.0mol/L+2,4-D0.5mol/L is the best medium MS1. The MS+BA1.0mol/L+2,4-D0.2mol/L is the best enrichment medium MS2. The optimum medium MS3of differentiation is MS+BA3.0mol/L. IBA and NAA can induce Chongming narcissus bulb take root, but NAA has actively promote role to induce Chongming narcissus rooting and stout root, MS4medium is1/2MS+NAA0.5mg/L+AC0.3%.(3) The analysis showed the factors have different influence of hexaploid induction, which was colchicine concentration> temperature> the processing time. Al、B1、C2respectively is the optimization levels of colchicine concentration, the processing time and temperature. In this experiment, the A1B1C2level combination is the optimal combination of Chongming narcissus hexaploid induction. (4) The234callus were treated by different colchicine concentration for1-4days and then transformed to MS medium.63bulbs were obtained and49bulbs among them were checked chromosome number from their root tips, found that colchicine can cause the extensive variations of root tip chromosome. Chongming narcissus hexaploid induction was not easy success, and aneuploidy mutation rate was high.18.4%bulbs (9/49) still kept triploidy and the most cells of its root tips contained30chromosomes.4.1%bulbs (2/49) displayed hexaploidy and contained60chromosomes.77.6%bulbs (38/49) displayed aneuploidy and the most cells of its root tips contained13-51chromosomes.