| Lily is a kind of perennial monocotyledonous bulbar flower which has high economical and ornamental value,and now has been one of the most important fresh flowers.But the bulbs degenerated severely,the pollens stained the petals,and the plant diseases and insect pests restricted the production and marketing remand of lily.so, cultivaring the innovative breeds which is high fastness,no pollens,no degeneration by some effective technology is very necessary.In present study,primary study was conducted for the lily innovative breeds by the technology of cross breeding and genetic transfomation.The results were as fllows:(1) Cross breedingTwo combinations of pollination was carried out in lilium longiflorum(♀)×Lilium lophophorum var.linearifolium(♂) and lilium longigflorum 'Gelria'(♀)×lilium longiflorum(♂),but no mature seeds were obtained from these two combinations in vivo culture.Ovules excised from each carpel 10 and 25 days after pollination(DAP) were cultured on B5 medium and half-strength B5 medium containing sucrose at different concentrations in vitro.In lilium longiflorum(♀)×Lilium lophophorum var.linearifolium (♂),1.17%of ovules excised at 10 DAP developed into seedlings.In lilium longiflorum'Gelria'(♀)×lilium longiflorum(♂),1.00%of ovules excised at 25 DAP developed into seedlings.In the present study,we observed normal and abnormal embryo development,and identified hybrids based on inter-simple sequence repeat(ISSR) analyse.(2) Genetic transfomationRegeneration system In present study,perfect regeneration system has been established,the regeneration rate of pseudo-bulblet,ledfstalk and leaf which come from Oriental hybrid lily(Lilium L.) cvs.'Sorbonne','Siberia','Acapulco'and lilium longiflorum can be reach to 95%,90%and 80%respectively.Transmit system Pseudo-bulblet,ledfstalk and leaf come from Oriental hybrid lily (Lilium L.) cvs.'Sorbonne','Siberia' and lilium longiflorum were utilized as explants in present study.The transmit system was studyed from basical culture medium,infaction time,co-culture time,Kan selecting concentration,bacterium selecting,hormone and scaring on surface of explants,then we obtained the optimal transmit system:improved MS culture medium(no NH4NO3),bacterium C58,infected 20 min,co-cultured 4 days, scared the explant surface,Kan 120,100,8凹?/L separate to pseudo-bulblet,ledfstalk and leaf.(3) Adjustment of the status of embryogenic callus and plant regeneration of Lilium longiflorumExperiments were performed in order to obtain embryogenic callus with better status.The effect of different concentrations of CoCl2,ABA and sucrose,in addition,the condition of illumination were studied.Enhanced proportion and dispersion of somatic embryos were achieved by the use of CoCl2 with the concentration of 0.05μM/L,and improved propagation volume and propagation coefficient of somatic embryos were obtained using 0.01μM/L and 0.02μM/L CoCl2;Some negative effect on growing status, water content,propagation volume and propagation coefficient of somatic embryos was observed when the concentration of ABA was gradually increased,while proportion and dispersion of somatic embryos were not affected by different ABA concentrations;It was beneficial for proportion and dispersion of somatic embryos by adding 6.0g/L or 9.0g/L sucrose and cultured in light,whereas,when somatic embryos were cultured at dark,only 6.0g/L sucrose could improve proportion and dispersion;Furthermore,the water content and dispersion of somatic embryos were not affected whether cultured in light or not.Considering all factors in present study,the results demonstrated that the most appropriate condition for the proliferation of somatic embryos was MS medium supplemented with NAA(5.4μM/L),TDZ(0.4μM/L),CoCl2(0.05μM/) and sucrose (6.0g/L) under light. |
