The Research Of Initiation And Subculture Of Embryogenesis Tissue Of Cunninghamialanceolata(Lamb.) Hook Based On The Immature Zygotic Embryo

The different developmental stages of immature cones of Chinese fir[Cunninghina lanceolata(Lamb.)Hook] were choosed as explants for inducing embryonic tissue.In the experiments,we studied the effects of auxin, cytokinin, abscisic acid and brassinolide on the intiation rate of embryonic tissue of Chinese fir.We also investigate the effect of different consistency of ABA and PEG4000 on the mature of somatic embryo.According to the results of our trials,we found the right developmental stages of immature cones and genetype and the optimal formula for inducing embryonic tissue.In our experiments,the intiation rate of embryonic tissue of Chinese fir incresed greatly.The main results our trials as follows:1.The developmental stages of immature cones of Chinese fir have great influnce on the intiation rate of embryonic tissue.The cones in the fertilization and prembryo stages are the optimal explant for inducing embryonic tissue of Chinese fir.Because of the lower intiation rate,the cones in the other stages are not appropriate for inducing embryonic tissue.2.Genetypes also have a great influnce on the intiation rate of embryonic tissue. In the stage of fertilization,the initiation rate of six genetype of Chinese fir as follows:己2(15.20%),洋21(11.38%),丁79 (10.40%),丁34(6.9%),洋38(6.04%),洋18(3.89%). In the stage of preembryo,the initiation rate of six genetype of Chinese fir as follows:己2(11.37%),丁79 (8.18%),洋38(8.09%),丁34(5.80%),洋21(4.47%),洋18(3.17%). In the stage of cleavage polyembryony,the initiation rate of six genetype of Chinese fir as follows:己2(4.02%),丁79 (3.44%),丁34(2.63%),洋21(2.47%),洋18(1.29%),洋38(1.17 %). In the stage of cleavage polyembryony,the initiation rate of six genetype of Chinese fir as follows::己2(1.63%),丁79 (1.40%),洋21(1.31%),丁34(1.05%),洋38(0.92%),洋18(0.88%).3.The combination of media and hormone also have a great influnce on the initiation rate. We choosed out the optimal combination of media and hormone accroding the different genetype.(1) For the genetype丁34, the best combination of media and hormone is DCR +NAA(1.0mg/L)+6BA(0.5mg/L)+KT(0.5mg/L)+ABA(0.5mg/L)+BL(0.05mg/L)in the stage of fertilization and preembryo.In the stage of cleavage polyembryony, the best combination of media and hormone is DCR+NAA(2.0mg/L)+6BA(0.5mg/L)+ KT(0.5mg/L)+ABA(1.0mg/L)+BL (0.025mg/L). In the stage of cylindrical embryo, DCR+NAA(2.0mg/L)+6BA(0.5mg/L)+ KT(0.5mg/L)+ABA(1.0mg/L)+BL(0.005mg/L).(2) For the genetype丁79, the best combination of media and hormone is LP+NAA(1.0mg/L)+6BA(0.5mg/L)+KT(0.5mg/L)+ABA(0.5mg/L)+BL(0.05mg/L)in the stage of fertilization and preembryo.In the stage of cleavage polyembryony, the best combination of media and hormone is DCR+NAA(2.0mg/L)+6BA(0.5mg/L)+ KT(0.5mg/L)+ABA (1.0mg/L)+BL(0.025mg/L).Inthe stage of cylindrical embryo, LP+NAA(2.0mg/L)+6BA (0.5mg/L)+ KT(0.5mg/L)+ABA(1.0mg/L)+BL(0.05mg/L).(3) For the genetype己2, the best combination of media and hormone is DCR+NAA(0.5mg/L)+6BA(0.5mg/L)+KT(0.5mg/L)+ABA(0.5mg/L)+BL(0.025mg/L)in the stage of fertilization and preembryo.In the stage of cleavage polyembryony, the best combination of media and hormone is DCR+NAA(1.0mg/L)+6BA(0.5mg/L)+ KT(0.5mg/L)+ABA(1.0mg/L)+BL (0.05mg/L). (4) For the genetype洋21, the best combination of media and hormone is LP+NAA(1.0mg/L)+6BA(0.5mg/L)+KT(0.5mg/L)+ABA(0.5mg/L)+BL(0.05mg/L)in the stage of fertilization and preembryo.In the stage of cleavage polyembryony, the best combination of media and hormone is DCR+NAA(1.0mg/L)+6BA(0.5mg/L)+ KT(0.5mg/L)+ABA(1.0mg/L)+BL (0.025mg/L).In the stage of cylindrical embryo, LP+NAA(2.0mg/L)+6BA(0.5mg/L)+ KT(0.5mg/L)+ABA(1.0mg/L)+BL(0.025mg/L).(5) For the genetype洋21, the best combination of media and hormone is LP+NAA(0.5mg/L)+6BA(0.5mg/L)+KT(0.5mg/L)+ABA(1.0mg/L)+BL(0.025mg/L)in the stage of fertilization and preembryo.In the stage of cleavage polyembryony, the best combination of media and hormone is DCR+NAA(1.0mg/L)+6BA(0.5mg/L)+ KT(0.5mg/L)+ABA(1.0mg/L)+BL (0.05mg/L). In the stage of cylindrical embryo, LP+NAA(2.0mg/L)+ KT(0.5mg/L)+ABA (1.0mg/L)+BL(0.025mg/L).(6) For the genetype洋38, in the stage of fertilization, the best combination of media and hormone is LP+NAA(0.5mg/L)+6BA(0.5mg/L)+KT(0.5mg/L)+ABA(1.0mg/L)+BL(0.025mg/L). In the stage of preembryo, the best combination of media and hormone is LP+NAA(0.5mg/L)+ 6BA(0.5mg/L)+KT(0.5mg/L)+ABA(1.0mg/L)+BL(0.025mg/L).In the stage of cleavag poly -embryony, the best combination of media and hormone is LP +NAA(2.0mg/L)+6BA(1.0mg/L) +ABA(0.5mg/L)+BL(0.05mg/L).In the stage of cylindrical embryo, the best combination of media and hormone is LP+NAA(2.0mg/L)+6BA(0.5mg/L)+KT(0.5mg/L)+ABA(1.0mg/L)+BL (0.05mg/L).4.The optimal media of subculture is DCR+NAA(0.1 mg/L)+KT(0.1 mg/L).