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【目的】培育耐寒性强的地被菊花[Dendranthemagrandiflorum(Ramat.)Kitamura]新材料。【方法】以FallColor品种的幼嫩叶片为外植体,探讨胚状体诱导所需的植物生长调节剂浓度和诱导培养时间等条件。【结果】叶片外植体在添加0.75mg·L-12,4-D的诱导培养基上诱导15d,再进行分生培养,不仅能够诱导胚性愈伤组织形成,还能够诱导胚状体发生,并进一步诱导芽再生,最终93%的供试外植体通过胚状体途径获得芽的再生。通过根癌农杆菌介导法将35S启动子驱动的逆境诱导转录因子DREB1A基因导入该品种。转化株在低温下的种子发芽率、扦插苗生长以及植株露地越冬生长状况等方面都明显优于对照。【结论】本研究成功地建立了地被菊花FallColor体细胞胚再生途径,并成功地获得了具有越冬耐性的地被菊花转化株系。
【Objective】 To cultivate the new material of Dendranthemagrandiflorum (Ramat.) Kitamura, which has high cold resistance. 【Method】 Young leaves of FallColor cultivars were used as explants to investigate the conditions of plant growth regulators and induction time required for embryoid induction. 【Result】 Leaf explants were cultured in induction medium supplemented with 0.75 mg · L-12,4-D for 15 days and then cultured in meristem, which not only induced the formation of embryogenic callus but also induced the embryogenesis , And further induced shoot regeneration. Finally, 93% of the tested explants achieved bud regeneration through the embryoid body pathway. The 35S promoter-driven stress-responsive transcription factor DREB1A gene was introduced into this variety by Agrobacterium tumefaciens-mediated method. Transformants in the low temperature seed germination rate, cuttings growth and plant overwintering winter growth conditions are significantly better than the control. 【Conclusion】 In this study, the regeneration pathway of FallColor somatic embryos was successfully established and the transformed chrysanthemum-transformed plants with overwintering tolerance were successfully obtained.