摘要
本研究通过农杆菌介导法将IPT基因转入到地被菊‘紫妍’中,并确立了最优遗传转化体系。具体操作为:将地被菊‘紫妍’的叶片在无菌条件下剪成0.5 cm×0.5 cm的小叶,在MS+6-BA 2.0 mg/L+NAA 1.0 mg/L培养基上预培养2 d,置于OD 600浓度为0.5的农杆菌菌液中侵染3 min,之后接种到共培养基(同预培养基)中,在28℃培养箱中暗培养36 h。再接种到Hyg浓度为3 mg/L、头孢浓度为500 mg/L的培养基(同预培养基)上诱导叶片分化和脱菌,待抗性愈伤组织分化出不定芽后,切下长约1.5 cm的不定芽,接种到1/2 MS+NAA 0.2 mg/L+Hyg 14 mg/L的培养基中进行生根培养。利用该体系对地被菊‘紫妍’叶片进行转化,可获得生根且长势良好的抗性植株。
Agrobacterium mediated method was used to transfer the IPT gene into Chrysanthemum grandiflorum‘Ziyan’and the optimal genetic transformation system was established.The specific operation was to cut the leaves of the Chrysanthemum grandiflorum‘Ziyan’into small leaves of 0.5 cm×0.5 cm under sterile conditions and pre-cultivated them on MS+6-BA 2.0 mg/L+NAA 1.0 mg/L medium for 2 days.Then,infected them in a agrobacterium bacterial solution with an OD 600 of 0.5 for 3 minutes and inoculated them onto co-culture medium(same as pre-culture medium)and black cultured at 28℃for 36 hours.Later,induced leaf differentiation and bacterial removal on medium(same as pre-culture medium)with a concentration of 3mg/L of Hyg and 500mg/L of Cephalosporin.After the resistant callus tissue differentiated into adventitious buds,cut off about 1.5 cm of adventitious buds and inoculated them into a 1/2 MS+NAA 0.2 mg/L+Hyg 14 mg/L medium for rooting culture.By using this system to transform the leaves of Chrysanthemum grandiflorum‘Ziyan’,resistant plants with good rooting and growth could be obtained.
作者
高松
Gao Song(National Forest Administration Bureau of Zhongtiao Mountain in Shanxi Province,Houma 043000,China)
出处
《山西林业科技》
2024年第2期7-10,共4页
Shanxi Forestry Science and Technology
关键词
地被菊‘紫妍’
IPT基因
遗传转化
抗性植株
Chrysanthemum grandiflorum‘Ziyan’
IPT gene
Genetic transformation
Resistant plants
