%0 Journal Article %T 农杆菌介导的胡麻转化因子的优化<br>Optimization of Flax (Linum usitatissimum L.) Transformation Factors Mediated by Agrobacterium %A 李淑洁 %A 罗俊杰 %A 林玉红 %A 裴怀弟 %A 李闻娟 %J 西北农业学报 %D 2018 %R 10.7606/j.issn.1004-1389.2018.07.007 %X 农杆菌介导法是植物遗传转化的重要方法之一，为了提高农杆菌介导的胡麻遗传转化效率，以‘陇亚10号’和‘定亚22号’下胚轴为基因转化受体，以pCAMBIA3301为转化质粒，分析草铵膦选择压、下胚轴预处理（剥表皮、预培养）、农杆菌菌株、侵染时间和乙酰丁香酮对gus表达率/频率和抗性苗率的影响，优化以上转化因子参数，建立高效的胡麻遗传转化体系，为后续的胡麻功能基因组学研究和遗传改良奠定基础。结果表明：下胚轴愈伤组织诱导、分化和再生苗生根阶段的草铵膦选择压分别是10 mg/L、5 mg/L和2 mg/L；下胚轴预处理对gus表达率/频率显著影响，却显著影响抗性苗分化率，不剥表皮和预培养3 d抗性苗率分别提高5倍和9倍；农杆菌菌株和侵染时间都显著影响gus表达率/频率和抗性苗率，C58c1侵染60 min时gus表达频率最高，为7.5，抗性苗率为60.3%。乙酰丁香酮能显著提高C58c1和GV3101介导转化时gus表达频率。<br>The transformation system mediated by Agrobacterium tumefaciens is one of the important methods in plants genetics and transformation. In order to improve the transformation efficiency, two common flax (Linum usitatissimum) domestic cultivar Longya-10 and Dingya-22 were used as transformation receptor in the study. Plasmid pCAMBIA3301 contain gusA gene as transformed vector, the effects of key parameters including phosphinothricin concentration, hypocotyl pretreatments (removal of epidermal cell or not, preculture length), Agrobacterium tumefaciens strain, inoculation time and acetosyringone (AS) on gus expression rate/frequency and regeneration ratio of phosphinothricin-resistant plantlet were analyzed. By optimizing the key parameters above, we established an efficient flax transformation system, and laid the foundation for the future functional genomics and genetic improvement in flax. The results indicated that appropriate concentration selection of phosphinothricin in callus induction, callus regeneration and rooting stage were 10 mg/L, 5 mg/L and 2 mg/L, respectively; Though the hypocotyl pretreatments had no great effects on gus expression, they dramatically effected regeneration ratio of phosphinothricin-resistant plantlet. Compared to the control, no-epidermis-removal and 3-day-preculture treatments exhibited 5 and 9 times increase, respectively. Both Agrobacterium tumefaciens strain and inoculation time dramatically effected gus expression and regeneration ratio of phosphinothricin-resistant plantlet. The highest gus expression frequency (7.5) and phosphinothricin-resistant plantlet ratio (60.3% ) were gained when hypocotyls were inoculation with C58c1 60 minutes. Acetosyringone with concentration of 100 μmol/L dramatically improved the gus expression frequency of C58c1 and GV3101. %K 胡麻 农杆菌介导法 转化因子 抗性苗率 gus表达率< %K br> %K Flax(Linum usitatissimum L.) Agrobacterium tumefaciens -mediated transformation Transformation factors Calli regeneration ratio gus expression efficiency %U http://xbnyxb.alljournals.cn/ch/reader/view_abstract.aspx?file_no=20180707&flag=1