CAJ | 학술논문

以花椰菜赛雪的带柄子叶为外植体,以Ms为基本培养基,GUS基因为报告基因,分析了遗传转化过程中的影响因子,如预培养时间、农杆菌菌液浓度、侵染时间、共培养时间、乙酰丁香酮浓度、延迟筛选时间等对外植体瞬间表达和稳定表达的影响.结果显示,以花椰菜的带柄子叶为外植体,预培养2d,农杆菌菌液为OD6000.3～0.4,侵染8min,共培养2d,乙酰丁香酮浓度为100μmol/L,延迟筛选7d,卡那霉素筛选压为5mg/L为最优的遗传转化方案,转化率最高可达35.7%.另外,GUS瞬间表达率和转化率并不存在绝对的相关性,但瞬间表达分析仍然可以作为外源基因进入受体细胞的指示.花椰菜农杆菌介导转化方案的优化研究为芸薹属蔬菜高效遗传转化提供了技术保障,有利于芸薹属蔬菜遗传育种与种质创新研究.
이화야채새설적대병자협위외식체,이Ms위기본배양기,GUS기인위보고기인,분석료유전전화과정중적영향인자,여예배양시간、농간균균액농도、침염시간、공배양시간、을선정향동농도、연지사선시간등대외식체순간표체화은정표체적영향.결과현시,이화야채적대병자협위외식체,예배양2d,농간균균액위OD6000.3～0.4,침염8min,공배양2d,을선정향동농도위100μmol/L,연지사선7d,잡나매소사선압위5mg/L위최우적유전전화방안,전화솔최고가체35.7%.령외,GUS순간표체솔화전화솔병불존재절대적상관성,단순간표체분석잉연가이작위외원기인진입수체세포적지시.화야채농간균개도전화방안적우화연구위예대속소채고효유전전화제공료기술보장,유리우예대속소채유전육충여충질창신연구.
An Agrobacterium-mediated transformation system,using transient and stable transformation assays,was used to evaluate some factors influencing transformation in cauliflower.These included the precuhivation time,the bacterial density.the inoculation duration time with Agrobacterium tumefaciens,the concentration of acetosyringone.the delay selection and the concentration of kanamycin in selection.Using cotyledons with 1-2mm petioles as explants,the best transformation parameters were:two-day pre-cultivation,OD6000.3～0.4 of bacterial density,eightminute infection with Agrobacterium tumefaciens,two-day cocuhivation,100μmol/L acetosyringone in the cocultivaiton medium and seven-day delay selection in kanamycin of 5mg/L.By optimizing the parameters on the procedure of Agrobacterium-mediated transformation,a high transformation efficiency(35.7%)was demonstrated.The GUS transient exzpression could be an important indicator for cauliflower transformation.The development of an efficient Agrobacterium-mediated transformation system opens up new opportunities for the functional characterization of genes and promotes the development of novel germplasm of Brassica vegetable genus.