| 摘要: |
| 为探究提高农杆菌侵染效率的最优制备方法，以玉米自交系‘B 104’为转化受体，分析比较YEP固体和液体培养基，AB液体培养基制备的农杆菌EHA 105(植物双元表达载体pCAMBIA 3301)瞬时侵染幼胚能力的差异。结果表明，AB液体培养基制备的农杆菌瞬时侵染率达到100%，染色面积超过30%的幼胚占比约为40%，显著高于YEP培养基制备的农杆菌(P<0.05)；AB液体培养基侵染的幼胚产生胚性愈伤的比率为49%，显著高于YEP培养基制备的农杆菌(P<0.05)；AB液体培养基侵染产生的转化苗阳性转化率显著高于YEP液体培养基的阳性转化率(P<0.05)。因此，AB液体培养基提高了农杆菌侵染幼胚的转化率，获得了更多的胚性愈伤，保障了较高的阳性遗传转化率，优化了玉米自交系‘B 104’遗传转化技术体系。 |
| 关键词: 玉米遗传转化 YEP培养基 AB液体培养基 瞬时侵染率 |
| DOI：10.11841/j.issn.1007-4333.2022.03.02 |
| 投稿时间：2021-03-15 |
| 基金项目:河南省教育厅高等学校重点科研项目(20A180006)；河南大学人才引进科研启动基金 |
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| Comparison of the efficiency of Agrobacterium prepared by three different methods infecting the immature embryo of a transient infecting maize inbred line ‘B 104' |
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FENG Xinlei,LI Jieping
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| (School of Life Science, State Key Laboratory of Crop Stress Adaptation and Improvement/State Key Laboratory of Cotton Biology, Henan University, Kaifeng 475004, China) |
| Abstract: |
| In order to investigate the difference in the infection efficiency of Agrobacterium prepared by different methods, a corn inbred line ‘B 104', which is used as receptor in maize genetic transformation, was selected as the study object. The transient infecting ability of Agrobacterium EHA 105(plant binary expression vector pCAMBIA 3301)prepared by YEP medium, solid and liquid culture methods and AB lipid medium were analyzed and compared. The data of GUS staining and embryogenic callus induced by selection medium showed that the method of preparing Agrobacterium using AB lipid medium displayed a transient infecting ratio of 100%, and the proportion of immature embryos with an infestation area over 30% is about 40%, indicating infection ability of Agrobacterium prepared with AB lipid medium significantly higher than YEP medium. At the same time, the ratio of embryogenic callus produced by the immature embryos infected by Agrobacterium prepared with AB lipid medium on the selective medium was 49%, which was significantly higher than that prepared with YEP medium. The rate of positive transformation induced by AB lipid medium was significantly higher than that of YEP lipid medium. Therefore, the method of preparing Agrobacterium using AB lipid medium can increase the transient infecting rate of immature embryos, ensure more embryogenic calluses can be obtained on the selective medium, and ultimately increase the positive genetic transformation rate. It is suitable to be used to optimize the transgenic transformation system of maize inbred line ‘B 104'. |
| Key words: maize genetic transformation YEP medium AB lipid medium transient transformation efficiency |
