| 基因枪转化小麦主要轰击参数的优化 |
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| 引用本文： | 闵东红,何莎,张彦,夏兰琴.基因枪转化小麦主要轰击参数的优化[J].作物学报,2013,39(1):60-67. |
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| 作者姓名： | 闵东红 何莎 张彦 夏兰琴 |
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| 作者单位： | 1西北农林科技大学农学院, 陕西杨凌 712100; 2中国农业科学院作物科学研究所, 北京100081 |
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| 基金项目： | 本研究由国家转基因生物新品种培育重大专项(2011ZX08010-004)资助。 |
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| 摘 要： | 基因枪转化是目前小麦遗传转化的主要方法。影响基因枪转化效率的主要参数有轰击距离、轰击压力、DNA总量及浓度、金粉颗粒大小及用量等。为改善目前小麦遗传转化效率偏低的现状, 以普通小麦品种科农199为受体材料, 设金粉大小(0.6 μm和1.0 μm)、轰击距离(5.5 cm和8.5 cm)及轰击压力(650、900和1100 psi)三因素共12个处理, 利用pAHC25质粒转化小麦幼胚, 以确定最佳轰击参数, 建立稳定高效的普通小麦基因枪转化体系。结果表明, 3个因素的不同组合对幼胚的愈伤诱导效率没有明显影响, 但对再生率影响较大, 以金粉0.6 μm、轰击距离5.5 cm的再生率最高。3个因素组合的不同处理对GUS瞬时表达有明显影响, 以金粉0.6 μm、轰击距离5.5 cm和轰击压力650 psi处理的GUS瞬时表达量最高。本试验最终获得28株转基因植株, 其中12株来自金粉0.6 μm、轰击距离5.5 cm和轰击压力650 psi的处理, 该处理的平均转化率达2.66%。
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| 关 键 词： | 小麦 基因枪转化 轰击参数 转化效率 |
| 收稿时间： | 2012-04-19 |
Optimization of Key Bombardment Parameters in Biolistic Mediated Transformation in Wheat |
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| MIN Dong-Hong,HE Sha,ZHANG Yan,XIA Lan-Qin.Optimization of Key Bombardment Parameters in Biolistic Mediated Transformation in Wheat[J].Acta Agronomica Sinica,2013,39(1):60-67. |
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| Authors: | MIN Dong-Hong HE Sha ZHANG Yan XIA Lan-Qin |
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| Institution: | 1.College of Agronomy, Northwest A＆F University, Yangling, Shanxi 712100, China;2.Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China |
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| Abstract: | Biolistic mediated method is an important approach in wheat genetic transformation. The objective of this study was to optimize the bombardment parameters and establish an efficient and stable biolistic transformation system using common wheat (Triticum aestivum L.) variety Kenong 199with 12 treatments of parameter combinations composed of gold particle size, target distance, and acceleration pressure. The results indicated that different combinations of parameters had different effects on the transient expression of GUS gene encoding β-glucuronidase, regeneration of calli, and final transformation efficiency. The parameter combination of 0.6 μm gold, 5.5 cm target distance, and 650 psi acceleration pressure exhibited good regeneration and the highest level GUS transient expression. Furthermore, stable expression of GUS gene in transgenic lines was confirmed by histochemical staining assay. Of the 28 transgenic T0 plants obtained, 12 were from the treatment with the above optimized parameter combination, with an average transformation efficiency of 2.66%. |
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| Keywords: | Wheat Biolistic mediated-transformation Bombardment parameter Transformation efficiency |
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