Production of transgenic orchardgrass via Agrobacterium-mediated transformation of seed-derived callus tissues (Retracted article. See vol. 198, pg. 110, 2013)
Production of transgenic orchardgrass via Agrobacterium-mediated transformation of seed-derived callus tissues (Retracted article. See vol. 198, pg. 110, 2013)
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DOI：
10.1016/j.plantsci.2006.05.006
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发表时间：
2006-09-01
期刊：
影响因子：
5.2
通讯作者：
Lee, Byung-Hyun
中科院分区：
文献类型：
--
作者：
Lee, Sang-Hoon;Lee, Dong-Gi;Lee, Byung-Hyun
采用农杆菌介导法对鸭茅进行遗传转化。已经从种子衍生的愈伤组织中发展出来。为了优化鸭茅遗传转化的条件，对影响农杆菌介导的DNA转移的几个因素进行了研究。种子胚性愈伤组织与A.含有双元载体pIG 121 Hm的根瘤菌菌株。GUS瞬时表达受A.根癌农杆菌菌株、共培养的持续时间以及在接种和共培养期间乙酰丁香酮的存在。在所测试的三个菌株中，EHA 101对瞬时GUS表达最有效，其次是GV 3101和LBA 4404。在接种和共培养培养基中加入200 μ M乙酰丁香酮，在目标愈伤组织中瞬时GUS表达从16.1%增加到56.1%。通过GUS活性证明了初级转化子的转基因性质。通过PCR和Southern印迹分析证实T-DNA整合到推定的转基因植物的基因组中。在分析的所有转基因植物中观察到一个或两个拷贝的转基因。转基因植株具有正常的表型，总转化效率为4.8%。因此，农杆菌介导的遗传转化可用于产生含有农艺学重要基因的转基因鸭茅。(c)2006爱思唯尔爱尔兰有限公司保留所有权利。
An Agrobacterium-mediated genetic transformation method for orchardgrass (Dactylis glomerata L.) has been developed from seed-derived callus. In order to optimize the conditions for orchardgrass transformation, several factors known to influence Agrobacterium-mediated DNA transfer were examined. Seed-derived embryogenic calli were co-cultivated with A. tumefaciens strains harboring the binary vector pIG 121 Hm. Transient GUS expression was greatly affected by the A. tumefaciens strain, the duration of co-cultivation, and the presence of acetosyringone during inoculation and co-cultivation. Among the three strains tested, EHA101 was most effective for transient GUS expression, followed by GV3101 and LBA4404. The inclusion of 200 mu M acetosyringone in both the inoculation and co-cultivation media increased transient GUS expression from 16.1 to 56.1% in targeted callus tissues. The transgenic nature of primary transformants was demonstrated by GUS activity. Integration of T-DNA into the genome of putative transgenic plants was confirmed by PCR and Southern blot analyses. One or two copies of the transgene were observed in all transgenic plants analyzed. The transgenic plants had a morphologically normal phenotype and the overall transformation efficiency was 4.8%. Thus, Agrobacterium-mediated genetic transformation can be used for the generation of transgenic orchardgrass containing genes of agronomic importance. (c) 2006 Elsevier Ireland Ltd. All rights reserved.