Zhang Chun-Yu, Li Hong-Yu, Liu Bin
College of Agriculture, Shenyang Agriculture University, Shenyang 110161, China.
Yi Chuan. 2012 Dec;34(12):1599-606. doi: 10.3724/sp.j.1005.2012.01599.
The decision of using selection marker is one of the key factors for success of plant genetic transformation and offspring screening. As two commonly used selection markers, hpt and bar genes are widely used in tissue culture-based rice transformation. To experimentally compare their performance, we investigated the efficiency of two transformation systems using Hygromycin and Bialaphos as the selection agents, respectively. The result indicated that the system using hpt gene as the selection marker saved 10 days and had double transformation efficiency and lower transgene copy number in comparison to the system using bar gene. Then, we assessed the feasibility of screening transgenic rice in the field by soaking the wild-type and transgenic seeds in a series of solutions containing step diluted hygromycin for two days. We targeted the suitable concentration for distinguishing the transgenic seeds from WT Kitaake seeds was 167 mg L(-1). However, the cost of screening by hygromycin is still much higher than that of Basta in field test. Therefore, this study experimentally demonstrated the advantages and disadvantages of the hpt and bar gene as the selection markers and thus provided a reference for choose of an appropriate selection marker according to the practical applications.
选择标记的使用决策是植物遗传转化和后代筛选成功的关键因素之一。作为两种常用的选择标记，潮霉素磷酸转移酶（hpt）基因和草丁膦抗性（bar）基因在基于组织培养的水稻转化中被广泛应用。为了通过实验比较它们的性能，我们分别研究了以潮霉素和双丙氨膦作为选择剂的两种转化系统的效率。结果表明，与使用bar基因的系统相比，使用hpt基因作为选择标记的系统节省了10天时间，转化效率提高了一倍，且转基因拷贝数更低。然后，我们通过将野生型和转基因种子在一系列含有逐步稀释的潮霉素的溶液中浸泡两天，评估了在田间筛选转基因水稻的可行性。我们确定区分转基因种子和野生型北陆种子的合适浓度为167毫克/升。然而，在田间试验中，用潮霉素筛选的成本仍远高于用草铵膦筛选的成本。因此，本研究通过实验证明了hpt和bar基因作为选择标记的优缺点，从而为根据实际应用选择合适的选择标记提供了参考。
