Gonzalez Padilla I M, Webb K, Scorza R
USDA-ARS Appalachian Fruit Research Station, 45 Wiltshire Rd, Kearneysville, WV 25430, USA.
Plant Cell Rep. 2003 Aug;22(1):38-45. doi: 10.1007/s00299-003-0648-z. Epub 2003 Jun 24.
We describe here an improved system for routinely developing transgenic plum plants (Prunus domestica L.) through the use of Agrobacterium tumefaciens. The production of non-transformed "escapes" has been virtually eliminated, and rates of plant establishment in the greenhouse have been dramatically improved. The system is based on the regeneration of shoots from hypocotyls extracted from mature seed. The shoot regeneration medium is Murashige and Skoog (MS) salts and vitamins supplemented with 7.5 microM thidiazuron and 0.25 microM indole-butyric acid. Transferring the explants after co-cultivation to shoot regeneration medium containing 80 mg l(-1) of kanamycin and 300 mg l(-1) of Timentin reduced the total number of regenerated shoots without affecting the transformation rate. Transformation rates using the described system averaged 1.2% of the hypocotyl slices producing transgenic plants, with a range of 0-4.2%. The transgenic shoots rooted at a rate of 90% on half-strength MS salts and vitamins supplemented with 5 microM alpha-naphthaleneacetic acid and 0.01 microM kinetin. Plantlets were transferred to a greenhouse directly from culture tubes with a 90% average survival.
我们在此描述一种改进的系统，用于通过根癌农杆菌常规培育转基因李属植物（欧洲李）。几乎消除了未转化的“逃逸植株”的产生，并且温室中植株的成活率得到了显著提高。该系统基于从成熟种子中提取的下胚轴再生芽。芽再生培养基是添加了7.5微摩尔噻苯隆和0.25微摩尔吲哚丁酸的Murashige和Skoog（MS）盐及维生素。共培养后将外植体转移至含有80毫克/升卡那霉素和300毫克/升替卡西林的芽再生培养基中，可减少再生芽的总数，而不影响转化率。使用所述系统，产生转基因植株的下胚轴切片的转化率平均为1.2%，范围为0 - 4.2%。转基因芽在添加了5微摩尔α - 萘乙酸和0.01微摩尔激动素的半强度MS盐及维生素上的生根率为90%。小植株直接从培养管转移至温室，平均成活率为90%。
