Hu Zhang-Hua, Chen Jin-Qing, Wu Guan-Ting, Jin Wei, Lang Chun-Xiu, Huang Rui-Zhi, Wang Fu-Lin, Liu Zhi-Hong, Chen Xiao-Yun
Institute for Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao. 2005 Apr;31(2):149-59.
An efficient and reproducible system has been developed for the production of transgenic plants in tall fescue (Festuca arundinacea Schreb.) using A. tumefaciens-mediated transformation. Two-months-old suspension cultures served as excellent explants for transformation. The explants were inoculated with an A. tumefaciens strain EHA105 carrying a plasmid pDBA121 containing genes for hygromycin phosphotransferase (hpt) and phosphinotricin acetyltransferase (bar). The commercial herbicide Basta was used as a selective agent. Inclusion of acetosyringone (ACS) 20 mg/L in the co-culture medium led to an increase in transformation efficiency. The efficiency of transformation was highly dependent on the genotype, the explant, the culture medium and selective agent used. Tall fescue transformation efficiency is 2.85-10.9 plants per gram fresh weight (FW) of suspension cultures. This is much higher than the corresponding values reported before (2-5 plants). So far more than 300 transgenic plants have been obtained, the fertility of some transgenic plants had decreased. Stable integration and high expression of the transgenes were confirmed by PCR analysis and Southern blot hybridization or herbicide Basta spraying test.
已开发出一种高效且可重复的系统，用于利用根癌农杆菌介导的转化方法在高羊茅（Festuca arundinacea Schreb.）中生产转基因植物。两个月大的悬浮培养物用作转化的优良外植体。将外植体用携带质粒pDBA121的根癌农杆菌菌株EHA105接种，该质粒含有潮霉素磷酸转移酶（hpt）和草丁膦乙酰转移酶（bar）基因。商业除草剂Basta用作选择剂。在共培养基中加入20 mg/L的乙酰丁香酮（ACS）可提高转化效率。转化效率高度依赖于基因型、外植体、所用培养基和选择剂。高羊茅的转化效率为每克悬浮培养物鲜重（FW）2.85 - 10.9株植物。这远高于之前报道的相应值（2 - 5株植物）。到目前为止，已获得300多株转基因植物，一些转基因植物的育性有所下降。通过PCR分析、Southern杂交或除草剂Basta喷洒试验证实了转基因的稳定整合和高表达。
