农杆菌介导的黄羊茅转化研究

Jun-Sheng Zhao, Da-Ying Zhi, Zhe-Yong Xue, Guang-Min Xia
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引用次数: 0

摘要

用2株农杆菌AGL1和GV3101转化4种高羊茅(羊茅)胚源愈伤组织。AGL1携带内含子atnhx1表达载体pROK2U,内含泛素启动子和npt II标记基因。GV3101携带内含子atnhx1表达载体pROK2,内含35S启动子和npt II基因。与AGL1或GV3101共培养后,用50 ~ 150 mg/L的paromomycine筛选愈伤组织,抗性愈伤组织再生抗性植株1126株。用10 ~ 20 mg/L卡那霉素进一步筛选,525株植株保持绿色。利用AtNHX1基因特异性引物和探针对抗性植株基因组DNA进行检测。PCR和Southern blot分析结果表明,外源靶基因AtNHX1基因已转移到不同品种的羊茅中。4个品种间的转化频率不同。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Research on Festuca arundinacea transformation mediated by Agrobacterium tumefaciens].

The embryo-derived calli from four types of tall fescues (Festuca arundinacea Schreb) were transformed with two Agrobactrium tumefaciens strains AGL1 and GV3101. AGL1 harbors an intron-AtNHX1 expression vector pROK2U containing ubiqutin promoter and npt II marker gene. GV3101 harbors an intron-AtNHX1 expression vector pROK2 containing 35S promoter and npt II gene. After infection and co-culture with AGL1 or GV3101, the embyogenic calli were selected with 50-150 mg/L paromomycine and 1126 resistant plants regenerated from the resistant calli. All plants were selected further with 10-20 mg/L Kanamycin and 525 of them remained green. Genome DNA of the resistant plants was checked with specific primers and probe from AtNHX1 gene. The results of PCR assay and Southern blot analysis indicted that exogenous target gene (AtNHX1 gene) had been transferred into different cultivars of Festuca arundinacea. Different transformation frequencies among the four cultivars were obtained.

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