| 摘要: |
| 在唐菖蒲‘Advanced Red’胚性愈伤组织受体系统的基础上,建立其遗传转化体系。采用根癌农杆菌(GV3101)介导法,研究了侵染液浓度、侵染时间、乙酰丁香酮(AS)浓度、负压和筛选方式等因子对唐菖蒲遗传转化效率的影响。结果表明:在遗传转化过程中,不经预培养,负压处理下,农杆菌菌液OD600为0.6~0.8,侵染时间15~20 min,添加100 μmol/L AS,共培养3 d,可获得较高的遗传转化效率。经过3~4个月选择培养,部分抗性植株经PCR和Southern杂交检测表明,目的基因gus已整合到唐菖蒲基因组中。 |
| 关键词: 唐菖蒲 gus 根癌农杆菌 遗传转化 |
| DOI：10.11841/j.issn.1007-4333.2011.06.011 |
| 投稿时间：2010-12-30 |
| 基金项目:农业部行业科技项目(200903020) |
|
| Establishment of agrobacterium-mediated genetictransformation system of Gladiolus |
|
ZHANG Zi-you, LIAN Qing-long, XIN Hai-bo, ZHONG Xiong-hui, LUO Xian, YI Ming-fang
|
| (College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China) |
| Abstract: |
| Genetic transformation system of Gladiolus cv.‘Advanced Red’ was established based on the regeneration system of embryogenic callus.Several factors were studied such as concentration of the infection,infection time,acetosyringone (AS) concentration,negative pressure and screening methods for genetic transformation of Gladiolus mediated by agrobac-terium tumefaciens(GV3101).The results showed that the highest rate of genetic transformation was obtained under the conditions of non pre-incubation,negative pressure treatment,agrobacterium concentration OD600 = 0.6-0.8,infection time 15-20 min,adding 100 μmol/LAS and 3 ds incuabtion.Following incubation for 3-4 months,the analysis of the resistant plants with PCR and southern blot indicated that the gus had been integrated into the genome of Gladiolus. |
| Key words: Gladiolus gus agrobacterium tumefaciens genetic transformation |
