High frequency shoot regeneration and Agrobacterium.mediated DNA transfer in Canola (Brassica napus)

被引：48

作者：

Natl. Agricultural Research Centre, Agricultural Biotechnology Institute, Park Road, Islamabad, Pakistan ^{[1
]}

不详 ^{[2
]}

机构：

[1] Natl. Agricultural Research Centre, Agricultural Biotechnology Institute, Islamabad, Park Road

[2] Department of Biological Sciences, Quaid-I-Azam University, Islamabad

来源：

Plant Cell, Tissue and Organ Culture | 2003年 / 75卷 / 03期

关键词：

Canola; Explant; In vitro; Regeneration; Transformation; Transgenic plants; Transient expression;

D O I：

10.1023/A:1025869809904

中图分类号：

学科分类号：

摘要：

Five different varieties of Brassica napus (Cyclone, Dunkled, Oscar, Rainbow and KS75) were tested for their regeneration response. Cyclone showed a very high frequency of regeneration (92%). The use of silver nitrate was a pre-requisite for efficient shoot regeneration. Hypocotyls were selected as the starting material for transformation experiments on the basis of high transient GUS expression. Explants were co-cultivated with Agrobacterium strain EHA101 harboring a binary vector pIG121Hm containing neomycin phosphotransferase II (NPTII) gene, conferring resistance to kanamycin, hygromycin phosphotransferase (HPT) gene, conferring resistance to hygromycin as selectable markers and β-glucuronidase (GUS) gene as a reporter. Acetosyringone promoted the transformation but was not an absolute requirement. A pre-selection period of 7 days after co-cultivation was essential for successful transformation. Kanamycin was efficient selective agent for selection and maximum transformation efficiency was 24%. GUS activity was evident in leaf tissues. All the transgenic plants have an expected band of 0.43 kb fragment by PCR analysis confirming the presence of foreign DNA into plant genome.

引用

页码：223 / 231

页数：8

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