Poplar Transgenic System by Using PMI as Biosafety Selective Marker

ZHA Lin; WANG Wei-dong; XU Chen; ZHUGE Qiang

[Objective]Using the PMI different from antibiotic marker gene as selective marker gene to study poplar transgenic system with two insect-resistant genes (Bt and CpTI) and provide experimental data for high-efficient and safe transgenic tree breeding. [Method] Populus × euramericana ‘Nanlin895’ genetically modified by CpTI obtained by using Kmr selective marker was chosen as a receipt, and the mannose sensitivity of poplar leaves, bud growth and stem rooting in a medium were tested for studying poplar transgenic system with two insect-resistant genes by way of Agrobacterum-mediated transformation. [Result] The suitable composition for genetic transformation of poplar leaves was: Mannose 8 g·L-1+ Sucrose 22 g·L-1 ; for bud growth:Mannose 10 g·L-1 + Sucrose 20 g·L-1 ; and for stem rooting: Mannose 8 g·L-1 +Sucrose 22 g·L-1. Based on the results, eight transgenic plants with Bt and CpTI genes we obtained. [Conclusion] The transgenic system of poplar was initially established by use of biosafety selective marker PMI. The best selection pressure was determined. The plant anti-insect expression vector containing PMI selective marker gene was established, and the transgenic poplar with two insect-resistant genes was obtained by Agrobacterum-mediated transformation.

1. Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, Jiangsu, China

2. Wuxi Forest Seed and Seedling Administration, Wuxi 214000, Jiangsu, China

3. Bioscience Department of Nanjing, Xiaozhuang University, Nanjing 210000, Jiangsu, China

Received Date: 2014-12-15

Abstract: [Objective]Using the PMI different from antibiotic marker gene as selective marker gene to study poplar transgenic system with two insect-resistant genes (Bt and CpTI) and provide experimental data for high-efficient and safe transgenic tree breeding. [Method] Populus × euramericana ‘Nanlin895’ genetically modified by CpTI obtained by using Kmr selective marker was chosen as a receipt, and the mannose sensitivity of poplar leaves, bud growth and stem rooting in a medium were tested for studying poplar transgenic system with two insect-resistant genes by way of Agrobacterum-mediated transformation. [Result] The suitable composition for genetic transformation of poplar leaves was: Mannose 8 g·L-1+ Sucrose 22 g·L-1 ; for bud growth:Mannose 10 g·L-1 + Sucrose 20 g·L-1 ; and for stem rooting: Mannose 8 g·L-1 +Sucrose 22 g·L-1. Based on the results, eight transgenic plants with Bt and CpTI genes we obtained. [Conclusion] The transgenic system of poplar was initially established by use of biosafety selective marker PMI. The best selection pressure was determined. The plant anti-insect expression vector containing PMI selective marker gene was established, and the transgenic poplar with two insect-resistant genes was obtained by Agrobacterum-mediated transformation.

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Reference (15)

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Poplar Transgenic System by Using PMI as Biosafety Selective Marker

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