利用PMI选择标记进行杨树转基因体系的研究
[目的]利用不同于抗生素的PMI为选择标记基因的遗传转化体系,对杨树进行双抗虫基因（Bt和Cp TI）的转基因研究。建立杨树以PMI为安全标记基因的转基因体系,为安全、高效的林木转基因育种研究提供实验依据。[方法]选择已有的转Cp TI抗虫基因（利用Kmr选择标记获得）的美洲黑杨杂种优良无性系南林895杨（Populus×euramericana‘Nanlin895’）为受体材料,对杨树叶片、叶片分化芽、茎段生根的甘露糖敏感性等筛选优化,采用农杆菌介导的方法进行双抗虫基因的研究。[结果]较为适合的杨树叶片筛选培养基为：甘露糖8 g·L^-1和蔗糖22 g·L^-1;叶片分化芽筛选培养基为：甘露...
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| Published in | 林业科学研究 Vol. 29; no. 2; pp. 221 - 226 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
无锡市林木种苗管理站，江苏 无锡 214000%南京林业大学南方现代林业协同创新中心,江苏 南京,210037%南京晓庄学院生命科学系,江苏 南京,210000
2016
南京林业大学南方现代林业协同创新中心，江苏 南京 210037 |
| Subjects | |
| Online Access | Get full text |
| ISSN | 1001-1498 |
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| Abstract | [目的]利用不同于抗生素的PMI为选择标记基因的遗传转化体系,对杨树进行双抗虫基因（Bt和Cp TI）的转基因研究。建立杨树以PMI为安全标记基因的转基因体系,为安全、高效的林木转基因育种研究提供实验依据。[方法]选择已有的转Cp TI抗虫基因（利用Kmr选择标记获得）的美洲黑杨杂种优良无性系南林895杨（Populus×euramericana‘Nanlin895’）为受体材料,对杨树叶片、叶片分化芽、茎段生根的甘露糖敏感性等筛选优化,采用农杆菌介导的方法进行双抗虫基因的研究。[结果]较为适合的杨树叶片筛选培养基为：甘露糖8 g·L^-1和蔗糖22 g·L^-1;叶片分化芽筛选培养基为：甘露糖10 g·L^-1和蔗糖20 g·L^-1;茎段生根筛选培养基：甘露糖8 g·L^-1和蔗糖22 g·L^-1。在此基础上,获得了转Bt和Cp TI双抗虫基因的植株8株。[结论]初步建立了以PMI为安全标记基因的杨树转基因体系：确定了PMI筛选的最适选择压,成功构建了含PMI选择标记基因的植物抗虫表达载体,通过农杆菌介导的遗传转化最终获得了转双抗虫基因植株。 |
|---|---|
| AbstractList | S792.119; [目的]利用不同于抗生素的 PMI 为选择标记基因的遗传转化体系，对杨树进行双抗虫基因（Bt 和 CpTI）的转基因研究。建立杨树以 PMI 为安全标记基因的转基因体系，为安全、高效的林木转基因育种研究提供实验依据。[方法]选择已有的转 CpTI 抗虫基因（利用 Kmr 选择标记获得）的美洲黑杨杂种优良无性系南林895杨（Populus × euramericana ‘Nanlin895’）为受体材料，对杨树叶片、叶片分化芽、茎段生根的甘露糖敏感性等筛选优化，采用农杆菌介导的方法进行双抗虫基因的研究。[结果]较为适合的杨树叶片筛选培养基为：甘露糖8 g·L －1和蔗糖22 g· L －1；叶片分化芽筛选培养基为：甘露糖10 g·L －1和蔗糖20 g·L －1；茎段生根筛选培养基：甘露糖8 g·L －1和蔗糖22 g·L －1。在此基础上，获得了转 Bt 和 CpTI 双抗虫基因的植株8株。[结论]初步建立了以 PMI 为安全标记基因的杨树转基因体系：确定了 PMI 筛选的最适选择压，成功构建了含 PMI 选择标记基因的植物抗虫表达载体，通过农杆菌介导的遗传转化最终获得了转双抗虫基因植株。 [目的]利用不同于抗生素的PMI为选择标记基因的遗传转化体系,对杨树进行双抗虫基因（Bt和Cp TI）的转基因研究。建立杨树以PMI为安全标记基因的转基因体系,为安全、高效的林木转基因育种研究提供实验依据。[方法]选择已有的转Cp TI抗虫基因（利用Kmr选择标记获得）的美洲黑杨杂种优良无性系南林895杨（Populus×euramericana‘Nanlin895’）为受体材料,对杨树叶片、叶片分化芽、茎段生根的甘露糖敏感性等筛选优化,采用农杆菌介导的方法进行双抗虫基因的研究。[结果]较为适合的杨树叶片筛选培养基为：甘露糖8 g·L^-1和蔗糖22 g·L^-1;叶片分化芽筛选培养基为：甘露糖10 g·L^-1和蔗糖20 g·L^-1;茎段生根筛选培养基：甘露糖8 g·L^-1和蔗糖22 g·L^-1。在此基础上,获得了转Bt和Cp TI双抗虫基因的植株8株。[结论]初步建立了以PMI为安全标记基因的杨树转基因体系：确定了PMI筛选的最适选择压,成功构建了含PMI选择标记基因的植物抗虫表达载体,通过农杆菌介导的遗传转化最终获得了转双抗虫基因植株。 |
| Abstract_FL | [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 ob-tained 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 1 0 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 vec-tor containing PMI selective marker gene was established,and the transgenic poplar with two insect-resistant genes was obtained by Agrobacterum-mediated transformation. |
| Author | 查琳 王伟东 续晨 诸葛强 |
| AuthorAffiliation | 南京林业大学南方现代林业协同创新中心,江苏南京210037 无锡市林木种苗管理站,江苏无锡214000 南京晓庄学院生命科学系,江苏南京210000 |
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| Author_FL | XU Chen ZHA Lin WANG Wei-dong ZHUGE Qiang |
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| Keywords | 选择标记 insect resistance Populus ×euramericana 'Nanlin895' 抗虫 transgenic system 转基因体系 PMI 南林 895 杨 selective marker |
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| Notes | 11-1221/S [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 Cp TI） and provide experimental data for high-efficient and safe transgenic tree breeding. [Method] Populus × euramericana ‘Nanlin895 ＇genetically modified by Cp TI obtained by using Kmrselective 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 Cp TI genes we obtained. [Conclusion]The transgenic system of poplar was initially established by use of bios |
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| Publisher | 无锡市林木种苗管理站，江苏 无锡 214000%南京林业大学南方现代林业协同创新中心,江苏 南京,210037%南京晓庄学院生命科学系,江苏 南京,210000 南京林业大学南方现代林业协同创新中心，江苏 南京 210037 |
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| Snippet | [目的]利用不同于抗生素的PMI为选择标记基因的遗传转化体系,对杨树进行双抗虫基因（Bt和Cp TI）的转基因研究。建立杨树以PMI为安全标记基因的转基因体系,为安全、高效的林木... S792.119; [目的]利用不同于抗生素的 PMI 为选择标记基因的遗传转化体系，对杨树进行双抗虫基因（Bt 和 CpTI）的转基因研究。建立杨树以 PMI 为安全标记基因的转基因体系，... |
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| SubjectTerms | PMI 南林895杨 抗虫 转基因体系 选择标记 |
| Title | 利用PMI选择标记进行杨树转基因体系的研究 |
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