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| Scarabaeid Larvae- and Herbicide-Resistant Transgenic Perennial Ryegrass (Lolium perenne L.) Obtained by Agrobacterium tumefaciens-Mediated Transformation of cry8Ca2, cry8Ga and bar Genes |
| WU Jin-xia, ZHANG Zhi-guo, ZHANG Qian, LANG Zhi-hong , SUN Xue-hui |
| 1.Biotechnology Research Institute, National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China |
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摘要 Insect pest and weeds are two major problems for forage and turf grasses. In this study, scarab larvae- and herbicideresistant transgenic perennial ryegrass (Lolium perenne L.) was obtained by transforming it with cry and bar genes simultaneously via the Agrobacterium-mediated method. To optimize the callus induction and plant regeneration conditions, various concentrations of 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine were assayed. The transformation efficiencies of different Agrobacterium suspension media, used during Agrobacterium-mediated transformation, were compared. Then, plasmids of pCAMBIA3301 containing cry gene (cry8Ca2 or cry8Ga) and bar gene, driven by ubiquitin promoter, were transformed into perennial ryegrass. The transformants were generated and confirmed by both Southern hybridization analysis and Western hybridization analysis. Further, the resistance of transgenic perennial ryegrass plants to scarab larvae and herbicide were analyzed. After 30 d of co-cultivation with scarab larvae, the damage to the root system of transgenic plants was less than that of non-transgenic control plants. Additionally, the leaves of transgenic plants were resistant to Basta®, while leaves of the wild plants wilted after Basta® spraying. These results show that cry gene and bar gene were successfully transferred into perennial ryegrass by the Agrobactgerium-mediated method, and convey resistance to scarab larvae and herbicide in transgenic perennial ryegrass plants.
Abstract Insect pest and weeds are two major problems for forage and turf grasses. In this study, scarab larvae- and herbicideresistant transgenic perennial ryegrass (Lolium perenne L.) was obtained by transforming it with cry and bar genes simultaneously via the Agrobacterium-mediated method. To optimize the callus induction and plant regeneration conditions, various concentrations of 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine were assayed. The transformation efficiencies of different Agrobacterium suspension media, used during Agrobacterium-mediated transformation, were compared. Then, plasmids of pCAMBIA3301 containing cry gene (cry8Ca2 or cry8Ga) and bar gene, driven by ubiquitin promoter, were transformed into perennial ryegrass. The transformants were generated and confirmed by both Southern hybridization analysis and Western hybridization analysis. Further, the resistance of transgenic perennial ryegrass plants to scarab larvae and herbicide were analyzed. After 30 d of co-cultivation with scarab larvae, the damage to the root system of transgenic plants was less than that of non-transgenic control plants. Additionally, the leaves of transgenic plants were resistant to Basta®, while leaves of the wild plants wilted after Basta® spraying. These results show that cry gene and bar gene were successfully transferred into perennial ryegrass by the Agrobactgerium-mediated method, and convey resistance to scarab larvae and herbicide in transgenic perennial ryegrass plants.
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Received: 26 January 2011
Accepted:
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| Fund: This work was supported by the National Basic Research Program of China (973 Program, 2007CB1089). |
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Corresponding Authors:
Correspondence SUN Xue-hui, Tel: +86-10-82106131, Fax: +86-10-82106132, E-mail: xhsun@caas.net.cn
E-mail: xhsun@caas.net.cn
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Cite this article:
WU Jin-xia, ZHANG Zhi-guo, ZHANG Qian, LANG Zhi-hong , SUN Xue-hui.
2012.
Scarabaeid Larvae- and Herbicide-Resistant Transgenic Perennial Ryegrass (Lolium perenne L.) Obtained by Agrobacterium tumefaciens-Mediated Transformation of cry8Ca2, cry8Ga and bar Genes. Journal of Integrative Agriculture, 12(1): 53-61.
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