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[SANET-MG] escape to the environment of transgenic Agrobacterium



Volume 26, Issue 11 , November 2006, Pages 3555-3561 Acta Ecologica Sinica
RESEARCH PAPER
Survival and escape of Agrobacterium tumefaciens in triploid hybrid lines of Chinese white poplar transformed with two insect-resistant genes

Minsheng Yanga, Dan Mia, D. Ewaldb, , , Ying Wangc, Haiyong Lianga and Zhixian Zhena

aAgricultural University of Hebei, Baoding 071000, China
bInstitute for Forest Genetics and Forest Tree Breeding, BFH, D-15377 Waldsieversdorf, Germany cInstitute of Tropical Biological Sciences, State Key Laboratory of Tropical Crops Biotechnology, Haikou 571101, China

Received 10 January 2006; accepted 30 August 2006. Available online 19 December 2006.


Abstract
Two partly modified insect-resistant genes (BtCryI Ac gene [Bt gene toxin against Lepidopterean insects] and API gene [arrowhead proteinase inhibitor]) were transferred to the triploid hybrid of Chinese white poplar ((Populus tomentosa Carr. × Populus bolleana Louche) × Populus tomentosa Carr.) mediated by A. tumefaciens. The survival of Agrobacterium in transgenic plants was examined during the processes of transplanting and subculturing on the nutrient medium. The results suggested that 80% of the plants, which were obtained by repeated selection on media added with 50 mg/L kanamycin and 300 mg/l carbenicillin, showed positive reactions after examination using molecular methods. The ELISA test indicated that the Bt toxoprotein was expressed in seven of the transgenic sub-clones. Leaves, stems, and roots of all the 28 transgenic plants were cultured on the YEB medium added with 50 mg/L kanamycin, and it was found that Agrobacterium survived in three sub-clones (33, 37, 5) and could have existed for 24 months in the bottle. These three transgenic sub-clones were transplanted and cultivated for one month in the room, and then the target Agrobacterium was found in rhizosphere of the sub-clone 33.
From conclusions:In the body of the transgenic plants could diffuse to the
soil after transplantation.
The results of the researches should be further tested in other kinds of plants to confirm the possibilities of the gene flow. Thus, if any plant transformation was performed with the help of Agrobacteria, tests to detect the engineered bacteria should be carried out before the transgenic plants would be released to the environment to avoid the escape of genes.



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