2005ACTA AGRONOMICA SINICARequires access

Screening of Wheat Genotypes Sensitive to Agrobacterium tumefaciens Infection and Transformation

Xing Ye

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Abstract

Thirty-five Chinese spring wheat genotypes were tested for their sensitivity to C58c1 Agrobacterium strain's (harboring a binary vector pPTN290 with npt Ⅱ gene and GUS gene) infection and final transformation. Six cultivars including PM97034, P187, Ba1401051, Yangmai 158, Xinchun 9 and Kefeng 6 were proved to be more sensitive than Bobwhite to the Agro-infection according to X-Gluc staining, with GUS gene transient expression frequency of 15.0% to 30.0%. Regeneration plants obtained from selection medium were identified by ELISA, PCR, leaf bleach and X-Gluc, and stable transformation events were obtained from Xinchun 9, PM97034 and Bobwhite by 3.3%, 2.0% and 1.1%, respectively. Southern blot analysis showed the alien genes integrated in most transgenic plants just by single copy. The alien genes in most T_1 transgenic lines segregated by Mendelian regulation, but the segregation ratio in a few T_1 transgenic lines was a little bit away from the expecting value, according to χ2_c test. Xinchun 9 and PM97034 are suggested to be used in wheat Agrobacterium mediated transformation to replace Bobwhite for better result.

About this research paper

What this paper is about

Thirty-five Chinese spring wheat genotypes were tested for their sensitivity to C58c1 Agrobacterium strain's (harboring a binary vector pPTN290 with npt Ⅱ gene and GUS gene) infection and final transformation. Six cultivars including PM97034, P187, Ba1401051, Yangmai 158, Xinchun 9 and Kefeng 6 were proved to be more sensitive than Bobwhite to the Agro-infection according to X-Gluc staining, with GUS gene transient expression frequency of 15.0% to 30.0%. Regeneration plants obtained from selection medium were identified by ELISA, PCR, leaf bleach and X-Gluc, and stable transformation events were obtained from Xinchun 9, PM97034 and Bobwhite by 3.3%, 2.0% and 1.1%, respectively. Southern blot analysis showed the alien genes integrated in most transgenic plants just by single copy. The alien genes in most T_1 transgenic lines segregated by Mendelian regulation, but the segregation ratio in a few T_1 transgenic lines was a little bit away from the expecting value, according to χ2_c test. Xinchun 9 and PM97034 are suggested to be used in wheat Agrobacterium mediated transformation to replace Bobwhite for better result.

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Available abstract

Thirty-five Chinese spring wheat genotypes were tested for their sensitivity to C58c1 Agrobacterium strain's (harboring a binary vector pPTN290 with npt Ⅱ gene and GUS gene) infection and final transformation. Six cultivars including PM97034, P187, Ba1401051, Yangmai 158, Xinchun 9 and Kefeng 6 were proved to be more sensitive than Bobwhite to the Agro-infection according to X-Gluc staining, with GUS gene transient expression frequency of 15.0% to 30.0%. Regeneration plants obtained from selection medium were identified by ELISA, PCR, leaf bleach and X-Gluc, and stable transformation events were obtained from Xinchun 9, PM97034 and Bobwhite by 3.3%, 2.0% and 1.1%, respectively. Southern blot analysis showed the alien genes integrated in most transgenic plants just by single copy. The alien genes in most T_1 transgenic lines segregated by Mendelian regulation, but the segregation ratio in a few T_1 transgenic lines was a little bit away from the expecting value, according to χ2_c test. Xinchun 9 and PM97034 are suggested to be used in wheat Agrobacterium mediated transformation to replace Bobwhite for better result.

Key concepts: Biology, Transformation (genetics), Agrobacterium tumefaciens, Agrobacterium, Gene, Genetically modified crops, Genetics, Molecular biology

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