Development and identification of transgenic rapeseed with a novel gene for glyphosate resistance

Glyphosate is the most widely used broad-spectrum herbicide in the world. However, at present there is no glyphosate-tolerant rapeseed variety with independent intellectual property rights in China. In the study, a novel glyphosate-resistant genes I. variabilis EPSPS was transferred to the Brassica napus pure line J9707 via the Agrobacterium tumefaciens-mediated hypocotyl method, and 126 T0-positive transgenic plants with 97.0% positive rate were generated. The T-DNA insertion with a single copy(44.8%) is dominant. The insertion locations of T-DNA in the lines of EPS-2, EPS-6, and EPS-7 were identified by inverse PCR method. The stability of the T-DNA insertion in these lines were further confirmed by insertion-specific PCR in their T0 to T3 plants. The gene expression analysis revealed that the I. variabilis EPSPS gene and its protein was stably expressed in different generations of transgenic lines in RNA and protein levels. Treatments with different doses of glyphosate indicated that the lines of EPS-1, EP