An efficient multiplex PCR assay for early detection of Agrobacterium tumefaciens in transgenic plant materials

Authors : Li YANG, Changchun WANG, Lihuan WANG, Changjie XU, Kunsong CHEN

Pages : 157-162

Doi:10.3906/tar-1009-1265

View : 12 | Download : 7

Publication Date : 2013-04-01

Article Type : Research Paper

Abstract :The multiplex polymerase chain reaction insert ignore into journalissuearticles values(MPCR); is a variant of conventional PCR in which 2 or more target sequences are amplified by more than 1 pair of primers in a single reaction mixture. In this study, an efficient MPCR assay was established and optimized for the quick detection of Agrobacterium-derived plant transformants. The neomycin phosphotransferase II gene fragment insert ignore into journalissuearticles values(nptII; target gene); and the chromosomal virulence gene ChvA fragment of Agrobacterium tumefaciens insert ignore into journalissuearticles values(external reference gene); were amplified simultaneously with 2 pairs of corresponding primers. The MPCR results showed that 9 out of 10 kanamycin-resistant lines tested were transformed with the foreign genes. This optimized MPCR assay, rapid, sensitive, and reliable, can facilitate the identification of Agrobacterium-derived plant transformants at the early stage of selection.
Keywords : Key words Agrobacterium tumefaciens, multiplex polymerase chain reaction, transgenic plant detection