The multiplex polymerase chain reaction (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. | Turkish Journal of Agriculture and Forestry Turk J Agric For (2013) 37: 157-162 © TÜBİTAK doi: Research Article An efficient multiplex PCR assay for early detection of Agrobacterium tumefaciens in transgenic plant materials 1, 1 1 1 2 2 Li YANG *, Changchun WANG , Lihuan WANG , Changjie XU , Kunsong CHEN College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua, Zhejiang 321004, . China 2 Department of Horticulture, Zhejiang University, Hangzhou 310029, . China Received: Accepted: Published Online: Printed: Abstract: The multiplex polymerase chain reaction (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 (nptII; target gene) and the chromosomal virulence gene ChvA fragment of Agrobacterium tumefaciens (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. Key words: Agrobacterium tumefaciens, multiplex polymerase chain reaction, transgenic plant detection 1. Introduction Since the first commercial release of genetically modified (GM) tomato in 1994 (Nap et al. 2003), the worldwide acreage devoted to GM crops has increased rapidly to an estimated 160 million ha in 2011. The main GM crops grown commercially are cotton, soybean, maize, oilseed rape, and potato, and many of these crops possess significant advantages .
