title: Detection of Bar Transgenic Sugarcane with a Rapid and Visual Loop-Mediated Isothermal Amplification Assay.
abstract: Genetic engineering offers an attractive alternative in sugarcane breeding for increasing cane and sugar yields as well as disease and insect resistance. Bar transgenic sugarcane employing the herbicide tolerance is a useful agronomical trait in weed control. In this study, a loop-mediated isothermal amplification (LAMP) assay for rapid detection of the bar gene in transgenic sugarcane has been developed and evaluated. A set of six primers was designed for LAMP-based amplification of the bar gene. The LAMP reaction conditions were optimized as follows: 5.25 mM of Mg(2+), 6:1 ratio of inner vs. outer primer, and 6.0 U of Bst DNA polymerase in a reaction volume of 25.0 μL. The detection limit of the recombinant plasmid 1Ac0229 was as low as 10 copies in the developed LAMP, which was 10-fold higher sensitive than that of conventional PCR. In 100 putative transgenic lines, the bar gene was detected in 100/100 cases (100%) by LAMP and 97/100 cases (97%) by conventional PCR, respectively. In conclusion, the developed LAMP assay is visual, rapid, sensitive, reliable, and cost-effective for detection of the bar specific transgenic sugarcane.
authors.affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Fujian Agriculture and Forestry University, Ministry of Agriculture Fuzhou, China.
authors.forename: Dinggang
authors.initials: D
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Fujian Agriculture and Forestry University, Ministry of Agriculture Fuzhou, China.
authors.forename: Chunfeng
authors.initials: C
authors.lastname: Wang
authors.affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Fujian Agriculture and Forestry University, Ministry of Agriculture Fuzhou, China.
authors.forename: Zhu
authors.initials: Z
authors.lastname: Li
authors.affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Fujian Agriculture and Forestry University, Ministry of Agriculture Fuzhou, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Fujian Agriculture and Forestry University, Ministry of Agriculture Fuzhou, China.
authors.forename: Shiwu
authors.initials: S
authors.lastname: Gao
authors.affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Fujian Agriculture and Forestry University, Ministry of Agriculture Fuzhou, China.
authors.forename: Jinlong
authors.initials: J
authors.lastname: Guo
authors.affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Fujian Agriculture and Forestry University, Ministry of Agriculture Fuzhou, China.
authors.forename: Wenying
authors.initials: W
authors.lastname: Lu
authors.affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Fujian Agriculture and Forestry University, Ministry of Agriculture Fuzhou, China.
authors.forename: Yachun
authors.initials: Y
authors.lastname: Su
authors.affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Fujian Agriculture and Forestry University, Ministry of Agriculture Fuzhou, China.
authors.forename: Liping
authors.initials: L
authors.lastname: Xu
authors.affiliation: Key Laboratory of Sugarcane Biology and Genetic Breeding, Fujian Agriculture and Forestry University, Ministry of Agriculture Fuzhou, China.
authors.forename: Youxiong
authors.initials: Y
authors.lastname: Que
keywords: PCR; bar; detection; genetically modified organism (GMO); loop-mediated isothermal amplification (LAMP); transgenic sugarcane