title: Development of selectable marker-free transgenic potato plants expressing cry3A against the Colorado potato beetle (Leptinotarsa decemlineata Say).
abstract: BACKGROUND
  Elimination of selectable marker genes (SMGs) is important for the safe assessment and commercial use of transgenic plants. The destructive and invasive Colorado potato beetle (CPB) poses a serious threat to potato production. In response to this need, selectable marker-free transgenic potato lines expressing cry3A were developed to control the damage and spread of CPB.
  
  
  RESULTS
  We simultaneously introduced cry3A and npt II genes harboured in different plasmids into the potato genome using the Agrobacterium-mediated cotransformation method. Four selectable marker-free transgenic potato (CT) lines expressing cry3A were developed by self-crossing segregation and molecular analyses, including Southern blot, western blot and enzyme-linked immunosorbent assay (ELISA) assays. CT lines were used in a resistance bioassay against CPB in the laboratory and field. In the laboratory, CT lines exhibited high resistance to CPB, and 100% mortality of first-instar larvae occurred 6 days after infestation. In the field, untransformed plant leaves were almost entirely consumed, with an average of 155 larvae present per plant 25 days after inoculation. However, CT lines showed no damage symptoms, with approximately 2.5 larvae surviving per plant.
  
  
  CONCLUSIONS
  We successfully eliminated SMGs from the transgenic potato lines expressing cry3A in order to decrease CPB damage, control the spread of this pest eastwards and alleviate the concern regarding the safe assessment of regulatory requirements. © 2015 Society of Chemical Industry.
authors.affiliation: State Key Laboratory of Plant Genome, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.|Institute of Plant Protection, Xinjiang Agricultural Academy of Sciences, Xinjiang, Urumqi, China.
authors.forename: Wen-chao
authors.initials: WC
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Plant Genome, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.|Institute of Cotton Research, Shanxi Agricultural Academy of Sciences, Shanxi, Yuncheng, China.
authors.forename: Zhi-an
authors.initials: ZA
authors.lastname: Wang
authors.affiliation: Institute of Cotton Research, Shanxi Agricultural Academy of Sciences, Shanxi, Yuncheng, China.
authors.forename: Xiao-li
authors.initials: XL
authors.lastname: Luo
authors.affiliation: State Key Laboratory of Plant Genome, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.|College of Biology Science and Technology, Gansu Agricultural University, Gansu, Lanzhou, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Plant Genome, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.|College of Biology Science and Technology, Gansu Agricultural University, Gansu, Lanzhou, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Chang
authors.affiliation: Institute of Plant Protection, Xinjiang Agricultural Academy of Sciences, Xinjiang, Urumqi, China.
authors.forename: Jiang
authors.initials: J
authors.lastname: He
authors.affiliation: Institute of Plant Protection, Xinjiang Agricultural Academy of Sciences, Xinjiang, Urumqi, China.
authors.forename: Er-xun
authors.initials: EX
authors.lastname: Tu
authors.affiliation: State Key Laboratory of Plant Genome, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Ying-chuan
authors.initials: YC
authors.lastname: Tian
authors.affiliation: College of Biology Science and Technology, Gansu Agricultural University, Gansu, Lanzhou, China.
authors.forename: Huai-jun
authors.initials: HJ
authors.lastname: Si
authors.affiliation: State Key Laboratory of Plant Genome, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Jia-he
authors.initials: JH
authors.lastname: Wu
mesh_terms: D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology*; D001517:Coleoptera / Q000187:drug effects* / Q000254:growth & development; D004731:Endotoxins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology*; D005819:Genetic Markers / Q000235:genetics; D006460:Hemolysin Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology*; D007814:Larva / Q000187:drug effects / Q000254:growth & development; D030821:Plants, Genetically Modified / Q000737:chemistry / Q000235:genetics; D011198:Solanum tuberosum / Q000235:genetics*
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D005819:Genetic Markers; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis
keywords: Colorado potato beetle; Cry3A; selectable marker free; selectable marker gene; transgenic potato