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Development of selectable marker‐free transgenic potato plants expressing cry3A against the Colorado potato beetle ( Leptinotarsa decemlineata Say)
Author(s) -
Guo Wenchao,
Wang Zhian,
Luo Xiaoli,
Jin Xin,
Chang Jing,
He Jiang,
Tu Erxun,
Tian Yingchuan,
Si Huaijun,
Wu Jiahe
Publication year - 2016
Publication title -
pest management science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.296
H-Index - 125
eISSN - 1526-4998
pISSN - 1526-498X
DOI - 10.1002/ps.4013
Subject(s) - colorado potato beetle , leptinotarsa , biology , selectable marker , inoculation , pest analysis , larva , transgene , horticulture , genetically modified crops , infestation , genetically modified organism , genetically modified tomato , gene , botany , genetics
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. CONCLUSION 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