Development of selectable marker-free transgenic potato plants expressing cry3A against the Colorado potato beetle (Leptinotarsa decemlineata Say).
Pest Manag Sci. 2016 Mar; 72(3):497-504.PM

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.

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.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Guo WC

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.

Wang ZA

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.

Luo XL

Institute of Cotton Research, Shanxi Agricultural Academy of Sciences, Shanxi, Yuncheng, China.

Jin X

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.

Chang J

He J

Institute of Plant Protection, Xinjiang Agricultural Academy of Sciences, Xinjiang, Urumqi, China.

Tu EX

Institute of Plant Protection, Xinjiang Agricultural Academy of Sciences, Xinjiang, Urumqi, China.

Tian YC

State Key Laboratory of Plant Genome, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.

Si HJ

College of Biology Science and Technology, Gansu Agricultural University, Gansu, Lanzhou, China.

Wu JH

State Key Laboratory of Plant Genome, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.

MeSH

AnimalsBacillus thuringiensisBacillus thuringiensis ToxinsBacterial ProteinsColeopteraEndotoxinsGenetic MarkersHemolysin ProteinsLarvaPlants, Genetically ModifiedSolanum tuberosum

Pub Type(s)

Journal Article

Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25820984

Citation

Guo, Wen-chao, et al. "Development of Selectable Marker-free Transgenic Potato Plants Expressing cry3A Against the Colorado Potato Beetle (Leptinotarsa Decemlineata Say)." Pest Management Science, vol. 72, no. 3, 2016, pp. 497-504.

Guo WC, Wang ZA, Luo XL, et al. Development of selectable marker-free transgenic potato plants expressing cry3A against the Colorado potato beetle (Leptinotarsa decemlineata Say). Pest Manag Sci. 2016;72(3):497-504.

Guo, W. C., Wang, Z. A., Luo, X. L., Jin, X., Chang, J., He, J., Tu, E. X., Tian, Y. C., Si, H. J., & Wu, J. H. (2016). Development of selectable marker-free transgenic potato plants expressing cry3A against the Colorado potato beetle (Leptinotarsa decemlineata Say). Pest Management Science, 72(3), 497-504. https://doi.org/10.1002/ps.4013

Guo WC, et al. Development of Selectable Marker-free Transgenic Potato Plants Expressing cry3A Against the Colorado Potato Beetle (Leptinotarsa Decemlineata Say). Pest Manag Sci. 2016;72(3):497-504. PubMed PMID: 25820984.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - Development of selectable marker-free transgenic potato plants expressing cry3A against the Colorado potato beetle (Leptinotarsa decemlineata Say). AU - Guo,Wen-chao, AU - Wang,Zhi-an, AU - Luo,Xiao-li, AU - Jin,Xin, AU - Chang,Jing, AU - He,Jiang, AU - Tu,Er-xun, AU - Tian,Ying-chuan, AU - Si,Huai-jun, AU - Wu,Jia-he, Y1 - 2015/04/20/ PY - 2014/12/11/received PY - 2015/03/11/revised PY - 2015/03/23/accepted PY - 2015/3/31/entrez PY - 2015/3/31/pubmed PY - 2016/11/1/medline KW - Colorado potato beetle KW - Cry3A KW - selectable marker free KW - selectable marker gene KW - transgenic potato SP - 497 EP - 504 JF - Pest management science JO - Pest Manag Sci VL - 72 IS - 3 N2 - 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. SN - 1526-4998 UR - https://www.unboundmedicine.com/medline/citation/25820984/Development_of_selectable_marker_free_transgenic_potato_plants_expressing_cry3A_against_the_Colorado_potato_beetle__Leptinotarsa_decemlineata_Say__ DB - PRIME DP - Unbound Medicine ER -

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Development of selectable marker-free transgenic potato plants expressing cry3A against the Colorado potato beetle (Leptinotarsa decemlineata Say).Pest Manag Sci. 2016 Mar; 72(3):497-504.PM