Gene-Splitting Technology: A Novel Approach for the Containment of Transgene Flow in Nicotiana tabacum

doi:10.1371/journal.pone.0099651

CORC > 中国农业科学院 > 作物科学研究所

	Gene-Splitting Technology: A Novel Approach for the Containment of Transgene Flow in Nicotiana tabacum
	Wang, Xu-Jing 1; Jin, Xi 1; Dun, Bao-Qing 2; Kong, Ning 1; Jia, Shi-Rong 1; Tang, Qiao-Ling 1; Wang, Zhi-Xing 1
刊名	PLOS ONE
	2014
卷号	9 期号:6 页码:-
ISSN号	1932-6203
DOI	10.1371/journal.pone.0099651
通讯作者	Wang, Xu-Jing
英文摘要	The potential impact of transgene escape on the environment and food safety is a major concern to the scientists and public. This work aimed to assess the effect of intein-mediated gene splitting on containment of transgene flow. Two fusion genes, EPSPSn-In and Ic-EPSPSc, were constructed and integrated into N. tabacum, using Agrobacterium tumefaciens-mediated transformation. EPSPSn-In encodes the first 295 aa of the herbicide resistance gene 5-enolpyruvyl shikimate-3-phosphate synthase (EPSPS) fused with the first 123 aa of the Ssp DnaE intein (In), whereas Ic-EPSPSc encodes the 36 C-terminal aa of the Ssp DnaE intein (Ic) fused to the rest of EPSPS C terminus peptide sequences. Both EPSPSn-In and Ic-EPSPSc constructs were introduced into the same N. tabacum genome by genetic crossing. Hybrids displayed resistance to the herbicide N-(phosphonomethyl)-glycine (glyphosate). Western blot analysis of protein extracts from hybrid plants identified full-length EPSPS. Furthermore, all hybrid seeds germinated and grew normally on glyphosate selective medium. The 6-8 leaf hybrid plants showed tolerance of 2000 ppm glyphosate in field spraying. These results indicated that functional EPSPS protein was reassembled in vivo by intein-mediated trans-splicing in 100% of plants. In order to evaluate the effect of the gene splitting technique for containment of transgene flow, backcrossing experiments were carried out between hybrids, in which the foreign genes EPSPSn-In and Ic-EPSPSc were inserted into different chromosomes, and non-transgenic plants NC89. Among the 2812 backcrossing progeny, about 25% (664 plantlets) displayed glyphosate resistance. These data indicated that transgene flow could be reduced by 75%. Overall, our findings provide a new and highly effective approach for biological containment of transgene flow.
学科主题	Multidisciplinary Sciences
语种	英语
出版者	PUBLIC LIBRARY SCIENCE
WOS记录号	WOS:000340947700124
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/4873]
专题	作物科学研究所
作者单位	1.Chinese Acad Agr Sci, Biotechnol Res Inst, Beijing 100193, Peoples R China 2.Chinese Acad Agr Sci, Inst Crop Sci, Natl Key Facil Crop Gene Resources & Genet Improv, Beijing 100193, Peoples R China
推荐引用方式 GB/T 7714	Wang, Xu-Jing,Jin, Xi,Dun, Bao-Qing,et al. Gene-Splitting Technology: A Novel Approach for the Containment of Transgene Flow in Nicotiana tabacum[J]. PLOS ONE,2014,9(6):-.
APA	Wang, Xu-Jing.,Jin, Xi.,Dun, Bao-Qing.,Kong, Ning.,Jia, Shi-Rong.,...&Wang, Zhi-Xing.(2014).Gene-Splitting Technology: A Novel Approach for the Containment of Transgene Flow in Nicotiana tabacum.PLOS ONE,9(6),-.
MLA	Wang, Xu-Jing,et al."Gene-Splitting Technology: A Novel Approach for the Containment of Transgene Flow in Nicotiana tabacum".PLOS ONE 9.6(2014):-.