Over-expression of Glutathione Synthetase Gene Enhances Cadmium Tolerance in Transgenic Tobacco Plant

doi:10.13523/j.cb.20141013

China Biotechnology

2014, Vol. 34

Issue (10): 79-86 DOI: 10.13523/j.cb.20141013

Over-expression of Glutathione Synthetase Gene Enhances Cadmium Tolerance in Transgenic Tobacco Plant

JIA Cui-cui¹, JI Jing¹, WANG Gang¹, TIAN Xiao-wei^2,3, DU Xi-long², GUAN Chun-feng¹, JIN Chao¹, WU Dian-yun²

1. School of Environmental Science and Engineering, Tianjin University, Tianjin 300072, China;
2. School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China;
3. College of Horticulture and Landscape, Tianjin Agricultural University, Tianjin 300384, China

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Abstract

Soil pollution, as a result of industrial development, is becoming a serious problem in China and raising increasing concerns from the public. Soil remediation through plant genetic engineering is a widely used method to tackle this problem. The theory is to provide a new genetic resource for heavy metal phytoremediation through the study and manipulation of heavy metal responsive rate-limiting enzymes in plants. The gene LcGS, coding glutathion synthetase (GS) of Lycium chinense was cloned using reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). Expression of LcGS in L.chinense under CdCl₂ stress over a 24h time period was monitored and analyzed using semi-quantitative RT-PCR. The expression of LcGS gene up-regulated initially following the increase of Cd stress time, then remained at a relatively high level from 9h, till the end of the monitoring period at 24h. The expression vector containing this gene was also constructed and named pCAMBIA2300-LcGS. Then the LcGS expression cassette was transformed into tobacco via Agrobacterium-mediated transformation. Integration of this foreign gene was confirmed using PCR. Under CdCl₂ stress, transgenic plants carrying LcGS gene contain higher contents of glutathione (GSH), phytochelatins (PCs) and chlorophyll than wild-type plants, confirming higher tolerance to CdCl₂. Therefore, over-expression of glutathion synthetase genes through genetic engineering can be a promising strategy for heavy metal phytoremediation.

Key words： Lycium chinense Glutathione synthetase CdCl₂ Stress PCs

Received: 16 July 2014 Published: 25 October 2014

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Cite this article:

JIA Cui-cui, JI Jing, WANG Gang, TIAN Xiao-wei, DU Xi-long, GUAN Chun-feng, JIN Chao, WU Dian-yun. Over-expression of Glutathione Synthetase Gene Enhances Cadmium Tolerance in Transgenic Tobacco Plant. China Biotechnology, 2014, 34(10): 79-86.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20141013 OR https://manu60.magtech.com.cn/biotech/Y2014/V34/I10/79

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