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Plant Molecular Biology Reporter

, Volume 29, Issue 1, pp 117–124 | Cite as

Regulation of Carotenoid Content in Tomato by Silencing of Lycopene β/ε-Cyclase Genes

  • Chao Ma
  • Binggang Ma
  • Juan He
  • Qingnan Hao
  • Xiaoyan Lu
  • Lei Wang
Article

Abstract

To conduct RNAi interference of Lyc-β and Lyc-ε genes, two plant expression vectors were constructed by inserting the intron fragments of the gusA gene into the two target gene fragments, which were designed in anti-sense directions. After the Agrobacterium tumefaciens-mediated transformation, 13 transgenic tomato plants (seven and six for Lyc-β and Lyc-ε, respectively) were obtained, which was further validated by PCR. Real-time PCR revealed that the messenger RNA abundance of Lyc-β gene and Lyc-ε gene in transgenic tomato plants was significantly reduced to 8.95% and 13.16%, respectively, of the level of the wild-type plant. Subsequent high-performance liquid chromatography analysis found that transgenic tomato plant had significantly increased lycopene content, with the highest value of 13.8 μg/g leaf dry weight, which was about 4.2-fold that of wild-type plant. Moreover, Lyc-β and Lyc-ε interference gene effects were observed on downstream products as well. β-Carotene and lutein contents decreased in Lyc-β RNAi lines, ranging from 40.7 to 117.3 μg/g and 4.9 to 23.5 μg/g leaf dry weight, respectively. In Lyc-ε RNAi lines, β-carotene content increased, ranging from 195.8 to 290.2 μg/g, while lutein content decreased, ranging from 3.7 to 11.3 μg/g. For total carotenoids, Lyc-β RNAi lines resulted in 2.9-fold decrease, while Lyc-ε RNAi lines yielded 1.7-fold increase in contents when compared to wild-type control. This study demonstrated that RNAi gene technology is an effective method for enhancing lycopene content in plants.

Keywords

β-Carotene Lutein Lycopene Lycopene β-cyclase Lycopene ε-cyclase RNAi 

Notes

Acknowledgements

This research was supported by National Natural Science Foundation of China (30460081) and Scientific Research Fund of Xinjiang BingTuan, China (ZD2007JC06).

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Chao Ma
    • 1
    • 2
  • Binggang Ma
    • 1
    • 2
  • Juan He
    • 2
  • Qingnan Hao
    • 2
  • Xiaoyan Lu
    • 1
    • 2
  • Lei Wang
    • 2
  1. 1.Key Laboratory of Oasis Ecology Agriculture of Xinjiang BingtuanShiheziPeople’s Republic of China
  2. 2.Department of Horticulture, Agricultural CollegeShihezi UniversityShiheziPeople’s Republic of China

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