Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (6): 1104-1113.doi: 10.3864/j.issn.0578-1752.2013.06.003

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Down-Regulation of the Expression of Grain ppo Genes to Improve Wheat Dough Whiteness by RNA Interference

 SUN  Zhong-Xia, YANG  Feng-Ping, ZHANG  Ting, SUI  Xiao-Yan, LIANG  Rong-Qi, LIU Qing , ZHANG  Xiao-Dong, LI  Bao-Yun   

  1. 1.College of Agronomy and Biotechnology, China Agricultural University/State Key Laboratory for Agrobiotechnology /Key Laboratory of Crop Genomics and Genetic Improvement, Ministry of Agriculture/Beijing Key Laboratory of Crop Genetic Improvement, Beijing 100193, China
    2.Beijing Agro-Biotech Research Center, Beijing Academy of Agriculture and Forestry, Beijing 100097, China
    3.Commonwealth Scientific and Industrial Research Organization, Plant Industry, P.O. Box 1600, Canberra, ACT 2601, Australia
    4.College of Life Science, Capital Normal University, Beijing 100037, China
  • Received:2012-12-30 Online:2013-03-15 Published:2013-02-27

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Abstract: 【Objective】 To develop the transgenic wheat germplasm with low flour PPO activity and high whiteness by RNA interference (RNAi) to specially down-regulate the expression of grain ppo genes. 【Method】 The RNAi vector pBAC47P-ppoIR, aimed at silencing wheat grain ppo genes, was transformed into the callus from Zhongyou9507 immature embryo by biolistic bombardment, and the transgenic plants were obtained after resistance selection, differentiation and regeneration, and then screened by PCR, Southern blotting, half-quantitative RT-PCR, northern blotting and enzyme activity analysis. 【Result】 A total of 27 transgenic T0 plants were obtained. Twelve transgenic lines of T1 generation confirmed by PCR and genomic Southern blotting showed successful integration of RNAi construct into the wheat genome. The expression of grain ppo genes of 20 T2 plants was significantly lower than that of control. Enzyme activity analysis of T2 grains at filling stage showed that the PPO activity of 6 lines reduced obviously. Dough whiteness analysis of T4 grains showed that the whiteness of 5 lines increased clearly and 2 lines improved significantly.【Conclusion】The RNAi vector pBAC47P-ppoIR could down-reglulate the expression of grain ppo genes, reduce the PPO activity, and increase the dough whiteness significantly, so this study will provide wheat breeder with breeding material for whiteness and quality improvement.

Key words: wheat , PPO , RNA interference(RNAi) , whiteness

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