Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (4): 791-800.doi: 10.16420/j.issn.0513-353x.2020-0265

• Research Papers • Previous Articles     Next Articles

Cloning of U6 Promoters and Establishment of CRISPR/Cas9 Mediated Gene Editing System in Eggplant

WANG Dan1,2, WANG Mi1,2, LIU Jun2, ZHOU Xiaohui2, LIU Songyu2, YANG Yan1,2,*(), ZHUANG Yong1,2,*()   

  1. 1. College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China
    2. Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement,Institute of Vegetable Crops,Jiangsu Academy of Agricultural Sciences, Nanjing 210014,China
  • Received:2021-10-20 Revised:2022-01-29 Online:2022-04-25 Published:2022-04-24
  • Contact: YANG Yan,ZHUANG Yong;


Seven U6 RNAs were identified from the eggplant(Solanum melongena L.)genome based on the conserved U6 RNA sequence in Arabidopsis and the responding promoters were cloned by PCR amplification. GUS staining results showed that four SmU6 promoters(SmU6-1P,SmU6-2P,SmU6-4P,SmU6-7P)were capable of transcriptional activity. The CRISPR/Cas9 vectors with AtU6-P/SmU6-1P driving SmWRKY4 sgRNA were constructed. T0 plants were obtained by Agrobacterium-mediated transformation of eggplant cotyledons. The DNA of T0 plants were extracted to amplify the target fragments,which were subsequently sequenced to analyze whether SmWRKY4 gene were edited or not. As a result,the CRISPR/Cas9 vector containing SmU6-1P could edit SmWRKY4 with an editing rate of 27.0%. However,no mutation was detected in T0 plants based on AtU6-P. These results indicated that the editing efficiency of eggplant U6 promoter was better than that of Arabidopsis U6 promoter in eggplant genetic transformation.

Key words: eggplant, U6 promoter, CRISPR/Cas9, gene editing

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