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ACTA HORTICULTURAE SINICA ›› 2014, Vol. 41 ›› Issue (6): 1105-1114.

• Vegetables • Previous Articles     Next Articles

Optimization of the Transgenic System and Genetic Transformation of SmARF8 Gene RNAi Vector in Red-purple Long Eggplant

BAO Chong-lai,DU Li-ming,HU Tian-hua,ZHU Qin-mei,HU Hai-jiao,and MAO Wei-hai *   

  1. (The Institute of Vegetable,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China)
  • Received:2014-02-13 Online:2014-06-25 Published:2014-06-25

Abstract: A high efficient genetic transformation system was developed using hypocotyl explants for producing transgenic red-purple long eggplant(Solanum melongena L.). To generate SmARF8 gene RNAi transgenic eggplant plants,the binary vector pCAMBIAI 35S-2300-SmARF8-INT with the selectable marker gene neomycin phosphotransferase(NPTⅡ)was constructed and transformed into eggplant hypocotyl explantsvia Agrobacterium tumefaciens-mediated infection. Transgenic plantlets were induced on MS selective media containing 2 mg · L-1 indole-3-acetic acid,0.5 mg · L-1 zeatin,25 mg · L-1 kanamycin and 500 mg · L-1 cefotaxime with regeneration ratio higher to 80%. Subsequently,the transgenic plantlets kept growing on same medium and then rooted efficiently on MS basic media to become transgenic plants. The RT-PCR and qRT-PCR analysis showed no or less transcript of SmARF8 in T0 transgenic generation. Further Southern blot analysis of selected T0 transgenic plants confirmed the presence of single T-DNA insertion. These results demonstrate that the endogenesis SmARF8 has been successfully knocked out in eggplant using the established transgenic system in this study.

Key words: eggplant, genetic transformation, RNA interference, Agrobacterium tumefaciens

CLC Number: