Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Production of transgenic cucumber expressing phytoene synthase-2A carotene desaturase gene

  • Jang, Hyun A (Department of Biology, Chungnam National Univ.) ;
  • Utomo, Setyo Dwi (Department of Agronomy and Horticulture, Faculty of Agriculture University of Lampung) ;
  • Kwon, Suk Yoon (Korea Research Institute of Bioscience and Biotechnology) ;
  • Ha, Sun-Hwa (Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University) ;
  • Xing-guo, Ye (Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement) ;
  • Choi, Pil Son (Department of Medicinal Plant Resources, Nambu University)
  • Received : 2016.08.09
  • Accepted : 2016.09.19
  • Published : 2016.09.30
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Abstract

The objectives of this study were to 1) evaluate the efficiency of the protocol of Agrobacterium-mediated transformation of cucumber to introduce phytoene synthase-2a carotene desaturase (PAC genes); 2) demonstrate the integration of PAC genes into the genome of putative transgenic cucumber based on growth on selection medium, PCR and Southern analysis; 3) evaluate the expression of PAC genes in transgenic cucumber based on the analysis of RT-PCR and Northern blot hybridization. Out of 5,945 cotyledonary-node explants inoculated with Agrobacterium, 65 (1.1%) explants produced 238 shoots. Integration of PAC genes into the genome of the cucumber was demonstrated based on the analysis of gDNA-PCR, 21 out of the 238 plants regenerated; while 6 plants proved positive for Southern blot hybridization. Transgene expression was demonstrated based on analysis of RT-PCR, 6 plants proved positive out of the 6 plants analyzed; while 4 plants out of 6 proved positive during Northern blot hybridization. This study successfully demonstrated the production of transgenic cucumber, integration, and expression of the PAC gene in cucumber.

Keywords

References

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