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Physiology and Molecular Biology of Plants

, Volume 24, Issue 4, pp 703–710 | Cite as

Elimination of macro elements from inoculation and co-cultivation media enhances the efficiency of Agrobacterium-mediated transformation in Petunia

  • Aso Nobakht Vakili
  • Hedayat Bagheri
  • Pejman Azadi
Research Article
  • 55 Downloads

Abstract

In order to evaluate the effect of inoculation and co-cultivation media elements on transformation frequency in Petunia hybrida, modified MS media with different elements were tested on Alvan and Large Flower Alvan (LF Alvan), two local cultivars. Leaf explants of both cultivars were inoculated with Agrobacterium tumefaciens strain LBA4404 (pBI121) containing neomycin phosphotransferase (nptII) and an intron-containing β-glucuronidase (gus) genes. When medium lacking KH2PO4, NH4NO3, KNO3, and CaCl2 was used as inoculation and co-cultivation medium, a higher frequency of transformation for Alvan (22%) and LF Alvan (16%) was obtained. Kanamycin resistant plantlets were stained blue by GUS assay. Furthermore, polymerase chain reaction (PCR) analysis revealed the presence of both gus and nptII genes in all putative transformants. Finally, southern blot hybridization confirmed insertion of 1–4 copies of gus gene in transgenic plants.

Keywords

Macro elements Co-cultivation Agrobacterium tumefaciens Transformation Petunia hybrida 

Abbreviations

LF Alvan

Large Flower Alvan

GUS

β-glucuronidase

CTAB

Cetyltrimethylammonium bromide

nptII

Neomycin phosphotransferase

CaMV

Cauliflower mosaic virus

Notes

Acknowledegments

This work was supported by Bu-ali Sina University, Hamedan, Iran, and Novin Giti Gene Biotech. Co. Biotechnology Incubator Center of National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.

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Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2018

Authors and Affiliations

  • Aso Nobakht Vakili
    • 1
  • Hedayat Bagheri
    • 2
  • Pejman Azadi
    • 3
  1. 1.Department of Biotechnology, Faculty of AgricultureBu Ali-Sina UniversityHamedanIran
  2. 2.Department of Plant Biotechnology, Faculty of AgricultureBu Ali-Sina UniversityHamedanIran
  3. 3.Department of Genetic Engineering, Agricultural Biotechnology Research Institute of Iran (ABRII)Agricultural Research, Education and Extension Organization (AREEO)KarajIran

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