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

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...

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Bibliographic Details
Published in:Physiology and molecular biology of plants 2018-07, Vol.24 (4), p.703-710
Main Authors: Nobakht Vakili, Aso, Bagheri, Hedayat, Azadi, Pejman
Format: Article
Language:English
Subjects:
Agrobacterium radiobacter
Ammonium nitrate
beta-glucuronidase
Biological and Medical Physics
Biomedical and Life Sciences
Biophysics
Calcium chloride
Cell Biology
coculture
Cultivars
Cultivation
Culture media
Explants
Genes
Genetic transformation
GUS gene
Hybridization
Inoculation
Kanamycin
kanamycin kinase
leaves
Life Sciences
Neomycin
Neomycin phosphotransferase
Petunia hybrida
Phosphotransferase
Plant Physiology
Plant Sciences
Plantlets
Polymerase chain reaction
potassium dihydrogen phosphate
potassium nitrate
Potassium phosphate
Potassium phosphates
Research Article
Southern blotting
Transgenic plants
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Summary: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 KH 2 PO 4 , NH 4 NO 3 , KNO 3 , and CaCl 2 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.
ISSN:0971-5894
0974-0430
DOI:10.1007/s12298-018-0553-0