Work

Parameters influencing Agrobacterium-mediated transformation system in safflower genotypes AKS-207 and PKV Pink

Year: 2016

Type: article

Abstract: Shoot regeneration in safflower ( Carthamus tinctorius ‘AKS 207’ and ‘PKV Pink’) genetically transformed using Agrobacterium was used for assessing various constraints to the efficiency of transformation including infection period, virulence induction medium, co-cultivation period, bacterial titre, selection regime, and the natural phenolic compound acetosyringone. Transformation frequency was promising with 8–10-day-old cotyledonary leaf explants. Therefore, explants of that age cultured on Agrobacterium minimal medium (AB) containing 100 µM acetosyringone were infected with Agrobacterium (cell titre 0.5 OD 600nm ) for 15 min followed by 48 h of co-cultivation on kanamycin-enriched medium (50 mg/L). Transformation of the shoots was confirmed using β-glucuronidase (GUS) histochemical assay and polymerase chain reaction (PCR). With the transformation protocol thus optimized, the transformation frequency as determined using GUS assays was 54.0 % for AKS 207 and 47.6 % for PKV Pink. The corresponding figures using PCR were 27.0 and 33.3 %. The transformed shoots required 10–14 weeks of culture initiation but produced very few roots.

Source: 3 Biotech

DOI: 10.1007/s13205-016-0497-4

Authors: Dipti Raghunath Dhumale, Prashant Raghunath Shingote, Mahendra Shankarrao Dudhare, Pravin Vishwanath Jadhav, Prashant Bhaskar Kale

Institutions: Dr Panjabrao Deshmukh Krishi Vidyapeeth, National Research Centre on Plant Biotechnology

Language: English

Topic: Plant tissue culture and regeneration

Subfield: Plant Science

Field: Agricultural and Biological Sciences

Domain: Life Sciences

SDG: Zero hunger

Open Access status: hybrid

Estimated APC paid: $4,090