Articles
Agrobacterium-mediated genetic transformation of 146-2 Russian apricot and plum rootstock with the gfp gene
Article number
1282_16
Pages
95 – 100
Language
English
Abstract
A protocol for Agrobacterium-mediated genetic transformation was developed for 146-2 Russian apricot rootstock.
Whole leaves from in vitro cultured shoots were used as explants for transformation by A. tumefaciens CBE21 strain containing binary vector pBINmGFP5ER. It has the nptII gene coding for neomycin phosphotransferase II as a plant-selectable marker under the control of the NOS (nopaline synthase) promoter and gfp reporter gene coding the green fluorescent protein under the control of the Cauliflower mosaic virus (CaMV) 35S promoter.
Integration of the nptII and gfp into transgenes was confirmed by PCR. The expression of the green fluorescent protein was observed by fluorescence microscopy.
Transformation efficiency based on PCR analysis of kanamycin resistant independent lines was 0.41-0.83%. All transgenic lines showed kanamycin resistance at concentration 40 mg L‑1. They were rooted and acclimatized to greenhouse conditions.
Whole leaves from in vitro cultured shoots were used as explants for transformation by A. tumefaciens CBE21 strain containing binary vector pBINmGFP5ER. It has the nptII gene coding for neomycin phosphotransferase II as a plant-selectable marker under the control of the NOS (nopaline synthase) promoter and gfp reporter gene coding the green fluorescent protein under the control of the Cauliflower mosaic virus (CaMV) 35S promoter.
Integration of the nptII and gfp into transgenes was confirmed by PCR. The expression of the green fluorescent protein was observed by fluorescence microscopy.
Transformation efficiency based on PCR analysis of kanamycin resistant independent lines was 0.41-0.83%. All transgenic lines showed kanamycin resistance at concentration 40 mg L‑1. They were rooted and acclimatized to greenhouse conditions.
Authors
L.Y. Mourenets, A.S. Pushin, S.V. Dolgov
Keywords
leaf explants, CBE21, A. tumefaciens, nptII, GFP
Groups involved
- Division Plant Genetic Resources, Breeding and Biotechnology
- Division Vine and Berry Fruits
- Division Temperate Tree Fruits
- Division Tropical and Subtropical Fruit and Nuts
- Division Horticulture for Human Health
- Division Ornamental Plants
- Division Vegetables, Roots and Tubers
- Commission Banana
- Working Group Biotechnology of Horticultural Species
- Division Temperate Tree Nuts
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