Stable genetic transformation of high oleic Helianthus annuus L. genotypes with high efficiency

• Published in: Plant science (Limerick) Vol. 171; no. 5; pp. 546 - 554
• Main Authors: Mohamed, Sh, Boehm, R., Schnabl, H.
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier Ireland Ltd 01-11-2006; Elsevier Science
• Subjects: Agrobacterium; Agronomy. Soil science and plant productions; beta-glucuronidase; Biological and medical sciences; Capella; Fundamental and applied biological sciences. Psychology; gene expression regulation; Genetic engineering applications; genetic transformation; genetic variation; Genetics and breeding of economic plants; green fluorescent protein; Gus gene; Helianthus; Helianthus annuus; Helianthus annuus L; inheritance (genetics); Meristem transformation; Mgfp5 gene; oil crops; Particle bombardment; Plant breeding: fundamental aspects and methodology; reporter genes; shoot meristems; transgenes; transgenic plants; Vacuum infiltration; hptII; gus; MS; Gus gene; Vacuum infiltration; rpm; nptII; gfp; BAP; Mgfp5 gene; nos; Meristem transformation; CaMV; 4-MU; Helianthus annuus L; MUG; Particle bombardment; Genetic transformation; Compositae; Meristem; Helianthus annuus; Gene; Dicotyledones; Angiospermae; Spermatophyta
• ISSN: 0168-9452; 1873-2259
• DOI: 10.1016/j.plantsci.2006.05.012

Two previously optimized procedures for genetic transformation of two economically important high oleic sunflower genotypes (
H. annuus cv. capella and SWSR2 inbred line) were used to generate stably transformed sunflower plants of these genotypes expressing
gus reporter gene:
Agrobacterium tumefaciens-mediated gene transfer to juvenile split apical meristems was applied to cv. capella, and a biolistic gene transfer method using the same target tissue was applied to SWSR2 inbred line. Out of 120 originally used explants of cv. capella, five transgenic regenerants could be identified by biochemical and molecular analysis. Thus, transformation efficiency amounted to 4.1%. For SWSR2 inbred line, 125 explants were used and six transgenic regenerants could be identified in an identical manner, resulting in a transformation efficiency of 4.8%. The highest expression level of the
gus gene resulted in a yield of recombinant protein of approx. 0.2% total soluble protein. Additionally, T
1 progeny was tested for
gus gene inheritance in both genotypes. From approx. 75% of the T
0 plants, the
gus gene was stably transmitted to the T
1 progeny and expressed in a detectable amount. To test the reproducibility of the transformation procedures, they were used to introduce another reporter gene,
mgf5, into both genotypes. The resulting transformation efficiencies were 3.3 for both genotypes.
gfp proved to be a suitable reporter gene to select transgenic regenerants in early developing stages. The data presented here demonstrate for the first time the reproducible transformation of high oleic sunflower genotypes with high frequency.