Optimization of Parameters Influencing Biolistic DNA Transformation of Common Bean (Phaseolus vulgaris L.)

in HortScience
View More View Less
  • 1 1Dept. of Horticulture, Univ. of Nebraska–Lincoln, Lincoln, NE 68583
  • 2 2Dept. of Plant Pathology, Univ. of Nebraska–Lincoln, Lincoln, NE 68583
  • 3 3Center for Biotechnology, Univ. of Nebraska–Lincoln, Lincoln, NE 68583

Optimization of parameters influencing biolistic transformation is a crucial stage towards repeatable transformation of common beans. However, there has been no published study on such optimization of this crop species in a helium particle delivery system (BioRad). Using an intron-containing β-glucuronidase (GUS) gene as a reporter, we optimized several critical parameters of biolistic PDS-1000/He delivery system for common bean transformation. The target explant tissues included cotyledons, zygotic embryos, and meristemic shoot tips suitable for organogenesis. Thus, pretreatment of target tissues with osmotic medium containing 0.15–0.25 m mannitol and 0.15–0.25 m sorbitol, positioning of target tissues in 4 cm microcarrier flying distance, the use of 1.6-μm gold particle and high concentration of coating DNA, and bombardment of young immature tissues twice at 2000 psi, etc., significantly increased transformation rate and achieved the best coverage and penetration of the meristemic areas involved in direct shoot organogenesis.

If the inline PDF is not rendering correctly, you can download the PDF file here.

All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 67 11 0
PDF Downloads 35 13 1