OPTIMIZATION OF BIOLISTICS PARAMETERS IN EXPLANTS OF RAPID-CYCLING FAST PLANTS (BRASSICA RAPA L.)

• Published in: Journal of the North Carolina Academy of Science Vol. 124; no. 2; pp. 58 - 60
• Main Authors: YOUNG, MARGARET, COGBILL, SALIMAH, JONES, GIEIRA, FAULCON, TRAVIS, GREEN, KALEENA, MCDANIEL, MOSES, HARMON, GARY, BLACKMON, RONALD
• Format: Journal Article
• Language: English
• Published: North Carolina Academy of Science, Inc 01-07-2008
• Subjects: Biolistics; Biology; DNA; Modeling; Photoperiod; Physiological stress; Plant cells; Plants; Plasmids; Seedlings
• ISSN: 2167-5872; 2167-5880

Rapid-cycling fast plants (Brassica rapa: RCBr) is a useful model plant system for teaching and plant research because of its short generation time (seed-to-seed in less than 40 days). Many of its mutants are commercially available and there are several simple experiments that can be set up to study physiological stresses. Unlike other model plants, there are no reported studies on biolistics transformation of this plant. The objectives were to determine if leaf expiants were amenable to bombardment by looking at transient GUS expression, and to optimize the parameters involved. Seeds of RCBr were surface sterilized and germinated in the dark on a Murashige and Skoog-based basal medium. After three days, the seedlings were transferred to the same medium and placed under a 16 hr photoperiod. Aseptically grown leaves were excised and placed on medium containing 1.0 mg l-1 BA (6-benzylaminopurine) and 1.0 mg l-1 NAA (naphthaleneacetic acid), one day prior to bombardment. The plasmid vector (pCAMBIA2301) containing the gus gene was adhered onto gold particles and shot into the leaf tissues. These leaves, along with controls, were assayed histochemically for the GUS protein, two days after bombardment. Optimized parameters were a rupture pressure of 900 psi, a target distance of 3 cm and shooting the leaf explants once with 1 μg of DNA. This is the first report indicating that RCBr is amenable to biolistics transformation.