TITLE:
Production of Transgenic Camelina sativa Plants via Agrobacterium-Mediated Transformation of Shoot Apical Meristems
AUTHORS:
Viji Sitther, Behnam Tabatabai, Oluwatomisin Enitan, Somayeh Gharaie Fathabad, Sadanand Dhekney
KEYWORDS:
Genetic Engineering, Green Fluorescent Protein, Micropropagation, Oilseed
JOURNAL NAME:
American Journal of Plant Sciences,
Vol.10 No.1,
January
4,
2019
ABSTRACT: A
method to produce transgenic Camelina sativa plants in cvs. PI650159
and PI650161 was developed. Micropropagated shoot meristem cultures were established
from in vitro germinated seedlings and used as target tissues for Agrobacterium-mediated
transformation. A plasmid harboring enhanced green fluorescent protein, β glucuronidase and neomycin phosphotransferase II genes were used to optimize
parameters for transgenic plant production. Kanamycin at 40 mg·l-1 was
effective in suppression of non-transformed cells while permitting growth of
transgenic tissues. Shoot apical meristems co-cultivated with Agrobacterium exhibited stable enhanced green fluorescence protein (EGFP) and β glucuronidase (GUS)
expression after culture on plant regeneration medium. We observed
transformation efficiencies of 53.33% in cv. PI650159 and 98.33% in cv.
PI650161. The presence of transgenes in both cultivars was confirmed by PCR,
while quantitative real-time PCR detected single copy integration in Pl650161
and two copy integration in Pl650159. Transgenic plants exhibited EGFP and GUS
expression in all tissues including shoots, leaves, buds, floral organs, seeds,
and pods. Our results demonstrate a simple and efficient technique using apical
shoot meristems for production of transgenic C. sativa plants that can be used
for transfer of desirable traits.