Agrobacterium-Mediated Transformation In The Green Alga Haematococcus Pluvialis (Chlorophyceae, Volvocales)

The first successful Agrobacterium-mediated transformation
of the green alga Haematococcus pluvialis
Flot. using the binary vectors hosting the genes
coding for GUS (b-glucuronidase), GFP (green fluorescent
protein), and hpt (hygromycin phosphotransferase)
is reported here. Colonies resistant to
hygromycin at 10 mg Æ L)1 expressed b-glucuronidase.
The greenish yellow fluorescence of GFP was
observed when the hygromycin-resistant cells were
viewed with a fluorescent microscope. PCR was used
to successfully amplify fragments of the hpt
(407 bp) and GUS (515 bp) genes from transformed
cells, while Southern blots indicated the integration
of the hygromycin gene into the genome of H. pluvialis.
SEM indicated that the cell wall of H. pluvialis
was altered on infection with Agrobacterium. The
transformation achieved here by Agrobacterium does
not need treatment with acetosyringone or the
wounding of cells. A robust transformation method
for this alga would pave the way for manipulation
of many important pathways relevant to the food,
pharmaceutical, and nutraceutical industries.