Improving the Agrobacterial Attachment to Monocot Cells Using a Cellulose Fibrous Network


Global Biotechnology Congress 2016, Boston, United States Of America, 22 - 25 August 2016, pp.183

  • Publication Type: Conference Paper / Full Text
  • City: Boston
  • Country: United States Of America
  • Page Numbers: pp.183
  • Istanbul University Affiliated: Yes


Agrobacterium tumefaciens is an important biotechnological tool that has been used extensively as a
vector for genetic transformation of plants. Cereal crops, such as barley, are not the natural hosts for
the Rhizobiaceae family and therefore recalcitrant for Agrobacterium-mediated transformation.
Modification of transformation procedures is still necessary due to difficulties in the inoculation step
towards the high production of transgenic plants. In this study, we have transformed barley tissues
with LBA4404 strain with pPZP201-GFP-PMI plasmid in the presence of fibrous cellulose in the inoculation medium.
Addition of cellulose increased the T-DNA transfer efficiency based on the number of GFP-expressing clusters on calli
counted after 10 days of transformation. Increased attachment to callus tissue was also confirmed by a binding assay
which revealed that 2-5 fold more bacteria stayed bound on the tissue after a sonication treatment. Cellulose fibrils
collected the free bacteria in the medium observed by phase-contrast microscopy. Bacterial cell attachment and
proliferation on cellulose fibrils were assessed using scanning electron microscopy. Average 40-120 bacteria attached to
cellulose fibrils with 50µm in size were observed. The fibrous cellulose network may be used for large-scale monocot
transformations by eliminating the genotype-dependency.