Agriculture Reference
In-Depth Information
target protein is done by using specific antibodies. Generally for the detection of
target protein, primary antibodies are the polyclonal antibodies raised in rabbit
against the N-terminal peptide of GUS reporter protein and secondary antibodies
are raised in donkey against rabbit IgG. The membranes with the primary and
secondary antibodies can be exposed to X-ray films, and the intensity of signals can
be quantified by using the ImageJ program (Rybicki and von Wechmar
1982
).
10.5 Conclusions and Future Prospects
It can be concluded that efficient transformation of plants with transgenes encap-
sulated in nanocarriers is relatively a new area in plant biotechnology. Genetic
engineering is today
s most important technique for fulfillment of the basic needs of
'
the world
s population. Till now, liposomes have been explored, and other nano-
technologies (nanoparticle, carbon nanotube, quantum dots, nanoemulsion, etc.)
will definitely be used as they were successfully explored in animal biotechnology.
Although nanotechnology-based biotransformation is relatively efficient, its com-
bination with commonly applied approaches in transgenic plant development will
surely improve in terms of efficiency and productivity, and the chances of transgene
silencing can also be minimized. Nanoparticles-mediated gene transfer depends not
only on the defined nanoparticle size and shape but also on surface functionalization
of NPs, nucleic acid protection ability, and biocompatibility. All these parameters
need to be fine-tuned for their use as carriers in gene transfer method. The
mechanism of biotransformation in nanotechnology is still not clearly understood,
but the possible role of endocytosis process as well as transcellular and paracellular
transport of the nanocarriers-encapsulated transgene cannot be ruled out. Different
kinds of nanoparticles can be loaded with nucleic acids (DNA or RNA), and cells
appear to be quite indifferent to the chemical nature of these nanoparticles when it
comes to an uptake by endocytosis.
On the ground of earlier reports that revealed the application of liposomal- and
nanoparticle-based gene transfer, various mechanistic ideas can be postulated to
improve the efficacy of this novel nonviral gene-protected biotransformation
approach. Modification of physicochemical properties of nanocarriers, protoplast,
and plasma membrane may significantly improve the liposomal- and nanoparticle-
encapsulated gene into the plant cell and establish these gene transfer methods as
sole process rather than adjuvant.
'
Acknowledgements The authors are thankful to the Department of Science and Technology
(Nano Mission Project) and to the University Grants Commission, New Delhi, for providing
financial assistance under the UGC-SAP program. MKR is thankful to FAPESP for the financial
assistance to visit the Institute of Chemistry, State University of Campinas, Brazil.
