Utilizing Science & Technology and Innovation
for Development
A novel approach for assessing the
impacts of metal oxide nano-particles on
the physiological, biochemical, and
ultrastructural characteristics in tobacco
plant and the systemic movement of plant
RNA viruses.
Marriott Hotel- Amman, August 12th, 2015
Project Team
•
Principle Investigator: Dr. Rami Alkhatib/Jordan University of
Science and Technology/Biotechnology and Genetic Engineering
Department /Jordan
•
Co-Investigator: Dr. Fares Howari/ Director Office of Sustainable
Eco Innovation, Zayed University/ UAE
•
Co-Investigator: Dr. M-Ali AL-Omari (AL-Akhras)/JUST/ Jordan
•
Co-Investigator: Dr. Borhan Albiss/ JUST/ Jordan
•
Co-Investigator: Dr. Laith AL-Eitan/ JUST/ Biotechnology and
Genetic Engineering Department /Jordan
•
Co-Investigator: Dr. Nour Abdo/ Public Health/ Jordan University
of Science and Technology/Jordan
Brief Description
Agricultural damages caused by pathogenic attack
(insect, viral, and fungal diseases) are enormous, and
because of the environmental concerns, the communities
avoid the total dependence on chemical applications.
 Our knowledge to explore the physiological and
metabolic impact of metal oxide nano-particles on
plant is poorly understood.
 The perception about the systemic movement of
plant RNA viruses and the factors involved in this
process is not fully elucidated.
Justifications
 To date, the physiological and biochemical effects of
different concentrations of metal oxide nano-particles in
plants are still not fully understood.
Using microscopy techniques will enable us to localize where metal oxide nanoparticles are accumulating in the plant organs.
To the best of our knowledge, this will be the first pioneer study to examine the
effect of metal oxide nano-particles on the systemic movement of plant RNA
viruses and the factors involved in this process.
Objectives
To examine the physiological (shoot height, root length, root and
shoot dry mass, photosynthetic rate, stomatal conductance,
transpiration rate, and chlorophyll content); and ultrastructural
(chloroplast morphology, thickness of cell wall, and localization of
metal oxide nano-particles in the plant different tissues).
 To determine the biochemical (carbohydrate content,
protein content, Proline concentration) effects of metal
oxide nano-particles.
 To elucidate the effects of of metal oxide nano-particles
on the nano-machine systemic movement of certain
plant RNA viruses (Turnip vein-clearing virus (TVCV),
Tobacco musaic virus (TMV), and Tobacco etch virus
(TEV) using microscopy and molecular techniques.
Scope of work/Duration
Estimated Budget
This Research proposal has three directions/ phases:
• Conducting a physiological and biochemical experiments as well as
analyzing all physiological and biochemical parameters to examine the
Phase I
effects of metal oxide-nano particles.
Phase II
• To localize where metal oxide-nano particles are accumulating in the
plant tissues using SEM and TEM.
• Test the effects of metal oxide nano-particles on the nano-machine
systemic movement of certain plant RNA viruses (Turnip vein-clearing
Phase IV virus (TVCV), Tobacco musaic virus (TMV), and Tobacco etch virus
(TEV) using microscopy and molecular techniques.
Duration: 36 months (each scope: 12 months)
Estimated Budget : 117,000 JD
Methodology of Implementation
Metal oxide nano-particles with diameters of
approximately 9 nm and18 nm will be prepared
according to the chemical co-precipitation method.
 Tobacco (Nicotiana tabacum var. Turkish) will be grown
hydroponically in a growth chamber at a photosynthetic photon
flux density (PPFD) of 250–300 µmol m-2 s-1 under controlled
temperature (23–30 ºC), and humidity (60 ± 5%).
 Different concentrations of Metal oxide nano-particles will be
added to the Hoagland solution.
Prepare metal oxide
nano-particles
Grow Tobacco
hydroponically
Treat Tobacco with
different
concentrations at
different conditions
Methodology of Implementation
 Physiological and biochemical parameters will be
measured using different techniques and kits (i.e.
carbohydrate conc., protein conc., and proline
conc.)
 Ultrastructural parameters will be examined using both TEM
and SEM devices.
 Viral presence or absence will be detected using
immunolabeling techniques in microscopy.
Test and measure
Physiological and
biochemical
parameters
Examine
Ultrastructural
parameters using
both TEM and SEM
devices.
Use immunolabeling
techniques in
microscopy to
investigate Viral
presence or absence
Expected Output
 Elucidate the physiological and biochemical
effects of metal oxide nano-particles in the plants
will shed more light about how these particles
enhance or inhibit the plant growth.
The ultrastructural study will allow us to localize where
these particles accumulate and show their role in the plant
growth.
Knowing the plant factors that may be responsible for the
systemic movement inhibition, if any, will allow us to
genetically engineer them and produce plants resistance to
viral infection.
Impact
 Enhancing seed germination and plant production
especially for concentrations improving the
physiological and biochemical factors in the plant.
Producing plants more resistance to biotic and abiotic stress
which minimize our dependence on chemical pesticides.
Preventing plant viral infections especially in crops which will
reduce the cost and loss of these crops.
Sustainability
 Based on the biochemical, physiological
parameters, and the toxicity of metal oxide
nanoparticles, other plants species (mainly crops)
will be used in the future.
 Identifying and characterizing the plant factors responsible for
the virus movement inhibition and genetically engineered them to
produce crops with highly virus resistance. We plan to secure fund
for this part from Genetic Engineering Companies.
 Appling the genetically engineered product to improve
agriculture in general.
Action Plan
This project will be conducted over a period of 36 months
after funding the proposal.
Project Direction A: Physiological and biochemical analyses
Stage 1: Measuring all physiological parameters (shoot height,
root length, dry mass for shoot and root, photosynthetic
parameters, and stomatal conductance)
Stage 2: Measuring all the biochemical parameters (carbohydrate
conc., protein conc., proline conc., and GABA conc.)
Stage 3: Statistical analyses for all parameters measured.
Stage 4: Finalizing the data and writing a manuscript.
Action Plan
In the first year after the proposal acceptance and receiving
the funds and Reagents, Kits, consumables
Project Direction B: Metal oxide nano-particles localization in
plant tissues
Stage 1: Conducting a microscopy study for localizing metal oxide
nano-particles accumulation in both root and shoot
Stage 2: Plant infection with different RNA viruses and examine
the effects of different concentrations of metal oxide nano-particles
on the systemic movement of these viruses using immunolabeling
technique.
Stage 3: Image processing and preparing them for publication.
Stage 4: Finalizing the data and writing a manuscript.
Action Plan
Project Directions C: Plant-virus interaction
Stage 1: Plant inoculation with RNA viruses, then
apply metal oxide nano-particles for 14 days.
Stage 2: Examine the presence or the absence of the
virus in the upper leaves using TEM.
Stage 3: Writing manuscript for publication.