Ma. Theresa Jonna A. Atienza
To assess the effect of nanomaterials on the survival of gusA-labelled PGPB inoculum strain in the soil and in the rhyzosphere.
1. Proper laboratory attire (lab gown, closed shoes, gloves, etc.) will be worn at all times.
2. The correct containers will be used in conducting transformation and conjugation experiments.
3. The isolates and other materials will be handled carefully to avoid spills.
4. Work surfaces will always be sterilized before and after every activity.
5. The researcher will wash and disinfect her hands after working with the isolates and before leaving the laboratory.
6. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human consumption are not permitted in the laboratory.
7. All materials that were in contact with the isolates will be disinfected and/or decontaminated.
8. The cultures and containers will be decontaminated once the project has been terminated.
1. The transformed PGPB inoculum strain will be placed in a durable, sealed and labeled container.
2. The sealed container will be placed in an appropriate secondary container during transport such as a cooler. Additionally, absorbent materials such as paper towels will be placed in the secondary container to absorb all free liquids in the event that primary containers rupture or break during transport.
3. The primary containers will be packed in the secondary container in a manner that will reduce shock, rupture, and/or breakage. Bubble wrap or similar shock-absorbing materials may also be used to minimize the potential for primary container rupture.
4. The secondary container will also be labeled with a bried description of the contents and an emergency contact name and phone number.
5. A private vehicle will be used for transport. The transport container will be secured in an upright position, and in a location in the vehicle whereby if an accident were to occur, the container or its contents will not be an exposure rist to the driver or to the environment, such as the backseat.
1. The ASC screenhouse is sturdy and can withstand strong wind and rain. Additional nets can be placed in the sides and top area to reduce impact in case a typhoon occurred.
2. Durable pots will be used to avoid breakage that may result to the escape of transconjugants to the environment.
3. The leachate from each will be collected regularly by placing collectors at the bottom of the pots. the leachate will be decontaminated prior to disposal.
4. All the materials that were in contact with the transformed isolates will be decontaminated when the project has been terminated.
a) Project activities shall be conducted at the Molecular Genetics Laboratory of the University of the Philippines Los Baños - National lnstitute of Molecular Biology and Biotechnology (UPLB-BIOTECH) and the UPLB Agricultural Science Cluster (UPLB-ASC) screenhouse.
b) ln view of the renovation of the UPLB-ASC screenhouse, the UPLB-IBC, prior to conduct of screenhouse activities, must carry-out and submit a report on the inspection of the said screenhouse confirming adequacy of the facility to effectively contain the transgenic materials.
c) The proponent shall adhere as closely as possible to the schedule of activities reflected in the submitted Gantt chart.
d) Any modifications in the schedule of activities shall be made with the concurrence of the UPLB-IBC and the DOST-BC.
e) Signage should be in place indicating that the facility is restricted/limited access;
f) The proponent must provide a logbook to record access of personnel to the experimental facility.
g) The proponent must take measures to ensure that only the DOST-BC authorized personnel are allowed inside the experimental facilities.
h) The experimental facilities must be kept under lock when not in use.
i) ln case of undue destruction of experimental materials resulting from unauthorized entry of personnel or breach of containment of experimental facilities, the proponent shall implement measures to prevent the inadvertent escape of any viable material within the facility.
j) The proponent and the supervising IBC shall be held accountable for the undue destruction of the experimental materials and the consequences that their inadvertent escape may cause to the surrounding environment
k) The DOST-BC should be informed in advance of any visitations by unauthorized persons.
l) The DOST-BC shall be informed immediately of any intrusion by unauthorized persons.
m) The proponent shall strictly observe proper disposal procedures for all materials.
n) Movement of all materials will be done in compliance with all relevant biosafety and phytosanitary requirements of the Philippines.
o) Any additional requirement that the DOST-BC may impose, as necessary, for the duration of the experiment shall be complied with.
p) The proponent shall submit through the IBC a completion report within 90 days after completion of the experiment.
q) The IBC shall submit to the DOST-BC a report on the completion of this project, in the prescribed format.
Developments in nanotechnology are leading to a rapid proliferation of new materials. Nanoparticles (NPs) have received a lot of attention and concern recently due to their rapidly increasing applications. These can be used in electronics, personal care products, biosensors, food additives, pigments and rubber manufacture. The elevated production and use of these nanomaterials will result in an increased NP release into the environment. However, there is very little understanding on the behavior of these nanomaterials in the environment. Although a lot of benefits are being realized, it is still possible that the accumulation of nanomaterials in the environment may cause toxicity. It is therefore important to determine the possible detrimental effect that the nanomaterials can cause on the environment. Assessment of the safety of nanomaterials should be started with the soil microorganisms. Microorganisms are of great environmental importance because they are the foundation of aquatic and terrestrial ecosystems and provide key environmental services ranging from primary productivity to nutrient cycling and waste decomposition.
This project will utilize advanced techniques in molecular biology to profile the soil microbial community and determine its response to the application of nanomaterials, including engineered metal NPs (ZnO and CuO), nano-fertilizers and nano-plant hormones. The effect of nanomaterials on the survival of gusA-labelled microbial inoculant strain will also be determined, including the effect of nanomaterials on the effectiveness of formulated microbial inoculants.
Activity 2 involves the insertion of the plasmid pCAM121 containing mTn5SSgusA21 to Escherichia coli strain S17-1 (λ-pir) followed by bacterial conjugation experiments to transfer the plasmid pCAM121 containing mTn5SSgusA21 to three PGPB inoculum strains. The transformation and bacterial conjugation experiments, and the characterization of the transconjugants will be performed at the Nano-Biotech Information and Translational Laboratory (NBITRL) in BIOTECH-UPLB from January to April, 2017. The transformed isolate will be used as an inoculant in a pot experiment with nanomaterials as treatments. The survival of PGPB isolates in the soil and in the rhizosphere will be monitored regularly. The pot experiment will be set-up at the Agricultural Systems Cluster Screenhouse from May to August, 2017.
Public Information Sheet
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