|Project Title||Evaluation of Coconut Wrinkled gene expression and its effect in oil biosynthesis in a Model Monocot Zea mays L.|
|Project Type||Confined Test|
|Name of Institution||University of the Philippines - Los Banos|
|Supervising IBC||University of the Philippines-Los Banos (UPLB)|
Antonio C. Laurena
|Experimental Facility/Site||lnstitute of Plant Breeding, UP Los Baños, College, Laguna, Region 4A|
|Purpose / Objectives||
To express selected coconut genes associated with oil biosynthesis in corn (Zea mays L.) and analyze its effects.
All activities concerning gene cloning, construction of transformation cassettes and tissue culture will be conducted inside a BSL2 facility. Likewise, Seedling will be regenerated inside a BL2 greenhouse. Access to both facilities will be restricted to authorized personnel only. All materials that will be used during and after each experiment wlll be sterilized and decontaminated using necessary procedures. All personnel that will be involved in the project will be informed and will adhere to Good Laboratory Practices (GLP).
|Conditions for Approval||
a) All activities shall be conducted at the P2 Facility (Biochemistry Laboratory) of the Institute of Plant Breeding, College of Agriculture and Food Science, University of the Philippines Los Baños, College, Laguna;
b) The proponent shall submit to the DOST-BC a modified Gantt chart and shall adhere as close as possible to the revised schedule of activities;
c) Any modifications in the schedule and additional activities must be communicated to the DOSTBC for information and/or approval;
d) All confinement measures should be in place prior to the commencement of the activities;
e) Signage should be in place indicating that the facility is a restricted/limited access facility and only those personnel approved by the DOST-BC maybe allowed access;
f) Only authorized personnel as indicated in the proposal shall be allowed access to the facility for the entire duration of the experiment;
g) Monitoring for biosafety compliance shall be conducted by the lnstitutional Biosafety Committee (lBC), report of which shall be submitted to the DOST-BC within five (5) working days after the conduct of the monitoring activity;
h) Measures to limit access of unauthorized persons should be strictly observed during conduct of the experiment and that the designated facility must be kept secured at all times;
i) The DOST-BC shall be informed in advance of any visitations by unauthorized personnel for approval.
j) The DOST-BC shall be informed immediately of any intrusion by unauthorized persons;
k) The proponent shall ensure that stray animals are excluded from the facilities at all times;
l) In case of incident or undue destruction in the facility resulting from unauthorized entry of personnel or breach in containment, the proponent shall implement the biosafety contingency measures to prevent the inadvertent escape of any viable materials to the outside environment;
m) 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; and
n) All viable materials within the experiment area shall be duly accounted for.
|Date of Approval (DD-MM-YYYY)||21-04-2018|
|Date of Completion (DD-MM-YYYY)||N/A|
One of the major goals of the agricultural sector worldwide is to meet the unceasing rise in demand for oil consumption. Studies have shown that Wrinkled1 gene (WRI1) act as a master regulator for plant oil biosynthesis and alteration in its expression may result in the improvement of oil content in crops. ln this project, we would like to express WRll in an experimental monocot model system, corn. The overarching goalof this activity is to evaluate the expression of WRI1 gene in maize tissues and to analyse its effect on its oil content.
A Phase 2 continuation of Project 5 of the Coconut Genomics Program is being implemented at the lnstitute of Plant Breeding, University of the Philippines Los Baños (lPB-UPLB). The project has a duration of two-years and has the following objectives to accomplish: biolistic transformation of corn lines with the transformation cassettes assembled during phase one of the project, regenerate transformed corn tissues into plantlets under contained laboratory conditions, analyse the gene expression through molecular screening and analyse the effect of the gene in oil biosynthesis.
A total of four constructs will be used in the biolistic transformation during the phase 2 of the project, namely, pCambia1302-GFP, pCambia1302-WRl1, pCambia1312-cocosin, and pCambia1302-cocosin-Wri1.The corn lines Pi 23 and CML 161 are still to be used as it has been determined to be the most suitable for transformation. The expression and effects of WRI1 gene on Z. mays will be analyzed through molecular screening (PCR and qPCR) and oil analysis (total fat content and fatty acid profiling).