1) To generate 40 transgenic events of wheat Lr67res in IR64;
2) to characterize and identify 5 homozygous lines with single insertion;
3) to evaluate the resistance of the transgenic plant against three fungal pathogens including rice blast, false smut and brown spot; and
4) to analyze other agronomic traits, e.g., yield and biomass of transgenic plants.
1. Access to the organism, and other contaminated materials will be limited to those who will be granted permission to the restricted containment facilities in the Institute.
2. Only personnel trained to work under aseptic conditions and with sufficient knowledge of molecular techniques and/or properly supervised personnel will be allowed to take part in the experiment.
3. Seeds of the engineered rice lines will be handled in compliance to all relevant biosafety and phyto-sanitary requirements of the Philippines.
4. Engineered rice lines will be grown and kept in isolation in either CL4 or transgenic screen houses (CS-01) to prevent access by unauthorized personnel.
5. Samples collected from the transformed plants will be placed in sealed plastic bags and then placed in a second sturdy, leak proof, plastic container which will be hermetically sealed.
6. Inoculated plants will be kept in isolation in the CL4 transgenic greenhouse to prevent access of unauthorized personnel.
7. Samples collected from the inoculated plants will be placed in sealed plastic bags and then placed in a second sturdy, leak proof, plastic container which will be hermetically sealed.
8. All the following other materials contaminated with the pathogen will be decontaminated by autoclaving at high pressure prior to disposal according to standard biosafety protocol
9. Plants and plant materials for disposal will be placed in sealed plastic bags and will be decontaminated by autoclaving and then disposed, following the standard procedures prescribed by the biosafety committee.
10. Precautionary measures will always be observed and good laboratory practices will be stringently imposed to prevent cross-over and dissemination transgenic plant materials.
a) Project activities shall be conducted at the Genetic Transformation and Plant Pathology Laboratories, CL4 transgenic glasshouse and at CS-01 transgenic screen house located at the lnternational Rice Research lnstitute in Los Banos, Laguna.
b) Prior to the initiation of the activities, the proponenUs shall discuss the monitoring schedule with the Department of Agriculture-Bureau of Plant lndustry (DA-BPl) to identify the activities that will requlre their presence. The proponenUs shall furnish the DOST-BC with a copy of the agreed monitoring schedule.
c) The proponent shall adhere as closely as possible to the schedule of activities reflected in the submitted Gantt chart.
d) The proponent shall be required to submit a new proposal should they decide to do confined testing.
e) Any modifications in the schedule of activities shall be made with the concurrence of the lRRl-lBC and the DOST-BC. Signage should be in place indicating that the facility is restricted/limited access; The proponent must provide a logbook to record access of personnel to the experimental facilities
h) The proponent must take measures to ensure that only the DOST-BC authorized personnel are allowed inside the experimental facilities
i) The experimental facilities must be kept under lock when not in use.
j) 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 materral within the facility
k) The proponent and the supervising IBC shall be held accountable forthe undue destruction of the experimental materials and the consequences that their inadvertent escape may cause to the surrounding environment.
l) The DOST-BC should be informed in advance of any visitations by unauthorized persons
m) The DOST-BC shall be informed immediately of any intrusion by unauthorized persons
n) The proponent shall strictly observe proper disposal procedures for all materials.
o) Movement of all materials will be done in compliance with all relevant biosafety and phytosanitary requirements of the Philippines.
p) Any additional requirement that the DOST-BC may impose, as necessary. for the duration of the experiment shall be complied with.
q) The proponent shall submit through the IBC a completion report within 90 days after completion of the experiment.
r) The IBC shall submit to the DOST-BC a report on the completion of this project, in the prescribed format.
The wheat gene Lr67 confers durable and broad-spectrum disease resistance against the four biotrophic fungal pathogens wheat leaf rust (Puccinia triticina), stripe rust (Puccinia striiformis f. sp. tritici), stem rust (Puccinia graminis f. sp. tritici) and powdery mildew (Blumeria graminis f. sp. tritici) (Herrera-Foessel et al., 2014). The gene has been used in wheat breeding for almost a century and no adaptation of the pathogens has been observed. Because of its durability and broad spectrum, Lr67 is increasingly being valued as a “backbone” gene to be used in combination with other genes towards achieving more durable resistance in breeding programs world-wide. Molecular analyses have shown that this multi-pathogen resistance is conferred by a single gene encoding a predicted hexose transporter. It was further found that the resistance allele (Lr67res) differed from the susceptible allele (Lr67sus) by two amino-acid residues. Lr67res exerts a dominant-negative effect through heterodimerization with Lr67sus to reduce glucose uptake (Moore et al., 2015). Intriguingly, transgenic sorghum with the wheat Lr67res gene showed enhanced resistance against sorghum rust (Lagudah, Dibley, Schnippenkoetter-CSIRO unpublished). It is thus interesting to test whether Lr67res could also function in rice for enhanced resistance against fungal pathogens. In this study, we will generate transgenic rice plants by introducing Lr67res from wheat into the susceptible rice variety IR64 and test the derived transgenic plants to three different rice fungal diseases including rice blast, false smut, and brown spot. Other agronomic traits of transgenic plants, e.g., yield will be also evaluated.
This project will be mainly conducted at GTL molecular laboratory, the plant pathology main laboratory, CL4 growth facility, and CS-01 at International Rice Research Institute. Through this project, we aim 1) to generate 40 transgenic events of wheat Lr67res in IR64; 2) to characterize and identify 5 homozygous lines with single insertion; 3) to evaluate the resistance of the transgenic plant against three fungal pathogens: rice blast, false smut and brown spot; and 4) to analyze other agronomic traits, e.g., yield and biomass of transgenic plants.
This project is to be completed in two years from the time of commencement. The developed 5 homozygous transgenic lines of IR64 will be kept for further research through project extension or for a new project after this project is completed.
Public Information Sheet
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