Project Information

Reference Number 20180319
Project Title Confined Test (CT) to assess the agronomic traits of IR64 and Ciherang-Sub1 CRISPR lines
Project Type Confined Test
Status Ongoing
Name of Institution International Rice Research Institute
Cooperating Institution Heinrich-Heine-Universitat Dusseldorf
Supervising IBC International Rice Research Institute (IRRI)
Project Leader(s)

Dr. Ricardo Oliva
Dr. Inez Slamet-Loedin

Experimental Facility/Site CT2 (Block F8 - 9), IRRI Experimental Stations, Los Baños, Laguna
Purpose / Objectives

To determine the growth performance and evaluate agronomic traits and yield of the selected SWEET mutant lines of T4 IR64-IRS1132 and T3 Ciherang-Sub1-IRS1132.

Biosafety measures

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 confined field trial 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 phytosanitary requirements of the Philippines.

4. Engineered rice lines will be kept in isolation in the designated confined site (CT2, F8-9), 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. Plants and plant materials for disposal will be placed in sealed plastic bags and will be ploughed under and buried within the test site following standard procedures prescribed by the biosafety committee.                 

10. Precautionary measures will always be observed and good practices will be stringently imposed to prevent cross-over and dissemination of transgenic plant materials.

Conditions for Approval

a) The activities shall be conducted at Block F8 and F9 (CT-2) confined test site inside the Transgenic Research Facility (TRF) of Zeigler Experimental Station (ZES) at the International Rice Research Institute (IRRI), Los Baños, Laguna under the supervision of the IRRI-IBC;

b) The DOST-BC approved Project Information Sheet (PIS) shall be posted for three (3) consecutive weeks in the public announcement board of the barangay(s) and city municipality where the proposed activities will be carried out;

c) Proof of posting of the PIS, which may either be a certification from the duly authorized Barangay leader(s) or any affidavit executed by the Proponent, should be submitted to the DOST-BC within ten (10) days from the last day of posting;

d) Only upon completion of the public comment period and determination that no comments were received or comments were satisfactorily addressed by the proponent shall the DOST-BC issue a letter to the IRRI-IBC informing that planting/transplanting in the confined test site can commence;

e) Prior to the initiation of the activities, the proponents shall discuss the monitoring schedule with the Department of Agriculture-Bureau of Plant Industry-National Plant Quarantine Service Division (DA-BPI-NPQSD) to identify the activties that will require their presence. The proponents shall furnish the DOST-BC with a copy of the agreed monitoring schedule;

f) All confinement measures should be in place prior to the actual transplanting of rice seedlings;

g) At least on hundred fifty (150) meters physical or temporal reproductive isolation of the GMO shall be observed at all times;

h) The proponent shall ensure that rodent control measures are in place and that stray animals are excluded from the confined test site while trials are being conducted;

i) The proponent shall adhere as closely as possible to the schedule of implementation and monitoring activites for this project;

j) Modifications in the planting schedule and other activities must be officially communicated by the IRRI-IBC to the DOST-BC for information and/or approval;

k) Monitoring for biosafety compliance during stages of the project implementation, such as, but not limited to, the movement of transgenic materials, transplanting, reproductive stage, harvest, post-harvest monitoring and termination, shall be conducted by the IRRI-IBC. Report of which shall be submitted to the DOST-BC within five (5) days after the conduct of the monitoring activity;

l) Movement of all transgenic materials should be done in compliance with the DOST-BC and the DA-BPI-NPQSD guidelines;

m) The proponent of all transgenic materials should be done in compliance with the DOST-BC and the DA-BPI-NPQSD guidelines;

n) The proponent shall inform the DOST-BC immediately of any intrusion by unauthorized persons;

o) The proponent shall ensure the security of the experiment, the field workers, and the monitors;

p) In case of undue destruction of the experimental materials resulting from unauthorized entry of personnel or breach of containment of the confined test site, the proponent should implement measures to prevent the inadvertent escape of any viable material to the outside environment;

q) The proponent and the supervising IBC shall be held accountable for the undue destruction of the experimental materials and the consequences that its inadvertent escape may cause to the surrounding environment;

r) The rice grains or any plant part shall not be eaten or fed to humans, wildlife and livestock;

s) The proponent shall provide a space within the test site for disposal of the used plant materials;

t) The harvested grains shall be properly labeled and stored at the designated seed storage facility where transgenic seeds are kept, while plant materials and other plant debris used within the test site should be buried in a pit within the test site or plowed under;

u) The proponent shall leave the test site fallow for at least thirty (30) days. During the fallow period, the proponent shall strictly monitor emergence of volunteer plants and plant pest/diseases within the site for two (2) consecutive weeks after harvest. If necessary, the proponent may irrigate the test site to induce germination of seeds. Any volunteer plants that will emerge, or any ratoon residues shall be uprooted and disposed either by plowing under or by burying in the pit intended for transgenic plant biomass from this confined field test;

v) The IBC shall determine whether the confined test site shall be released from post-harvest restrictions. This shall form part of the collated post-harvest monitoring report of the project to be submitted by the IBC;

w) The proponent shall comply with additional requirements that the DOST-BC may impose, as necessary, during the conduct of the experiment;

x) The proponent shall submit, through the IBC, a completion report within ninety (90) days after the official completion of the confined test and after all post harvest activities have been completed; and

y) The IRRI-IBC shall submit a report on all its monitoring activites upon completion of this project by submitting an IBc completion report in the prescribed format; and

z) Should there be a change in project leadership, the proponent, thru the IRRI-IBC should officially notify the DOST-BC and submit the undertaking signed by both parties regarding the change in leadership and acceptance of the latter of the responsibilities to take over the project.

Date of Approval (DD-MM-YYYY) 19-01-2019
Date of Completion (DD-MM-YYYY) N/A
Executive Summary

The project aims to develop elite rice lines carrying robust and broad-spectrum resistance to different strains of bacterial leaf blight pathogen. We have used CRISPR-Cas9 technology to introduce site-directed mutagenesis in the target promoter regions of three sugar transporters, called Sweet genes. In particular, we edited the Sweet 11 (Os8N3), Sweet 13 (Os12N3) and SWEET 14 (Os11N3) promoter regions in IR64 and Ciherang-Sub1 backgrounds (DOST-BC ref. no. 2015-0287) which are common targets of many strains of Xanthomonas oryzae pv. oryzae (Xoo) collected worldwide. We have generated 39 advanced lines without Cas9 transgene but with mutations that confers broad-spectrum resistance to different Xoo strains (DOST-BC ref. no. 2016-0305). The transformation work was approved by the Philippines government under DOST-BC ref. no.: 2015-0287 and continued our work under DOST-BC ref no.: 2016-0305 for further advancement and selection of the SWEET mutant lines and resistance screening against the bacterial blight disease. From our screen house experiments, we have identified promising lines: eight T4 lines of IR64-IRS1132 and five T3 lines of Ciherang-Sub1-IRS1132. We will subject these selected lines for field evaluation or confined test to determine if there are any yield penalty brought about by mutagenesis. We will assess the growth performance and quantify yield of the 13 SWEET mutant lines in comparison with the non-transformed counterparts. We expect to produce a number of lines with optimized mutations that retain the resistance trait but have no trade-off in paddy field conditions. The CT will be conducted over two wet seasons of 2019 and 2020 in transgenic confined test site, CT 2 (F8-9), at the Transgenic Research Facility of the International Rice Research Institute.