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Project Information

Reference Number :
20160305
Project Title :
Targeted mutagenesis in the promoter region of the sugar transporter genes in rice for bacterial disease resistance
Project Type :
Contained Use (Greenhouse)
Status :
Ongoing
Cooperating Institution:
Carnegie Institute for Science and Iowa State University, USA
Supervising IBC:
International Rice Research Institute (IRRI)
Project Leader(s) :
Dr. Ricardo Oliva
Dr. Inez Hortense Slamet-Loedin
Experimental Facility/Site:
IRRI
Purpose / Objectives :

To produce a broadly effective and durable, long lasting resistance to bacterial blight of rice. Specifically, it aims to:

1.     Generate Samba Mahsuri, Ciherang-Sub1  and MTU1010 mutant lines on the sugar transporter promoters (SWEET11?, SWEET13?, and SWEET14?) using genome editing technology

2.     Produce individual quintuple mutant lines

3.     Test the resistance of the mutant lines that will be generated in this project including the previously generated IR64 mutants against Xoo pathogens and

4.     Evaluate the agronomic traits and yield penalty of the mutant rice lines produced by genome editing technology under screenhouse condition. 

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 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 phytosanitary requirements of the Philippines.

4. Engineered rice lines will be kept in isolation in Bay7, CL4 and transgenic screen houses (CS-01), respectively, 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 by 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 temperature and high pressure to kill the Xoo inoculumprior to disposal according to standard biosafety protocol.

                  - pots and soil used to grow the uninoculated plants

                  - culture media, diluents and buffers

                  - syringe, gloves  and non-disposable laboratory materials

9. Plants and plant materials for disposal will be placed in sealed plastic bags and will be decontaminated by autoclaving and then disposed of following 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.

Conditions for Approval:

a) Project activities shall be conducted at the BL2 lsolatron Room at Molecular Genetics Laboratory of the University of the Philippines Los Bafros - National lnstitute of Molecular Biology and Biotechnology (UPLB-BTOTECH).

b) The proponent shall adhere as closely as possible to the schedule of activities reflected in the submitted Gantt chart.

c) Any modifications in the schedule of activities shall be made with the concurrence of the UPLB-IBC and the DOST-BC.

d) Signage should be in place indicating that the facility is restricted/limited access.

e) The proponent must provide a logbook to record access of personnel to the experimental facility.

f) The proponent must take measures to ensure that only the DOST-BC authorized personnel are allowed inside the experimental facilities.

g) The experimental facilities must be kept under lock when not in use.

h) 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.

i) 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.

j) The DOST-BC should be informed in advance of any visitations by unauthorized persons.

k) The DOST-BC shall be informed immediately of any intrusion by unauthorized persons.

l) The proponent shall strictly observe proper disposal procedures for all materials.

m) Movement of all materials will be done in compliance with all relevant biosafety and phytosanitary requirements of the Philippines.

n) Any additional requirement that the DOST-BC may impose, as necessary, for the duration of the experiment shall be complied with.

o) The proponent shall submit through the IBC a completion report within 90 days after completion of the experiment.

p) The IBC shall submit to the DOST-BC a report on the completion of this project, in the prescribed format.

Date of Approval (DD-MM-YYYY):
21-01-2017
Date of Completion (DD-MM-YYYY):
Executive Summary:

Bacterial blight disease in rice caused by Xanthomonas oryzae pv. oryzae (Xoo) is increasingly affecting yield of rice farmers in Asia and Africa. The pathogen uses a suite of genes known as transcription-activator like effectors (TALe) to specifically induce host genes essential for pathogen’s proliferation and disease progression (Bogdanove et al., 2010; Verdier et al., 2012; Schornack et al., 2013; Streubel et al., 2013). Recently, reports have identified the three susceptibility (S) genes, namely, SWEET11, SWEET13 and SWEET14, in rice which are target of a number ofXoo strains that are collected in different locations worldwide. With this knowledge and with the development of a novel tool, CRISPR/Cas9 system for mutagenesis, it is possible to alter the recognition specificity of the target S gene and therefore, it can render resistance to rice plants (Jiang et al., 2013; Cong et al., 2013; Jiang et al., 2014; Shan et al., 2014).In this study, we will engineer relevant rice breeding lines to produce a durable, long lasting resistance to bacterial blight by introducing mutations in specific TALe target sites of SWEET genes promoter. We will do crossing activities for the generated lines to achieve the desired quintuple mutant rice lines. Ultimately, we will allow the generated SWEET mutant lines for infections with Xoo pathogens in contained trials to test if the mutations can create resistance. Also, we will examine any trade-off associated with the mutagenesis by measuring the agronomic and yield characteristics of the mutants compared with the wild-type parentals. This study is the second phase of DOST-BC ref. no. 2015-0287 project wherein transgenic facilities (CL4 and CS-01) at IRRI will be used for the growth of the engineered plants. The expected duration of the project is four (4) years. 

Public Information Sheet

Title Date of Approval Date of Posting File Attachment