Project Information

Reference Number 20190320
Project Title Gene validation of candidate genes for high out-crossing trait using transgenic approaches (Phase II)
Project Type Contained Use (Laboratory)
Status Ongoing
Name of Institution International Rice Research Institute
Cooperating Institution N/A
Supervising IBC International Rice Research Institute (IRRI)
Project Leader(s)

Dr. Sung-Ryul Kim
Dr. Inez H. Slamet-Loedin

Experimental Facility/Site Genetic Transformation Laboratory (GTL), Genomics and Genetics Laboratory (GGL) and the CL4 glass bays at the International Rice Research Institute
Purpose / Objectives

To validate gene function of newly identified candidate genes and promoters involved in increasing cross pollination rate in rice which can be used in hybrid rice breeding.

Biosafety measures

Appropriate biosafety measures as implemented by IRRI, and the requirements of the DOST-BC will be strictly observed and followed.  Transgenic and untransformed control plants will only be grown in CL4 glass bays, a well-contained facility.

Biosafety measures that will be taken:

  • All vectors will be kept in the plasmid collection of Genetic Transformation Laboratory (GTL) after construction. 
  • The tested plants will be kept in isolation in the CL4 glass bays to prevent access of unauthorized personnel.
  • Plant materials for disposal will be placed in sealed plastic bags and disposed of in dedicated disposal pit for transgenic materials.
  • Plants grown in CL4 will be destroyed and the pots and soil will be autoclaved at high temperature and high pressure.
  • All materials movement and disposal will be done only under supervision of the IRRI-IBC and the Bureau of Plant Industry-Plant Quarantine Service (BPI-PQS)
  • Access to GTL, GGL and CL4 facilities are for authorized staff only.
  • A barcode entry system is applied in the facilities to record the entry of personnel; a log book is available for visitors not having IRRI access card system.

Transgenic plants from CL4 will be destroyed after the completion of the experiment. The seeds from transgenic plants during the conduct of the study will be kept at CL4 seed storage room. And after completion of the project, seeds will be kept for additional 3 years for publication purpose as an evidence of gene validation tests.

Conditions for Approval

a) All activities shall be conducted at the GTL, the Genetics and Genomics Laboratory and the CL4 Transgenic Glasshouse at IRRI, Los Baños, Laguna and measures to limit access to authorized personnel should be strictly observed;

b) Thorough cleaning of vents and drainage of the CL4 Transgenic Glasshouse shall be conducted prior to the start of the activity;

c) 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;

d) Any modifications in the schedule and addiional activities must be communicated to the DOST-BC for information and/or approval;

e) All confinement measures should be in place prior to the commencement of the activities;

f) Signages should be in place indicating that the facility is a restricted/limited access facility and only those personnel approved by the DOST-BC and the IRRI-IBC, in case of changes in personnel during the course of the project, maybe allowed access;

g) Any changes in the list of authorized personnel shall be approved by the IRRI-IBC and notification on the same shall be communicated to the DOST-BC for record purposes;

h) Monitoring for biosafety compliance shall be supervised by the IRRI-IBC;

i) The IRRI-IBC shall approve request for visitation of unauthorized personnel and notification on the same shall be sent to the DOST-BC;

j) The proponent and the IRRI-IBC shall ensure that there are no breach in the biosafety protocol imposed on the project;

k) 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; and

l) All viable materials within the experiment area shall be duly accounted for.

Date of Approval (DD-MM-YYYY) 25-05-2019
Date of Completion (DD-MM-YYYY) N/A
Executive Summary

Rice (Oryza sativa L.) is the most important food crop in the world, feeding over half of the global population. Hybrid rice varieties have some strength, especially in high grain yield. However, high F1 seed cost is a major limiting factor of the adoption of hybrid rice cultivars to rice farmers in South and Southeast Asian countries. A practical way to improve hybrid seed production is to increase the cross pollination rate. For this, near isogenic lines (NILs) having long-exserted stigma (increased stigma length) in the popular cultivar IR64 and the popular commercial hybrid parental  backgrounds (IR58025A/B and IR68897A/B) were developed using the wild rice relative, O. longistaminata (OL) which has naturally long-exserted stigma. To identify the gene controlling high out-crossing (HO) trait derived by the long exserted stigma phenotype, were QTL analysis and fine mapping conducted using BC6F2 and BC6F3 populations derived from IR64 x OL cross. Further gene validation experiments using transgenic approaches were conducted (DOST-BC Ref No. 2017-0309, Phase I). Further precise fine mapping using the populations derived from the crosses OL x IR58025B and OL x IR68897B was conducted and located the HO gene(s) on the ~151kb size of OL segment containing ~25 genes. This project aims to validate the potential candidate genes for the long-exserted stigma which influences high cross pollination in rice. The studies will be conducted in DOST-BC approved facilities at IRRI which are the Genetic Transformation Laboratory, The Genetics and Genomics Laboratory and the CL4 glasshouses. The project is expected to run for three years.