Reynante L. Ordonio, PhD.

Generally, our objective is to produce improved lines of popular released varieties that carry a combination of useful traits through CRISPR-CAS technology. Specifically, we aim to:

1. Identify and assemble suitable recipient popular indica varieties that respond well to tissue culture;

2. To design appropriate constructs for gene editing by CRISPR-Cas9 technology involving the following traits: rice tungro spherical virus (RTSV) resistance, bacterial leaf blight (BLB) resistance, lower amylose content than the original (if needed) and aroma; and

3. To screen for successfully edited plants by DNA sequencing, expression analysis, phenotyping and PCR

The BL2 Screenhouse is within the premises and under total control of PhilRice-CES.

Living modified organisms (E coli and Agrobacterium) will be used in the BL2 laboratory under aseptic conditions by trained personnel. Any implements that come into contact with them will be autoclaved and disposed properly. The use of floral dip will also be done in the laboratory under aseptic conditions, after which, the plants will be transported back to the BL2 screenhouse under IBC monitoring.

Access to the BL2 screenhouse will be given to project staff and authorized personnel and to DOST-BC-approved visitors. Only one entry point will be provided, which will be secured at all times. Movement of people within the area will also be documented. A logbook will be maintained in the CL2 screenhouse to record the entry of personnel and their activities. Moreover, the CL2 screenhouse facility will be inspected regularly to ensure adequate protection against rats, birds, etc.

Gene-edited seeds harvested from the BL2 screenhouse will be kept in sealed/locked containers separate from other ordinary materials. Movement of all materials will be done in compliance with all the relevant biosafety and phytosanitary requirements of the Philippine Government.

Collection and transport of plant materials from the BL2 screenhouse to the BL2 Isolation Room will be supervised by an IBC representative. Access to the Isolation Room will be restricted to authorized personnel only. Living tissues will be autoclaved before they are discarded. Transport of seedling regenerants from the BL2 laboratory to the BL2 screen house will also be implemented under IBC supervision.

Plant materials for disposal will be placed in sealed plastic bags. CL2-grown plants for disposal will be autoclaved and then put in the concrete compost bin near the area.

Any unplanned activity will be monitored and reported. There will be a logbook and other monitoring systems. Accidental release will be taken care of by cleaning the contaminated area. A thorough investigation of accidental release will be conducted to prevent similar incident from happening in the future. In the extreme event of an accident arising at the field site, all plant materials in the experimental plots will be plowed back into the soil. Any case of sabotage will be immediately reported to the appropriate authorities.