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Project Information

Reference Number :
Project Title :
Confined test of transgenic biofortified rice with increased concentration of Fe and Zn in the grain
Project Type :
Confined Test
Status :
Cooperating Institution:
Supervising IBC:
International Rice Research Institute (IRRI)
Project Leader(s) :
Dr. Inez H. Slamet-Loedin
Experimental Facility/Site:
Blocks F1-F2 of the Experimental Station at the International Rice

Research Institute (IRRI), Los BaƱos, Laguna.
Purpose / Objectives :

1. To evaluate the agronomic performance and grain Fe and Zn concentrations of promising transgenic events containing genes encoding for iron storage-protein from soybean and other dicot crops [Phaseolus vulgaris, Pisum sativum and apple] (DOST-BC 2014-0281), and for iron transporter/chelator from rice (DOST-BC- 2007-0221) in BR28, IRRI 156 and IR64 genetic background in a replicated yield trial.

2. To evaluate the progenies of crosses between transgenic lines having soybean ferritin with iron transporter/chelator gene from rice in BR29, Ciherang, Ciherang-sub1 and IR69428 backgrounds.

3. To evaluate the agronomic performance in the ‘observation nurseries’ of backup events containing genes encoding for iron storage protein and chelator from soybean and intragenic/cisgenic events expressing iron chelator and/or storage genes solely from rice.

Biosafety measures:

1. The site is inside the identified Confined Test Area of the IRRI experiment station. This test area has been used previously for confined test of iron-biofortified materials (DOST-BC- 2014-0282) for two seasons.

2. The confined test area will be accessed only by the authorized staff who are directly involved in the experiment (Annex 7). For additional labor and activities done by IRRI Experimental Farm or other contract workers, a regular, authorized personnel will be present to ensure adherence to biosafety protocols. Only DOST-BC- approved visitors could access the site.

3. Temporal and physical isolation of the materials from the cultivated rice planted in nearby fields will be observed.

4. The harvested transgenic seeds will be packed in hermetically-sealed aluminum foil packages and stored in a locked seed room dedicated to the storage of transgenic seeds only. This transgenic seed storage room is located in K.J. Lampe Bldg., IRRI. Transfer will be in accordance with IRRI-IBC protocols.

5. Transgenic seeds for discarding will be autoclaved prior to burying in the transgenic pit.

6. All transgenic rice straws/stubbles and non-transgenic border plants will be buried in the transgenic pit located inside the confined site or plowed under the trial site.

7. The trial site will be fallowed for 30-45 days after harvest and volunteer plants will be buried in the transgenic pit inside the confined site or plowed under the trial site. Postharvest monitoring will be conducted by the monitoring team (DOST-BC, IRRI IBC, BPI and SHU personnel).

8. Plant material movements will strictly follow the relevant biosafety and phytosanitary requirements of the Philippine Government, IRRI-IBC, and other monitoring bodies.

9. Accidental release will be taken cared of by cleaning the site affected by accidental release. This will be conducted under the supervision of the proper authorities.

Conditions for Approval:

a) Before commencing with the activities, the DOST-BC shall inspect and approve the proposed site for the confined test (CT) located at Block F1-2 of the IRRI Experimental Station, Los Baños, Laguna.

b) Prior to the initiation of the activities, the proponent/s 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 activities that will require their presence. The proponent/s shall furnish the DOST-BC with a copy of the agreed monitoring schedule.

c) 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 IBC. Report of which shall be submitted to the DOST-BC not more than five (5) days after the conduct of the monitoring activity.

d) 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 or municipality where the proposed activities will be conducted.

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

f) A letter will be issued to the IBC informing that planting/transplanting on the confined test site can commence

g) The proponent shall adhere as closely as possible to the schedule of implementation and monitoring activities for this activity.

h) Modifications in the planting schedule and other activities must be communicated to the DOST-BC for information and/or approval.

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

j) At least 150 meters physical or temporal reproductive isolation of the GMO shall be observed at all times.

k) All confinement measures should be in place before the actual transplanting of rice seedlings.

l) The proponent shall ensure that only authorized personnel are allowed inside the test site.

m) The DOST-BC shall be informed in advance of any visitations by unauthorized personnel for approval.

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

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

p) The proponent shall ensure the security of the trial, the field workers, and the monitors.

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

r) 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.

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

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

u) 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.

v) The proponent shall leave the test site fallow for at least 30 days. During the fallow period, the proponent shall strictly monitor emergence of volunteer plants and plant pests/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.

w) 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 IBC.

x) The proponent shall comply with additional requirements that the DOST-BC may impose, as necessary, during the experiment period.

y) The proponent shall submit through the IBC a completion report ninety (90) days after the official completion of the confined test after completion of all post-harvest monitoring of volunteers.


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

Iron (Fe) and zinc (Zn) deficiencies are the most common micronutrient deficiencies globally

affecting more than 2 billion people. Biofortification of rice to increase micronutrient content,

particularly iron, is considered one of the options to complement existing approaches to alleviate this

‘hidden hunger’. Rice grains of commonly cultivated varieties contain low levels of Fe when milled,

while in unpolished state (brown rice), Fe and Zn in the bran are mostly unavailable for absorption

by the human body. Polished (milled) rice grains of commonly cultivated and consumed rice

varieties only contain approximately 2 ppm Fe and 16 ppm Zn. The HarvestPlus rice breeding

programs for biofortified rice target 13 ppm Fe and 28 ppm Zn in milled rice grains to reach

approximately 30% of the estimated average requirement (EAR) in human (Bouis et al., 2011).

We developed transgenic indica rice lines (using cultivars IR64 and IR69428 as backgrounds)

harboring iron storage protein ferritin gene from soybean (NCBP ref. no. 1999-0119) and/or iron

transporter/chelator genes from rice (NCBP ref no 2007-0221). Two events reached the Fe target

level and also exceeded the Zn target level in polished grains when evaluated under confined tests

(DOST-BC- Ref. 2011-0264, DOST-BC- Ref. 2013-0276, DOST-BC- Ref. No. 2014-0282) (Trijatmiko

et al., 2016). We recently developed back-up events using dicot ferritins genes in combination with

rice iron homeostasis genes or solely rice genes in the background varieties of IRRI 156 (NSIC Rc

238) and BRRI Dhan 28 (DOST-BC- Ref. No. 2014-0281). The back- up events have been evaluated

in the transgenic screen house. Selected promising events with Fe concentration in the polished

grains >10 ppm and Zn concentration > 35 ppm are proposed to be evaluated in this confined test

in comparison to the previous best event. The increase of grain Zn was made possible because the

inserted rice gene proved to chelate and translocate to the grains both Fe and Zn, but not the

unwanted heavy metals (i.e. Cadmium). In addition, the current conventionally-bred materials have a

maximum amount of Zn in polished grains of 24 ppm, which is still is below the target proposed by

nutritionists. Hence, transgenic breeding lines reaching both Fe and Zn nutrition targets are


This proposed confined test consists of Lead Event Nursery (LEN) and Observation Nursery (ON).

The LEN includes 17 independent events and 3 stacking crosses, 28 non-transgenic controls (17

null azygous, 3 crosses null, 4 wild types and 4 tissue culture controls). The (ON) consists of 8

independent transgenic events and 13 controls (8 azygous nulls, 3 wild types and 2 tissue culture

controls. The backup events will be compared with the best identified event (IR64-IRS495- 274) in

the previous three CTs. The ON is conducted for events with limited seeds. The LEN will have

four replications while the ON will have two replications.

Agronomic characteristics, yield and Fe and Zn concentrations will be analyzed for all the entries.


Public Information Sheet

Title Date of Approval Date of Posting File Attachment