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Development of Transgenic Rice

Dr.Md Enamul Hoque
CSO and Head (In Charge)
Biotechnology Division,BRRI-Gazipur

Dr.Md.Shamsher Ali
CSO
Biotechnology Division, BRRI-Gazipur

Dr.Nilufar Yasmin Shaikh
SSO
Biotechnology Division,BRRI-Gazipur

S M Hisam Al Rabbi
SO
Biotechnology Division, BRRI-Gazipur

S A Rezvi
SO
Biotechnology Division, BRRI-Gazipur

Ripon Kumar Roy
SO
Biotechnology Division, BRRI-Gazipur

Abstract/ Executive Summary:

Agricultural productivity is severely affected by soil salinity. It is known that medium level of soil salinity can reduce crop yield by 20-40 %. But high soil salinity is not suitable at all for crop growing. It is estimated that about 1.0 million hectare of land in Bangladesh is affected by various degrees of soil salinity. Planting transgenic rice having salt tolerance have the potential of decreasing the annual rice yield loss. Such varieties that can tolerate high soil salinity level would be a practical solution. Therefore, attempts are undertaken to develop salt tolerant rice variety through transformation. The study is ongoing and still trying to optimize various conditions for transformation. GUS expression was found in putative transgenic calli. Unfortunately no regenerant was recovered.

Keywords: Calli,Transformation, Salt tolerant, Gene

Location: Biotechnology Division, BRRI-Gazipur

Start Date: 01-01-2008

End Date:

Program Area: Variety and Species

Commodity: Rice

Objectives:

To develop salt tolerant rice genotypes and to insert salt tolerance gene into widely adapted elite rice variety.

Methodology:

BRRI dhan28 and BRRI dhan29 were chosen for present transformation work. The GlyI and GlyII gene construct in Agrobacterium (LBA4404 strain) was used for transformation studies using hygromycin base selection. Both mature and immature embryos were used as explants. To get embryogenic calli, media compositions as well as environmental condition were optimized. After sterilization mature seed were plated on callus induction medium (CI)consisting of MS basal salt supplemented with 2.0 mg/L 2,4-D, 0.3 g/L casamino acid, 0.5 mg/L proline, 0.5 mg/L glutamine, 7.0 g/L agar maintaining pH 5.8. Incubated seeds were then kept for 3- 7 days at 28 ± 1^oC temperature under continuous light for callus induction. Then these calli were placed into the inoculation media (LB medium supplemented with 50 mg/L kanamycin and 100 μM acetosyringone) for 3 minutes. Then the calli were transferred to co- cultivation (cc) media consisting of MS basal salt supplemented with 10 mg/L glucose, 0.3 g/L casamino acid, 100 μM acetosyringone, 30.0 g/L sucrose and 7.0 g/L agar with pH 5.2 and kept for three days in dark at 25^oC. The calli were then transferred to the selection media containing MS basal salt supplemented with 0.5 g/L proline, 0.5 g/L glutamine, 0.3 g/L casamino acid, 7.0 g/L agar, 50 mg/L hygromycin B, 250 mg/L cefotaxime and pH 5.8. Cultures were then incubated in dark at 28 ± 1^oC for 4 weeks (two cycles, each cycle consist of 2 weeks). After antibiotic selection, putative transgenic calli weretransferred into the regeneration media containing MS basal salt supplemented with 2.0 mg /L BAP,1.0 mg/L NAA, 1.0 mg/L kinetine, 50 mg/L hygromycin B, 250 mg/L cefotaxime, 30.0 g/L sucrose,7.0 g/L agar, pH 5.8 and cultures were kept at 27 ± 1^oC under continuous light. In case of immature embryos, the explants were kept dark at 27±1°C temperature. CI media (MS salts, supplemented with 30.0 g/L sucrose, 2.0 mg/L 2,4-D, 0.3 g/L casamino acid, agar 7.0 g/L and pH 5.8). 5 days old calli were used for bacterial infection. The next steps were same as mature embryos.

Source of Information: Proceedings of the BRRI Annual Internal Review for 2008-09

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The study is ongoing and still trying to optimize various conditions for transformation. GUS expression was found in putative transgenic calli. Unfortunately no regenerant was recovered.

Knowledge Management: Report/Proceedings