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Development of Drought and Salt of Tolerant Transgenic Rice Lines

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

Shahanaz Sultana
SSO
Biotechnology Division, BRRI, Gazipur

Ripon Kumar Roy
SO
Biotechnology Division, BRRI, Gazipur

shampa Das Joya
SO
Biotechnology Division, BRRI, Gazipur

Abstract/ Executive Summary:

Drought and soil salinity are the major environmental stresses affecting the plant growth adversely and reduces the grain yield. For drought almost 2.0 million 13 hectare of land and for soil salinity about 1.0 million hectare of land in Bangladesh is affected by varying degrees of soil salinity. Therefore, attempts are undertaken to develop suitable drought and salt tolerant transgenic rice variety through transformation.Ten putative transgenic plantlets were regenerated from BRRI dhan29 and transferred to the earthen pot.Further confirmation will be done by PCR amplification

Keywords: Drought tolerant, Calli, Soil Salinity, Transformation

Location: Biotechnology Division, BRRI-Gazipur

Start Date: 01-01-2012

End Date:

Program Area: Variety and Species

Commodity: Rice

Objectives:

To make BRRI dhan29 drought and salt tolerant through genetic transformation.

Methodology:

BRRI dhan29 was used as plant material and gene construct having TPSP gene was used for genetic transformation. Dehausked sterilized seeds were placed on callus induction medium (MS + 30 g/l maltose+ 3 mg/l 2,4-D + 0.25 mg/l BAP + 0.6 g/l proline+ 0.3 g/l cassamino acid +0.1 g myoinositol) and kept 21 days for calli induction at 25⁰C temperature in dark. Then calli were subcultured on same callus induction media and kept 5 days at 25⁰C temperatur in dark for actively growing calli. After that calli were infected with agrobacterium containing TPSP gene and cocultivated on cocultivaton media (Callus induction media + 10g/l glucose + 100 μM acetosyringone) for 48 hrs at 25⁰C temperature in dark.Then washed with sterile distilled water containing cefotaxime (250 mg/lit). After that these calli placed on selection media containing hygromycine (50 mg/l) and cefotaxime (250 mg/l) for 15 days for first selection. Then the survived calli were transferred to newly prepared selection media (as first selection) for second selection for another 15 days. Again the survived calli from second selection were transferred to newly prepared 12 selection media (as first selection) for third selection for another 15 days. Then survived micro calli were transferred to regeneration media I (Callus induction media+ 4 mg/l BAP + 1.2 mg/l Kinetine + 0.5 mg/l NAA + 250 mg/l cefataxime + 50 mg/l Hygromycine+ 40 g/l, Sucrose + 4 g/l phytagel) and incubated 12 days at 25⁰C temperature under light (16 h light/ 8 h dark). After that regenerated plantlets were transferred to rooting medium (half strength MS + 40 g/l, Sucrose + 4 g/l phytagel) for rooting and incubated 21 days at 25⁰C temperature in light (16 h light/ 8 h dark). Then plantlets were transferred to the test tubes containing half strength Youshida medium (Hydrophonic culture medium) and kept it 15 days for hardening. During hardening half strength Youshida media changed after 2-3 days interval. Finally these plantlets were shifted to earthen pot and kept there until maturity .

Source of Information: Proceedings of the BRRI Annual Internal Review for 2012-13

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Ten putative transgenic plantlets were regenerated from BRRI dhan29 and transferred to the earthen pot. Further confirmation will be done by PCR amplification.Experiment is to be done for developing BRRI dhan29 drought and salt tolerant through genetic transformation.

Knowledge Management: Report/Proceedings