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        In vitro Propagation and Genetic Transformation System Using Immature Embryo in Elite Rice (Oryza sativa L.) Cultivars

        ( Md Mahmudul Islam ),( Zahida Yesmin Roly ),( Young Sook Lee ),( Md Khalekuzzaman ) 한국육종학회 2014 Plant Breeding and Biotechnology Vol.2 No.1

        Rice (Oryza sativa L.), as a cereal grain, is the most widely consumed staple food for a large part of the world`s human population especially in Asia. Three indica rice (Oryza sativa L.) cultivars namely BRRI dhan29, BRRI dhan56 and BRRI dhan57 were used in this study to investigate the regeneration ability in rice. Regeneration potential was found to be highest (80.00%) for BRRI dhan29 and lowest (70.00%) for BRRI dhan57. Consequently, the highly regenerating indica rice cultivar BRRI dhan29 was used for genetic transformation. Embryogenic calli induced after 20 days were used for genetic transformation in the experiment. Agrobacterium strain LBA4404 was transformed with pBI121 binary vector which contains kanamycin resistance gene as a selectable marker gene and GUS as a reporter gene. Calli infected with this strain were analyzed by using 5-bromo 4-chloro 3-indolyl- D glucuronide (X-gluc) as a substrate. Expression of gene was determined by using the transformed shoots and roots. Stable integration and expression of GUS gene were also confirmed by using PCR analysis. The frequency of transformation in terms of transient GUS assay was found to be 35.0 ± 2 S.E. This study will provide valuable information for genetic transformation in cereal crops.

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