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Regeneration of transgenic plants from two indica rice (Oryza sativa L.) cultivars using shoot apex explants

  • Genetic Transformation and Hybridization
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Abstract

We have established a reproducible procedure for transformation of shoot apices and regeneration of transgenic plants for two indica rice cultivars, white ponni (WP) and Pusa Basmathi 1 (PB 1). Four-day-old shoot apex explants were transformed by cocultivation with Agrobacterium tumefaciens strain EHA 101 harbouring a binary plasmid pRIT1. The vector contained an improved hygromycin phosphotransferase (hpt) gene for hygromycin resistance driven by actin 1 promoter and the reporter gene β-glucuronidase intron (INT-GUS) controlled by CaMV 35S promoter. Rice shoots were induced on media containing 0.1 mg/l napthalene acetic acid (NAA), 1.0 mg/l kinetin (kn), 1.0 mg/l N 6-benzyleaminopurin (BAP), 300 mg/l casaminoacid, 500 mg/l proline, 50 mg/l hygromycin and 500 mg/l cefotaxime. Transgenic plants were raised in pots and seeds were collected. Histochemical and polymerase chain reaction (PCR) analyses of field established transgenic rice plants and their offsprings confirmed the presence of GUS gene. Integration of T-DNA into the genome of putative transgenics was further confirmed by southern analysis. The transformation efficiency of WP was found to be ranging from 5.6 to 6.2% whereas in the case of PB1, it was from 7 to 8%. Progeny analysis of these plants showed a pattern of classical Mendelian inheritance for both hpt and GUS gene.

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Abbreviations

BAP:

N 6-Benzyleaminopurine

CaMV 35S:

Cauliflower mosaic virus promoter

CB-INT:

Catalase bean intron

GUS:

β-Glucuronidase

hpt:

Hygromycin phosphotransferase

kn:

Kinetin

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Acknowledgments

We thank Dr. R. Terada, National Institute of Basic Biology, Japan, for providing Agrobacterium strain and a binary vector and Professor K. Veluthambi, Madurai Kamaraj University, Madurai, India, for pIG221 vector and for helping us to carry out some of the experiments in his laboratory.

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  1. Entomology Research Institute, Loyola College, Chennai, 600034, India

    S. Arockiasamy & S. Ignacimuthu

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  1. S. Arockiasamy
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  2. S. Ignacimuthu
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Correspondence to S. Ignacimuthu.

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Communicated by P. Kumar.

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Arockiasamy, S., Ignacimuthu, S. Regeneration of transgenic plants from two indica rice (Oryza sativa L.) cultivars using shoot apex explants. Plant Cell Rep 26, 1745–1753 (2007). https://doi.org/10.1007/s00299-007-0377-9

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  • DOI: https://doi.org/10.1007/s00299-007-0377-9

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