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Genetic transformation of cultivated jute (Corchorus capsularis L.) by particle bombardment using apical meristem tissue and development of stable transgenic plant

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Abstract

An in vitro plant regeneration and genetic transformation protocol was established in jute (Corchorus capsularis L. var JRC321). One-day-old apical, meristematic tissues of germinating seedlings were used as explants. Multiple shoots were regenerated from each explant using Murashige and Skoog basal medium containing 1.78 µM benzylamino purine and 4.92 µM indole-3-butyric acid. Transformation was carried out in three independent sets (each set comprising of three independent experiments each comprising three replications with 35 explants per replication) using the bialaphos resistance gene (bar), synthetically designed for high level plant expression. The positive transformants containing the bar gene were selected in growth medium containing 2.5 mg/l bialaphos. Polymerase chain reaction (PCR), Southern and northern blots, real-time quantitative PCR, western blot and enzymatic assay of five putative transformants from three independent sets provided evidence for full-length gene integration into the genomic DNA of transformed jute, as well as high level expression of the transgene. Analysis of the T1 plants revealed a stable inheritance of the transgene through the progenies. The data presented in this report showed considerable advancement in jute transformation and should improve future genetic engineering strategies to be employed for improvement of this very important fibre crop.

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Acknowledgments

The authors extend their sincere appreciation to Shila Bhattacharjee for her extensive assistance with the tissue culture, Sudarshan Maity and Uttam Dogra for maintenance and rearing of transgenic jute lines, and Meghnath Prasad for assistance in preparing this manuscript. Finally, the authors would like to extend their sincere thanks and gratitude to the distinguished associate editor for her extensive and incisive comments to improve the clarity of the manuscript. The authors are also thankful to the distinguished reviewers for their critical comments which helped to improve the quality of the manuscript.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Advanced Laboratory for Plant Genetic Engineering, Indian Institute of Technology, Kharagpur, 721302, India

    Jagannath Bhattacharyya, Anirban Chakraborty, Souri Roy, Subrata Pradhan, Joy Mitra, Monami Chakraborty, Anulina Manna, Narattam Sikdar, Saikat Chakraborty & Soumitra Kumar Sen

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  1. Jagannath Bhattacharyya
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  2. Anirban Chakraborty
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  3. Souri Roy
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  4. Subrata Pradhan
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  5. Joy Mitra
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  6. Monami Chakraborty
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  7. Anulina Manna
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  8. Narattam Sikdar
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  9. Saikat Chakraborty
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  10. Soumitra Kumar Sen
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Correspondence to Soumitra Kumar Sen.

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Souri Roy, Subrata Pradhan and Joy Mitra have contributed equally to this work.

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Fig. 1

The standard curve generated by serial dilution of synthetic bar gene expression cassette and quantification of mRNA copy numbers of the synthetic bar gene in jute transformants based on the extrapolated standard curve (TIFF 20 kb)

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Bhattacharyya, J., Chakraborty, A., Roy, S. et al. Genetic transformation of cultivated jute (Corchorus capsularis L.) by particle bombardment using apical meristem tissue and development of stable transgenic plant. Plant Cell Tiss Organ Cult 121, 311–324 (2015). https://doi.org/10.1007/s11240-014-0702-2

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