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Microprojectile bombardment mediated genetic transformation of embryo axes and plant regeneration in cumin (Cuminum cyminum L.)

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Abstract

Pre-cultured cumin embryos were bombarded under 27 inches Hg vacuum, 25 mm distance from rupture disc to macrocarrier, 10 mm macrocarrier flight distance using 1100 psi rupture disc and 9 cm microprojectile travel distance. An average of 110 embryos was used per shot and 91% embryos showed transient GUS expression after 24 h. Shoot tips and roots of T0 plantlets exhibited GUS expression done after 3 months of bombardment. Transformation was confirmed with PCR amplification of 0.96 and 1.3 kb band of hptII and gus genes respectively from T0 transgenics and southern blot analysis using PCR amplified DIG labeled hptII gene as probe. It is the first successful attempt of transformation of cumin plant through direct gene transfer using particle gene gun and adequately exhibiting the possibility of stable transformation in cumin.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid.

B5:

Gamborg’s medium.

BA:

6-Benzyladenine.

GUS:

β-Glucuronidase.

HPT:

Hygromycin-phosphotransferase.

IAA:

Indole-3-acetic acid.

MS:

Murashige and Skoog basal salt media.

NAA:

α-Naphthaleneacetic acid.

PDS:

Particle delivery system.

PGR:

Plant growth regulator.

TDZ:

1-Phenyl-3-(1,2,3-thiadiazol-5-yl) urea (thidiazuron).

X-Gluc:

5-Bromo, 4-chloro, 3-indolyl, β-d-glucuronide.

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Acknowledgment

The financial support received from CSIR (Network Project NWP-20) is thankfully acknowledged. Authors are thankful to Mr. Adarsh Kishore Sharma for providing technical assistance.

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Authors and Affiliations

  1. Discipline of Marine Biotechnology and Ecology, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR), G. B. Marg, Bhavnagar, Gujarat, 364002, India

    Natwar Singh, Avinash Mishra, Mukul Joshi & Bhavanath Jha

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  1. Natwar Singh
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  2. Avinash Mishra
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  3. Mukul Joshi
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  4. Bhavanath Jha
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Correspondence to Bhavanath Jha.

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Singh, N., Mishra, A., Joshi, M. et al. Microprojectile bombardment mediated genetic transformation of embryo axes and plant regeneration in cumin (Cuminum cyminum L.). Plant Cell Tiss Organ Cult 103, 1–6 (2010). https://doi.org/10.1007/s11240-010-9746-0

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