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Chloroplast transformation in new cultivars of tomato through particle bombardment

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Abstract

A protocol has been established for genetic transformation of the chloroplasts in two new cultivars of tomato (Solanum lycopersicum L.) grown in India and Australia: Pusa Ruby and Yellow Currant. Tomato cv. Green Pineapple was also used as a control that has previously been used for establishing chloroplast transformation by other researchers. Selected tomato cultivars were finalized from ten other tested cultivars (Green Pineapple excluded) due to their high regeneration potential and better response to chloroplast transformation. This protocol was set up using a chloroplast transformation vector (pRB94) for tomatoes that is made up of a synthetic gene operon. The vector has a chimeric aadA selectable marker gene that is controlled by the rRNA operon promoter (Prrn). This makes the plant or chloroplasts resistant to spectinomycin and streptomycin. After plasmid-coated particle bombardment, leaf explants were cultured in 50 mg/L selection media. Positive explant selection from among all the dead-appearing (yellow to brown) explants was found to be the major hurdle in the study. Even though this study was able to find plastid transformants in heteroplasmic conditions, it also found important parameters and changes that could speed up the process of chloroplast transformation in tomatoes, resulting in homoplasmic plastid-transformed plants.

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Data availability

The entire dataset is available as supplemental material to this publication.

Abbreviations

Prrn:

rRNA Pomoter

IAA:

Indole 3-acetic Acid

BAP:

N6-benzylamino Purine

LCY:

Lycopene Cyclase

BHY:

β-Carotene Hydroxylase

BKT:

β-Carotene Ketolase

PRM:

Plant Regeneration Media

CTAB:

Cetyltrimethylammonium Bromide

GP:

Green Pineapple

YC:

Yellow Currant

PR:

Pusa Ruby

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Acknowledgements

We thank Prof. Dr. Ralph Bock (Max Planck Institute for molecular plant physiology) for providing pRB94 chloroplast specific vector and Deakin University Australia for partially supporting this research and providing postgraduate research scholarship to NT.

Funding

Deakin University Australia is acknowledged for partially supporting this research and providing postgraduate research scholarship to the first author.

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Author notes

  1. Sangram K. Lenka

    Present address: Department of Plant Biotechnology, Gujarat Biotechnology University, Gujarat International Finance Tec (GIFT)-City, North Gate, Gandhinagar, 382355, Gujarat, India

Authors and Affiliations

  1. TERI-Deakin Nano-Biotechnology Centre, The Energy Resources Institute (TERI), New Delhi, 110003, India

    Neha Tanwar, Binod K. Mahto, Kailash C. Bansal & Sangram K. Lenka

  2. University Department of Botany, Ranchi University, Ranchi, Jharkhand, 834008, India

    Binod K. Mahto

  3. School of Life and Environmental Sciences, Deakin University, Waurn Ponds Campus, Geelong, VIC, 3216, Australia

    Neha Tanwar, James E. Rookes & David M. Cahill

  4. National Academy of Agricultural Sciences, New Delhi, 110012, India

    Kailash C. Bansal

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  1. Neha Tanwar
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Correspondence to Kailash C. Bansal or Sangram K. Lenka.

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Tanwar, N., Mahto, B.K., Rookes, J.E. et al. Chloroplast transformation in new cultivars of tomato through particle bombardment. 3 Biotech 14, 120 (2024). https://doi.org/10.1007/s13205-024-03954-3

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