Chloroplast transformation in new cultivars of tomato through particle bombardment

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 c...

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Bibliographic Details
Published in:3 Biotech 2024-04, Vol.14 (4), p.120-120, Article 120
Main Authors: Tanwar, Neha, Mahto, Binod K., Rookes, James E., Cahill, David M., Bansal, Kailash C., Lenka, Sangram K.
Format: Article
Language:English
Subjects:
Agriculture
Australia
Bioinformatics
Biomaterials
Biotechnology
Bombardment
Cancer Research
Chemistry
Chemistry and Materials Science
Chloroplasts
Coated particles
Cultivars
currants
Explants
Expression vectors
genetic markers
Genetic transformation
India
leaves
operon
Particle bombardment
Pineapples
Protocols and Methods
rRNA
Solanum lycopersicum
Spectinomycin
Stem Cells
Streptomycin
synthetic genes
Tomatoes
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Summary: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.
ISSN:2190-572X
2190-5738
2190-5738
DOI:10.1007/s13205-024-03954-3