Abstract
Efficient transformation system for genetic improvement is essential in Crocus sativus, as it lacks sexual reproduction. This is the first report wherein an efficient protocol is developed for the transformation of Crocus sativus L. by Agrobacterium rhizogenes strain ARqua1 with a transformation efficiency of 78.51%. The ARqua1 strain harboring both Ri plasmid and binary vector plasmid pSITE-4NB, and marker genes for red fluorescent protein (RFP) and a β-glucuronidase (GUS) reporter gene were used for selection. Transformation was confirmed by RFP signal, GUS reporter assay and polymerase chain reaction (PCR) analysis of the test samples after 21 days post inoculation. These results confirm the establishment of protocol for hairy root transformation in C. sativus that can be further used for gene transfer or gene editing in Crocus for its genetic improvement.
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Abbreviations
- RFP:
-
Red fluorescent protein
- CaMV:
-
Cauliflower mosaic virus
- AS:
-
Acetosyringone
- GUS:
-
β-Glucuronidase
- MS media:
-
Murashige and Skoog media
- PCR:
-
Polymerase chain reaction
- T-DNA:
-
Transfer DNA
- DNA:
-
Deoxyribonucleic acid
- Vir genes:
-
Virulence genes
- LB broth:
-
Luria–Bertani broth
- IBA:
-
Indol-3-Butyric acid
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Acknowledgments
We kindly thank Dr. Praveen Verma, senior scientist, NIPGR, India for providing the guidance and resources in the development of the protocol.
Funding
This work was supported by UGC-DAE (University Grants Commision and Consortium of Scientific Research, University of Jammu, J&K and NIPGR, Delhi.
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SS designed and performed the experiments and wrote the manuscript. YS revised and updated the manuscript. PKV supervised the experiments. JV supervised the experiments and edited the manuscript.
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Sharma, S., Singh, Y., Verma, P.K. et al. Establishment of Agrobacterium rhizogenes-mediated hairy root transformation of Crocus sativus L.. 3 Biotech 11, 82 (2021). https://doi.org/10.1007/s13205-020-02626-2
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DOI: https://doi.org/10.1007/s13205-020-02626-2