NDLI: Agrobacterium-mediated genetic transformation of safflower (Carthamus tinctorius L.)

Content Provider	Springer Nature Link
Author	Sri Shilpa, K. Dinesh Kumar, V. Sujatha, M.
Copyright Year	2010
Abstract	Reproducible and highly efficient protocols for shoot regeneration and genetic transformation mediated by Agrobacterium have been established for safflower (Carthamus tinctorius L.). Agrobacterium tumefaciens strain LBA 4404 with gus reporter gene and hygromycin (hpt gene) as plant selection marker was used as the plant transformation vector. Genetic transformation experiments were carried out to evaluate the efficacy of various parameters such as genotype, seedling age, co-cultivation period, bacterial titer, enzymatic pre-treatment of target tissues, use of compounds that induce vir-gene enhancer, acetosyringone (AS), explant type and explant injury to enhance transformation efficiency. Transformation frequency was high when root and hypocotyl explants of 8-day-old seedlings of safflower cv. HUS-305 were co-cultivated with bacterial cell density of 0.5 OD$_{600}$ during a period of 2 days followed by selection regime of 10–15–15 mg/l hygromycin. The frequency of rooting of the primary transformants was low (18.0%) when compared with the regenerated shoots (70.0%), and seven shoots survived on transfer to soil. The putative transformants were confirmed by β-glucuronidase (GUS) histochemical assay, polymerase chain reaction (PCR), reverse-transcriptase PCR (RT-PCR) and Southern blot analysis. With the optimized transformation protocol, putative transformed shoots were obtained with frequency of 51.0% within 8–10 weeks of culture initiation.
Starting Page	387
Ending Page	401
Page Count	15
File Format	PDF
ISSN	01676857
Journal	Plant Cell, Tissue and Organ Culture (PCTOC)
Volume Number	103
Issue Number	3
e-ISSN	15735044
Language	English
Publisher	Springer Netherlands
Publisher Date	2010-07-11
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Agrobacterium-mediated transformation Carthamus tinctorius L. Co-cultivation Tissue culture Transgene expression Plant Pathology Plant Genetics & Genomics Plant Physiology Plant Sciences
Content Type	Text
Resource Type	Article
Subject	Horticulture

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