NDLI: A rapid and efficient method for in vitro shoot organogenesis and production of transgenic Bacopa monnieri L. mediated by Agrobacterium tumefaciens

Content Provider	Springer Nature Link
Author	Mahender, Aileni Mallesham, Bulle Srinivas, Kota Kranthi Kumar, Gadidasu Venugopal Rao, Kokkirala Rajesh, Yarra Zhang, Peng Sadanandam, Abbagani
Copyright Year	2011
Abstract	Efficient shoot regeneration and Agrobacterium-mediated genetic transformation systems were developed for Bacopa monnieri L. (Scrophulariaceae), a plant well known for its medicinal properties. Leaf explants were cultured on Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP), and in combination with either indole-3-acetic acid (IAA) or napthalene-3-acetic acid. A combination of BAP (17.80 μM) and IAA (2.28 μM) maximized shoot initiation (85.2 ± 2.43) with greatest shoot length (2.8 ± 0.22), and was obtained directly from leaf explants without an intervening callus phase. Leaf segments from in vitro grown plants were co-cultivated with Agrobacterium tumefaciens LBA4404 harboring pCAMBIA1301 with ß-glucuronidase (uidA) and hygromycin phosphotransferase (hpt) genes. The co-cultivated explants were transferred to selective shoot induction and elongation medium. The elongated hygromycin-resistant shoots were subsequently rooted on MS medium supplemented with 4.9 μM indole-3-butyric acid and 25 mg/l hygromycin (SSRM). Successful transformation was confirmed by monitoring histochemical GUS activity during shoot elongation and PCR analyses using uidA- and hpt-specific primers. Integration of hpt into the genome of transgenic plants was also verified by Southern blot analysis. The highest transformation efficiency achieved was 70.6%, with an average of 10.4 ± 0.15 transgenic plantlets per explant using the present transformation system. Therefore, these highly efficient and rapid regeneration and transformation systems create significant potential for engineering of B. monnieri with a view to detailed biomolecular analyses or for further enhancement of its medicinal properties.
Starting Page	153
Ending Page	159
Page Count	7
File Format	PDF
ISSN	10545476
Journal	In Vitro Cellular & Developmental Biology - Plant
Volume Number	48
Issue Number	2
e-ISSN	14752689
Language	English
Publisher	Springer-Verlag
Publisher Date	2012-01-25
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Bacopa monnieri Genetic transformation Organogenesis Cell Biology Plant Sciences Plant Genetics & Genomics Plant Breeding/Biotechnology Developmental Biology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Developmental Biology Plant Science Biotechnology

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