Effect of over-expression of Linum usitatissimum PINORESINOL LARICIRESINOL REDUCTASE (LuPLR) gene in transgenic Phyllanthus amarus
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
View Archive Info| Field | Value | |
| Title |
Effect of over-expression of Linum usitatissimum PINORESINOL
LARICIRESINOL REDUCTASE (LuPLR) gene in transgenic
Phyllanthus amarus
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| Creator |
Banerjee, Anindita
Chattopadhyay, Sharmila |
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| Subject |
Drug Development/Diagnostics & Biotechnology
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| Description |
Shoot tip explants of Phyllanthus amarus were
cocultivated with Agrobacterium tumefaciens strain LBA
4404 carrying plasmid pCAMBIA 2301 harbouring genes
coding for betaglucuronidase (gus), kanamycin (kan), and
neomycin phosphotransferase II (nptII) along with a gene
coding for Linum usitatissimum PINORESINOL LARICIRESINOL
REDUCTASE (Lu-PLR). Transformed shoot
tip explants were maintained in a Murashige and Skoog
(MS) medium containing TDZ 1.54 mg l-1, kan 50 mg l-1
and cephotaxime 62.5 mg l-1. The optimum medium for
regeneration of multiple shoots was MS supplemented with
TDZ 1.54 mg l-1, kan 50 mg l-1. Efficient and effective
rooting of plantlets was achieved by culturing the in vitro
regenerated shoots on liquid � MS medium containing
0.7 mg l-1 indole 3-butyric acid (IBA) and 5 mg l-1 kan.
Rooted plants were acclimatized in the mixtures of vermiculite
and soil. The transformation of kan-resistant
plantlets regenerated from shoot-tip explants was confirmed
by GUS and polymerase chain reaction (PCR) analysis.
Southern blot and reverse transcribed PCR (RT-PCR)
analysis confirmed successful integration and expression of
Lu-PLR gene. Quantitative analysis of phyllanthin performed
on transgenic and wild plants using high-performance
liquid chromatography (HPLC) revealed that
transgenic lines contained higher phyllanthin content (0.3–
0.81% w/w) than wild plants (0.09% w/w). The highest
yield of phyllanthin was detected in transgenic lines was upto 1.16, 1.22 and 1.23 folds higher than that of wild plant.
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| Date |
2010
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Identifier |
http://www.eprints.iicb.res.in/101/1/PLANT_CELL_TISSUE_AND_ORGAN_CULTURE_103(3)315%2D323;2010[11].pdf
Banerjee, Anindita and Chattopadhyay, Sharmila (2010) Effect of over-expression of Linum usitatissimum PINORESINOL LARICIRESINOL REDUCTASE (LuPLR) gene in transgenic Phyllanthus amarus. Plant Cell Tiss Organ Cult, 103 (3). pp. 315-323. |
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| Relation |
http://dx.doi.org/10.1007/s11240-010-9781-x
http://www.eprints.iicb.res.in/101/ |
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