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Indian Journal of Biotechnology (IJBT) >
IJBT Vol.11 [2012] >
IJBT Vol.11(1) [January 2012] >
| Title: | In vitro regeneration of Hypericum perforatum L. using thidiazuron and analysis of genetic stability of regenerants |
| Authors: | Banerjee, Arpita
Bandyopadhyay, Subhendu
Raychaudhuri, Sarmistha Sen |
| Keywords: | Hypericum Perforatum L.
In Vitro Regeneration
PCR-RFLP
RAPD
Thidiazuron |
| Issue Date: | Jan-2012 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | In the present study, in vitro regeneration method for Hypericum
perforatum L. using different plant growth regulators has been developed.
Callus was induced from hypocotyl explants on MS medium using 0.5 mg L-1
2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg L-1 kinetin (Kn).
Shoots were developed on the medium containing 1 mg L-1 thidiazuron
and rooting was achieved with 2 mg L-1 indoleacetic acid (IAA).
Random amplified polymorphic DNA (RAPD) and restriction fragment length
polymorphism (RFLP) were used to assess the genetic stability of the
regenerated plants. Among a large number of regenerated plantlets, 10 plants
were randomly selected for molecular analysis. 15 RAPD primers generated
monomorphic banding pattern among the mother and regenerated plants. PCR-RFLP
profiles of nuclear ribosomal internal transcribed spacer (nrITS) region using EcoRV, ClaI and HpaII also did
not show any variation in banding pattern. The entire ITS region of the mother
plant, and a randomly selected regenerated plant, when cloned and sequenced
showed query coverage of 99% when analyzed through CLUSTAL W. These molecular
analyses showed that the regenerated plants produced were similar to the mother
plant both in phenotypic and genotypic characters. |
| Page(s): | 92-98 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-0967 (Online); 0972-5849 (Print) |
| Source: | IJBT Vol.11(1) [January 2012]
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