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In vitro regeneration of Anethum graveolens, antioxidative enzymes during organogenesis and RAPD analysis for clonal fidelity

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Biologia Plantarum

Abstract

An efficient in vitro regeneration protocol was developed for medicinally important aromatic plant Anethum graveolens. Nodal segments were cultured onto Murashige and Skoog (MS) basal medium supplemented with different auxins and cytokinins singly as well as in combinations. The optimum callus induction (93.33 %) was obtained on medium fortified with 2.2 μM N6-benzyladenine (BA) and 0.21 μM α-naphthaleneacetic acid. The best shoot regeneration (85.7 %) with 12.86 shoots per explant was achieved in two weeks when callus was subcultured on MS medium amended with 2.2 μM BA and 1.85 μM kinetin. The average length of regenerated shoots varied from 3.15 to 4.8 cm. The rooting of regenerated shoots was nearly 100 % on ¼ MS augmented with 4.9 μM indolebutyric acid with a maximum root length of 5.1 cm. Plantlets were successfully acclimatized with 60 % survival rate. During organogenesis, catalase and ascorbate peroxidase activity increased while superoxid dismutase activity decreased. Clonal fidelity of in vitro raised plants has been checked by random amplified polymorphic DNA using 10 selected decamer primers. It has been found that regenerated plants are true to type plants.

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Abbreviations

APX:

scorbate peroxidase

BA:

N6-benzyladenine

CAT:

catalase

CTAB:

cetyltrimethylammonium bromide

2,4-D:

2,4-dichlorophenoxyacetic acid

EtBr:

ethidium bromide

IAA:

indoleacetic acid

IBA:

indolebutyric acid

Kin:

kinetin

MS:

Murashige and Skoog

NAA:

α-naphthaleneacetic acid

PCR:

polymerase chain reaction

RAPD:

random amplified polymorphic DNA

SOD:

superoxidase dismutase.

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Acknowledgements

The financial support provided by Department of Science and Technology, New Delhi for the Project “Banasthali Centre for Education and Research in Basic Science” under their CURIE Programme is gratefully acknowledged. G.S. Shekhawat also acknowledges financial support received from the Department of Science and Technology, Rajasthan, Jaipur.

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Correspondence to G. S. Shekhawat.

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Jana, S., Shekhawat, G.S. In vitro regeneration of Anethum graveolens, antioxidative enzymes during organogenesis and RAPD analysis for clonal fidelity. Biol Plant 56, 9–14 (2012). https://doi.org/10.1007/s10535-012-0009-2

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  • DOI: https://doi.org/10.1007/s10535-012-0009-2

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