Morphogenetic and biochemical studies in callus cultures of devil’s thorn (Tribulus terrestris Linn.)
Loading...
Date
2013
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
CCSHAU
Abstract
Present investigation was carried out on Tribulus terrestris Linn. for the selection of suitable nutritive medium and explant for callus induction, growth and differentiation. In vitro studies were conducted with employing different explants viz. leaf, internode and node used for callogenesis and growth, node explant was found to be the best for callus induction and growth. Best callus growth was obtained on MS medium fortified with 2,4-D (2.0 mg/l) in combination with Kn (0.5 mg/l) depending on fresh and dry weight of callus. This callus selected for further studies of biochemicals and protein profile during differentiation.
MS medium supplemented with BAP (3.0 mg/l) + NAA (1.5 mg/l) enabled the early regeneration of shoot from callus. Early root emergence from callus obtained on medium MS+NAA (2.0 mg/l).
Biochemicals like starch and total soluble sugars decreased while reducing sugar, total phenol and total protein increased during root differentiation from callus. Enzyme activity viz. α- amylase, acid invertase, peroxidase and acid phosphatase increased during root differentiation. In shoot differentiating calli, all these metabolites were increased and enzyme activity of α-amylase and acid phosphatase increased while acid invertase and peroxidase activity decreased during shoot differentiation.
SDS-PAGE studies revealed that there was synthesis of three root specific polypeptide bands
with (MW 40.64, 19.23 and 11.81 kDa) and three shoot specific bands (MW 28.10, 17.02 and 12.46 kDa) during differentiation. MW 24.96 kDa and 21.20 kDa bands were common bands which disappeared during root and shoot differentiation.
TLC analysis showed that the diosgenin and hecogenin were present in the calli as well as in
natural fruits. They were further confirmed by IR spectroscopy. The incorporation of cholesterol in the
medium was observed to be effective for the increased steroidal sapogenins (hecogenin and diosgenin) production in calli than natural fruits.
Description
Keywords
Planting, Vegetative propagation, Biological development, Organic compounds, Auxins, enzymes, Biological phenomena, Regeneration, Proteins, Tissue culture