In vitro propagation and genetic fidelity studies in chrysanthemum (Chrysanthemum morifolium Ramat.)
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Date
2011
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Publisher
CCSHAU
Abstract
The present study was conducted on the three cultivars of chrysanthemum (Mayur, Dolly
Pink, Royal Purple). The objective of the study was to developed an efficient protocol for direct
regeneration, multiplication and rooting in vitro conditions and checked the genetic fidelity of the
micropropagated plants. Nodal segment and shoot tip explants, after being sterilized with 0.1% of
mercuric chloride and 70% ethanol, were inoculated in Murashige and Skoog (MS) media with varied
concentrations of indole acetic acid (IAA), benzyl amino purine (BAP), napthelic acetic acid
(NAA)and their combinations. Different parameters including shoot regeneration percentage,
multiplication average number of shoots per explant, length of shoots (cm), number of leaves per shoot,
number of days taken for root initiation, number of roots per shoot, length of root (cm), were studied
during the course of study. Intermediate concentrations of benzyladenine purine (BAP 1.0 mg/l) was
found superior to all the other BAP concentration. MS media fertified with 1.0 mg/l BAP had produced
the maximum shoots (90.53 per cent in shoot tip and 90.80 per cent in nodal segment), shoots per
explant (4.06), length of shoot (3.90 cm), number of leaves (4.93). Similarly, when the combination of
different concentration of IAA and BAP were used, significant results regarding the regeneration in
chrysanthemum plantlets were achieved. Satisfactory rooting response was obtained with (2.0 mg/l and
1.5 mg/l IBA0. In vitro raised plantlets were transferred to potted soil for the hardening and after
hardening the leaves were collected from the potted chrysanthemum plants for DNA extracts. The
maximum survival was found (100%) in potting mixture sand +soil+FYM (1:1:1) and
sand+soil+vermicompost (1:1:1). Twenty primers were used for DNA amplification. Out of twenty
primers screened, thirteen primers produced amplification. Reproducible and clear banding patterns
were obtained in a reaction mixture of 10μl containing 50 ng template DNA, 100 μM of dNTPs mix, 1
μM of primer, 1.5 Mm MgCl2 , 1μl of 10 X Taq DNA polymerase buffer and 2.0 units of Taq DNA
polymerase. All the bands were similar and no polymorphism was to be found, which showed that all
the plants raised through micro propagation were true to type or identical to the mother plants.
Description
Keywords
Chrysanthemum, DNA amplification, BAP, IBA, Micropropagation