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LNS Silva, PCD Perea, SGJN Senanayake and Nilanthi Dahanayake*
Department of Agricultural Biology, Faculty of Agriculture, University of Ruhuna, Mapalana,
Kamburupitiya, Sri Lanka
Abstract
Calotropis gigantea (Wara) is one of the least concerned native plant in Sri Lanka. Because of its high pharmaceutical
application and other usage, this species has been over exploited and included in the Red list since year 2012.
The present study was carried out to develop a protocol to produce healthy plantlets of C. gigantea. Murashige
and Skoog medium (MS) (0, ¼, ½, ¾ and full MS) were tested to select the best medium for in vitro seed germination.
To determine the best explant and 6-Benzyl amino purine (BAP) concentration for micro propagation, in
vitro shoot tip, stem without node, stem with node, leaf and root were tested with different concentrations of BAP
(0, 1 mg/l, 2 mg/l, 3 mg/l, 4 mg/l, 5 mg/l and 6 mg/l) and 0.1 mg/l NAA. Agar and Corn flour were used to gelatinize
the medium. To determine the proper maturity level of plantlet, 1, 1 ½, 2, 2 ½ and 3 months old plants were
tested. Then regenerated shoots were transferred to different IBA (Indole 3-butyric acid) concentrations (0, 0.5
mg/l, 1 mg/l and 2 mg/l) for rooting. Ten replicates per each treatment were used in every experiment. Data were
analyzed according to 2 factor factorial Completely Randomized Design (CRD) using SAS statistical software.
The medium with agar (without MS) showed the highest seed germination of 72% while the medium containing
full MS showed the lowest seed germination of 34%. Significantly higher number of shoots (2.4) was observed
from MS medium with 2 mg/l BAP + 0.1 NAA + Agar within 11days, showing 100% regeneration from stem with
node. When corn flour was used instead of agar for the above medium, shoots regeneration was 2.7 within 17
days. One month old explant (stem with node) showed significantly higher number of shoots (2.0) whereas three
month old explant showed lowest number of shoots (0.8). The highest number of roots (3.4) was observed from
1mg/l IBA +MS + Agar. However, with corn flour instead of agar, average number of roots was 4.4. Considering
these observations it could be concluded that, Agar is suitable as a medium for seed germination, and stem with
node is the best explant to produce large number of healthy shoots. The best BAP concentration for shoot regeneration
was 2mg/l BAP+0.1mg/l NAA, while 1mg/l IBA was good for rooting. One month old explants were better
than matured explants and corn flour was better compared with agar in increasing the growth and development
of C. gigantea.
Key words:
Calotropis gigantea, Micropropagation, MS Medium, BAP, NAA
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* - Corresponding Author
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