Crocus sativus L. (Saffron): In vitro studies with special reference to karyotyping and antioxidant potential
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Date
2008
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Birsa Agricultural University, Kanke, Ranchi, Jharkhand
Abstract
Crocus sativus L. is grown in small area with a limitation for healthy planting material.
The plant is sterile, so its improvement through breeding is quite difficult. Use of saffron
stigma in medicinal purpose is well reported. In the present work, efficient in vitro
methods for microcorm production via shoot tips and corm of Crocus sativus L. was
developed. Best survival percentage during surface sterilization of Crocus sativus L.
explants were achieved by treating shoot tips with 0.2% of HgCl2 for 25 minutes and
corms with 0.2% HgCl2 for 40 minutes. Media was prepared on the basis of earlier
reports on in vitro propagation of saffron with some modification as well as new
compositions. Out of these media C11 was found to be the best among all used during
experiments. Bud breaking from corm was induced on C11 media (MS media
supplemented with 2.0 mg/L of NAA, 5.0 mg/L of BAP, 100.0 mg/L of AdSO4 and
sucrose 4%). Highest mean number of buds (2.4) was observed after 150 days of
inoculation in the same media. From corm, highest mean number of minicorm per
explant (4.3) was observed after 150 days of inoculation in the same media. When apical
buds was inoculated on C11, kept at culture room condition (light 3000 lux and
temperature 25 ± 2º C) highest mean number of microcorm was 34.6 which was reduced
to 32.8 when kept at dark at 4-8º C after 125 days. Initial induction of microcorm in dark
condition was more rapid than culture room condition but after 100 days of interval
induction rate was reduced, in comparison to culture kept under light. Moreover, mean
weight of microcorm/ culture bottle kept at dark was 1.085 times more than that of kept
at culture room condition. After emergence of apical bud from microcorm on C11 media,
microcorms from C11 media were transferred to different media. Maximum mean
number of buds/ culture bottle was 24.2, which was observed on C11 media.
Multiplication of microcorms from single microcorm was observed in the same media.
Overall study revealed that out of different media, C11 was the best for inducing in vitro
response. The combination of phytohormones used in this media was not reported
earlier. Callus growth was observed on C11 and C31 (MS media supplemented with 2,4-
D 1.0 mg/L, kinetin 1.0 mg/L and sucrose 3%) media. Later when the callus which was
proliferating on C31 media was subcultured to C31 and C30 media (MS media
supplemented with 2,4-D 0.5 mg/L, kinetin 1.0 mg/L and sucrose 3%), somatic embryos
was induced. Leaf was emerged from apical buds, when inoculated on different media
with or without section of corm. Abnormalities in leaf were also observed. No positive
results were obtained, when stigma was used as explant and inoculated on MS basal with
various concentrations and combinations of hormones and other additives. However,
some of the stigma was found alive after 200 days of inoculation. Attempts were made to
harden plant but no positive results were obtained.
The sterility of Crocus sativus L. is well reported; to study its chromosome, karyotyping
was done. During this study, somatic chromosome number was found 3X=2n= 24, range
of chromosome length was 4.16-9.17 μm, total volume of chromosome was 312.57 μm3,
absolute chromosome length was 158.67 μm and karyotype formula was A3B15C6.
Antioxidant activity by DPPH and ABTS assay showed that 200 μg/ml of crude
methanolic extract of Crocus sativus was able to inhibit the DPPH radical formation by
42.8% and 99.01 μg/ml of plant extract was able to inhibit ABTS radical formation by
36.84% respectively. Addition of 10, 20 and 30 μg/μl of plant extract to the reaction
mixture of H2O2 indicated the significant protection against damage of pUC 18.
Description
Crocus sativus L. (Saffron): In vitro studies with special reference to karyotyping and antioxidant potential
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