Shoot induction from Clinacanthus nutans (Burm.f.) lindau nodal in response to plant growth regulators and media composition on growth and antioxidant activities
Clinacanthus nutans, locally known as Sabah Snake Grass or Belalai Gajah, is one of the most promising medicinal plants in Malaysia’s herbal industry which posses numerous biological properties. Besides that, C. nutans extract produce a wide range of bioactive compounds including several specific...
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| Format: | Thesis |
| Language: | English |
| Published: |
2020
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/90215/1/FP%202020%2019%20ir.pdf |
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| Summary: | Clinacanthus nutans, locally known as Sabah Snake Grass or Belalai Gajah, is
one of the most promising medicinal plants in Malaysia’s herbal industry which
posses numerous biological properties. Besides that, C. nutans extract produce
a wide range of bioactive compounds including several specific compounds such
as clinamides, entadamide, clinacoside and cycloclinacoside which only can be
found in C. nutans extract. Due to high therapeutic potentials, C. nutans has
been overexploited to satisfy the demands from pharmaceutical industry.
Conventionally propagated of C. nutans by stem cutting is known to produce
inconsistent secondary metabolites due to external factors in the environment
such as climate, pests and diseases and fertilizer application. In the present
study, plant tissue culture technique was adopted for shoot induction and
biomass production of C. nutans for extraction of secondary metabolites. The
nodal segments of C. nutans were used as explants and Murashige and Skoog
(MS medium) was used as basal medium. The shoot induction of C. nutans
nodal segments was carried out by supplementing the MS medium with
cytokinins; 6-benzylaminopurine (BAP) or kinetin at concentration 4, 8, 12 and 16
μM each. The best treatment was recorded from MS medium supplemented with
12 μM BAP with 100% shoot induction, 1.30 shoots, 2.29 cm length of shoots,
4.78 leaves and 16.00 mg of leaves fresh weight after four weeks of incubation.
In subsequent experiments, MS medium was supplemented with auxins; indole
butyric acid (IBA) and naphthalene acetic acid (NAA) at concentration 2, 4, 6 and
8 μM each, followed by modification of MS medium strength into half, full and
double strength. Lastly, sucrose concentration was tested at 20, 25, 30, 35, 40,
45 and 50 g/L, respectively. Based on the results obtained, full-strength MS
medium supplemented with 12 μM BAP and 30 g/L sucrose without addition of
auxin produced an optimum growth with 100% shoot induction, 1.67 shoots, 3.23
cm length of shoots, 7.33 leaves and 21.57 mg of leaves fresh weight after six
weeks of incubation. Subsequently, analyses of phenolics content and
antioxidant properties between tissue-cultured and conventionally propagated leaves were conducted. The leaves from both sources were extracted using
aqueous at temperature 25 and 100ºC. In the study on phenolics content, the
highest total polyphenols, phenolic acids and falvonoids content were recorded
from 100ºC aqueous extract of tissue-cultured leaf with 2.77 mg GAE/g DW, 4.29
mg GAE/g DW and 21.76 mg RE/g DW, respectively. In antioxidant properties
analysis, 100ºC aqueous extract of tissue-cultured leaf was exhibited the highest
DPPH free radical scavenging activity (3.20 mg TE/g DW), ABTS scavenging
activity (1.50 mg TE/g DW), ferric reducing antioxidant power (9.76 mg TE/g DW)
and superoxide anion radical scavenging activity (53.74%). Meanwhile, the
highest iron (II) chelating activity was recorded from 25ºC aqueous extract of
conventionally propagated leaf with 69.24% chelating activity. In the present
study, shoot induction from C. nutans nodal segments was successfully
conducted and bioactive compounds analysis showed that tissue-cultured leaf
was able to produce higher phenolics content and antioxidant properties
compared to conventionally propagated leaf. |
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