Establishment of in vitro culture of clinacanthus nutans (Burm. f.) lindau and antibacterial assay of different crude methanolic extracts
Clinacanthus nutans (Burm f.) Lindau or Sabah Snake Grass from the Acanthaceae family is a well-known medicinal plant and has been widely used in many Asian countries especially in Malaysia, Thailand, Indonesia, and China. The present study was aimed to establish axenic in vitro cultures, induce...
| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
Universiti Malaysia Sarawak, (UNIMAS)
2015
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| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/10808/ http://ir.unimas.my/id/eprint/10808/1/Cherrie.pdf |
| _version_ | 1848836865470234624 |
|---|---|
| author | Lim, Cherrie Han Rou |
| author_facet | Lim, Cherrie Han Rou |
| author_sort | Lim, Cherrie Han Rou |
| building | UNIMAS Institutional Repository |
| collection | Online Access |
| description | Clinacanthus nutans (Burm f.) Lindau or Sabah Snake Grass from the Acanthaceae family is a
well-known medicinal plant and has been widely used in many Asian countries especially in
Malaysia, Thailand, Indonesia, and China. The present study was aimed to establish axenic in vitro
cultures, induce shoot and callus from axenic explants, evaluate antibacterial activities of the
plant’s crude methanolic extracts, and analyze the phytochemical components present in the plant
extracts. Results showed that explants soaked in 10% Clorox® for 15 minutes produced the highest
percentage of axenic leaf explants (98%). Treatment with 15% Clorox® for 20 minutes and
additional initial step of washing explants in diluted dish wash liquid was the best to produce
contamination-free internodal (96.67%) and nodal explants (100%). The efficiency of plant
regeneration via direct organogenesis from axenic nodal explants (source: mother plant and in vitro
plantlet) was tested on Murashige and Skoog (MS) medium supplemented with BAP, IBA or TDZ
at different concentrations (0.0- 2.0 mg/L) either alone and in combinations. Control medium
without plant growth regulator (PGR) was found to be the best for shoot proliferation as it
successfully produced healthier plantlets with longer shoots and vigorous roots within eight weeks
when compared to other treatments. The effect of BAP alone, BAP in combination with 2, 4-D, and
BAP in combination with NAA (0.0- 4.0 mg/L) was evaluated on the callus induction and
formation. Explants cultured on the medium including 2, 4-D or without any PGR failed to form
callus after eight weeks. The results showed that 100% of internodal explants culturing on MS
medium supplemented with BAP alone or in combination with NAA produced callus. Leaf
explants was not suitable for callus induction due to the poor performance of callus growth, hence,
only internodal explants cultured on the medium fortified with 1.0 mg/L BAP under light condition
was chosen for mass production of callus for the use of successive experiments. Subsequently,
callus culture, leaf and stem from both in vitro plantlets and potted grown plants were extracted
using methanol. The percentage of extraction yields for in vitro cultures (8- 19%) were higher than
potted grown plant (< 5%). The crude methanolic extracts of C. nutans were then screened for its
antibacterial activity and phytochemical composition by gas chromatography-mass spectrometry
(GC-MS) analysis. Plant extracts were tested against Bacillus cereus, Listeria monocytogenes,
Salmonella typhi and Echerichia coli using agar disc diffusion, and broth dilution to determine
minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The
results showed that all extracts exhibited bacteria inhibition. B. cereus was inhibited by all plant
crude methanolic extracts with larger inhibition zone compared to other bacteria while potted
grown leaf extract exhibited the greatest inhibition diameter in agar disc diffusion (19.38 mm). In
broth dilution test, callus and potted grown leaf extracts again presented the best bioassay activity
against B. cereus, with the MIC and MBC value at 3.13 mg/mL. From the GC-MS analysis, the
components revealed from the crude methanolic extracts of C. nutans are mainly composed of
hydrocarbon, followed by ester compounds. Alcohol, aromatic compounds, and aldehyde were also
found. Several bioactive compounds that may have contributed to the antibacterial activities were
detected. |
| first_indexed | 2025-11-15T06:30:33Z |
| format | Thesis |
| id | unimas-10808 |
| institution | Universiti Malaysia Sarawak |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T06:30:33Z |
| publishDate | 2015 |
| publisher | Universiti Malaysia Sarawak, (UNIMAS) |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | unimas-108082023-07-03T07:58:14Z http://ir.unimas.my/id/eprint/10808/ Establishment of in vitro culture of clinacanthus nutans (Burm. f.) lindau and antibacterial assay of different crude methanolic extracts Lim, Cherrie Han Rou SB Plant culture Clinacanthus nutans (Burm f.) Lindau or Sabah Snake Grass from the Acanthaceae family is a well-known medicinal plant and has been widely used in many Asian countries especially in Malaysia, Thailand, Indonesia, and China. The present study was aimed to establish axenic in vitro cultures, induce shoot and callus from axenic explants, evaluate antibacterial activities of the plant’s crude methanolic extracts, and analyze the phytochemical components present in the plant extracts. Results showed that explants soaked in 10% Clorox® for 15 minutes produced the highest percentage of axenic leaf explants (98%). Treatment with 15% Clorox® for 20 minutes and additional initial step of washing explants in diluted dish wash liquid was the best to produce contamination-free internodal (96.67%) and nodal explants (100%). The efficiency of plant regeneration via direct organogenesis from axenic nodal explants (source: mother plant and in vitro plantlet) was tested on Murashige and Skoog (MS) medium supplemented with BAP, IBA or TDZ at different concentrations (0.0- 2.0 mg/L) either alone and in combinations. Control medium without plant growth regulator (PGR) was found to be the best for shoot proliferation as it successfully produced healthier plantlets with longer shoots and vigorous roots within eight weeks when compared to other treatments. The effect of BAP alone, BAP in combination with 2, 4-D, and BAP in combination with NAA (0.0- 4.0 mg/L) was evaluated on the callus induction and formation. Explants cultured on the medium including 2, 4-D or without any PGR failed to form callus after eight weeks. The results showed that 100% of internodal explants culturing on MS medium supplemented with BAP alone or in combination with NAA produced callus. Leaf explants was not suitable for callus induction due to the poor performance of callus growth, hence, only internodal explants cultured on the medium fortified with 1.0 mg/L BAP under light condition was chosen for mass production of callus for the use of successive experiments. Subsequently, callus culture, leaf and stem from both in vitro plantlets and potted grown plants were extracted using methanol. The percentage of extraction yields for in vitro cultures (8- 19%) were higher than potted grown plant (< 5%). The crude methanolic extracts of C. nutans were then screened for its antibacterial activity and phytochemical composition by gas chromatography-mass spectrometry (GC-MS) analysis. Plant extracts were tested against Bacillus cereus, Listeria monocytogenes, Salmonella typhi and Echerichia coli using agar disc diffusion, and broth dilution to determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The results showed that all extracts exhibited bacteria inhibition. B. cereus was inhibited by all plant crude methanolic extracts with larger inhibition zone compared to other bacteria while potted grown leaf extract exhibited the greatest inhibition diameter in agar disc diffusion (19.38 mm). In broth dilution test, callus and potted grown leaf extracts again presented the best bioassay activity against B. cereus, with the MIC and MBC value at 3.13 mg/mL. From the GC-MS analysis, the components revealed from the crude methanolic extracts of C. nutans are mainly composed of hydrocarbon, followed by ester compounds. Alcohol, aromatic compounds, and aldehyde were also found. Several bioactive compounds that may have contributed to the antibacterial activities were detected. Universiti Malaysia Sarawak, (UNIMAS) 2015 Thesis NonPeerReviewed text en http://ir.unimas.my/id/eprint/10808/1/Cherrie.pdf Lim, Cherrie Han Rou (2015) Establishment of in vitro culture of clinacanthus nutans (Burm. f.) lindau and antibacterial assay of different crude methanolic extracts. Masters thesis, Universiti Malaysia Sarawak, (UNIMAS). |
| spellingShingle | SB Plant culture Lim, Cherrie Han Rou Establishment of in vitro culture of clinacanthus nutans (Burm. f.) lindau and antibacterial assay of different crude methanolic extracts |
| title | Establishment of in vitro culture of clinacanthus nutans (Burm. f.) lindau and antibacterial assay of different crude methanolic extracts |
| title_full | Establishment of in vitro culture of clinacanthus nutans (Burm. f.) lindau and antibacterial assay of different crude methanolic extracts |
| title_fullStr | Establishment of in vitro culture of clinacanthus nutans (Burm. f.) lindau and antibacterial assay of different crude methanolic extracts |
| title_full_unstemmed | Establishment of in vitro culture of clinacanthus nutans (Burm. f.) lindau and antibacterial assay of different crude methanolic extracts |
| title_short | Establishment of in vitro culture of clinacanthus nutans (Burm. f.) lindau and antibacterial assay of different crude methanolic extracts |
| title_sort | establishment of in vitro culture of clinacanthus nutans (burm. f.) lindau and antibacterial assay of different crude methanolic extracts |
| topic | SB Plant culture |
| url | http://ir.unimas.my/id/eprint/10808/ http://ir.unimas.my/id/eprint/10808/1/Cherrie.pdf |