Analysis of bioactive pigments in coloured callus of <i>Azadirachta indica</i> for possible use as functional natural colourants

Ashokhan, Sharmilla; Srinivasan, Ramasamy; Karsani, Saiful Anuar; Othman, Rashidi; Yaacob, Jamilah Syafawati

doi:10.1108/prt-11-2017-0095

Cited by 4 publications

(9 citation statements)

References 53 publications

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“…Based on the results, it was observed that green callus extract contained the highest level of AZA (214.53 ± 33.63 mg g -1 DW), followed by brown callus extract (51.56 ± 7.22 mg g -1 DW) and cream callus extract (28.49 ± 4.66 mg g -1 DW). These results are parallel to previously reported biochemical activities, which showed that green callus extracts possessed the highest bioactive compound content, antioxidant and cytotoxic potentials [38].…”

Section: Extraction and Quantification Of Azadirachtin (Aza) From Colsupporting

confidence: 91%

“…Total AZA was extracted using absolute methanol, followed by liquid-liquid extraction using dichloromethane according to the protocol described by Singh and Chaturvedi [35]. We had previously reported that the colored callus of A. indica exhibits a wide range of biological actions such as radical scavenging and cytotoxic activities [38]. Due to these promising bioactivities, the colored callus obtained in this study (green, brown and cream) were evaluated for the presence of AZA, which is the most abundant bioactive compound of this species.…”

Section: Extraction and Quantification Of Azadirachtin (Aza) From Colmentioning

confidence: 99%

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Effect of Plant Growth Regulators on Coloured Callus Formation and Accumulation of Azadirachtin, an Essential Biopesticide in Azadirachta indica

Ashokhan

Othman

Rahim

et al. 2020

Plants

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For centuries, Azadirachta indica or neem has been utilized as a primary source of medicine due to its antimicrobial, larvacidal, antimalarial and antifungal properties. Recently, its potential as an effective biopesticide has garnered attention, especially towards efficient and continuous production of its bioactive compounds. The present study investigated the effect of the plant growth regulators (PGRs) thiadiazuron (TDZ) and 2,4-dichlorophenoxyacetic acid (2,4-D) on the induction of colored callus formation and subsequent accumulation of azadirachtin (AZA) in A. indica. An efficient protocol was established for micropropagation and colored callus production of this species, followed by quantification of AZA (a mixture of azadirachtin A and B) and its safety assessment. For induction of the callus, leaf and petiole explants obtained from a young growing neem plant were excised and cultured on Murashige and Skoog (MS) medium supplemented with TDZ (0.2–0.6 mg L−1) and 2,4-D (0.2–0.6 mg L−1), either applied singly or in combination. Callus was successfully induced from both explant types at different rates, where media with 0.6 mg L−1 of TDZ resulted in the highest fresh weight (3.38 ± 0.08 g). In general, media with a single hormone (particularly TDZ) was more effective in producing a high mass of callus compared to combined PGRs. A culture duration of six weeks resulted in the production of green, brown and cream colored callus. The highest callus weight and accumulation of AZA was recorded in green callus (214.53 ± 33.63 mg g−1 dry weight (DW)) induced using TDZ. On the other hand, small amounts of AZA were detected in both brown and cream callus. Further experimentation indicated that the green callus with the highest AZA was found to be non-toxic (LC50 at 4606 µg mL−1) to the zebrafish animal model. These results suggested that the addition of different PGRs during in vitro culture could prominently affect callus and secondary metabolite production and can further be manipulated as a sustainable method for the production of a natural and environmentally friendly pesticide.

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Section: Extraction and Quantification Of Azadirachtin (Aza) From Colsupporting

confidence: 91%

Section: Extraction and Quantification Of Azadirachtin (Aza) From Colmentioning

confidence: 99%

Effect of Plant Growth Regulators on Coloured Callus Formation and Accumulation of Azadirachtin, an Essential Biopesticide in Azadirachta indica

Ashokhan

Othman

Rahim

et al. 2020

Plants

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“…All extraction procedures were conducted in complete darkness. After homogenizing the samples, they were soaked in 60 mL of absolute methanol and incubated at 4 • C for 48 h. The filtrate was collected and stored at −20 • C. Re-extraction and filtration of the residue were performed and the extracts were combined and centrifuged for 5 min at 9000 rpm, 4 • C. A portion of these supernatants was immediately used for subsequent pigment analysis [80] to determine the total chlorophyll, carotenoid, and anthocyanin contents, while the remaining supernatants were concentrated to dryness using a rotavapor at 45 • C. The solvent-free extract was adjusted to a concentration of 10 mg/mL with absolute methanol and stored at −20 • C in an airtight glass vial until further analysis. The flow chart diagram of experimental outline is represented in Figure 3.…”

Section: Experimental Design and Sample Preparationmentioning

confidence: 99%

Organic Amendments Effects on Nutrient Uptake, Secondary Metabolites, and Antioxidant Properties of Melastoma malabathricum L.

Rusli

Abdullah

Yaacob

et al. 2022

Plants

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Amelioration of soil acidity can boost soil fertility, hence increasing nutrient uptake, secondary metabolite, and its antioxidant potential. In the present study, the effectiveness of food waste compost and palm kernel biochar was assessed as soil amendments for Melastoma malabathricum L. grown in acidic soil conditions. A six-month greenhouse study was conducted using completely randomized design (CRD) with three treatment groups, including control plants (T1), plants amended with palm kernel biochar (T2), and plants amended with food waste compost (T3). Data analysis revealed that Melastoma malabathricum L. amended with T3 recorded the highest total chlorophyll content (433.678 ± 13.224 µg g−1 DW), followed by T2 and T1. The increase in chlorophyll content was contributed by the increase in soil pH. This was shown by the positive significant correlations between soil pH and chlorophyll a (r2 = 0.96; p ≤ 0.01) and chlorophyll b (r2 = 0.778; p ≤ 0.01). In addition, the same treatment exhibited the highest total anthocyanin content (leaves; 36.1 × 10−2 ± 0.034 mg/g DW and root extract; 8.9 × 10−2 ± 0.020 mg/g DW), total phenolic content (stem extract; 4930.956 ± 16.025 mg GAE/g DE), and total flavonoid content (stem extract; 209.984 ± 0.572 mg QE/g DE). Moreover, this study also found that the highest antioxidant potential against 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2-Azinobis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals was exhibited by samples supplemented with food waste compost (T3), followed by palm kernel biochar (T2). This indicates that the soil amendments have the capacity to enhance the secondary metabolites that protect plants, therefore ameliorating Melastoma malabathricum L.’s response towards acidic stress, and resulting in better antioxidant properties. Furthermore, this study also recorded better nutrient uptake in T3. With the significantly higher levels of macronutrient in the soil, the food waste compost could enhance the nutrient properties, secondary metabolites, and antioxidant capacity of Melastoma malabathricum L. grown in acidic soil conditions.

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“…CM media consisted of 4.4 g of MS (Murashige and Skoog, 1962), 30 g/L of sucrose and 2 g/L of gelrite and supplemented with 0.6 mg/L TDZ, while MSO was similar to CM media but devoid of any plant growth regulators. The pH of both media (CM and MSO) were adjusted to 5.8 using 0.1 M NaOH or 0.1 M HCl (Ashokhan et al, 2018). The callus cultures were maintained in the culture room at 25 ± 2 • C with a 16 h photoperiod and 1000 lux of light intensity.…”

Section: Callus Induction and Proliferationmentioning

confidence: 99%

“…Meanwhile, the remaining portion of the supernatants were concentrated under reduced pressure at 60 • C using rotary evaporator (Rotavapor R-3 BÜCHI Labortechnik, 9230 Switzerland) to remove excess solvent. The resultant dried extracts were weighed and re-dissolved in absolute methanol to a concentration of 20 mg/mL and used for subsequent total phenolics, total flavonoids, phytochemical and antioxidants analysis (Ashokhan et al, 2018).…”

Section: Extraction Of Bioactive Pigments From Green Callus Of a Indicamentioning

confidence: 99%

Evaluation of the effect of jasmonic acid elicitation on composition of pigments and biological activities in green callus of neem (Azadirachta indica)

Fauzi

Rahim²,

Majid³

et al. 2022

Front. Sustain. Food Syst.

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This study was carried out with the aim of determining the effects of jasmonic acid (JA) elicitation on the bioactive pigments' biosynthesis and the antioxidant activities in green callus of Azadirachta indica of two different ages (4- and 8-week-old). Plant tissue culture technique was employed to induce the formation of green callus from leaf explants of A. indica on Murashige and Skoog (MS) medium supplemented with 0.6 mg/L thidiazuron (CM) and three different concentrations of JA (2, 4, and 6 mg/L). The methanolic extracts from the green callus were used for determination of total chlorophyll content (TCh), total carotenoid content (TC), total anthocyanin content (TAC), total phenolic content (TPC), and total flavonoid content (TFC) through colorimetric and HPLC analyses. The highest amount of yield was obtained from CM and 2 mg/L JA (2JA) extracts for 4- and 8-week-old samples, respectively. Phytochemical screening revealed the presence of alkaloids, flavonoids, phenols, tannins, and terpenoids in all 4- and 8-week-old samples elicited with 2, 4 and 6 mg/L JA. The highest value for TAC, TCh, TC, TPC, and TFC of 4- and 8-week-old samples were from callus cultured on media supplemented with 6 mg/L JA (6JA) and 4 mg/L JA (4JA), respectively. The lowest IC50 values were found to be 8.29 ± 0.10 mg/mL (6JA) for 4-week-old and 7.73 ± 0.03 mg/mL (4JA) for 8-week-old samples. The highest Ferric Reducing Antioxidant Power (FRAP) values obtained in this study were 90.60 ± 1.55 g/g (6JA), and 74.59 ± 3.91 g/g (4JA), respectively, for 4- and 8-week-old samples. Moreover, Pearson's correlation analysis revealed a significant correlation between TAC, TCh, TC, TPC, and TFC with ABTS and FRAP assays. In addition, PCA analysis revealed that 83.5% of the information (variances) contained in the data were retained by the first two principal components. Overall, these findings suggested that JA supplementation into the culture media significantly increase the chlorophyll, carotenoid, anthocyanin, phenolic and flavonoid contents and JA concentrations at 6 mg/L JA and 4 mg/L JA yielded the highest pigments content in 4- and 8-weeks-old callus, respectively.

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Analysis of bioactive pigments in coloured callus of Azadirachta indica for possible use as functional natural colourants

Cited by 4 publications

References 53 publications

Effect of Plant Growth Regulators on Coloured Callus Formation and Accumulation of Azadirachtin, an Essential Biopesticide in Azadirachta indica

Effect of Plant Growth Regulators on Coloured Callus Formation and Accumulation of Azadirachtin, an Essential Biopesticide in Azadirachta indica

Organic Amendments Effects on Nutrient Uptake, Secondary Metabolites, and Antioxidant Properties of Melastoma malabathricum L.

Evaluation of the effect of jasmonic acid elicitation on composition of pigments and biological activities in green callus of neem (Azadirachta indica)

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