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Fulltext Comparative studies on cellular behaviour of carnation (Dianthus caryophyllus Linn. cv. Grenadin) grown in vivo and in vitro for early detection of somaclonal variation

Yaacob JS, Taha RM, Khorasani Esmaeili A

ScientificWorldJournal, 2013;2013:686752.
PMID: 23766703 DOI: 10.1155/2013/686752

The present study deals with the cytological investigations on the meristematic root cells of carnation (Dianthus caryophyllus Linn.) grown in vivo and in vitro. Cellular parameters including the mitotic index (MI), chromosome count, ploidy level (nuclear DNA content), mean cell and nuclear areas, and cell doubling time (Cdt) were determined from the 2 mm root tip segments of this species. The MI value decreased when cells were transferred from in vivo to in vitro conditions, perhaps due to early adaptations of the cells to the in vitro environment. The mean chromosome number was generally stable (2n = 2x = 30) throughout the 6-month culture period, indicating no occurrence of early somaclonal variation. Following the transfer to the in vitro environment, a significant increase was recorded for mean cell and nuclear areas, from 26.59 ± 0.09 μm² to 35.66 ± 0.10 μm² and 142.90 ± 0.59 μm² to 165.05 ± 0.58 μm², respectively. However, the mean cell and nuclear areas of in vitro grown D. caryophyllus were unstable and fluctuated throughout the tissue culture period, possibly due to organogenesis or rhizogenesis. Ploidy level analysis revealed that D. caryophyllus root cells contained high percentage of polyploid cells when grown in vivo and maintained high throughout the 6-month culture period.
Exploring the potential of vermicompost as a sustainable strategy in circular economy: improving plants' bioactive properties and boosting agricultural yield and quality

Kamar Zaman AM, Yaacob JS

Environ Sci Pollut Res Int, 2022 Feb;29(9):12948-12964.
PMID: 35034296 DOI: 10.1007/s11356-021-18006-z

Consumption of natural resources and waste generation continues to rise as the human population increases. Ever since the industrial revolution, consumers have been adopting a linear economy model based on the 'take-make-dispose' approach. Raw materials are extracted to be converted into products and finally discarded as wastes. Consequently, this practice is unsustainable because it causes a massive increase in waste production. The root problems of the linear system can be addressed by transitioning to a circular economy. Circular economy is an economic model in which wastes from one product are recycled and used as resources for other processes. This literature review discovers the potential of vermicompost as a sustainable strategy in circular economy and highlights the benefits of vermicompost in ensuring food security, particularly in improving agricultural yield and quality, as well as boosting crop's nutritional quality. Vermicompost has the potential to be used in a variety of ways in the circular economy, including for agricultural sustainability, managing waste, pollutant remediation, biogas production and animal feed production. The recycling of organic wastes to produce vermicompost can benefit both the consumers and environment, thus paving the way towards a more sustainable agriculture for the future.
Fulltext Protein profiling and histone deacetylation activities in somaclonal variants of oil palm (Elaeis guineensis Jacq.)

Yaacob JS, Loh HS, Mat Taha R

ScientificWorldJournal, 2013;2013:613635.
PMID: 23844406 DOI: 10.1155/2013/613635

Mantled fruits as a result of somaclonal variation are often observed from the oil palm plantlets regenerated via tissue culture. The mantling of fruits with finger-like and thick outer coating phenotypes significantly reduces the seed size and oil content, posing a threat to oil palm planters, and may jeopardize the economic growth of countries that depend particularly on oil palm plantation. The molecular aspects of the occurrence of somaclonal variations are yet to be known, possibly due to gene repression such as DNA methylation, histone methylation and histone deacetylation. Histone deacetylases (HDACs), involved in eukaryotic gene regulation by catalyzing the acetyl groups are removal from lysine residues on histone, hence transcriptionally repress gene expression. This paper described the total protein polymorphism profiles of somaclonal variants of oil palm and the effects of histone deacetylation on this phenomenon. Parallel to the different phenotypes, the protein polymorphism profiles of the mantled samples (leaves, fruits, and florets) and the phenotypically normal samples were proven to be different. Higher HDAC activity was found in mantled leaf samples than in the phenotypically normal leaf samples, leading to a preliminary conclusion that histone deacetylation suppressed gene expression and contributed to the development of somaclonal variants.
Fulltext Antibacterial Properties and Effects of Fruit Chilling and Extract Storage on Antioxidant Activity, Total Phenolic and Anthocyanin Content of Four Date Palm (Phoenix dactylifera) Cultivars

Samad MA, Hashim SH, Simarani K, Yaacob JS

Molecules, 2016 Mar 26;21(4):419.
PMID: 27023514 DOI: 10.3390/molecules21040419

Phoenix dactylifera or date palm fruits are reported to contain natural compounds that exhibit antioxidant and antibacterial properties. This research aimed to study the effect of fruit chilling at 4 °C for 8 weeks, extract storage at -20 °C for 5 weeks, and extraction solvents (methanol or acetone) on total phenolic content (TPC), antioxidant activity and antibacterial properties of Saudi Arabian P. dactylifera cv Mabroom, Safawi and Ajwa, as well as Iranian P. dactylifera cv Mariami. The storage stability of total anthocyanin content (TAC) was also evaluated, before and after storing the extracts at -20 °C and 4 °C respectively, for 5 weeks. Mariami had the highest TAC (3.18 ± 1.40 mg cyd 3-glu/100 g DW) while Mabroom had the lowest TAC (0.54 ± 0.15 mg cyd 3-glu/100 g DW). The TAC of all extracts increased after storage. The chilling of date palm fruits for 8 weeks prior to solvent extraction elevated the TPC of all date fruit extracts, except for methanolic extracts of Mabroom and Mariami. All IC50 values of all cultivars decreased after the fruit chilling treatment. Methanol was a better solvent compared to acetone for the extraction of phenolic compounds in dates. The TPC of all cultivars extracts decreased after 5 weeks of extract storage. IC50 values of all cultivars extracts increased after extract storage except for the methanolic extracts of Safawi and Ajwa. Different cultivars exhibited different antibacterial properties. Only the methanolic extract of Ajwa exhibited antibacterial activity against all four bacteria tested: Staphylococcus aureus, Bacillus cereus, Serratia marcescens and Escherichia coli. These results could be useful to the nutraceutical and pharmaceutical industries in the development of natural compound-based products.
Fulltext Vermicompost Supplementation Improves the Stability of Bioactive Anthocyanin and Phenolic Compounds in Clinacanthus nutans Lindau

Yusof Z, Ramasamy S, Mahmood NZ, Yaacob JS

Molecules, 2018 Jun 04;23(6).
PMID: 29867000 DOI: 10.3390/molecules23061345

This project studied the effect of vermicompost application on the composition of bioactive anthocyanin and phenolic compounds, and the antioxidant activity of Clinacanthus nutans. The correlation between the bioactive constituents and antioxidant capacity was also evaluated. In this project, a field study was conducted using a randomized complete block design (RCBD) with four treatment groups, including control plants (CC), plants supplied with chemical fertilizer (CF), plants supplied with vermicompost (VC), and plants supplied with mixed fertilizer (MF). The leaves of C. nutans from all treatment groups were harvested, subjected to solvent extraction, and used for quantification of total anthocyanin content (TAC), total phenolic content (TPC), and total flavonoid content (TFC). The initial antioxidant activity of the extracts was evaluated using 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays, as well as after two and four weeks of storage at -20 °C and 4 °C. Data analysis showed that CC plants contained the highest TAC (2180.14 ± 338.43 µg/g dry weight) and TFC (276.25 ± 3.09 mg QE/g dry weight). On the other hand, CF plants showed the highest TPC (181.53 ± 35.58 mg GAE/g dry weight). Moreover, we found that CC plants had the highest antioxidant potential against DPPH radicals whereas MF plants showed the lowest antioxidant potential. After four weeks of extract storage at -20 °C and 4 °C, the TPC, TFC, TAC, and antioxidant potential of the extracts decreased. Extracts from VC showed the lowest percentage of total phenolic and total flavonoid loss after extract storage at -20 °C and 4 °C compared with other plant extracts. At this juncture, it could be deduced that the application of vermicompost had little effect on the expression of phenolics, flavonoids, or anthocyanin in C. nutans. However, the extract from plants treated with vermicompost (VC and MF) showed better stability compared with CC and CF after extract storage at different temperatures.
Fulltext Optimization of culture conditions (sucrose, pH, and photoperiod) for in vitro regeneration and early detection of somaclonal variation in ginger lime (Citrus assamensis)

Yaacob JS, Mahmad N, Mat Taha R, Mohamed N, Mad Yussof AI, Saleh A

ScientificWorldJournal, 2014;2014:262710.
PMID: 24977187 DOI: 10.1155/2014/262710

Various explants (stem, leaf, and root) of Citrus assamensis were cultured on MS media supplemented with various combinations and concentrations (0.5-2.0 mg L(-1)) of NAA and BAP. Optimum shoot and root regeneration were obtained from stem cultures supplemented with 1.5 mg L(-1) NAA and 2.0 mg L(-1) BAP, respectively. Explant type affects the success of tissue culture of this species, whereby stem explants were observed to be the most responsive. Addition of 30 gL(-1) sucrose and pH of 5.8 was most optimum for in vitro regeneration of this species. Photoperiod of 16 hours of light and 8 hours of darkness was most optimum for shoot regeneration, but photoperiod of 24 hours of darkness was beneficial for production of callus. The morphology (macro and micro) and anatomy of in vivo and in vitro/ex vitro Citrus assamensis were also observed to elucidate any irregularities (or somaclonal variation) that may arise due to tissue culture protocols. Several minor micromorphological and anatomical differences were observed, possibly due to stress of tissue culture, but in vitro plantlets are expected to revert back to normal phenotype following full adaptation to the natural environment.
Fulltext Berberine Inhibits Telomerase Activity and Induces Cell Cycle Arrest and Telomere Erosion in Colorectal Cancer Cell Line, HCT 116

Samad MA, Saiman MZ, Abdul Majid N, Karsani SA, Yaacob JS

Molecules, 2021 Jan 13;26(2).
PMID: 33450878 DOI: 10.3390/molecules26020376

Colorectal cancer (CRC) is the most common cancer among males and females, which is associated with the increment of telomerase level and activity. Some plant-derived compounds are telomerase inhibitors that have the potential to decrease telomerase activity and/or level in various cancer cell lines. Unfortunately, a deeper understanding of the effects of telomerase inhibitor compound(s) on CRC cells is still lacking. Therefore, in this study, the aspects of telomerase inhibitors on a CRC cell line (HCT 116) were investigated. Screening on HCT 116 at 48 h showed that berberine (10.30 ± 0.89 µg/mL) is the most effective (lowest IC50 value) telomerase inhibitor compared to boldine (37.87 ± 3.12 µg/mL) and silymarin (>200 µg/mL). Further analyses exhibited that berberine treatment caused G0/G1 phase arrest at 48 h due to high cyclin D1 (CCND1) and low cyclin-dependent kinase 4 (CDK4) protein and mRNA levels, simultaneous downregulation of human telomerase reverse transcriptase (TERT) mRNA and human telomerase RNA component (TERC) levels, as well as a decrease in the TERT protein level and telomerase activity. The effect of berberine treatment on the cell cycle was time dependent as it resulted in a delayed cell cycle and doubling time by 2.18-fold. Telomerase activity and level was significantly decreased, and telomere erosion followed suit. In summary, our findings suggested that berberine could decrease telomerase activity and level of HCT 116, which in turn inhibits the proliferative ability of the cells.
Resource recovery from hydroponic wastewaters using microalgae-based biorefineries: A circular bioeconomy perspective

Ajeng AA, Rosli NSM, Abdullah R, Yaacob JS, Qi NC, Loke SP

J Biotechnol, 2022 Dec 10;360:11-22.
PMID: 36272573 DOI: 10.1016/j.jbiotec.2022.10.011

As the world's population grows, it is necessary to rethink how countries throughout the world produce food in order to replace the conventional and unsustainable agricultural techniques. Microalgae cultivation using a nutrient-rich solution from hydroponic systems not only presents a novel approach to solving problems pertaining to the impact of the discharges on the natural environment but also provides a plethora of other biotechnological applications particularly in the productions of high value-added products and plants growth stimulants, which can be potentially assimilated into the circular bioeconomy (CBE) in the hydroponic sector. In this review, the potential and practicability of microalgae to be merged into hydroponics CBE are reviewed. Overall, the integration of microalgal biorefineries in hydroponics systems can be realized after considering their Technology Readiness Level and System Readiness Level beforehand. Several suggestions on strains and hydroponics system improvement using existing biotechnological tools, Artificial Intelligence (AI) and nanobiotechnology in support of the CBE will be covered.
Magnesium oxide nanoparticles improved vegetative growth and enhanced productivity, biochemical potency and storage stability of harvested mustard seeds

Gautam A, Sharma P, Ashokhan S, Yaacob JS, Kumar V, Guleria P

Environ Res, 2023 Jul 15;229:116023.
PMID: 37121351 DOI: 10.1016/j.envres.2023.116023

A field study was conducted to investigate the influence of MgO-NPs priming on growth and development of mustard. Priming of mustard seeds before sowing with MgO-NPs at concentration 10, 50, 100, and 150 μg/ml enhanced the vegetative parameters of plants, with considerable increase in leaf area. MgO-NPs exposure increased the photosynthetic pigment accumulation in mustard that led to increase in biomass, carbohydrate content, and the yield in terms of total grain yield. Increased chlorophyll has simultaneously increased the oxidative stress in plants, and hence stimulated their antioxidant potential. A consistent increase was observed in the content of mustard polyphenols and activity of SOD, CAT, and APX on MgO-NPs exposure. MgO-NPs induced oxidative stress further reduced the protein content and bioavailability in mustard. We further, evaluated the influence of MgO-NPs on the quality of mustard harvested seeds. The seeds harvested from nanoprimed mustard possessed increased antioxidant potential and reduced oxidative stress. The carbohydrate and protein accumulation was significantly enhanced in response to nanopriming. Reduced chlorophyll content in seeds obtained from nanoprimed mustard indicated their potential for disease resistance and stability on long term storage. Therefore, the seeds harvested from MgO-NPs primed mustard were biochemically rich and more stable. Therefore, MgO-NPs priming can be potentially used as a novel strategy for growth promotion in plants where leaves are economically important and a strategy to enhance the seed quality under long term storage conditions.
Fulltext Effect of Plant Growth Regulators on Coloured Callus Formation and Accumulation of Azadirachtin, an Essential Biopesticide in Azadirachta indica

Ashokhan S, Othman R, Abd Rahim MH, Karsani SA, Yaacob JS

Plants (Basel), 2020 Mar 11;9(3).
PMID: 32168737 DOI: 10.3390/plants9030352

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.
Inhibitory impact of MgO nanoparticles on oxidative stress and other physiological attributes of spinach plant grown under field condition

Gautam A, Sharma P, Ashokhan S, Yaacob JS, Kumar V, Guleria P

Physiol Mol Biol Plants, 2023 Dec;29(12):1897-1913.
PMID: 38222280 DOI: 10.1007/s12298-023-01391-9

Green synthesis of NPs is preferred due to its eco-friendly procedures and non-toxic end products. However, unintentional release of NPs can lead to environmental pollution affecting living organisms including plants. NPs accumulation in soil can affect the agricultural sustainability and crop production. In this context, we report the morphological and biochemical response of spinach nanoprimed with MgO-NPs at concentrations, 10, 50, 100, and 150 µg/ml. Nanopriming reduced the spinach root length by 14-26%, as a result a reduction of 20-74% in the length of spinach shoots was observed. The decreased spinach shoot length inhibited the chlorophyll accumulation by 21-55%, thus reducing the accumulation of carbohydrates and yield by 46 and 49%, respectively. The reduced utilization of the total absorbed light further enhanced ROS generation and oxidative stress by 32%, thus significantly altering their antioxidant system. Additionally, a significant variation in the accumulation of flavonoid pathway downstream metabolites myricitin, rutin, kaempferol-3 glycoside, and quercitin was also revealed on MgO-NPs nanopriming. Additionally, NPs enhanced the protein levels of spinach probably as an osmoprotectant to regulate the oxidative stress. However, increased protein precipitable tannins and enhanced oxidative stress reduced the protein digestibility and solubility. Overall, MgO-NPs mediated oxidative stress negatively affected the growth, development, and yield of spinach in fields in a concentration dependent manner.
SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-023-01391-9.
Harnessing the potential of mutation breeding, CRISPR genome editing, and beyond for sustainable agriculture

Nor A'azizam NM, Chopra S, Guleria P, Kumar V, Abd Rahim MH, Yaacob JS

Funct Integr Genomics, 2024 Feb 29;24(2):44.
PMID: 38421529 DOI: 10.1007/s10142-024-01325-y

By 2050, the global population is projected to exceed 9.5 billion, posing a formidable challenge to ensure food security worldwide. To address this pressing issue, mutation breeding in horticultural crops, utilizing physical or chemical methods, has emerged as a promising biotechnological strategy. However, the efficacy of these mutagens can be influenced by various factors, including biological and environmental variables, as well as targeted plant materials. This review highlights the global challenges related to food security and explores the potential of mutation breeding as an indispensable biotechnological tool in overcoming food insecurity. This review also covers the emergence of CRISPR-Cas9, a breakthrough technology offering precise genome editing for the development of high-yield, stress-tolerant crops. Together, mutation breeding and CRISPR can potentially address future food demands. This review focuses into these biotechnological advancements, emphasizing their combined potential to fortify global food security in the face of a booming population.
Berberine and RNAi-Targeting Telomerase Reverse Transcriptase (TERT) and/or Telomerase RNA Component (TERC) Caused Oxidation in Colorectal Cancer Cell Line, HCT 116: An Integrative Approach using Molecular and Metabolomic Studies

Samad MA, Saiman MZ, Abdul Majid N, Karsani SA, Yaacob JS

Cell Biochem Biophys, 2024 Mar;82(1):153-173.
PMID: 38198024 DOI: 10.1007/s12013-023-01210-8

Colorectal cancer (CRC) is the most common cancer in both men and women and is associated with increased telomerase levels and activity. The potential downstream effects of TERT and/or TERC downregulation by berberine (a telomerase inhibitor) or RNA interference (RNAi) on various target RNAs, proteins, relative telomerase activity (RTA), relative telomere length (RTL), hydrogen peroxide concentration [H2O2], percentage of cell cycle distribution, cell size and granularity as well as cellular metabolites were explored in HCT 116 cell line. Knockdown of TERT decreased TERC. The downregulation of TERT and/or TERC caused increment of [H2O2], G0/G1 phase arrest in addition to decreased S and G2/M phases, as well as diminished cell size. RTL was later reduced as a result of TERT, TERT and/or TERC downregulation which decreased RTA. It was discovered that xanthine oxidase (XO) was significantly and positively correlated at FDR-adjusted p value
Fulltext Shading Effects on Leaf Gas Exchange, Leaf Pigments and Secondary Metabolites of Polygonum minus Huds., an Aromatic Medicinal Herb

Mohd Yusof FF, Yaacob JS, Osman N, Ibrahim MH, Wan-Mohtar WAAQI, Berahim Z, et al.

Plants (Basel), 2021 Mar 23;10(3).
PMID: 33806923 DOI: 10.3390/plants10030608

The growing demand for high value aromatic herb Polygonum minus-based products have increased in recent years, for its antioxidant, anticancer, antimicrobial, and anti-inflammatory potentials. Although few reports have indicated the chemical profiles and antioxidative effects of Polygonum minus, no study has been conducted to assess the benefits of micro-environmental manipulation (different shading levels) on the growth, leaf gas exchange and secondary metabolites in Polygonum minus. Therefore, two shading levels (50%:T2 and 70%:T3) and one absolute control (0%:T1) were studied under eight weeks and 16 weeks of exposures on Polygonum minus after two weeks. It was found that P. minus under T2 obtained the highest photosynthesis rate (14.892 µmol CO2 m-2 s-1), followed by T3 = T1. The increase in photosynthesis rate was contributed by the enhancement of the leaf pigments content (chlorophyll a and chlorophyll b). This was shown by the positive significant correlations observed between photosynthesis rate with chlorophyll a (r2 = 0.536; p ≤ 0.05) and chlorophyll b (r2 = 0.540; p ≤ 0.05). As the shading levels and time interval increased, the production of total anthocyanin content (TAC) and antioxidant properties of Ferric Reducing Antioxidant Power (FRAP) and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) also increased. The total phenolic content (TPC) and total flavonoid content (TFC) were also significantly enhanced under T2 and T3. The current study suggested that P.minus induce the production of more leaf pigments and secondary metabolites as their special adaptation mechanism under low light condition. Although the biomass was affected under low light, the purpose of conducting the study to boost the bioactive properties in Polygonum minus has been fulfilled by 50% shading under 16 weeks' exposure.
Genetic and phenotype recovery of Ananas comosus var. MD2 in response to ionizing radiation

Ma S, Mohd Raffi AN, Rosli MA, Mohd Zain NA, Ibrahim MH, Karsani SA, et al.

Sci Rep, 2023 Jan 05;13(1):182.
PMID: 36604574 DOI: 10.1038/s41598-022-26745-3

Due to their sessile nature, plants are exposed to various environmental stressors such as exposure to high levels of harmful ultraviolet (UV), ionizing, and non-ionizing radiations. This exposure may result in various damages, ranging from DNA and chromosomal aberrations to phenotypic abnormalities. As an adaptation, plants have evolved efficient DNA repair mechanisms to detect and repair any damage caused by exposure to these harmful stressors to ensure their survival. In this study, the effects of gamma radiation (as a source of ionizing radiation) on clonal Ananas comosus var. MD2 was evaluated. The morphology and physiology of the clonal plantlets before and after exposure to gamma radiation were monitored at specific time intervals. The degree of genetic variation between the samples pre- and post-irradiation was also analyzed by using inter-simple sequence repeat (ISSR) markers. The resulting data revealed that the heights of the irradiated plantlets were significantly reduced (compared to control), but improved with the recovery period. Irradiated samples also exhibited relatively good photosynthetic efficiency that further improved as the plantlets recover. These observations were supported by the ISSR analysis, where the genetic dissimilarities between the irradiated samples and control were reduced by 0.1017, after 4 weeks of recovery. Overall, our findings suggested that the phenotype recovery of the clonal A. comosus var. MD2 plantlets was contributed by their ability to detect and repair the DNA lesions (as exemplified by the reduction in genetic dissimilarity after 4 weeks) and hence allow the plantlets to undergo phenotype reversion to normal plant stature.

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