REVIEW ARTICLE ON ESTABLISHMENT OF CELL SUSPENSION CULTURE OF IXORA COCCINEA AND INVESTIGATION OF ITS BIOCHEMICAL COMPOUNDS

REVIEW ARTICLE ON ESTABLISHMENT OF CELL SUSPENSION CULTURE OF IXORA COCCINEA AND INVESTIGATION OF ITS BIOCHEMICAL COMPOUNDS

Muhammed Rashad ^*, Salman Fayis. P. K, Shabeeh Bin Anwar M. T, Aashiq Hussain D. A.R, Basit Rashid Bhat, Celestin Baboo R. V and Sirajudheeen M. K

Jamia Salafiya Pharmacy College, Malappuram, Kerala, India.

ABSTRACT: This literature review comprehensively explores the scientific foundation for establishing cell suspension cultures of Ixora coccinea and investigating its biochemical compounds. Ixora coccinea, a species native to India and utilized extensively in traditional Ayurvedic and folk medicine, is recognized for its diverse pharmacological activities attributed to a rich array of secondary metabolites. The plant's classification as a threatened species underscores the imperative for sustainable production methods, which plant cell suspension culture effectively addresses. This review details the botanical and phytochemical profile of I. coccinea, elucidates the principles and methodologies of plant cell suspension culture, examines the current status of Ixora in-vitro studies and relevant examples from the Rubiaceae family, and outlines qualitative and quantitative analytical techniques for biochemical compound identification. While offering significant advantages such as scalability and controlled production, plant cell culture presents challenges including genetic instability and contamination. The successful implementation of this project holds substantial promise for the sustainable bioproduction of valuable pharmaceutical compounds, contributing to both natural product research and biodiversity conservation.

Keywords: Ixora coccinea, Cell culture, Biochemical compounds

INTRODUCTION:

Overview of Medicinal Plants and Secondary Metabolites: Medicinal plants have historically formed the bedrock of traditional medicine systems globally, including Ayurveda in India, leveraging various plant parts for therapeutic interventions ¹. The profound efficacy of these botanical resources is primarily due to specialized organic compounds known as secondary metabolites. Unlike primary metabolites, which are essential for growth, secondary metabolites fulfill crucial roles in plant survival, defense, and ecological interactions ⁵.

These compounds, typically categorized into alkaloids, terpenes, and phenolics, exhibit diverse biological properties ⁵. Secondary metabolites are highly sought after in pharmaceuticals, food processing, cosmetics, and dye manufacturing ¹. However, their natural production in intact plants is low, necessitating large-volume harvesting ⁵. Traditional sourcing is also vulnerable to geopolitical instabilities, environmental fluctuations, and slow growth rates ⁵. This creates a bottleneck in the sustainable supply of these valuable compounds, making alternative, controlled, and efficient production methods imperative.

Rationale for In-vitro Culture in Natural Product Research: Plant tissue culture, a biotechnological approach, offers a compelling solution to challenges in natural product sourcing. This technique is based on "tot potency," the principle that a single plant cell can regenerate into a complete plant under appropriate conditions ⁶. It enables aseptic cultivation of isolated plant cells, tissues, or organs in a sterile, controlled growth medium, detached from native environmental conditions ⁶.

A significant advantage is the "large-scale production of specific secondary metabolites" ¹. This controlled environment ensures a standardized, continuous, and reliable supply, free from seasonal or climatic variations ⁵. Plant tissue culture also plays a vital role in mitigating the "continuous depletion of plants" and their habitats, contributing directly to the "conservation of wild populations" ⁵.

Beyond supply and conservation, in vitro plant cell culture provides a platform for bioprospecting and drug discovery by enabling precise manipulation of metabolite profiles. Control over the culture medium, growth regulators, and elicitors allows researchers to induce or enhance biosynthesis of specific secondary metabolites ⁵.

This facilitates systematic investigation into compound production mechanisms and their applications, positioning cell culture as a dynamic tool for pharmaceutical innovation.

Botanical and Phytochemical Profile of Ixora coccinea:

Taxonomy and Distribution: Ixora coccinea belongs to the Rubiaceae family (Madder family) ¹. The genus Ixora comprises approximately 500 species, primarily in tropical Asia and Africa ³. I. coccinea is indigenous to Karnataka and Kerala in India ¹ and widely cultivated as an ornamental plant across India and South Asia ⁴.

Morphologically, I. coccinea is a "dense, multi-branched evergreen shrub," typically 1.2–2 meters tall, though it can reach 3.6 meters ². It has a rounded form, often spreading wider than its height ².

Its adaptability to diverse soil types and climates ¹⁶, despite its classification as a "threatened species" in some contexts, suggests genetic robustness. This hardiness indicates its amenability to in-vitro environments, simplifying initial explant collection and callus induction.

FIG. 1: IXORA COCCINEA

Traditional uses and Ethno-botanical Significance: For centuries, various parts of Ixora coccinea (flowers, leaves, roots, stem) have been integrated into traditional Indian medicine, Ayurveda, and folk practices ¹. Its traditional applications are broad, treating hepatic disorders, cancers, microbial infections, pain, inflammatory conditions, nausea, hiccups, anorexia, sores, chronic ulcers, sprains, eczema, boils, contusions, diarrhea, fever, bronchitis, dysentery, hypertension, irregular menstruation, female reproductive organ infections, hemoptysis, and asthma ^{2, 4}. Ripe fruits are also consumed ¹. Its traditional use in wound healing is particularly well-documented ⁴.

The broad spectrum of traditional medicinal uses strongly suggests a complex array of bioactive compounds, each potentially contributing to different pharmacological effects. This extensive traditional knowledge provides a robust rationale for investigating the "useful biochemical compounds... qualitatively and quantitatively" from I. coccinea suspension culture ¹.

Major Phytochemical Constituents: Phytochemical investigations reveal Ixora coccinea is rich in secondary metabolites, including triterpenoids (lupeol, ursolic acid, oleanolic acid, betunolic acid, amyrins, ixorene), aromatic acrid oils, tannins, saponins, carbohydrates, fatty acids, and flavonoids (rutin, formononetin, β-sitosterol, quercetin, kaempferol). Sterols, lecocyanadin, anthocyanins, and proanthocyanidins have also been identified ¹. Essential oil from I. coccinea flowers predominantly contains triterpenes (62.60%), monoterpenes (31.73%), sesquiterpenes (3.35%), and esters (2.29%) ². Major triterpenesare ursolic acid (27.34%), oleanolic acid (20.16%), and lupeol (15.10%), classifying I. coccinea flower as an "ursolic acid chemotype" ².

Quantitative analysis shows significant amounts of alkaloids, phenols, flavonoids, saponins, and tannins ¹⁹.

The dominance of specific triterpenes like ursolic acid, oleanolic acid, and lupeol, combined with documented pharmacological activities, suggests these are keyactive principles. For this project, optimizing conditions in cell suspension culture to maximize these high-value triterpenes is crucial.

Documented Pharmacological Activities: Ixora coccinea exhibits a broad spectrum of scientifically documented pharmacological activities, including antioxidant, anti-inflammatory, antimicrobial, hepatoprotective, gastroprotective, anti-diarrheal, anti-nociceptive, anti-mutagenic, anti-neoplastic, chemopreventive, and wound healing effects ².

Specific studies highlight its potent antioxidant activity against reactive oxygen species (ROS), with an IC₅₀ of approximately 2.8 µg/mL ⁹. Its anti-inflammatory properties are evidenced by membrane stabilization ¹². I. coccinea enhances cutaneous wound healing by upregulating collagen and basic fibroblast growth factor expression ¹¹. Its anti-asthmatic properties are linked to mitigating airway inflammation ¹².

The table below summarizes key phytochemicals and their documented pharmacological activities:

TABLE 1: KEY PHYTOCHEMICALS AND DOCUMENTED PHARMACOLOGICAL ACTIVITIES OF IXORA COCCINEA

Conservation Status and Importance of Sustainable Sourcing: The project protocol explicitly states that Ixora coccinea is considered a "threatened species" ¹. This underscores the critical need for sustainable production methods for its valuable biochemical compounds. The broader context of plant conservation highlights that "continuous depletion of plants poses a significant threat to plant species and their natural habitats, leading to the extinction of many valuable and endemic species" ⁵. Plant cell cultures are a "promising alternative" for producing high-value secondary metabolites "without needing entire plants" ⁵, directly contributing to the "preservation of wild populations" ⁵. The threatened status of I. coccinea elevates this research to one with ecological and ethical implications, aligning it with global conservation efforts by offering a pathway to harness the plant's medicinal properties sustainably.

Principles and Methodology of Plant Cell Suspension Culture:

Fundamentals of Plant Tissue Culture and Cell Totipotency: Plant tissue culture is a biotechnological technique rooted in the fundamental principle of "totipotency" ⁶. This principle states that a single plant cell, given appropriate cues, possesses the genetic capability to differentiate and regenerate into a complete plant organism ⁶. This attribute forms the theoretical basis for in-vitro plant propagation. The technique involves aseptic cultivation of isolated plant cells, tissues, or organs in a sterile, controlled growth medium, detached from their natural environment ⁶. A key application is "rapid and disease-free micropropagation of plantlets," independent of external seasonal, climatic, or geographic factors, ensuring a consistent supply of plant material ⁶.

Establishment of Cell Suspension Cultures: Establishing cell suspension cultures typically begins with selecting a suitable explant, a small piece of plant tissue, ideally froma prolific producer of the desired metabolite ⁵. The explant undergoes thorough surface-sterilization to eliminate microbial contamination ¹. Sterile explants are then aseptically placed on a solid nutrient medium (agar-based) to induce callus formation ¹. Callus is an undifferentiated, proliferating mass of plant cells ⁵. For suspension cultures, "friable callus" which readily breaks into single cells or small aggregates is chosen due to its dissociation propensity in liquid media ¹. Small pieces of friable callus are transferred into a liquid nutrient medium in a flask ¹. The flask is placed on an orbital or rotary shaker for continuous agitation, ensuring aeration, uniform cell and nutrient distribution, and efficient gaseous exchange, all vital for cell proliferation ¹. As cells divide, they disperse, forming the cell suspension culture ⁵.

To sustain robust growth and large-scale production, "periodically transfer a portion of the suspension culture to fresh liquid medium" via subculturing ¹. During subculturing, larger aggregates are discarded, maintaining a uniform, actively growing population ⁵. Cell growth can be monitored by direct cell counting (haemocytometer) or measuring packed cell volume (PCV) ¹.

Optimization of Culture Conditions for Cell Growth and Metabolite Production: Optimizing culture conditions is critical for enhancing cell proliferation and secondary metabolite accumulation in plant cell suspension cultures ⁵.

Medium Composition: The liquid medium must be "supplemented with necessary nutrients and growth regulators (hormones)" ¹. Adjustments to nutrient composition, carbon source, and osmotic stabilizers can "simulate conditions like drought," influencing metabolic processes and leading to biomass and secondary metabolite buildup ⁵.

Plant Growth Regulators (PGRs): Judicious addition of auxins (e.g., 2,4-D, NAA, IAA, IBA) and cytokinins (e.g., BAP, KIN, TDZ) significantly impacts callus induction, shoot regeneration, and secondary metabolite synthesis ⁵. For Ixora species, Murashige and Skoog (MS) medium with BAP and NAA has been effective for shoot regeneration and callus induction ¹⁵.

Carbon Source: The type and concentration of the carbon source (e.g., glucose-fructose mixtures, saccharides) are paramount for maximizing productivity, as high starting cell density and increased saccharide levels can boost metabolite synthesis ⁵.

Nitrogen Source: The nitrogen source and its concentration (e.g., varying NO3-/NH4+ ratios) also influence cell proliferation and metabolite accumulation ⁵.

Environmental Factors: Maintaining "optimal conditions like temperature, light, and humidity" is essential and species-specific ¹. Environmental stressors (heat, salinity, dryness, light intensity) can significantly influence secondary metabolite accumulation ⁵. Light irradiation can stimulate anthocyanin synthesis, with "light-grown cultures often producing more phenolic compounds" ⁵. Proper temperature treatment is also necessary ⁵.

Elicitation: Elicitors trigger specific signal transduction cascades in plant cells, activating defense systems and enhancing secondary metabolite production ⁵. These include chemical elicitors (e.g., jasmonic acid, methyl jasmonate, synthetic analogs of salicylic acid) and biological elicitors (e.g., fungal components) ⁵. "Jasmonic acid and methyl jasmonate are critical regulators of secondary metabolite synthesis" ⁸. The variability of secondary metabolite production implies that investigating biochemical compounds will require a systematic optimization strategy. Simply establishing a culture does not guarantee desired yields; understanding and manipulating these cues is vital. This highlights the need for a dedicated optimization phase, potentially involving factorial experimental design, to identify favorable conditions for specific compound biosynthesis.

Advantages of Plant Cell Suspension Cultures as Bio-factories: Plant cell cultures offer compelling advantages as bio-factories, being a "cost-effective alternative to traditional cultivation methods" and "more reliable than collecting plants from the wild" ⁸.

Scalability: Cultures can be "scaled up for large-scale production" in bioreactors, providing "ideal conditions for large-scale plant production for commercial purposes" ⁵.

Controlled Conditions: Production occurs under highly controlled, sterile conditions, "unaffected by soil or climate changes" ⁵. This ensures a "constant supply of product" with standardized levels, minimizing variability ⁶.

Genetic Amenability: Plant cells are amenable to "genetic transformation" and "metabolic engineering," enhancing secondary metabolite production ⁵. They perform crucial "post- translational modifications" and possess Cytochrome P450 (CYP450) enzymes vital for biosynthesis, offering an advantage over microbial systems ⁸.

Conservation: This method significantly contributes to "conservation of wild populations," especially for threatened species like I. coccinea, by reducing reliance on wild harvesting ⁵.

Safety and Environmental Friendliness: Plant cell cultures offer a "safe, low cost, and environmentally friendly manner" for producing valuable metabolites, aligning with sustainability goals ⁸. Transgenic plant suspension cultures generally face “fewer regulatory issues compared with whole plants" ⁸.

Challenges and Limitations in Cell Suspension Culture for Secondary Metabolite Production: Despite advantages, industrial application of plant cell suspension cultures faces significant challenges:

Efficiency and Growth: Issues include "poor cell efficiency," "slow growth" rates, and "inadequate regulation of cellular differentiation," limiting productivity and economic viability ⁵.

Genetic Instability: "Genetic instability of high-producing cell lines" can lead to a "decrease" in product synthesis after passages, necessitating continuous screening and selection of stable lines ⁵.

Nutrient Requirements: Cultures often "require a carbon source and sometimes precursors" for optimal metabolite production, adding to medium cost and complexity ⁸.

Complex Genetics: For "non-model plant suspension culture[s]," complex genetic content can make genetic manipulation challenging ⁸.

Product Inhibition and Toxicity: Accumulation of the desired product or toxic intermediates can lead to "production inhibition" or cell death ⁸.

Browning of Medium: A common problem in Ixora micropropagation is "exudation of phenolics," causing "browning of medium" ¹⁵. This can be mitigated by "repeated subculturing" ¹⁵.

Contamination: High microbial contamination persists in liquid cultures, requiring stringent "proper surface sterilization of the explant" and aseptic conditions ¹⁵.

These challenges underscore that establishing I. coccinea suspension culture is an initial step; long-term viability and high-yield production demand continuous monitoring and adaptive strategies, including ongoing cell line selection, optimization of subculturing, and addressing product toxicity.

Secondary Metabolite Production in Ixora Species and Related Rubiaceae Family via Cell Culture:

General Overview of Secondary Metabolite Biosynthesis in Plants: Plants produce a vast array of secondary metabolites, which are not essential for primary metabolic functions but are crucial for plant-environment interactions ⁶. These compounds accumulate in response to biotic (bacteria, viruses, fungi, insects, herbivores) and abiotic factors (climatic variations, soil/water parameters) ⁶.

Biosynthesis pathways are mediated by complex cellular messengers, metabolic changes, gene activation, and signaling cascades ⁶. Secondary metabolites are broadly categorized into phenolics, terpenes, and alkaloids ⁵. They exhibit immense chemical and biological diversity and are often "species- and organ-specific" ⁵. Understanding these pathways is crucial for manipulating and enhancing secondary metabolite production in-vitro ¹⁷.

Current Status of Ixora Species Cell Culture: While direct evidence of large-scale secondary metabolite production from Ixora coccinea cell suspension culture is limited in the provided literature, existing research on in-vitro propagation and callus induction of Ixora species, including I. coccinea, provides a crucial foundation.

Micropropagation of Ixora typically involves culturing shoot tips on Murashige and Skoog (MS) medium with cytokinin ¹⁵. Callus initiation in I. coccinea has been successful from sterile leaves or flower petals, responding to 2,4-D and TDZ ¹⁵. Optimal shoot regeneration conditions involve BAP (4.0 mg/l) ¹⁵. Subculture on MS medium with BAP (1.0 mg/l) and NAA (0.5 mg/l) achieved high survival (80.8%) and rooting (30.5%) ratios ¹⁵. Calli have been produced from various explants on media containing KIN, NAA, or 2,4-D ¹⁵.

Since callus induction is demonstrated for Ixora, the initial step for establishing suspension culture is supported by existing protocols. This foundational success suggests I. coccinea cells are amenable to in vitro manipulation, a prerequisite for initiating suspension cultures. This allows leveraging established methods, potentially reducing time for preliminary optimization.

Common challenges in Ixora micropropagation include "browning of medium" due to phenolic exudation, mitigated by "repeated subculturing," and "high degree of contamination," requiring rigorous "proper surface sterilization of the explant" ¹⁵. Addressing these known challenges is critical for successful establishment and maintenance of I. coccinea cell suspension cultures.

Examples of Secondary Metabolite Production from Rubiaceae Family in Cell Culture: The Rubiaceae family, which includes Ixora coccinea, is known for its diverse bioactive metabolites ¹⁰. Studies on various genera within this family demonstrate the feasibility of producing high-value secondary metabolites through cell culture. These compounds often serve as chemotaxonomic markers ¹⁰. The Rubiaceae family produces iridoids, indole alkaloids, anthraquinones, terpenoids (diterpenes and triterpenes), flavonoids, and other phenolic derivatives, with an emphasis on bioactive alkaloids ¹⁸. For example, Uncaria, Psychotria, Hedyotis, and Ophiorrhiza general produce iridoids, anthraquinones, triterpenes, and various alkaloid subclasses ¹⁸.

Morinda species are known for anthraquinones like alizarin ¹⁸. Significant examples of secondary metabolite production from Rubiaceae species via cell culture include:

Cinchona species: Historical source of quinine, an anti-malarial alkaloid. Cell culture offers a controlled environment for production ¹⁸.

Psychotria species: Psychotria viridis produces β-carboline indole alkaloids (harmine, harmaline, tetrahydroharmine) with CNS activity ¹⁸.

Cephaelis ipecacuanha: An important source of emetine, analkaloid with emetic, antihelminthic, and expectorant effects ¹⁸.

Genipa americana: Yields genipin, aniridoid with antiangiogenic, anti-inflammatory, and antioxidant activity ¹⁸.

The success in producing various secondary metabolites from diverse Rubiaceae species in-vitro provides a strong precedent for the potential of Ixora coccinea cell suspension cultures. The family's capacity for producing a wide range of pharmacologically active compounds through plant cell culture supports the feasibility of the proposed project for I. coccinea.

Potential for Biochemical Compound Production from Ixora coccinea Suspension Culture: Given the rich phytochemical profile of Ixora coccinea and the documented success of cell culture in producing secondary metabolites from other Rubiaceae species, there is substantial potential for bioproduction of valuable biochemical compounds from I. coccinea cell suspension cultures. The plant contains significant amounts of triterpenoids (e.g., ursolicacid, oleanolic acid, lupeol), flavonoids (e.g., rutin, quercetin, kaempferol, anthocyanins), alkaloids, and phenolics, all with documented pharmacological activities like antioxidant, anti-inflammatory, antimicrobial, and wound-healing effects ¹.

Plant cell suspension cultures offer a controlled environment where production of these metabolites can be optimized and scaled up ¹. This approach mitigates issues with traditional cultivation, such as low natural yields, environmental variability, and the species' threatened status ⁵. By manipulating culture conditions (nutrient composition, plant growth regulators, elicitors), biosynthesis and accumulation of desired compounds can be significantly enhanced ⁵. The project's aim to "investigate the presence of useful biochemical compounds of suspension culture of Ixora coccinea qualitatively and quantitatively" ¹ is well-justified and holds promise for discovering and producing high- value compounds for pharmaceutical applications.

Qualitative and Quantitative Analysis of Biochemical Compounds: The comprehensive investigation of biochemical compounds from Ixora coccinea cell suspension cultures requires robust extraction procedures followed by qualitative screening and advanced quantitative analytical techniques.

Extraction Procedures: Efficient extraction is the initial step. Various methods include:

Hydroalcoholic Extraction and Fractionation: Macerating powdered plant material in a hydroalcoholic solvent (e.g., EtOH–H2O 70:30) at room temperature, followed by filtration, concentration, and sequential extraction with solvents of increasing polarities (e.g., chloroform, ethyl acetate, n-butanol) ²⁰. Column chromatography can further isolate compounds ²⁰.

Supercritical Fluid Extraction (SFE): Uses supercritical carbon dioxide as a solvent, efficient for non-polar to moderately polar compounds under controlled temperature and pressure (e.g., 40 °C, 10-30 MPa). It is environmentally friendly and leaves no solvent residues ²⁰.

Microwave-Assisted Extraction (MAE): Employs microwave irradiation to heat solvents and plant tissues, increasing extraction kinetics, reducing time and solvent waste, and promoting higher extraction rates. Performed in sealed vessels at specific temperatures and durations (e.g., 40-120 °C for 5-15 minutes) ²⁰.

The choice of extraction method significantly influences the yield and profile of recovered compounds ²⁰.

Qualitative Phytochemical Screening Methods: Qualitative screening involves rapid, cost-effective tests to identify major secondary metabolite classes, typically relying on color reactions or precipitate formation with specific reagents ²¹:

Alkaloids: Brown color with Dragendroff’s reagent ²¹.

Tannins: Brownish-black color with ferric chloride solution ²¹.

Flavonoids: Reddish color with magnesium ribbon fragments and concentrated hydrochloric acid ²¹.

Sterols and Triterpenoids: Reddish (steroids) or yellow (triterpenoids) lower layer with concentrated sulfuric acid ²¹.

Carbohydrates: Absence of red precipitates with Benedict’s reagent upon boiling ²¹.

Fats and Oils: Formation of soap or partial neutralization with alcoholic potassium hydroxide and phenolphthalein upon heating ²¹.

Glycosides: Pink ammonia cal layer after hydrolysis, benzene extraction, and ammonia treatment ²¹.

Saponins: Persistent foam upon vigorous shaking with water ²¹.

These tests provide initial insights into the broad chemical composition, guiding subsequent quantitative analyses.

Advanced Quantitative Analytical Techniques: For precise qualitative identification and quantitative determination, advanced analytical techniques are indispensable due to their high sensitivity, specificity, and resolution:

High-Performance Liquid Chromatography (HPLC): Often coupled with a photodiode array (PDA) detector, HPLC is widely used for simultaneous estimation of phenolic compounds and other metabolites. Compounds are identified by retention time and UV-Vis spectra, compared with standards ²⁰.

Gas Chromatography-Mass Spectrometry (GC-MS): Effective for volatile and semi- volatile compounds, including essential oil components like triterpenes, monoterpenes, and sesquiterpenes found in I. coccinea ². Compounds are separated in the gas phase and identified by mass fragmentation patterns, often using spectral libraries ²³.

Liquid Chromatography-Mass Spectrometry (LC-MS): A powerful technology for plant metabolomics, known for high sensitivity. It couples LC separation with MS identification, allowing comprehensive characterization of specialized metabolites like flavonoids, phenylpropanoids, and oxylipins ²².

Nuclear Magnetic Resonance (NMR) Spectroscopy: Increasingly used for metabolite fingerprinting of natural extracts, offering direct quantitative information without prior chromatographic separation ²². It is widely employed for characterization and structure determination of natural products, providing accurate qualitative and quantitative data from complex mixtures ². For example, 1D and 2D-NMR elucidated the structure of ixorenefrom I. coccinea leaves ².

UV-Vis Spectrophotometry: Measures light absorption in UV and visible regions. Commonly used for quantitative determination of total phenolic content, total flavonoid content, and specific pigments based on characteristic absorption spectra ²⁰.

Integrating these techniques will enable thorough and precise investigation of biochemical compounds from Ixora coccinea cell suspension culture, fulfilling the project's objective.

Potential Applications of Ixora coccinea Biochemical Compounds: The biochemical compounds from Ixora coccinea cell suspension culture hold significant potential for various industrial applications, particularly in pharmaceuticals. The plant's documented pharmacological activities (antioxidant, anti-inflammatory, antimicrobial, wound-healing) suggest a wide range of therapeutic uses for its isolated metabolites ².

High-value compounds like ursolic acid, oleanolic acid, and lupeol, major triterpenes in I. coccinea, are known for their anti-inflammatory, antioxidant, and anti-cancer properties ². These could be developed into novel drug candidates or active pharmaceutical ingredients. Flavonoids (rutin, quercetin, kaempferol) offer antioxidant and anti-inflammatory benefits, suitable for nutraceuticals or functional food additives ². Beyond pharmaceuticals, I. coccinea secondary metabolites could find applications in cosmetics (antioxidant, skin- healing) and food (natural colorants, preservatives) ⁸.

CONCLUSION: The establishment of Ixora coccinea cell suspension culture and subsequent investigation of its biochemical compounds represent a scientifically sound and critically important endeavor. Ixora coccinea is a traditionally significant medicinal plant, rich in diverse secondary metabolites with documented pharmacological activities, including potent antioxidant, anti- inflammatory, and wound-healing properties. Its classification as a threatened species underscores the imperative for sustainable and controlled production methods.

Plant cell suspension culture, leveraging the principle of totipotency, offers a viable and advantageous alternative to traditional sourcing, providing a scalable, consistent, and environmentally responsible platform for producing these valuable compounds. While challenges such as genetic instability and contamination exist, these can be systematically addressed through rigorous optimization, careful cell line management, and advanced biotechnological strategies. The existing success in in-vitro propagation of Ixora species and the proven capacity of other Rubiaceae members to produce secondary metabolites in cell culture provide a strong foundation for this project. The qualitative and quantitative analysis of the compounds produced will not only validate the efficacy of the suspension culture but also pave the way for the development of novel pharmaceutical agents, contributing significantly to both natural product research and the conservation of this valuable species. The successful execution of this project promises to unlock the full therapeutic potential of Ixora coccinea through sustainable bioproduction.

ACKNOWLEDGEMENT: Nil

CONFLICT OF INTEREST: Nil

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    How to cite this article:

    Rashad M, Fayis PKS, Binanwar MTS, Hussain DARA, Bhat BR, Baboo RVC and Sirajudheeen MK: Review article on establishment of cell suspension culture of Ixora coccinea and investigation of its biochemical compounds. Int J Pharmacognosy 2025; 12(8): 621-29. doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.12(8).621-29.

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