Ocimum Sanctum Research Articles

Disinfection Ability of Ocimum sanctum Extract against Candida albicans Inoculated on Denture Resin and Soft Liner: An In Vitro Study

Disinfection Ability of Ocimum sanctum Extract against Candida albicans Inoculated on Denture Resin and Soft Liner: An In Vitro Study

Impact of Treated Castor Leaves on Growth and Productivity of Eri Silkworms: A Bioassay Study

One of the most common foliar diseases in castor cultivation for ericulture is Cercospora leaf spot, caused by Cercospora ricinella Sacc. & Berl. which seriously damages leaves meant as food for eri silkworms. Deterioration in the quality or quantity of castor leaves primarily hampers the health of eri silkworm, which ultimately affects the economic parameters of the cocoons produced by them. A bioassay study was conducted during 2019-2021 at College of Sericulture, Assam Agricultural University, Jorhat, Assam, to check the effects of feeding castor leaves, treated with treatments found promising for eco-friendly management of Cercospora leaf spot of castor. Eight treatments with three replications were taken for the experiment which included a phytoextract of Tulsi (Ocimum sanctum) @ 5%, a bio agent (Trichoderma viride) @ 5% and a fungicide (Copper oxychloride) @ 0.2 % and their combinations, applied as foliar spray at 15 days interval after 30 days of transplanting. These treated leaves were fed as first feed to the 3rd instar silkworms after maintaining 25-30 days as safe period. Silkworms fed with castor leaves treated with T.viride + Tulsi extract showed the best results in overall health (ERR 91.66%, Fecundity 391, Shell ratio 13.44%) followed by worms fed with leaves treated with Tulsi extract + T. viride + COC (ERR 80.55%, Fecundity 370, Shell ratio 12.59%).

In vitro efficacy of essential oils against various Candida species

Candida species are responsible for the most common fungal infections worldwide. We studied the in vitro antifungal activity of a large panel of essential oils (EOs) against various Candida species. The EOs activity against Candida spp. was tested using a gradient microdilution assay ranging from 4% to 0.008% (v/v). After a preliminary screening including 31 EOs, seven selected EOs were tested against 13 clinical isolates and four reference strains belonging to six Candida species. Cinnamomum zeylanicum and Cymbopogon giganteus EOs exhibited the best antifungal activity against all clinical and reference strains, with MIC ranges of 0.015%–0.25% (v/v). EOs from Litsea citrata, Backhousia citriodora and Ocimum sanctum presented MIC ranges of 0.03%–0.5% (v/v). The antifungal efficacy of EOs was independent of the susceptibility of Candida strains to usual antifungal agents. These EOs could have a promising antifungal action.

Enhanced In Vitro Plant Morphogenesis of Tobacco: Unveiling Indoleamine-Modulated Adaptogenic Properties of Tulsi (Ocimum sanctum L.).

The medicinal plant tulsi (Ocimum sanctum L.) is acknowledged for its invigorating and healing properties that enhance resilience to stress in various human and animal models by modulating antioxidant compounds. While extensive research has documented these effects in humans, the adaptogenic potential of tulsi in stressful in vitro plant systems has not been explored. This study aimed to elucidate the adaptogenic properties of tulsi leaf extract on the in vitro regeneration of tobacco leaf explants through an investigation of the indoleamines at different developmental stages. Shoot regeneration from leaf explants on the medium supplemented with tulsi extract (20%) was compared to the control, and the differences in indoleamine compounds were analyzed using ultra-performance liquid chromatography. Treatment of the explants with the extract resulted in an almost two-fold increase in the number of regenerants after four weeks of culture, and 9% of the regenerants resembled somatic embryo-like structures. The occurrence of browning in the extract-treated explants stopped on day 10, shoots began to develop, and a significant concentration of tryptamine and N-acetyl-serotonin accumulated. A comparative analysis of indoleamine compounds in intact and cut tobacco leaves also revealed the pivotal role of melatonin and 2-hydroxymelatonin functioning as antioxidants during stress adaptation. This study demonstrates that tulsi is a potent adaptogen that is capable of modulating plant morphogenesis in vitro, paving the way for further investigations into the role of adaptogens in plant stress biology.

Application and evaluation of plant-based edible active coatings to enhance the shelf-life and quality attributes of Jara lebu (Citrus medica)

Edible coatings for fruits and vegetables are the subject of intensive agro-based research. These coatings provide value to the product due to their multifunctionality and sustainability. The current study focuses on the development and evaluation of plant-based edible active coatings for Jara lebu (Citrus medica), with the aim of determining the effectiveness of these coatings in extending the shelf-life and preserving the quality attributes. Different blends of corn starch and various plant extracts were formulated and all formulations were applied by immersion onto the fruit surface. The study had five treatment groups: T0, T1, T2, T3, and T4. T0 served as the control group, while T1 consisted of a mixture of 2% corn starch and 0.5% glycerol. T2 included a combination of 2% corn starch, 0.5% glycerol, and 1.5% holy basil. T3 comprises 2% corn starch, 0.5% glycerol, and 1.5% wild turmeric. Lastly, T4 consisted of 2% corn starch, 0.5% glycerol, and 1.5% Indian pennywort. Control and coated samples were kept under the same conditions for 35 days before being evaluated for changes in their physiological, physicochemical, and sensory qualities. Coated sample T2 significantly prolonged the shelf-life of Jara lebu samples, having the least weight reduction (26.25%) and retaining most of the essential nutrients (TSS = 7.09%, pH = 3.0, vitamin C = 22.03 mg/100 g, TPC = 44.57 mg GAE/g DW, TFC = 45.24 mg QE/g DW, antioxidant = 86.09%). This sample received the highest overall acceptability score, a maximum of 8.24. Sensory evaluations revealed no adverse effects on taste, aroma or appearance, suggesting these coatings can be an eco-friendly and efficient method for preserving the freshness and quality of Jara lebu and potentially other citrus fruits.

Ethnomedicinal knowledge and utilization of the medicinal plant resources by the tribal people of the Jhargram District, West Bengal, India

The study strives to investigate and document the traditional ethnomedicinal knowledge of indigenous plant species used by the tribal people of Jhargram district, West Bengal, India. It has been stated by the 142 informants that, out of a total of 80 plant species, 45 classes, 46 orders, 46 families and 75 genera are generally used as medicinal plants to cure several diseases. The most dominant medicinal plant families found in this region are Fabaceae, Apocynaceae, Anacardiaceae, Lamiaceae, Acanthaceae and Malvaceae. The most important species based on use value (UV) are Azadirachta excelsa, Cissus quadrangularis, Tylophora indica and Ocimum tenuiflorum. The informant consensus factor (Fic) values range from 0.46 to 0.71. The fidelity level (FL) value in the present study ranges from 25.58% to 93.42%. The study makes it clear that the aged informants have contributed more to the information about the value of the ethnomedicinal plants. Less information from the younger generation proves significant erosion of the traditional knowledge of this precious store. Some ethnomedicinal species with high UV and FL need to be reassessed for their phytochemical and pharmacological importance.

Evaluation of the Effects of Edible Coatings of Ocimum sanctum and Aloe vera on Jaggery Shelf Life

Background: In western Uttar Pradesh, jaggery is a widespread cottage enterprise based on agriculture, and farmers are forced to sell their product at a lesser price when it is still fresh. Therefore, it was thought to be desirable to create better storage techniques in order to extend its shelf life. Aloe vera, and Ocimum sanctum, often known as "Tulsi," are frequently utilised as antimicrobial food additives because they offer a host of other health advantages in addition to their well-known antibacterial qualities. Because the edible coatings made of these herbs provide a semi-permeable barrier to gases and water vapours, they may prolong the shelf life of jaggery by preventing degradation. Objective: The goal of the current study was to assess the ability of edible coatings of common Indian herbs, such as tulsi and Aloe vera,, to extend the shelf life of jaggery while maintaining attributes that are equal to those of fresh jaggery, in accordance with the guidelines set forth by the Food Safety and Standards Authority of India (FSSAI). Methodology: The physicochemical characteristics, antioxidant activity, total viable count, and antibacterial activity of tulsi-Aloe vera, coated (TAC), Ocimum sanctum, and Aloe vera, (AC) jaggery were assessed and compared with non-coated control. The physicochemical properties were ascertained using standard methodology for measurement of reducing sugars, proteins, phenols, saponins, tannin, alkaloids, and flavonoids. The antimicrobial activity was ascertained by means of the agar double diffusion method. According to established protocol, antioxidant activity was assessed using the DPPH radical scavenging assay and the reducing power assay. Results: According to the research, there is no discernible microbiological deterioration and the edible coatings containing tulsi and Aloe vera, extend the shelf life of jaggery during storage. When compared to uncoated jaggery, coatings were efficient at preventing the growth of both Gramme positive and Gramme negative microorganisms. Over the course of six months, the herb-infused coatings also retained their phenolic, flavonoid, and tannin contents, which improved their anti-oxidant efficacy when compared to the untreated control group.

Exploring ZnO nanoparticles: UV–visible analysis and different size estimation methods

The study investigates several theoretical and empirical methods for determining the size of zinc oxide nanoparticles (ZnO NPs) and studies their characteristics by using solely UV–visible spectroscopy. The investigation begins with the green synthesis of three samples of ZnO NPs using three different reducing agents Aspergillus Niger fungal biomass (AN), Aloe barbadensis Mille (ABM) and Ocimum tenuiflorum (OT) leaves, respectively with Zinc Sulphate as a precursor involving precipitation method. The study analyses the UV–visible data of each sample to obtain optical bandgap, Urbach energy, refractive index, optical conductivity. and apply several methods or Models such as Effective Mass Approximation (EMA) or Brus Equation (BE), Hyperbolic Band Model (HBM), Meulenkamp's Expression (ME), Tight Binding Model (TBM) and a simplified version of EMA (SEMA) to estimate the size of ZnO NPs. The effectiveness of size-determining models was assessed using two sets of one-way analysis of variance (ANOVA) for each ZnO sample and models Followed by Tukey post-hoc analysis to identify the exact sample that is different from other samples.

Phytomediated stress modulates antioxidant status, induces overexpression of CYP6M2, Hsp70, α-esterase, and suppresses the ABC transporter in Anopheles gambiae (sensu stricto) exposed to Ocimum tenuiflorum extracts.

The incorporation of phytoactive compounds in the management of malarial vectors holds promise for the development of innovative and efficient alternatives. Nevertheless, the molecular and physiological responses that these bioactive substances induce remain underexplored. This present study investigated the toxicity of different concentrations of aqueous and methanol extracts of Ocimum tenuiflorum against larvae of Anopheles gambiae (sensu stricto) and unraveled the possible underlying molecular pathways responsible for the observed physiological effects. FTIR and GCMS analyses of phytoactive compounds in aqueous and methanol crude extracts of O. tenuiflorum showed the presence of OH stretching vibration, C = C stretching modes of aromatics and methylene rocking vibration; ring deformation mode with high levels of trans-β-ocimene, 3,7-dimethyl-1,3,6-octatriene in aqueous extract and 4-methoxy-benzaldehyde, 1,3,5-trimethyl-cyclohexane and o-cymene in methanol extract. The percentage mortality upon exposure to methanol and aqueous extracts of O. tenuiflorum were 21.1% and 26.1% at 24 h, 27.8% and 36.1% at 48 h and 36.1% and 45% at 72 h respectively. Using reverse transcription quantitative polymerase chain reaction (RT-qPCR), down-regulation of ABC transporter, overexpression of CYP6M2, Hsp70, and α-esterase, coupled with significantly increased levels of SOD, CAT, and GSH, were observed in An. gambiae (s.s.) exposed to aqueous and methanol extracts of O. tenuiflorum as compared to the control. Findings from this study have significant implications for our understanding of how An. gambiae (s.s.) larvae detoxify phytoactive compounds.

Comparative evaluation of bio efficacy of Neem (Azadirachta indica) and Tulsi (Ocimum tenuiflorum) extracts in controlling termites

Comparative evaluation of bio efficacy of Neem (Azadirachta indica) and Tulsi (Ocimum tenuiflorum) extracts in controlling termites

Structural, optical, thermal, and dielectric studies of urea-doped potassium hydrogen phthalate single crystals: effect of Ocimum Tenuiflorum extract

Structural, optical, thermal, and dielectric studies of urea-doped potassium hydrogen phthalate single crystals: effect of Ocimum Tenuiflorum extract

Antifungal Potential of Ocimum sanctum Linn. Essential Oil against Fungal Pathogens Causing Economic Losses in Cymbopogon flexuosus Nees Ex Steud. (Lemongrass)

Cymbopogon flexuosus Nees Ex Steud. (Commonly known as lemongrass, family Poaceae) is one of the commercially important aromatic plants having medicinal properties. India is one of the largest producers and exporters of lemongrass essential oil in the world; in the year 2021-22, India exported about 610 tons of lemongrass oil, contributing more than 4 lakh US $ million in Gross Domestic Productivity (GDP). However, despite a good national and international market, this crop suffers quantitative and qualitative losses due to the leaf spot and leaf blight diseases caused by the fungi Curvularia trifolii and Rhizoctonia solani, respectively; therefore, an attempt has been made to manage these fungal pathogens. The essential oil of basil (Ocimum sanctum Linn.) was extracted by hydro-distillation method using Clevenger apparatus. The oil thus extracted was used for in vitro antifungal efficacy against both the test fungi Curvularia trifolii and Rhizoctonia solani, using the poison food technique. The results show that the minimum inhibitory concentration (MIC) of the basil oil against C. trifolii 2000 ppm and R. solani was 1600 and however, it was cidal in nature at 3200 and 2400 ppm against both fungi, respectively. The current study’s findings show that O. sanctum oil could be an alternative to synthetic fungicides after detailed investigations.

A COMPARATIVE PHARMACOGNOSTIC EVALUATION OF THE LEAVES OF TULSI (OCIMUM BASALICUM) AND (OCIMUM SANCTUM)

The pharmacognostic evaluation of Ocimum basilicum (Sweet Basil) and Ocimum sanctum (Holy Basil) involved detailed macroscopic and microscopic analyses of their leaves. Ocimum basilicum leaves typically measure 4-8 cm in length, displaying an elliptic ovate shape, whereas Ocimum sanctum leaves are relatively smaller, measuring about 2-2.5 cm in length and exhibiting an ovate shape. Microscopic examination further distinguished between the two species: O. sanctum displayed a square-shaped epidermis characterized by parenchymatous cells and sclerenchyma patches, whereas O. basilicum showcased a dorsiventral mesophyll structure with stomata and glandular trichomes. These distinctive features contribute significantly to the standardization, authentication, and quality control processes essential for ensuring the efficacy and safety of herbal medicine formulations containing Ocimum basilicum and Ocimum sanctum.

Antibacterial Activity of Silver Nanoparticles Synthesized from Endophytic Streptomyces sp. CR 13 Isolated from the Roots of Ocimum tenuiflorum L.

In this study, Streptomyces sp. CR 13 from the roots of Ocimum tenuiflorum L. was isolated and characterized using biochemical, morphological and genomic approaches. Silver nanoparticles (AgNPs) were synthesized extracellularly using cell-free extract and characterized with UV-vis spectroscopy, Fourier transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM), X-ray diffraction (XRD), EDX, DLS and zeta potential techniques. The maximum peak of UV-visible spectroscopy was observed at 425 nm and the TEM images revealed that the nanoparticles were spherical in shape and ranged in size from 18 to 21 nm. DLS analysis revealed particles with an average diameter of 147.9 nm. The zeta potential analysis of biosynthesized AgNPs showed a peak at -35.2 mV, indicating their high stability. Moreover, AgNPs synthesized by endophytic Streptomyces showed significant antimicrobial activity against Escherichia coli, Klebsiella pneumonia and Stphylococcus aureus.

Effects of AgO incorporation with 2D rGO nanocomposite characteristics and its beneficial electrochemical detection of acetaminophen

In this study, the AgO/rGO nanocomposite was facile synthesized via green molecules controllable route using Ocimum tenuiflorum leaf extract. And, we delve into the electrochemical sensing capacity of nanocomposite for the detection of both acetaminophen and 4-nitrophenols. XRD study revealed that the wider (002) rGO diffraction plane integrates with the Ag crystalline characteristic, with a nanocomposite crystallite size of 10.3 nm. Ocimum tenuiflorum biomolecules effectively reduced the metal ions as well as graphene oxide to the expected elemental compound. The incorporation of various leaf biomolecules such as poly-phenolic and alkaloids was bonded with rGO sheet position in the form of C-OH, C–H, N–H, with C = C molecular presence. The efficient interfacial contact between Ag, O and C (rGO) were investigated using XPS, which claimed that the biomolecular group phenol (O–H) with metallic compound as C-O-Ag around 533.9 eV, which has a significant witness for biomolecules incorporations. An identifiable count of elongated spherical-shaped Ag nanoparticles was stacked as well as incorporated between the thin rGO layers with 31.5 nm of particle size was confirmed by HR-TEM analysis. The electrochemical sensing parameters for acetaminophen the proposed sensor exhibits the lowest limit of detection of 0.25 mM for the linear range of concentrations 0.005 to 0.3 mM and 4-aminophenol exhibits the lowest limit of detection of 0.012 mM for the linear range of concentrations 0.001 to 0.3 mM. Overall, this research underscores the potential of AgO/rGO as a robust and efficient platform for achieving enhanced sensitivity in electrochemical sensing applications.

Magnesium Oxide Nanoparticles: Biogenic Synthesis and Biomedical Applications

AbstractCurrently, nanotechnology is developing at an exponential rate because of its widespread applications in various fields of life sciences. Traditionally, nanoparticles are fabricated through a variety of physical and chemical approaches but because of their limited efficiency in biomedical applications, biogenic synthesis of nanoparticles has gained much attention. Biogenically fabricated magnesium oxide nanoparticles (MgONPs) possess unique features that enabled their use in therapeutics and environmental biotechnology. Biogenically MgONPs can be fabricated using plant extracts (Manihot esculenta, Rhizophora lamarckii, Ocimum sanctum, rosemary, Bauhinia purpurea, and others), bacteria (S. coelicolor, A. johnsonii RTN), fungi (Aspergillus flavus TFR‐12, Aspergillus brasiliensis TFR 2, white‐button mushroom's) and algae (S. whigti). These biogenically synthesized MgONPs are being reported as antifungal, antioxidant, antibacterial, antipyretic, anti‐inflammatory, and anticancer agents. Moreover, their photocatalytic activity is far being discovered against certain organic dyes. This review focuses on the biogenic synthesis of MgONPs, their applications in therapeutics, and as antimicrobial agents as well as future prospects associated with their applications in the biomedical field.

A comprehensive investigation on alleviating oxidative stress and inflammation in hyperglycaemic conditions through in vitro experiments and computational analysis

Protein glycation, hyper-inflammatory reactions, and oxidative stress play a crucial role in the pathophysiology of numerous diseases. The current work evaluated the protective ability of ethyl alcohol extract of leaves from holy basil (Ocimum sanctum Linn) against inflammation, oxidative stress, glycation and advanced glycation endproducts formation.Various in vitro assays assessed prementioned properties of holy basil. In addition, molecular docking was conducted. The highest hydrogen peroxide reduction activity (72.7 %) and maximum percentage of DPPH scavenging (71.3 %) depicted its vigorous antioxidant abilities. Furthermore, it showed the most excellent protection against proteinase activity (67.247 %), prevention of denaturation of egg albumin (65.29 %), and BSA (bovine serum albumin) (68.87 %) with 600 µg/ml. Percent aggregation index (57.528 %), browning intensity (56.61 %), and amyloid structure (57.0 %) were all reduced significantly using 600 μg/ml of extract. Additionally, the antimicrobial potential was also confirmed. According to a molecular docking study, active leaf extract ingredients were found to bind with superoxide dismutase, catalase, and carbonic anhydrase. As a conclusion, O. sanctum has a variety of health-promoting properties that may reduce the severity of many diseases in diabetic patients. However, in order to ascertain the mechanisms of action of the components of its leaves in disease prevention, more thorough research based on pharmacological aspects is needed.

Ocimum sanctum-mediated co/cu/zn-doped magnesium oxide nanoparticles: Photocatalytic, antibacterial, and antioxidant properties for environmental remediation

This research intends to formulate Ocimum sanctum mediated pure and transition metal (Co2+/Cu2+/Zn2+) doped magnesium oxide nanoparticles (MgO NPs). The cubic structure of so synthesized MgO NPs was confirmed by XRD analysis. As per UV–visible spectral analysis, band gap (Eg) is tuned upon doping; doping with Co, Cu and Zn, their respective Eg are 1.65 eV, 1.96 eV and 3.66 eV in relation to MgO, Eg = 3.50 eV. The average particle size decreased from 17 nm for MgO NPs to 8–13 nm for doped MgO NPs, based on TEM analysis. EDX and XPS analysis verified the presence of Mg, O and dopants, Co, Cu and Zn in synthesized NPs. SEM investigations showed that NPs are agglomerated and spherical. Zn–MgO NPs outperform Co/Cu doped MgO NPs in their photocatalytic efficacy and comparative degradation of methyl orange (MO) and malachite green (MG) dyes confirmed that they are more efficient in degrading MG. Antibacterial investigation revealed that Cu and Zn doped MgO NPs are efficient antibacterial agents against B. subtilis and E. coli than MgO NPs. In this study, Co–MgO NPs were ineffective as antibacterial agents. S. aureus was found resistant to all NPs. Using the DPPH radical scavenging technique, these biosynthesized nanoparticles displayed 62–69 % antioxidant activity.

Bioactive compounds from Ocimum tenuiflorum and Poria cocos: A novel natural Compound for insomnia treatment based on A computational approach

Insomnia, a widespread public health issue, is associated with substantial distress and daytime functionality impairments and can predispose to depression and cardiovascular disease. Cognitive Behavioral Anti-insomnia therapies including benzodiazepines often face limitations due to patient adherence or potential adverse effects. This study focused on identifying novel bioactive compounds from medicinal plants, aiming to discover and develop new therapeutic agents with low risk-to-benefit ratios using computational drug discovery methods. Through a systematic framework involving compound library preparation, evaluation of drug-likeness and pharmacokinetics, toxicity prediction, molecular docking, and molecular dynamic simulations, two natural compounds such as 2-(4-hydroxy-3-methoxyphenyl)-8-methoxy-6-prop-2-enyl-3,4-dihydro-2H-chromen-3-ol from Ocimum tenuiflorum and 7-(2-hydroxypropan-2-yl)-1,4a-dimethyl-9-oxo-3,4,10,10a-tetrahydro-2H-phenanthrene-1-carboxylic acid from Poria cocos exhibited high binding affinity with orexin receptor type 1 (OX1R) and type 2 (OX2R), surpassing commercial drugs used in insomnia treatment. Additionally, they showed interactions with critical amino acid residues within the receptors that play crucial roles in competitive inhibitor activity, like commercial drugs such as Suvorexant, Lemborexant, and Daridorexant. Further, molecular dynamics simulations of the protein-ligand complexes under conditions that mimic the in vivo environment revealed both compounds' sustained and robust interactions with the OX1R and OX2R, reinforcing their potential as effective therapeutic candidates. Furthermore, upon evaluating both compounds' drug-likeness, pharmacokinetics, and toxicity profiles, it was discerned that they displayed considerable drug-like properties and favorable pharmacokinetics, along with diminished toxicity. The research provides a solid foundation for further exploring and validating these compounds as potential anti-insomnia therapeutics.

Effect of Time of Transplanting and Spacing on Tulsi (Ocimum sanctum)

An experiment investigating the impact of transplanting time and spacing on Tulsi (Ocimum sanctum) was conducted over three Kharif seasons from 2018-19 to 2020-21 at AICRP on Medicinal, Aromatic Plants and Betelvine. Twenty treatment combinations were arranged in a FRBD design. Transplanting times varied from June to August, with spacing options ranging from 60x45 cm to 75 x 60 cm. Recorded observations included plant height, primary branches per plant, fresh herbage yield, and dry herbage yield. Results indicated significant differences across all parameters based on transplanting time and spacing. Transplanting during the first fortnight of July (D3) resulted in the highest values for plant height (98.20 cm), branches per plant (16.63), fresh herbage yield (99.98 q/ha), and dry herbage yield (18.91 q/ha). The treatment of transplanting during the second fortnight of June (D2) showed comparable results. Among spacing options, 60 x 45 cm (S1) demonstrated the most favorable outcomes. Notably, the combined treatment of transplanting during the first fortnight of July with 60 x 45 cm spacing (D3S1) yielded the highest plant height (99.21 cm), fresh herbage yield (122.54 q/ha), and dry herbage yield (23.02 q/ha). Economically, this treatment also proved superior, with a net monetary return of ₹91453/- and a B:C ratio of 1.99. Overall, findings suggest that transplanting Tulsi seedlings between the second fortnight of June and the first fortnight of July, coupled with 60 x 45 cm spacing, maximizes productivity and economic returns.