Sweet basil (Ocimum basilicum) leaf and seed extracts alleviate neuronal dysfunction in aluminum chloride-induced neurotoxicity in Drosophila melanogaster Meigen model.

Ocimum basilicum is an important medicinal plant and culinary herb generally known as sweet basil (SB). These plants are effective radical scavengers, that have been employed in treatment of nervous system disorders, and thus, could be beneficial for the management of neurodegenerative diseases (NDs). Current clinical treatments for NDs present several side effects, therefore, there is need to develop new treatments that can mitigate these deadly diseases. Hence, this study investigated the neuroprotective activities of SB leaf and seed in aluminum chloride (AlCl3)-induced toxicity in Drosophila melanogaster. HPLC characterization of the leaves and seeds were carried out. AlCl3-diet was used to induce neurodegeneration and treated flies received SB leaf and seed extracts-supplemented diet. Survival and locomotor performance activities/levels of oxidative biomarkers [reactive oxygen species (ROS), thiobarbituric acid reactive species (TBARS), total thiol, catalase, superoxide dismutase (SOD) and glutathione-S-transferase (GST)], enzymes linked with neurodegeneration (acetylcholinesterase (AChE) and monoamine oxidase (MAO)) were investigated. SB leaf had significantly (p < 0.05) higher polyphenol contents; gallic acid and P-coumaric acid were the most abundant polyphenol in the leaf and seed respectively. Percentage survival and locomotor rates, level/activities of total thiol, catalase, SOD and GST were significantly (p < 0.05) reduced while ROS, TBARS, AChE and MAO activities were significantly (p < 0.05) increased in AlCl3-diet-fed flies. Treatment with SB leaf and seed diet lessened these observed impairments. However, SB leaf had better neuroprotective activities that could be related to the observed higher phenolic constituents. Hence, SB leaf diet may offer improved therapeutic effect in NDs.

Long-term food supplementation with sweet basil (Ocimum basilicum L.) prevents age-associated cognitive decline in female mice.

Background: Mild cognitive decline, a common issue in aging, affects memory, learning, and attention. Nutrition can influence cognition, and research indicates that Ocimum sp. (holy basil and sweet basil) leaf extracts may enhance cognition in rodents and humans. However, these studies do not address whether these benefits extend to fresh or dry leaves consumed in typical human diets, along with physiological aging. Aim: To investigate the effects of sweet basil supplementation on cognition in mature and aged female mice. Methods: Female C57bl mice were divided into four groups: 8-month-old mature adults and 18-month-old aged adults, each receiving either a control or supplemented diet. The supplemented diet included a mix of standard chow and fresh basil leaves, administered for 2-8 months. Cognitive and behavioral assessments were conducted using the novel object recognition (NOR), Morris water maze (MWM), and elevated plus maze (EPM) tasks, focusing on memory, learning, and anxiety. Results: No cognitive improvement was observed in mature mice. However, aged mice receiving long-term basil supplementation showed enhanced discrimination in NOR and stayed closer to the absent platform in MWM compared to nonsupplemented controls. While aging mice exhibited reduced anxiety-like behavior in EPM, basil supplementation prevented this reduction. Conclusion: Basil supplementation appears beneficial in elderly mice, potentially preventing age-related cognitive decline and behavioral changes. These findings support the benefits of basil consumption in cognition and underscore its potential role in promoting healthy aging. Incorporating basil into the diet at a younger age may preserve memory and mitigate behavioral changes as individuals age.

Morphological and molecular characterization of downy mildew on sweet basil (Ocimum basilicum) caused by Peronospora belbahrii in Turkiye.

BACKGROUND: Sweet basil (Ocimum basilicum) is one of the most significant aromatic plants in Turkiye. Recently, a new pathogen induced symptoms were discovered and identified as basil downy mildew caused by Peronospora belbahrii Thines. The pathogen has been introduced into the country and it has quickly become the most damaging disease in basil cultivation. The purpose of this study was to investigate the molecular and morphological properties of the causal organism of downy mildew observed on sweet basil and determine the disease incidence and prevalence in Antalya province. METHODS AND RESULTS: According to morphological characteristics (conidia, conidiophores) disease was determined as downy mildew caused by P. belbahrii. Pathogenicity tests were performed by spraying with a sporangial suspension of P. belbahrii (1 × 105 sporangia/mL). After 1 week, all inoculated plants exhibited characteristic downy mildew symptoms on their leaves, whereas non-inoculated control plants remained disease-free. All molecular analyses involving the internal transcribed spacer region were amplified using Nested PCR with primer pairs ITS4 and ITS6 for the first round and ITS4 and DC6 for the second round. Resulting sequences of all the nested PCR products had 99% similarity with P. belbahrii isolates. Disease incidence was 22.4-70.2% of sweet basil cultivation area in Antalya province. CONCLUSIONS: Based on the molecular analysis, morphological characteristics and pathogenicity tests the pathogen was identified as P. belbahrii. To our knowledge, this is the first report of downy mildew caused by P. belbahrii on sweet basil in Turkiye.

Optimization of the Extraction Conditions of Bioactive Compounds from Ocimum basilicum Leaves Using Ultrasound-Assisted Extraction via a Sonotrode.

Sweet basil (Ocimum basilicum) leaves are rich in bioactive compounds that present therapeutic benefits for human health. Ultrasonic-assisted extraction (UAE) is frequently used to obtain phenolic compounds from plants/herbal sources. However, few works have developed multi-variable studies to find the optimal conditions to extract the maximum amount of compounds, especially when applied to UAE via a sonotrode. The purpose of this work was to perform a multi-variable study by employing a Box-Behnken design to collect the highest active compound content from Ocimum basilicum leaves. The efficacy of the design was endorsed by ANOVA. The studied parameters for UAE via a sonotrode were the ethanol/water ratio, amplitude, and time. The analyzed responses were the rosmarinic acid, the sum of phenolic acids, and the sum of phenolic compounds content. The optimal conditions were found to be 50% ethanol/water, 50% amplitude, and 5 min. Twenty bioactive compounds were identified by HPLC-ESI-TOF-MS when the extract was collected by applying the optimal conditions. Ocimum basilicum may be appreciated as a valuable source of important bioactive substances for pharmaceutical use.

First Report of Downy Mildew on Sweet Basil Caused By Peronospora belbahrii in India.

Sweet basil (Ocimum basilicum L.; Family Lamiaceae) is an annual aromatic and medicinal plant grown in tropical and subtropical regions of the world. In India, it is cultivated as a commercial crop on ~8,000 ha. Aerial plant parts and essential oil of sweet basil are used in pharmaceutical, perfumery, food industries and in different formulations of traditional Ayurvedic and Unani medicines (Shahrajabian et al. 2020). The leaves have the highest concentrations of secondary metabolites such as terpenes and phenylpropanoids which provide the distinctive aroma (Viuda-Martos et al. 2011). During October 2020, severe foliar disease was observed in experimental fields of sweet basil at Council of Scientific and Industrial Research (CSIR)-Central Institute of Medicinal and Aromatic Plants (CIMAP) in Lucknow, India. Initial symptoms included large, interveinal chlorotic lesions on the adaxial surface of the leaves and black sporulation on the abaxial surface. Within a few days, the abaxial side of leaves turned necrotic, and leaf senescence and defoliation occurred on plants with severe symptoms. Disease incidence was 20 to 30% of plants. The pathogen was characterized morphologically using a light microscope. Sporangiophores were hyaline, dichotomously branched, 186.9 to 423.07 × 6.85 to 9.06 µm and, branched 3 to 5 times with each branch, terminating in two slightly curved branchlets, the longer one 7.05 to 25.31 µm and the shorter one 4.98 to 15.92 µm. Each branchlet had a single sporangium at the tip. Conidia were ellipsoidal to sub-globose, olive-brown in color, and typically measured 25.21 to 33.86 × 17.92 to 26.24 µm, each, without a pedicel. Based on these morphological characteristics, the foliar disease was identified as downy mildew was caused by Peronospora belbahrii (Thines et al. 2009). Eight symptomatic and two asymptomatic plant samples were collected from different locations in the field, and genomic DNA was extracted from the conidia of the eight naturally infected tissues of sweet basil samples as well as leaf tissues from two asymptomatic plants, using the CTAB method. The internal transcribed spacer region was amplified using ITS1 and ITS4 primers. Only eight infected samples amplified products of expected size (~ 700 bp) and two asymptomatic samples showed no amplification. Only five amplified PCR products were sequenced (White et al. 1990). All five sequences were identical and were a 98.1% match with five P. belbahrii isolates (MN450330.1, MN308051.1, MH620351.1, KJ960193, and MF693898). The consensus sequence was deposited into the NCBI database (GenBank Accession No. MW689257). Downy mildew caused by P. belbahrii previously has been reported on sweet basil from several countries (Wyenandt et al. 2015). To confirm the pathogenicity of these isolates on sweet basil (cv. CIM-Saumya), 25 - day-old sweet basil plants were sprayed with a suspension (1 × 105 sporangia/ml) of P. belbahrii. All plants were kept in a growth chamber with a 23/18°C diurnal cycle with 65 to 85% relative humidity for 24 h. Non-inoculated plants treated with sterile water served as a control treatment. After 8 days, typical symptoms of downy mildew appeared on all the inoculated plants while non-inoculated plants remained asymptomatic. Inoculated leaves with symptoms consistent of downy mildew were collected and the causal agent again identified as P. belbahrii on the basis of microscopic examination and ITS rDNA sequence data. To our knowledge, this is the first report of downy mildew caused by P. belbahrii on sweet basil in India. The pathogen has a broad host range and may pose a serious threat to the cultivation of this valuable crop in India. Thus, it is pertinent to develop effective control measures to avoid further spread and mitigate economic loss. References: Shahrajabian, M. H., et al. 2020. Int. J. Food Prop. 23:1961-1970. Wyenandt, C. A., et al. 2015. Phytopathology 105:885. Thines, M., et al. 2009. Mycol. Res. 113:532. White, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Viuda-Martos, M., et al. 2011. Food Control. 22:1715.

Sweet Basil Has Distinct Synthases for Eugenol Biosynthesis in Glandular Trichomes and Roots with Different Regulatory Mechanisms.

Production of a volatile phenylpropene; eugenol in sweet basil is mostly associated with peltate glandular trichomes (PGTs) found aerially. Currently only one eugenol synthase (EGS), ObEGS1 which belongs to PIP family is identified from sweet basil PGTs. Reports of the presence of eugenol in roots led us to analyse other EGSs in roots. We screened for all the PIP family reductase transcripts from the RNA-Seq data. In vivo functional characterization of all the genes in E. coli showed their ability to produce eugenol and were termed as ObEGS2-8. Among all, ObEGS1 displayed highest expression in PGTs and ObEGS4 in roots. Further, eugenol was produced only in the roots of soil-grown plants, but not in roots of aseptically-grown plants. Interestingly, eugenol production could be induced in roots of aseptically-grown plants under elicitation suggesting that eugenol production might occur as a result of environmental cues in roots. The presence of ObEGS4 transcript and protein in aseptically-grown plants indicated towards post-translational modifications (PTMs) of ObEGS4. Bioinformatics analysis showed possibility of phosphorylation in ObEGS4 which was further confirmed by in vitro experiment. Our study reveals the presence of multiple eugenol synthases in sweet basil and provides new insights into their diversity and tissue specific regulation.

Effect of basil use in meatball production on heterocyclic aromatic amine formation.

Herein, the effects of basil usage in meatball production on various quality criteria and heterocyclic aromatic amine (HAA) formation were investigated. The use of basil at every rate caused a significant reduction in TBARS value compared to control group. Cooking caused an increase in pH and TBARS values. IQx, IQ, AαC and MeAαC compounds could not be detected, while various amounts of MeIQx, MeIQ, 7,8-DiMeIQx, 4,8-DiMeIQx and PhIP were determined in the samples. Total HAA contents were determined up to 1.61 ng g-1 and increasing of cooking temperature increased total HAA content, except for meatball with 1% basil. The reducing or enhancing effect of the use of basil in meatball production on the formation of HAA varied depending on the usage rate and cooking temperature. It was determined that even if 100 g of the meatballs containing 0.5% basil cooked at 250 °C whose total amount of HAA content was the highest, is eaten, the intake amount is far below 1 µg.

Characterization of a sweet basil acyltransferase involved in eugenol biosynthesis.

Sweet basil (Ocimum basilicum) plants produce its characteristic phenylpropene-rich essential oil in specialized structures known as peltate glandular trichomes (PGTs). Eugenol and chavicol are the major phenylpropenes produced by sweet basil varieties whose synthetic pathways are not fully elucidated. Eugenol is derived from coniferyl acetate by a reaction catalysed by eugenol synthase. An acyltransferase is proposed to convert coniferyl alcohol to coniferyl acetate which is the first committed step towards eugenol synthesis. Here, we perform a comparative next-generation transcriptome sequencing of different tissues of sweet basil, namely PGT, leaf, leaf stripped of PGTs (leaf-PGT), and roots, to identify differentially expressed transcripts specific to PGT. From these data, we identified a PGT-enriched BAHD acyltransferase gene ObCAAT1 and functionally characterized it. In vitro coupled reaction of ObCAAT1 with eugenol synthase in the presence of coniferyl alcohol resulted in eugenol production. Analysis of ObCAAT1-RNAi transgenic lines showed decreased levels of eugenol and accumulation of coniferyl alcohol and its derivatives. Coniferyl alcohol acts as a common substrate for phenylpropene and lignin biosynthesis. No differences were found in total lignin content of PGTs and leaves of transgenic lines, indicating that phenylpropene biosynthesis is not coupled to lignification in sweet basil.

Silicon supplementation mitigates salinity stress on Ocimum basilicum L. via improving water balance, ion homeostasis, and antioxidant defense system.

Salinity is a key worldwide ecological restriction to sustainable crop production and food security. Various methods were used for inducing salinity tolerance including biotechnological approaches or application of stress tolerance-inducing substances. Silicon supplementation has a decisive role in alleviating of salinity injury, however, the definite mechanisms behind stay scantily understood, and must be examined. The imperative roles of sodium metasilicate (Si, 100 ppm) application methods (foliar spraying at 100 mg/l; soil additive at 100 mg/kg soil; foliar spraying at 100 mg/l plus soil additive at 100 mg/kg soil), in improving growth and essential oil yield, maintaining water status, activating antioxidant system, and keeping ion homeostasis of salt affected-sweet basil (6000 mg NaCl/kg soil) were studied. Salinity induced a notable increase in oxidative biomarkers, coupled with higher osmolyte concentration and osmotic potential (OP) values, as well as increased superoxide dismutase and peroxidase activities. Alternatively, sweet basil growth, essential oil yield, and catalase activity were reduced under salinity. Furthermore, salinity aggravated ion imbalance, decreased photosynthetic pigment and disrupted the plants' water status. Silicon application drastically increased osmolyte accumulation associated with sustained water status, increased OP, and improved osmotic adjustment (OA) capacity. Additionally, Si application enhanced antioxidant aptitude associated with decreased oxidative biomarkers and improved growth, photosynthetic pigment, and essential oil yield. Greater outcomes were achieved with the foliar spraying method, compared with other application methods. Salinity stress evoked modification in protein assimilation capacity and possibly will withdraw protein biosynthesis and reduce total protein band number; however, Si application may adjust the expression of salinity inducible proteins. Foliar spraying of Si with or without soil additive accelerates the expression of peroxidase isozyme over salinized or control plants. Collectively, Si foliar spraying alleviated salinity-related injuries on sweet basil by maintaining water status, increasing osmolyte assimilation, improving OA, enhancing redox homeostasis, and antioxidant capacity.

Biological value and chemical components of essential oils of sweet basil (Ocimum basilicum L.) grown with organic fertilization sources.

BACKGROUND: Environmental safety of the agricultural utilization of livestock and poultry manures from intensive farming is attracting great attention and is effective as a good source of nitrogen for sustainable crop production. The present study aimed to determine the effects of organic poultry manure applications on the biological value and essential oil content of sweet basil (Ocimum basilicum L.). In this context, different doses of chicken, kibele (fermented chicken manure) and turkey manure (750, 1000, 1250 and 1500 kg da-1 ) with a control (no manure) were applied by sowing. RESULTS: The essential oils consisted of p-Allyl-anisole as the most abundant component (5.65-17.90%), followed by nerol (6.69-16.11%), linalool (5.10-10.81%) and z-citral (5.23-10.73%). The contents of most of the chemical constituents varied significantly (P < 0.05) with organic poultry manure applications. The highest concentration for essential oils components was found in the chicken manure application. Both the antioxidant activity and total phenolic amounts of the basil extracts varied significantly as the organic poultry manure applications changed. The highest antioxidant activity and total phenolic amounts were obtained in the treatments with chicken manure doses of 1000 and 1250 kg da-1 , which were significantly different compared to the other treatments and the control treatment. Evaluation of antimicrobial activity of the essential oils against bacterial strains: Escherichia coli ATCC 35218, Staphylococcus aureus ATCC 43300 and Candida albicans ATCC 10231 was assessed by the disc diffusion method and determination of the minimum inhibitory concentration. The results of antimicrobial assays indicated that all of the tested microorganisms were affected. The highest antibacterial activity against all tested microbial species was observed with 1250 kg da-1 chicken manure applications. CONCLUSION: Among the organic poultry fertilizations, chicken fertilization was of particular interest. Especially, chicken manure doses of 1000-1250 kg da-1 had the highest antibacterial and antioxidant activity, as well as total phenolics, flavonoids and essential oil components. © 2018 Society of Chemical Industry.

Ocimum basilicum improve chronic stress-induced neurodegenerative changes in mice hippocampus.

Alzheimer's disease (AD), one of the progressive neurodegenerative diseases might be associated with exposure to stress and altered living conditions. This study aimed to evaluate the effectiveness of Ocimum basilicum (OB) essential oils in improving the neurodegenerative-like changes induced in mice after exposed to chronic unpredictable mild stress (CUMS). Forty male Swiss albino mice divided into four groups (n = 10); the control, CUMS, CUMS + Fluoxetine, CUMS + OB were used. Behavioral tests, serum corticosterone level, hippocampus protein level of the glucocorticoid receptors (GRs) and brain-dreived neurotropic factor (BDNF) were determined after exposure to CUMS. Hippocampus was histopathologically examined. Data were analyzed using statistical package for the social sciences (SPSS) and P value of less than 0.05 was considered significant. OB diminished the depression manifestation as well as impaired short term memory observed in the mice after exposure to the CUMS as evidenced by the forced swimming and elevated plus maze test. OB also up-regulated the serum corticosterone level, hippocampal protein level of the glucocorticoid receptor and the brain-derived neurotropic factor and reduced the neurodegenerative and atrophic changes induced in the hippocampus after exposure to CUMS. Essential oils of OB alleviated the memory impairment and hippocampal neurodegenerative changes induced by exposure to the chronic unpredictable stress indicating that it is the time to test its effectiveness on patients suffering from Alzheimer disease.

Two CYP716A subfamily cytochrome P450 monooxygenases of sweet basil play similar but nonredundant roles in ursane- and oleanane-type pentacyclic triterpene biosynthesis.

The medicinal plant sweet basil (Ocimum basilicum) accumulates bioactive ursane- and oleanane-type pentacyclic triterpenes (PCTs), ursolic acid and oleanolic acid, respectively, in a spatio-temporal manner; however, the biosynthetic enzymes and their contributions towards PCT biosynthesis remain to be elucidated. Two CYP716A subfamily cytochrome P450 monooxygenases (CYP716A252 and CYP716A253) are identified from a methyl jasmonate-responsive expression sequence tag collection and functionally characterized, employing yeast (Saccharomyces cerevisiae) expression platform and adapting virus-induced gene silencing (VIGS) in sweet basil. CYP716A252 and CYP716A253 catalyzed sequential three-step oxidation at the C-28 position of α-amyrin and ß-amyrin to produce ursolic acid and oleanolic acid, respectively. Although CYP716A253 was more efficient than CYP716A252 for amyrin C-28 oxidation in yeast, VIGS revealed essential roles for both of these CYP716As in constitutive biosynthesis of ursolic acid and oleanolic acid in sweet basil leaves. However, CYP716A253 played a major role in elicitor-induced biosynthesis of ursolic acid and oleanolic acid. Overall, the results suggest similar as well as distinct roles of CYP716A252 and CYP716A253 for the spatio-temporal biosynthesis of PCTs. CYP716A252 and CYP716A253 might be useful for the alternative and sustainable production of PCTs in microbial host, besides increasing plant metabolite content through genetic modification.

Can Ocimum basilicum relieve chronic unpredictable mild stress-induced depression in mice?

BACKGROUND: Depression is one of the important world-wide health problems. OBJECTIVES: This study aimed to assess the ameliorative effect of Ocimum basilicum (OB) essential oil on the behavioral, biochemical and histopathological changes resulted from exposure to chronic unpredictable mild stress (CUMS). It also aimed to investigate the underlying mechanism in an animal model of depression. MATERIALS AND METHODS: Forty male Swiss albino mice were divided into four groups (n=10): control, CUMS (exposed to CUMS for 4weeks), CUMS plus fluoxetine, and CUMS plus OB. At the end of the experiment, behavioral changes, serum corticosterone level, protein and gene expressions of brain derived neurotropic factor (BDNF) and glucocorticoid receptors (GR) in the hippocampus was all assessed. Immunoexpression of surface makers of glial fibrillary acidic protein (GFAP), Ki67, Caspase-3, BDNF and GR in the hippocampus were estimated. Data were analyzed by using the statistical package for the social sciences (SPSS). RESULTS: OB alleviated both behavioral and biochemical changes recorded in mice after exposure to CUMS. It also reduced neuronal atrophy observed in the hippocampal region III cornu ammonis (CA3) and dentate gyrus and restored back astrocyte number. OB decreased apoptosis in both neurons and glial cells and increased neurogenesis in the dentate gyrus in a pattern comparable to that of fluoxetine. Increased BDNF and GR gene and protein expressions seems to be behind the antidepressant-like effect of OB. CONCLUSION: Ocimum basilicum ameliorates the changes induced after exposure to the chronic stress. Assessing Ocimum basilicum efficacy on human as antidepressant is recommended in further studies.

Evaluation of the chemical composition, antioxidant and anti-inflammatory activities of distillate and residue fractions of sweet basil essential oil.

In this study, the chemical composition and antioxidant and anti-inflammatory activities of sweet basil (Ocimum basilicum L. Lamiaceae family) were evaluated. Sweet basil is a food-related plant that is widely used in traditional Chinese medicine. Sweet basil crude oil was processed via molecular distillation and further characterized using gas chromatography-mass spectrometry (GC-MS) to screen for new compounds. The GC-MS analysis identified thirty-eight compounds. The major constituents of the residue fraction were estragole (17.06%), methyl eugenol (11.35%) and linoleic acid (11.40%), while the distillate fraction primarily contained methyl eugenol (16.96%), α-cadinol (16.24%) and α-bergamotene (11.92%). The antioxidant (DPPH and ABTS assays) and anti-inflammatory (in Raw264.7 cells) activities were evaluated. The residue fraction markedly scavenged the DPPH (IC50 = 1.092 ± 0.066 mg/mL) and ABTS (IC50 = 0.707 ± 0.042 mg/mL) radicals. Meanwhile, the distillate fraction distinctly suppressed the production of cytokines (TNF-α, IL-ß, IL-6) and their gene expression in LPS-induced Raw264.7 cells and suppressed NO and iNOS in an in vitro model when compared with the crude oil. In conclusion, the fractions obtained from sweet basil crude oil showed different antioxidant and anti-inflammatory properties, and they could be used as an effective source of natural antioxidant and anti-inflammatory agents after molecular distillation. Thus, the properties of essential oils in natural herbal medicines may be maximized to provide a valuable therapeutic strategy for treating various disorders caused by extreme oxidative stress.

Metabolic engineering of terpene biosynthesis in plants using a trichome-specific transcription factor MsYABBY5 from spearmint (Mentha spicata).

In many aromatic plants including spearmint (Mentha spicata), the sites of secondary metabolite production are tiny specialized structures called peltate glandular trichomes (PGT). Having high commercial values, these secondary metabolites are exploited largely as flavours, fragrances and pharmaceuticals. But, knowledge about transcription factors (TFs) that regulate secondary metabolism in PGT remains elusive. Understanding the role of TFs in secondary metabolism pathway will aid in metabolic engineering for increased yield of secondary metabolites and also the development of new production techniques for valuable metabolites. Here, we isolated and functionally characterized a novel MsYABBY5 gene that is preferentially expressed in PGT of spearmint. We generated transgenic plants in which MsYABBY5 was either overexpressed or silenced using RNA interference (RNAi). Analysis of the transgenic lines showed that the reduced expression of MsYABBY5 led to increased levels of terpenes and that overexpression decreased terpene levels. Additionally, ectopic expression of MsYABBY5 in Ocimum basilicum and Nicotiana sylvestris decreased secondary metabolite production in them, suggesting that the encoded transcription factor is probably a repressor of secondary metabolism.

Integrating non-invasive VIS-NIR and bioimpedance spectroscopies for stress classification of sweet basil (Ocimum basilicum L.) with machine learning.

Plant stress diagnosis is essential for efficient crop management and productivity increase. Under stress, plants undergo physiological and compositional changes. Vegetation indices obtained from leaf reflectance spectra and bioimpedance spectroscopy provide information about the external and internal aspects of plant responses, respectively. In this study, bioimpedance and vegetation indices were noninvasively acquired from sweet basil (Ocimum basilicum L.) leaves exposed to three types of stress (drought, salinity, and chilling). Integrating the vegetation index, a novel approach, contains information about the surface of plants and bioimpedance data, which indicates the internal changes of plants. The fusion of these two datasets was examined to classify the types and severity of stress. Among the eight supervised machine learning models (three linear and five non-linear), the support vector machine (SVM) exhibited the highest accuracy in classifying stress types. Bioimpedance spectroscopy alone exhibited an accuracy of 0.86 and improved to 0.90 when fused with vegetation indices. Additionally, for drought and salinity stresses, it was possible to classify the early stage of stress with accuracies of 0.95 and 0.93, respectively. This study will allow us to classify the different types and severity of plant stress, prescribe appropriate treatment methods for efficient cost and time management of crop production, and potentially apply them to low-cost field measurement systems.

Comparative Transcriptome Analysis Revealed Candidate Gene Modules Involved in Salt Stress Response in Sweet Basil and Overexpression of ObWRKY16 and ObPAL2 Enhanced Salt Tolerance of Transgenic Arabidopsis.

Sweet basil (Ocimum basilicum L.) is an important aromatic plant with high edibility and economic value, widely distributed in many regions of the tropics including the south of China. In recent years, environmental problems, especially soil salinization, have seriously restricted the planting and spread of sweet basil. However, the molecular mechanism of the salt stress response in sweet basil is still largely unknown. In this study, seed germination, seedling growth, and chlorophyll synthesis in sweet basil were inhibited under salt stress conditions. Through comparative transcriptome analysis, the gene modules involved in the metabolic processes, oxidative response, phytohormone signaling, cytoskeleton, and photosynthesis were screened out. In addition, the landscape of transcription factors during salt treatment in sweet basil was displayed as well. Moreover, the overexpression of the WRKY transcription factor-encoding gene, ObWRKY16, and the phenylalanine ammonia-lyase-encoding gene, ObPAL2, enhanced the seed germination, seedling growth, and survival rate, respectively, of transgenic Arabidopsis, suggesting that they might be important candidates for the creation of salt-tolerant sweet basil cultivars. Our data enrich the study on salt responses in sweet basil and provide essential gene resources for genetic improvements in sweet basil in the future.

Suppression of essential oil biosynthesis in sweet basil cotyledons under hypergravity conditions.

The mechanism through which gravity influences the biosynthesis of essential oils in herbs is an important issue for plant and space biology. Sweet basil (Ocimum basilicum L.) seedlings were cultivated under centrifugal hypergravity conditions at 100 g in the light, and the growth of cotyledons, development of glandular hairs, and biosynthesis of essential oils were analyzed. The area and fresh weight of the cotyledons increased by similar amounts irrespective of the gravitational conditions. On the abaxial surface of the cotyledons, glandular hairs, where essential oils are synthesized and stored, developed from those with single-cell heads to those with four-cell heads; however, hypergravity did not affect this development. The main components, methyl eugenol and 1,8-cineole, in the essential oils of cotyledons were lower in cotyledons grown under hypergravity conditions. The gene expression of enzymes in the phenylpropanoid pathway involved in the synthesis of methyl eugenol, such as phenylalanine ammonia lyase (PAL) and eugenol O-methyltransferase (EOMT), was downregulated by hypergravity. Hypergravity also decreased the gene expression of enzymes in the 2C-methyl-d-erythritol 4-phosphate (MEP) pathway involved in the synthesis of 1,8-cineole, such as 1-deoxy-d-xylulose-5-phosphate synthase (DXS) and 1,8-cineole synthase (CINS). These results indicate that hypergravity without affecting the development of glandular hairs, decreases the expression of genes related to the biosynthesis of methyl eugenol and 1,8-cineole, which may cause a decrease in the amounts of both essential oils in sweet basil cotyledons.

Application of ZnO NPs, SiO2 NPs and Date Pollen Extract as Partial Substitutes to Nitrogen, Phosphorus, and Potassium Fertilizers for Sweet Basil Production.

The reduction in mineral fertilizer usage is crucial to the production of medicinal and aromatic products for safety and health purposes. Presently, nanotechnology and the utilization of natural extracts have been extensively studied due to their significant contribution. Ocimum basilicum is commonly employed for various medicinal and aromatic applications. Therefore, randomized complete block design field experiments containing 10 treatments were conducted during the 2021 and 2022 seasons to investigate the effect of nanoparticles (NPs) of ZnO (1.5 and 2.0 g/L) and SiO2 (100 and 150 mg/L) and date palm pollen extract (DPPE) at 10 and 20 g/L either alone or in combination with the ¾ or ½ NPK recommended dose (RD). The NPK RD was served as a control treatment on basil plant production in each season. The effectiveness of ZnO NPs, SiO2 NPs, and DPPE for the decrease in NPK utilization was evaluated. Meanwhile, the most effective treatment for vegetative traits (except for plant height), essential oil %, and yield was ½ NPK RD + 20 g/L DPPE + 2.0 g/L ZnO NPs. Such a treatment increased the branch number/plant, main stem diameter, relevant chlorophyll content, fresh weight/plant, dry weight/plant, essential oil %, and essential oil yield/plant by 21.00 and 9.94%, 58.70 and 40.00%, 20.69 and 15.83%, 68.83 and 58.28%, 48.70 and 56.16%, 45.71 and 35.53%, and 113.22 and 110.32% over the control in the two seasons, respectively. For total phenol and antioxidant activity, the most effective treatments were the ¾ NPK +1.5 g/L ZnO NPs and ½ NPK +2.0 g/L ZnO NPs, respectively. Simultaneously, essential oil composition (with their compound numbers identified (11-29 for control and ¾ NPK RD + 1.5 g/L ZnO NPs)) and the percentage of total compounds, monoterpene hydrocarbons, sesquiterpene hydrocarbons, and oxygenated hydrocarbons were varied among the used applications. The major observed compounds (>8%) estragole, methyl eugenol, linalool, cineole, and caryophyllene were found in different treatments. Thus, the findings of this study indicate the favorable utilization of ZnO NPs, SiO2 NPs, and DPPE in reducing the application of NPK, which may present a novel strategy and beneficial approach.

Qualitative Profiling, Antioxidant and Antimicrobial Activities of Polar and Nonpolar Basil Extracts.

Basil (Ocimum basilicum L.) is a widely used culinary herb. In this study, ethanol, dichloromethane, and sunflower oil were used separately as solvents with distinct polarities for the extraction of basil aerial parts to simulate the different polarity conditions in domestic food processing. The oil extract (OE) was re-extracted with acetonitrile, and the chemical composition, antioxidant potential, and antimicrobial activities of the ethanol (EE), dichloromethane (DCME), and acetonitrile (ACNE) extracts were determined. A total of 109 compounds were tentatively identified in EE, DCME, and ACNE by HPLC-DAD/ESI-ToF-MS. Fatty acids were present in all extracts. Phenolic acids and flavonoids dominated in EE. DCME was characterised by triterpenoid acids, while diterpenoids were mainly found in ACNE. The extracts were analysed for their antioxidant capacity using the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) assay. EE and DCME showed significant radical scavenging potential. Antimicrobial activity was explored in eight bacterial, two yeast, and one fungal species. All extracts exhibited high antifungal activity, comparable to or better than that of the commercial drug nistatin. Antibacterial activities were notable for EE and ACNE, while DCME showed no activity against bacteria in the applied concentration ranges. The different polarities of the solvents led to distinctive phytochemical compositions and bioactivities in the extracts.