Bridging fungal resistance and plant growth through constitutive overexpression of Thchit42 gene in Pelargonium graveolens.

KEY MESSAGE: Thchit42 constitutive expression for fungal resistance showed synchronisation with leaf augmentation and transcriptome analysis revealed the Longifolia and Zinc finger RICESLEEPER gene is responsible for plant growth and development. Pelargonium graveolens essential oil possesses significant attributes, known for perfumery and aromatherapy. However, optimal yield and propagation are predominantly hindered by biotic stress. All biotechnological approaches have yet to prove effective in addressing fungal resistance. The current study developed transgenic geranium bridging molecular mechanism of fungal resistance and plant growth by introducing cassette 35S::Thchit42. Furthermore, 120 independently putative transformed explants were regenerated on kanamycin fortified medium. Primarily transgenic lines were demonstrated peak pathogenicity and antifungal activity against formidable Colletotrichum gloeosporioides and Fusarium oxysporum. Additionally, phenotypic analysis revealed ~ 2fold increase in leaf size and ~ 2.1fold enhanced oil content. To elucidate the molecular mechanisms for genotypic cause, de novo transcriptional profiles were analyzed to indicate that the auxin-regulated longifolia gene is accountable for augmentation in leaf size, and zinc finger (ZF) RICESLEEPER attributes growth upregulation. Collectively, data provides valuable insights into unravelling the mechanism of Thchit42-mediated crosstalk between morphological and chemical alteration in transgenic plants. This knowledge might create novel opportunities to cultivate fungal-resistant geranium throughout all seasons to fulfil demand.

Acyclic monoterpenoid-rich essential oil of Cymbopogon distans mitigates skin inflammation: a chemico-pharmacological study.

The topical application of essential oils is considered an effective treatment for skin diseases. Cymbopogon distans (Nees ex Steud.) Wats (Poaceae) is a promising aromatic grass widespread in the Himalayan temperate zone. Therefore, using in-vitro and in-vivo bioassays, we examined the chemical and pharmacological characteristics of essential oil hydro-distilled from C. distans coded as CDA-01, specifically concerning skin inflammation. Characterization using GC-FID and GC-MS provided a chemical fingerprint for CDA-01, enabling the identification of 54 compounds; amongst them, citral (34.3%), geranyl acetate (21.2%), and geraniol (16.4%) were the most abundant. To examine the anti-inflammatory potential, CDA-01 treatment on LPS-stimulated macrophage cells in addition to 12-O-tetradecanoylphorbol-13-acetate (TPA) generated cutaneous inflammatory reaction in the mouse ear was assessed through quantification of the inflammatory markers. Consequently, CDA-01 demonstrated protection against inflammation caused by LPS by lowering the pro-inflammatory cytokines (IL-6 and TNF-α) level in HaCaT cells with negligible cytotoxicity. Consistent with the in-vitro findings, CDA-01 treatment reduced pro-inflammatory mediators (TNF-, IL-6, and NO) and lipid peroxidation in an in-vivo investigation. Subcutaneous inflammation in TPA-treated mice ears was similarly decreased, as evidenced by the histological and morphological studies. As a result of our findings, it is possible that CDA-01 could be an effective treatment for skin inflammation disorders.

Sesquiterpenoids isolated from davana (Artemisia pallens Wall. ex DC) mitigates parkinsonism in Caenorhabditis elegans disease model.

Parkinson's disease (PD) is a multifactorial ailment that severely affects the viability of dopaminergic neurons leading to progressive loss of motor control. The current regimen for PD treatment includes synthetic drugs that lack efficacy and cause serious side effects. Consequently, recent drug development studies are focusing on alternative medicines from plant sources. Artemisia pallens Wall. ex DC, commonly known as davana, is an annual aromatic herb cultivated in southern India. Given the diverse traditional and scientifically documented therapeutic effects of A. pallens, the pharmacological potential of the isolates of the plant, namely bicyclogermacrene (D1), cis-davanone (D3), and cis-hydroxy davanone (D5), was tested for anti-Parkinson's activity in Caenorhabditis elegans model. The tested compounds alleviated α-synuclein (α-syn) aggregation and maximum decline was observed in 25 µM D1 supplemented worms. Additionally, D1 modulated dopamine regulated nonanol-1 repulsion and locomotory behaviour of C. elegans validating its future use as a dopamine-enhancing agent. The genetic regulation mediating the above effects validated through the qPCR study showed that D1 supplementation displayed its anti-Parkinson's effect through upregulation of the antioxidant defence system genes (superoxide dismutase (sod)-1, sod-2, and sod-4) and PD associated pdr-1 gene that maintains the mitochondrial proteostasis. The molecular docking studies of C. elegans PDR-1 with D1 further confirmed its contribution in D1 induced abridgment of Parkinson disease linked pathologies in C. elegans disease model. Hence, this article proposes D1 as an effective regimen for curtailing the Parkinson disease linked pathologies through mechanism of maintaining cellular redox state and proteostasis.

p-Menthadienols-rich essential oil from Cymbopogon martini ameliorates skin inflammation.

Cymbopogon martini variety sofia, commonly known as ginger-grass, is an important aromatic crop used by the perfumery, medicinal and cosmetic industries worldwide. This study explores the chemical and possible pharmacological profile of hydro-distilled essential oil of C. martini variety sofia against skin inflammation. The essential oil extracted by the hydrodistillation process was analyzed by gas chromatography (GC), gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy (NMR) to identify its constituents, and was coded as CMA-01 for further in vitro and in vivo pharmacological study related to skin inflammation. The chemical fingerprint revealed that CMA-01 oil has (E)-p-mentha-2,8-dien-1-ol (21.0%), (E)-p-mentha-1(7),8-dien-2-ol (18.1%), (Z)-p-mentha-1(7),8-dien-2-ol (17.4%), (Z)-p-mentha-2,8-dien-1-ol (9.0%), limonene (7.7%), and (E)-carveol (5.7%) as major components. The pre-treatment of CMA-01 showed significant inhibition of pro-inflammatory markers in activated HaCat cells without cytotoxic effect. The in vivo study revealed the ameliorative impact of CMA-01 against skin inflammation induced by TPA in mouse ears as evidenced by a reduction of ear edema, pro-inflammatory mediators (IL-6, TNF-α), oxidative stress markers (malondialdehyde and nitric-oxide) and histological changes in ear tissues without any skin irritation response on rabbit skin. These findings suggest the suitability of CMA-01 as a valuable therapeutic candidate for the treatment of skin inflammation.

New Insights into the Chemical Composition, Pro-Inflammatory Cytokine Inhibition Profile of Davana (Artemisia pallens Wall. ex DC.) Essential Oil and cis-Davanone in Primary Macrophage Cells.

Artemisia pallens Wall. ex DC., popularly known as davana, has gained considerable attention because of its unique fragrance, high economic value, and pharmacological properties. The compositional complexity of davana essential oil (DO) has been a challenge for quality control. In this study, the chemical profile of DO was developed using polarity-based fractionation and a combination of gas chromatographic (GC-FID), hyphenated chromatographic (GC/MS), and spectroscopic (Fourier-Transform Infra-Red, 1D, 2D-Nuclear Magnetic Resonance) techniques. The analysis led to the identification of ninety-nine compounds. Major components of the DO were cis-davanone (D3, 53.0 %), bicyclogermacrene (6.9 %), trans-ethyl cinnamate (4.9 %), davana ether isomer (3.4 %), spathulenol (2.8 %), cis-hydroxy davanone (2.4 %), and trans-davanone (2.1 %). The study led to identifying several co-eluting novel minor components, which could help determine the authenticity of DO. The rigorous column-chromatography led to the isolation of five compounds. Among these, bicyclogermacrene, trans-ethyl cinnamate, and spathulenol were isolated and characterized by spectroscopic methods for the first time from DO. Pharmacological profile revealed that the treatment of DO and D3 inhibited the production of pro-inflammatory cytokines (TNF-α, IL-6) induced by lipopolysaccharide (LPS) in primary macrophages without any cytotoxic effect after administration of their effective concentrations. The result of this study indicates the suitability of DO and D3 for further investigation for the treatment of chronic skin inflammatory conditions.

Intercropping of aromatic crop Pelargonium graveolens with Solanum tuberosum for better productivity and soil health.

Farmers in hilly regions experience low production potential and resource use efficiency due to low valued crops and poorsoil health. Geranium (Pelargonium graveolens L.) is a vegetatively propagated initially slow growing, high value aromatic crop. Potato (Solanum tuberosum L.) is also vegetatively propagated high demand cash crop. A field experiment was carried out in temperate climate to investigate the influence of geranium intercropping at different row strips (1:1 and 1:2) and plant density (60 x 45, 75 x 45 and 90 x 45 cm) with potato intercrop on biomass, oil yield, monetary advantage and soil quality parameters. The row spacing 60x45cm and row strip 1:1 was found to be superior and produced 92 t ha(-1) and 14 kg ha(-1) biomass and oil yield, respectively. The row strip 1:2 intercrop earned a maximum $2107, followed by $1862 with row strip 1:1 at 60 x 45 cm plant density. Significant variations were noticed in soil organic carbon (Corg), total N (Nt), available nutrients, soil microbial biomass (Cmic) and nitrogen (Nmic) content. Maximum improvement of Corg (41.0%) and Nt (27.5%)with row strip 1:1 at 75 x 45 cm plant density. While higher soil respiration rate, Cmic, Nmic, and qCO2 was found with 1:2 row strip at 60 x 45 plant density. The buildup of Corg and Cmic potato intercrop can promote long term sustainability on productivity and soil health.

Neuroinflammation and acetylcholinesterase inhibition potentials of acyclic monoterpenoids isolated from Cymbopogon distans (Nees ex Steud.) Will. Watson.

Cymbopogon distans (Nees ex Steud.) Will. Watson (Poaceae) is a promising aromatic plant distributed in the Himalayas. In this study, five acyclic monoterpenoids, namely geranyl acetate (RS1), neral (RS2), geranial (RS3), citral (RS4) and geraniol (RS5) were isolated from the essential oil of C. distans. The isolated compounds were tested for in-vitro neuroinflammation inhibitory potential in LPS-stimulated BV2 microglial cells. RS1-RS4 exhibited significant neuroinflammation inhibition without any cytotoxic effect at the dose of 10 µM. RS4, the most active anti-neuroinflammatory compound (TNF-α 31.48 ± 1.00%; IL-6 24.02 ± 0.63%; IL-1ß 42.15 ± 1.76%) was also able to inhibit acetylcholinesterase (AChE) in a dose-dependent manner. The results showed that RS4 (an isomeric mixture of neral and geranial) has the potential to inhibit neuroinflammation and AChE, which are the biomarkers of neurodegenerative disorders.

Vasorelaxant property of 2-phenyl ethyl alcohol isolated from the spent floral distillate of damask rose (Rosa damascena Mill.) and its possible mechanism.

ETHNOBOTANICAL RELEVANCE: Rosa damascena Mill. (Rosaceae), commonly known as damask rose, is an ancient medicinal and perfumery plant used in Traditional Unani Medicine due to various therapeutic effects, including cardiovascular benefits. AIM OF THE STUDY: This study aimed to evaluate the vasorelaxant effect of the 2-phenyl ethyl alcohol (PEA) isolated from the spent flowers of R. damascena which remain after the extraction of essential oil. MATERIALS AND METHODS: The freshly collected flowers of R. damascena were hydro-distilled in a Clevenger's type apparatus to extract the rose essential oil (REO). After removing the REO, the spent-flower hydro-distillate was collected and extracted with organic solvents to yield a spent-flower hydro-distillate extract (SFHE), which was further purified by column chromatography. The SFHE and its isolate were characterized by gas chromatography (GC-FID), gas chromatography-mass spectrometry (GC-MS), and nuclear magnetic resonance (NMR) techniques. The PEA, isolated from SFHE, was evaluated for vasorelaxation response in conduit blood vessels like rat aorta and resistant vessels like mesenteric artery. The preliminary screening of PEA was done in aortic preparation pre-constricted with phenylephrine/U46619. Further, a concentration-dependent relaxation response to PEA has been elicited in both endothelium-intact and endothelium-denuded arterial rings, and the mode of action was explored. RESULTS: The SFHE revealed the presence of PEA as the main constituent (89.36%), which was further purified by column chromatography to a purity of 95.0%. The PEA exhibited potent vasorelaxation response both in conduit vessels like the rat aorta and resistance vessels like the mesenteric artery. The relaxation response is mediated without any involvement of vascular endothelium. Further, TEA sensitive BKCa channel was found to be the major target for PEA-induced relaxation response in these blood vessels. CONCLUSIONS: The spent flowers of R. damascena, which remain after the extraction of REO, could be used to extract PEA. The PEA possessed marked vasorelaxation properties in both aorta and mesenteric artery and showed promise for development into an herbal product against hypertension.

Variation in the volatile terpenoids of two industrially important basil (Ocimum basilicum L.) cultivars during plant ontogeny in two different cropping seasons from India.

BACKGROUND: Two Ocimum basilicum cultivars, 'Vikarsudha' and 'CIM-Saumya', grown in the Kumaon region of western Himalaya were evaluated for their essential oil yield and composition at different stages of plant growth during two distinct cropping seasons (spring-summer and rain-autumn). RESULTS: The highest yield of essential oil was obtained at full bloom stage in both cultivars in both cropping seasons. The essential oils obtained from different stages in two cropping seasons were analysed by capillary gas chromatography with flame ionisation detection, and gas chromatography-mass spectrometry. The major component of cultivar 'Vikarsudha' was methyl chavicol (84.3-94.3%), while for cultivar 'CIM-Saumya' the main components were methyl chavicol (62.5-77.6%) and linalool (14.4-34.1%). CONCLUSION: This study clearly indicated that cultivar, cropping season, plant ontogeny and plant part had significant effects on the yield and quality of the essential oil of O. basilicum. Further, the amount of methyl chavicol in the cultivars grown in this region was higher than in cultivars from other parts of India.

Chemical composition and antimicrobial activity of the leaf essential oil of Ravenia spectabilis Engl. (Rutaceae).

Ravenia spectabilis Engl. belongs to the family Rutaceae is known to possess several biologically active phytomolecules. This study was planned to investigate the chemical composition and antimicrobial activity of the leaf essential oil of R. spectabilis. The hydrodistillation of fresh leaves of R. spectabilis gave 0.19 ± 0.02% essential oil. The resulting essential oil was analysed by gas chromatography-flame ionization detector (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Altogether, thirty-one constituents forming 97.6 ± 1.72% of the total oil composition were identified. Major components of the oil were sabinene (60.8 ± 0.36%), α-pinene (5.4 ± 0.30%), myrcene (4.8 ± 0.25%), δ-3-carene (4.7 ± 0.62%) and ß-pinene (4.3 ± 0.17%). The in-vitro antimicrobial potential of the oil was examined against eight human pathogenic bacterial and fungal strains. The essential oil showed significant activity against Staphylococcus aureus, Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus, methicillin-resistant Staphylococcus epidermidis, Candida albicans, and Candida kefyr. This is the first report on R. spectabilis leaf essential oil composition and its antimicrobial activity. The essential oil could be a promising natural source of sabinene and antimicrobial for developing new phytotherapeutics.

Chemical composition, bactericidal kinetics, mechanism of action, and anti-inflammatory activity of Isodon melissoides (Benth.) H. Hara essential oil.

Present study was aimed to investigate the antibacterial activity, bactericidal mechanism of action, killing kinetics and anti-inflammatory activity of Isodon melissoides (Benth.) H. Hara essential oil. The gas chromatography-flame ionization detector (GC-FID) and gas chromatography-mass spectrometry (GC-MS) analysis revealed the presence of carvacrol (45.4%), p-cymene (11.6%) and thymol (11.3%) as major constituents of the oil. The oil displayed broad spectrum significant antibacterial activity (MIC: 0.13-8.33 ppm; MBC: 0.13->33.34 ppm) against test strains. The oil exhibited a time and dose-dependent bactericidal effect. The oil disrupted the cell membrane by changing the cell membrane permeability. The essential oil significantly decreased the overproduction of proinflammatory cytokines in LPS-induced inflammation in HaCaT cells without any cytotoxic effect. I. melissoides essential oil can be a promising alternative antimicrobial agent for the control of methicillin resistant staphylococci and other pathogenic bacteria tested, and also useful for the topical anti-inflammatory properties.

Chemical composition and antimicrobial activity of the essential oil of senescent leaves of guava (Psidium guajava L.).

The aim of the study was to investigate the chemical composition and antimicrobial activity of the essential oil of the senescent leaves of Psidium guajava L. (Myrtaceae). The hydrodistilled essential oil of the leaves was analysed by GC-FID and GC-MS. The antimicrobial activity of the oil was assessed against human and plant pathogenic microorganisms by disc diffusion, microdilution broth and poison food techniques. A total of forty-six constituents, representing 90.9% of the total oil composition were identified. Major constituents of the oil were limonene (29.1%), (E)-caryophyllene (15.7%), caryophyllene oxide (8.8%), caryophylla-4(12),8(13)-dien-5-ol (6.5%), (E)-nerolidol (4.0%), α-cadinol (3.4%), and muurola-4,10(14)-dien-1-ß-ol (2.5%). The chemical composition of the examined essential oil was quite different from those reported earlier. The essential oil showed significant inhibition of human pathogenic bacteria (MIC: 0.065-0.261 mg/mL) and plant pathogenic fungi, namely Curvularia lunata (86.02%) and Fusarium chlamydosporum (82.80%).

Gamma irradiation-induced variability in morpho-agronomic and oil quality traits of Mentha piperita L.

PURPOSE: To develop elite mutant in Mentha piperita having morphotype and oil content of Mentha arvensis, and oil quality of M. piperita. MATERIALS AND METHODS: The dormant runner of menthol rich genotype MPK-5 were subjected to different doses of γ- irradiation (10, 20, 30, 50, 70, 90, and 110 Gy) at a dose rate of 55 Gy/min to induce the genetic variability for herb and oil yield as well as oil quality. A wide spectrum of variability for agro-morphological traits, herb yield, and quality profile was observed among the mutants. RESULTS: The developed and selected superior mutants viz. MPK-5(1) and MPK-5(3) have a mean herb yield potential of 23.923 and 21.503 Kg/9 m2; mean oil yield of 92.953 and 80.047 ml/plot; mean menthol content of 69.012% and 69.160% with mean menthofuran content of 1.554% and 0.531%, respectively. CONCLUSIONS: Mutational breeding through γ- irradiation is considered complementary to the conventional breeding method, to broaden the spectrum of genetic variability. The developed and selected mutants namely viz. MPK-5(1) and MPK-5(3) identified as promising mutants, based on herb yield, oil yield, and essential oil quality, could be used as a parental line for exploitation in hybridization program/recombinant breeding.

Volatile constituents of Origanum vulgare L., 'thymol' chemotype: variability in North India during plant ontogeny.

Essential oils derived from six different phenophases, namely early vegetative stage, late vegetative stage, early flowering stage, full flowering stage (FFS), late flowering stage and seed shattering stage of Origanum vulgare L. grown in Kumaon region of Uttarakhand, India were investigated by GC and GC-MS. A total of 38 constituents, representing 97.4-99.7% of the total oil composition, were identified. Major components of oils were thymol (40.9-63.4%), p-cymene, (5.1-25.9%), γ-terpinene (1.4-20.1%), bicyclogermacrene (0.2-6.1%), terpinen-4-ol (3.5-5.9%), α-pinene (1.6-3.1%), 1-octen-3-ol (1.4-2.7%), α-terpinene (1.0-2.2%), carvacrol (<0.1-2.1%), ß-caryophyllene (0.5-2.0%) and ß-myrcene (1.2-1.9%). Thymol, terpinen-4-ol, 3-octanol, α-pinene, ß-pinene, 1,8-cineole, α-cubebene and (E)-ß-ocimene were observed to be higher during FFS. The study showed that plant stage had a significant effect on the essential oil content and composition of O. vulgare grown in the hilly tracks of Northern India.

Volatile constituents of essential oil and rose water of damask rose (Rosa damascena Mill.) cultivars from North Indian hills.

Rosa damascena Mill. is an important aromatic plant for commercial production of rose oil, water, concrete and absolute. The rose water and rose oil produced under the mountainous conditions of Uttarakhand were investigated for their chemical composition. The major components of rose water volatiles obtained from the bud, half bloom and full bloom stages of cultivar 'Ranisahiba' were phenyl ethyl alcohol (66.2-79.0%), geraniol (3.3-6.6%) and citronellol (1.8-5.5%). The rose water volatiles of cultivar 'Noorjahan' and 'Kannouj' also possessed phenyl ethyl alcohol (80.7% and 76.7%, respectively) as a major component at full bloom stage. The essential oil of cultivar 'Noorjahan' obtained from two different growing sites was also compared. The major components of these oils were citronellol (15.9-35.3%), geraniol (8.3-30.2%), nerol (4.0-9.6%), nonadecane (4.5-16.0%), heneicosane (2.6-7.9%) and linalool (0.7-2.8%). This study clearly showed that the flower ontogeny and growing site affect the composition of rose volatiles. The rose oil produced in this region was comparable with ISO standards. Thus, it was concluded that the climatic conditions of Uttarakhand are suitable for the production of rose oil of international standards.