Conserved Role of Heterotrimeric G Proteins in Plant Defense and Cell Death Progression.

Programmed cell death (PCD) is a critical process in plant immunity, enabling the targeted elimination of infected cells to prevent the spread of pathogens. The tight regulation of PCD within plant cells is well-documented; however, specific mechanisms remain elusive or controversial. Heterotrimeric G proteins are multifunctional signaling elements consisting of three distinct subunits, Gα, Gß, and Gγ. In Arabidopsis, the Gßγ dimer serves as a positive regulator of plant defense. Conversely, in species such as rice, maize, cotton, and tomato, mutants deficient in Gß exhibit constitutively active defense responses, suggesting a contrasting negative role for Gß in defense mechanisms within these plants. Using a transient overexpression approach in addition to knockout mutants, we observed that Gßγ enhanced cell death progression and elevated the accumulation of reactive oxygen species in a similar manner across Arabidopsis, tomato, and Nicotiana benthamiana, suggesting a conserved G protein role in PCD regulation among diverse plant species. The enhancement of PCD progression was cooperatively regulated by Gßγ and one Gα, XLG2. We hypothesize that G proteins participate in two distinct mechanisms regulating the initiation and progression of PCD in plants. We speculate that G proteins may act as guardees, the absence of which triggers PCD. However, in Arabidopsis, this G protein guarding mechanism appears to have been lost in the course of evolution.

Anti-oxidative and anti-inflammatory effects of Tagetes minuta essential oil in activated macrophages.

OBJECTIVE: To investigate antioxidant and anti-inflammatory effects of Tagetes minuta (T. minuta) essential oil. METHODS: In the present study T. minuta essential oil was obtained from leaves of T. minuta via hydro-distillation and then was analyzed by gas chromatography-mass spectrometry. The anti-oxidant capacity of T. minuta essential oil was examined by measuring reactive oxygen, reactive nitrogen species and hydrogen peroxide scavenging. The anti-inflammatory activity of T. minuta essential oil was determined through measuring NADH oxidase, inducible nitric oxide synthase and TNF-α mRNA expression in lipopolysacharide-stimulated murine macrophages using real-time PCR. RESULTS: Gas chromatography-mass spectrometry analysis indicated that the main components in the T. minuta essential oil were dihydrotagetone (33.86%), E-ocimene (19.92%), tagetone (16.15%), cis-ß-ocimene (7.94%), Z-ocimene (5.27%), limonene (3.1%) and epoxyocimene (2.03%). The T. minuta essential oil had the ability to scavenge all reactive oxygen/reactive nitrogen species radicals with IC50 12-15 µg/mL, which indicated a potent radical scavenging activity. In addition, T. minuta essential oil significantly reduced NADH oxidase, inducible nitric oxide synthaseand TNF-α mRNA expression in the cells at concentrations of 50 µg/mL, indicating a capacity of this product to potentially modulate/diminish immune responses. CONCLUSIONS: T. minuta essential oil has radical scavenging and anti-inflammatory activities and could potentially be used as a safe effective source of natural anti-oxidants in therapy against oxidative damage and stress associated with some inflammatory conditions.

Anti-inflammatory effect of Mentha longifolia in lipopolysaccharide-stimulated macrophages: reduction of nitric oxide production through inhibition of inducible nitric oxide synthase.

Mentha longifolia is an aromatic plant used in flavoring and preserving foods and as an anti-inflammatory folk medicine remedy. The present study assessed the effects of M. longifolia extracts, including essential oil and crude methanol extract and its fractions (ethyl acetate, butanol and hexane), on nitric oxide (NO) production and inducible NO synthase (iNOS) mRNA expression in lipopolysaccharide (LPS)-stimulated J774A.1 cells using real-time polymerase chain reaction (PCR). The cytotoxic effects of the extracts on the cells were examined and non-cytotoxic concentrations (<0.2 mg/ml) were used to examine their effects on NO production and iNOS mRNA expression. Only the hexane fraction that contained high levels of phenolic and flavonoid compounds at concentrations from 0.05-0.20 mg/ml significantly reduced NO production in LPS-stimulated cells (p < 0.001). Real-time PCR analysis indicated the ability of this fraction at the same concentrations to significantly decrease iNOS as well as TNFα mRNA expression in the cells (p < 0.001). All extracts were able to scavenge NO radicals in a concentration-dependent manner. At concentrations greater than 0.2 mg/ml, total radicals were 100% scavenged. In conclusion, M. longifolia possibly reduces NO secretion in macrophages by scavenging NO and inhibiting iNOS mRNA expression, and also decreases TNFα pro-inflammatory cytokine expression, thus showing its usefulness in the inflammatory disease process.

Antioxidant, nitric oxide scavenging and malondialdehyde scavenging activities of essential oils from different chemotypes of Zataria multiflora.

The antioxidant, nitric oxide (NO) scavenging and malondialdehyde (MDA) scavenging activities of different Zataria multiflora (ZM) chemotype essential oils (EOs) were investigated. The main components are: ZM1 (carvacrol, p-cymene), ZM2 (carvacrol, p-cymene), ZM3 (carvacrol, p-cymene), ZM4 (linalool), ZM5 (carvacrol, p-cymene, thymol), ZM6 (thymol, carvacrol, p-cymene, γ-terpienene), ZM7 (thymol, p-cymene, γ-terpienene) and ZM8 (carvacrol, linalool, p-cymene, thymol). The antioxidant capacities were estimated to be 863 ± 55, 619 ± 27, 876 ± 32, 38 ± 9, 649 ± 50, 595 ± 40, 696 ± 41 and 618 ± 9 µg ascorbic acid equivalents per millilitre for ZM1 to ZM8, respectively. The NO scavenging values were estimated to be 54 ± 1.2, 50 ± 1.4, 63 ± 1, 0.60 ± 0.1, 53 ± 0.7, 53 ± 1.5, 38 ± 1.1 and 46.5 ± 3 µg ascorbic acid equivalents per millilitre for ZM1 to ZM8, respectively. The MDA scavenging values were estimated to be 19 ± 1, 9 ± 1, 24 ± 1, 1.6 ± 0.6, 12 ± 1, 11.7 ± 1, 10 ± 1 and 12.5 ± 1.3 µg ascorbic acid equivalents per millilitre for ZM1 to ZM8, respectively. Among these EOs, ZM3 with carvacrol and p-cymene had higher antioxidant, NO scavenging and MDA scavenging properties.