Assessment of selection criteria using multi-year study for effective breeding program of Zingiber officinale L.

Background: Ginger has been an important cash crop with numerous applications since ancient times. As the demand for ginger is ever-growing and being a seasonal crop, a high-yielding variety of ginger would be economically profitable. Methods: In this study, 150 germplasm were collected from different regions of NE India and evaluated for three years in CRBD design with three replications. The present study thus focused on the variability, association, and diversity studies for the first time on 150 ginger germplasm from across North East India. The genotypic and phenotypic coefficient of variation, heritability, correlation, and path analysis were evaluated for the germplasm. Results: Analysis of variance (ANOVA) revealed considerable differences among the studied germplasm for studied characters, revealing sufficient variability in the materials. The Mahalanobis D2 and Tocher methods grouped the 150 ginger germplasm into ten clusters. Based on the results of the path coefficient analysis determined for essential oil yield and rhizome yield per plant, it can be concluded that the characters' initial rhizome weight, the weight of mother rhizome, and weight of secondary rhizome were the most important and appeared promising in improving the overall yield potential of ginger rhizome and essential oil yield. Thus, selection based on the identified traits would lead to an effective ginger breeding program for higher rhizome and essential oil yield.

Stability estimation through multivariate approach among solasodine-rich lines of Solanum khasianum (C.B. Clarke): an important industrial plant.

Solanum khasianum is a medicinally important plant that is a source of steroidal alkaloids 'solasodine.' It has various industrial applications, including oral contraceptives and other pharmaceutical uses. The present study was based on 186 germplasm of S. khasianum, which were analyzed for the stability of economically important traits like solasodine content and fruit yield. The collected germplasm was planted during Kharif 2018, 2019, and 2020 in RCBD with three replications at the experimental farm of CSIR-NEIST, Jorhat, Assam, India. A multivariate approach for stability analysis was applied to identify stable germplasm of S. khasianum for economically important traits. The germplasm was analyzed for additive main effects and multiplicative interaction (AMMI), GGE biplot, multi-trait stability index, and Shukla's variance which were evaluated for three environments. The AMMI ANOVA revealed significant GE interaction for all the studied traits. The stable and high-yielding germplasm was identified from the AMMI biplot, GGE biplot, Shukla's variance value, and MTSI plot analysis. Lines no. 90, 85, 70, 107, and 62 were identified as highly stable fruit yielders while, lines no. 1, 146, and 68 were identified as stable high solasodine lines. However, considering both traits, i.e., high fruit yield and solasodine content, MTSI analysis was performed which showed that lines 1, 85, 70,155, 71, 114, 65, 86, 62, 116, 32, and 182 could be used in a breeding program. Thus, this identified germplasm can be considered for further varietal development and could be used in a breeding program. The findings of the present study would be beneficial for the S. khasianum breeding program.

Novel ethyl p-methoxy cinnamate rich Kaempferia galanga (L.) essential oil and its pharmacological applications: special emphasis on anticholinesterase, anti-tyrosinase, α-amylase inhibitory, and genotoxic efficiencies.

Background: Kaempferia galanga (L.) is one of the prospective therapeutic plants with an aromatic rhizome, and belongs to the Zingiberaceae family. This herb is commonly used by local practitioners in traditional Asian medicine. Methods: In the present investigation, the novel Kaempferia galanga rhizome essential oil rich in ethyl p-methoxy cinnamate (EMCKG) was evaluated using GC/MS for chemical composition analysis. EMCKG was analyzed for its possible antimicrobial, neurodegenerative inhibitory, acetylcholinesterase, anti-inflammatory, and antioxidant activities as well as for the genotoxic effects using the standard methodologies. ANOVA and post hoc was performed to test the statistical significance of the study. Results: GC/MS analysis identified ethyl p-methoxy cinnamate as the major component of EMCKG essential oil with an area percentage of 66.39%. The EMCKG exhibited moderate (DPPH assay IC50 = 15.64 ± 0.263 µg/mL; ABTS assay IC50 = 16.93 ± 0.228 µg/mL) antioxidant activity than standard ascorbic acid (DPPH assay IC50 = 21.24 ± 0.413 µg/mL; ABTS assay IC50 = 21.156 ± 0.345 µg/mL). Similarly, EMCKG showed comparable activity in albumin denaturation (IC50 = 2.93 ± 0.59 µg/mL) and protease inhibitor assay (IC50 = 17.143 ± 0.506 µg/mL) to that of standard sodium diclofenac (IC50 = 23.87 ± 0.729 µg/mL and IC50 = 19.18 ± 0.271 µg/mL, respectively). The EMCKG exhibited a dose-dependent antimicrobial activity pattern with the highest inhibitory activity at 500 µg/mL against Staphylococcus aureus and considerable anticholinesterase activities (IC50 = 21.94 ± 0.109 µg/mL) compared to the standard galanthamine (IC50 = 27.18 ± 0.511 µg/mL). EMCKG also showed strong anti-diabetic activity (IC50 = 18.503 ± 0.480 µg/mL) and anti-tyrosinase activity (IC50 = 14.756 ± 0.325 µg/mL) as compared to the standards used (acarbose IC50 = 20.39 ± 0.231 µg/mL and kojic acid IC50 = 17.73 ± 0.192 µg/mL) in the study. Genotoxicity analysis of EMCKG revealed that at 1 µg/mL concentration has no toxic effects in mitosis of Allium cepa roots (Mitotic Index MI = 13.56% and chromosomal aberration CA = 07.60%). The ANOVA confirmed that except for the anticholinesterase activity, there is insignificant difference for essential oil and standards used for all the other bioactivities thus confirming their interchangeable applicability. Conclusions: Current research provides the basis for the fact that besides being a rich source of ethyl p-methoxycinnamate, EMCKG has the potential for future formulation and development of an inexpensive skin-care agent and for the preparation of anti-diabetic drugs.

Anethole rich Clausena heptaphylla (Roxb.) Wight & Arn., essential oil pharmacology and genotoxic efficiencies.

Anethole, a widely used industrial flavoring agent is majorly sourced from anise and star anise. The present study is aimed to the in-depth pharmacological analysis i.e. anti-diabetic, skin whitening, neurodegenerative disorder inhibitory activities of anethole-rich Clausena heptaphylla leaf essential oil (ARCHEO) (88.59%) as revealed by the Gas Chromatography/Mass Spectrometry (GC/MS) analysis and further confirmed by proton nuclear magnetic resonance 1H-NMR as well as to compare with standard compound anethole. ARCHEO (ABTS EC50 6.97 ± 0.004 µg/mL; Protease assay 4.51 ± 0.004 µg/mL) outperformed the standard compound anethole (ABTS EC50 9.48 ± 0.048 µg/mL; Protease assay EC50 22.64 ± 0.016 µg/mL) in antioxidant and anti-inflammatory experiments. ARCHEO was also shown to be more effective than the reference compound anethole in terms of anti-diabetic activity (EC50 22.35 ± 0.121 µg/mL), tyrosinase inhibitory activity (EC50 16.45 ± 0.012 µg/mL), and anti-cholinesterase activity (EC50 22.32 ± 0.016 µg/mL). However, ARCHEO exhibited lower antimicrobial activity towards all the tested microbes compared to standard compound anethole and as for the MIC, ARCHEO was effective only towards Salmonella typhimurium (60 µg/mL), Streptococcus mutans (20 µg/mL), and Aspergillus fumigatus (75 µg/mL). ARCHEO (11.11%) and anethole (12.33%) showed no genotoxic effect based on Allium cepa assay mitotic index value. Thus, ARCHEO could be a commercially viable and widely available cheaper source of anethole, which has buoyant demand in the field of food flavoring, fragrance, and pharmaceutical industries.

Evaluation of ultra high-performance liquid chromatography (uHPLC) assisted capsaicinoids content in four different extracts of (Capsicum chinense Jacq.): their pharmacological potentials and genotoxicity study.

The present study determined the capsaicin, dihydrocapsaicin and nordihydrocapsaicin of Capsicum chinense Jacq., ethanol, methanol, acetonitrile and dry acetone extracts using ultra high-performance liquid chromatography (uHPLC) technique. Highest capsaicin (2.84%) and nordihydrocapsaicin (0.56%) content was recorded in ethanol extract, while methanol extracts constituted highest dihydrocapsaicin (1.27%). Strong anti-inflammatory activity was shown by ethanol extract. All the extracts were found to be weak anti-diabetic, skin whitening and neurodegenerative agent. In genotoxicity test, ethanol extract showed mitotic index (MI) of 13.16% which was close to distilled water 25.72%. The chromosomal aberration of 8.0% was shown by ethanolic extract. The mitotic index value and chromosome aberration percentages of all the four extracts were similar but far from positive mutagenic agent ethyl methane sulfonate. Ethanol extract proved to be the best solvent for capsaicinoids extraction in comparison to the other three solvents and emerged as a potential pharmacological candidate having excellent anti-inflammatory ability.

Chemical composition of Callistemon citrinus (Curtis) Skeels aerial part essential oil and its pharmacological applications, neurodegenerative inhibitory, and genotoxic efficiencies.

Callistemon citrinus aerial part essential oil (CCEO) was analyzed for chemical compositions using GC/MS. Pharmacological activities such as neurodegenerative inhibitory, antioxidant, anti-inflammatory, antimicrobial, and acetylcholinesterase activities were evaluated using the standard methodologies. Genotoxicity was investigated using Allium cepa assay. GC/MS analysis identified 27 compounds; eucalyptol (55.40%) was the major component. Radical scavenging activity showed IC50 value of 16.71 µg/mL. Protein denaturation assay showed IC50 value of 21.19 µg/mL and 19.53µg/mL in protease inhibitor activity. MIC assay revealed antimicrobial potential of CCEO against microbial strains B. cereus at 2.00 mg/mL, S. typhimurium at 4.50 mg/mL, S. mutans at 2.50 mg/mL, C. albicans at 4.00 mg/mL, and S. cerevisiae at 4.50 mg/mL concentrations. Mitotic index value (MI) of CCEO showed negligible genotoxicity with MI 17.25% close to distilled water 18.22%. Acetylcholinesterase strong inhibitory activity of CCEO was observed from IC50  = 6.335 µg/mL. CCEO could be a cheap and easy source for the extraction of the pure compound eucalyptol and possess various biological activities which increase its pharmacological value as well as its applicability in the field of flavor and fragrance industries. PRACTICAL APPLICATIONS: Eucalyptol is the major component of many pharmaceutical and food/flavor industries. The present investigation provides a new source for the isolation of pure compound eucalyptol in cost-effective way. Additionally, the essential oil could also be used for the pharmaceutical formulation of antioxidant, inflammation inhibitory, and neurodegenerative inhibitory drug preparation under the safety range concentration.

Comparative Analysis of In-Vitro Biological Activities of Methyl Eugenol Rich Cymbopogon khasianus Hack., Leaf Essential Oil with Pure Methyl Eugenol Compound.

BACKGROUND: The essential oil of methyl eugenol rich Cymbopogon khasianus Hack. was evaluated and its bioactivities were compared with pure methyl eugenol. So far, methyl eugenol rich essential oil of lemongrass was not studied for any biological activities; hence, the present study was conducted. OBJECTIVE: This study examined the chemical composition of essential oil of methyl eugenol rich Cymbopogon khasianus Hack., and evaluated its antioxidant, anti-inflammatory, antimicrobial, and herbicidal properties and genotoxicity, which were compared with pure compound, methyl eugenol. MATERIAL AND METHODS: Methyl eugenol rich variety of Cymbopogon khasianus Hack., with registration no. INGR18037 (c.v. Jor Lab L-9) was collected from experimental farm CSIR-NEIST, Jorhat, Assam (26.7378°N, 94.1570°E). The essential oil wasobtained by hydro-distillation using a Clevenger apparatus. The chemical composition of the essential oil was evaluated using GC/MS analysis and its antioxidant (DPPH assay, reducing power assay), anti-inflammatory (Egg albumin denaturation assay), and antimicrobial (Disc diffusion assay, MIC) properties, seed germination effect and genotoxicity (Allium cepa assay) were studied and compared with pure Methyl Eugenol compound (ME). RESULTS: Major components detected in the Essential Oil (EO) through Gas chromatography/mass spectroscopy analysis were methyl eugenol (73.17%) and ß-myrcene (8.58%). A total of 35components were detected with a total identified area percentage of 98.34%. DPPH assay revealed considerable antioxidant activity of methyl eugenol rich lemongrass essential oil (IC50= 2.263 µg/mL), which is lower than standard ascorbic acid (IC50 2.58 µg/mL), and higher than standard Methyl Eugenol (ME) (IC50 2.253 µg/mL). Methyl eugenol rich lemongrass EO showed IC50 38.00 µg/mL, ME 36.44 µg/mL, and sodium diclofenac 22.76 µg/mL, in in-vitro anti-inflammatory test. Moderate antimicrobial activity towards the 8 tested microbes was shown by methyl eugenol rich lemongrass essential oil whose effectiveness against the microbes was less as compared to pure ME standard. Seed germination assay further revealed the herbicidal properties of methyl eugenol rich essential oil. Moreover, Allium cepa assay revealed moderate genotoxicity of the essential oil. CONCLUSION: This paper compared the antioxidant, anti-inflammatory, antimicrobial, genotoxicity and herbicidal activities of methyl eugenol rich lemongrass with pure methyl eugenol. This methyl eugenol rich lemongrass variety can be used as an alternative of methyl eugenol pure compound. Hence, the essential oil of this variety has the potential of developing cost-effective, easily available antioxidative/ antimicrobial drugs but its use should be under the safety range of methyl eugenol and needs further clinical trials.

Study of Anti-oxidant, Anti-inflammatory, Genotoxicity, and Antimicrobial Activities and Analysis of Different Constituents found in Rhizome Essential Oil of Curcuma caesia Roxb., Collected from North East India.

BACKGROUND: This investigation was designed to evaluate the chemical composition, antioxidant, anti-inflammatory, genotoxicity, and antimicrobial activities of Curcuma caesia Roxb rhizome essential oil. METHODS: Gas Chromatography/Mass Spectroscopy (GC/MS) analysis was performed to determine the chemical composition, standard antioxidative test DPPH assay, reducing power assay, in vitro antiinflammatory activity (egg albumin denaturation, protease inhibitory assay) by using standard methods. Similarly, antimicrobial activity was tested using the disc diffusion method, minimum inhibitory concentration ability (MIC); while to test genotoxicity, Allium cepa assay was used. RESULTS: GC/MS analysis revealed eucalyptol (28.55%), epicurzerenone (19.62%), and camphor (21.73%) as the major components of C. caesia rhizome essential oil. Potent antioxidant (IC50= 48.08±0.003 µg/mL), anti-inflammatory (IC50= 121.7±0.0013 µg/mL), and antimicrobial activities of the essential oil were recorded better than the standard drugs Fluconazole for fungus and Ciprofloxacin for bacteria. The essential oil also possessed a strong antibacterial effect against two tested bacterial strains B. subtilis and B. cereus with 7.5 µg/mL MIC value, while for fungal strains the essential oil was most effective against S. cereviaceae with an MIC value of 2.5 µg/mL. All the data were recorded in triplicates. Allium cepa assay revealed minor genotoxicity with mitotic index, MI= 27.70%; chromosome aberration, A= 1.1% of C. caesia rhizome essential oil. CONCLUSION: C. caesia rhizome essential oil possesses potent antioxidant, anti-inflammatory, and antimicrobial properties with negligible genotoxicity. Hence, the present study is highly significant for the utilization of rhizome of C. caesia, a high-value ethnopharmacological plant for advanced R & D and commercial application.