Targeting Y220C mutated p53 by Foeniculum vulgare-derived phytochemicals as cancer therapeutics.

CONTEXT: The mutations in the TP53 gene are the most frequent (50-60% of human cancer) genetic alterations in cancer cells, indicating the critical role of wild-type p53 in the regulation of cell proliferation and apoptosis upon oncogenic stress. Most missense mutations are clustered in the DNA-binding core domain, disrupting DNA binding ability. However, some mutations like Y220C occur outside the DNA binding domain and are associated with p53 structure destabilization. Overall, the results of these mutations are single amino acid substitutions in p53 and the production of dysfunctional p53 protein in large amounts, consequently allowing the escape of apoptosis and rapid progression of tumor growth. Thus, therapeutic targeting of mutant p53 in tumors to restore its wild-type tumor suppression activity has immense potential for translational cancer research. Various molecules have been discovered with modern scientific techniques to reactivate mutant p53 by reverting structural changes and/or DNA binding ability. These compounds include small molecules, various peptides, and phytochemicals. TP53 protein is long thought of as a potential target; however, its translation for therapeutic purposes is still in its infancy. The study comprehensively analyzed the therapeutic potential of small phytochemicals from Foeniculum vulgare (Fennel) with drug-likeness and capability to reactivate mutant p53 (Y220C) through molecular docking simulation. The docking study and the stable molecular dynamic simulations revealed juglalin (- 8.6 kcal/mol), retinol (- 9.14 kcal/mol), and 3-nitrofluoranthene (- 8.43 kcal/mol) significantly bind to the mutated site suggesting the possibility of drug designing against the Y220C mutp53. The study supports these compounds for further animal based in vivo and in vitro research to validate their efficacy. METHODS: For the purposes of drug repurposing, recently in-silico methods have presented with opportunity to rule out many compounds which have less probability to act as a drug based on their structural moiety and interaction with the target macromolecule. The study here utilizes molecular docking via Autodock 4.2.6 and molecular dynamics using Schrodinger 2021 to find potential therapeutic options which are capable to reactive the mutated TP53 protein.

Characterization of fennel germplasm for physiological persistence and drought recovery: Association with biochemical properties.

In the perennial medicinal plant fennel (Foeniculum vulgare), persistence over years, production stability, and successful post-drought recovery are as important as plant productivity. Characterization of productivity, post-drought recovery, persistency and their association with phytochemical properties has not yet been performed in fennel. In this study, 64 fennel genotypes (from 23 different countries) from four subspecies/varieties including F. vulgare ssp. piperitum, F. vulgare var. vulgare and F. vulgare var. dulce and F. vulgare var. azoricum were evaluated in the field over four years (2015-2018), then was assessed for post-drought recovery over the next two years (2019-2020). High genotypic variation was observed among genotypes and subspecies. Based on the GC-MS analysis, trans-anethole (22.4-90.6%), estragole (2.1-25.8%), fenchone (4.9-19.8%), and limonene (0.5-11.9%) are major components in the essential oils of the studied germplasm. The highest persistence and the lowest average post-drought recovery belonged to ssp. piperitum. This subspecies also had the highest amount of limonene, fenchone, and estragole and the lowest amount of trans-anethole compared to other subspecies. The highest essential oil content and stability was observed in var. dulce. Seed yield and persistence were found to be negatively associated, suggesting that selection for more productivity may indirectly promote less persistent genotypes. Post-drought recovery was negatively associated with two main phytochemical compounds of essential oil, fenchone and estragole, but positively associated with trans-anethole. Persistence had negative correlation with estragole content. Results indicated that indirect selection for post-drought recovery and persistence may be possible through selection for phytochemical properties. The negative correlation between some essential oil components may indicate that some are isomers, which can limit the ability to select for certain combinations. Superior genotypes identified in this study can be used to construct populations for future studies and as parents of crosses to develop new varieties.

Correlation between insulin-like growth factor 1 gene expression and fennel (Foeniculum vulgare) seed powder consumption in muscle of sheep.

It has been shown that addition of fennel in the diets of domestic animals has positive and beneficial effects on growth and meat production traits. Thus, the purpose of current study was to investigate the effect of adding fennel in the ration on growth characteristics and on insulin-like growth factor 1 (IGF1) gene expression in muscle tissue of Kermani lamb. Feeding of animals were performed with three levels of fennel including zero, 10 and 20 g/kg dry matter (DM) for 90 days. After slaughter, small pieces of tissues were removed and rapidly transferred to a nitrogen tank. Then, total RNA extracting and the Real-Time PCR reaction was performed. Results showed that as the level of fennel in the diet increases the amount of IGF1 gene expression also increases significantly in humeral muscle and femur (leg) muscle tissues (p < 0.05). In animals fed with fennel, femur muscle weight, back muscle weight, lean meat weight, final weight, warm carcass weight and live daily gain were greater than in animals fed with diet without fennel (p < 0.05). According to the findings of this investigation, it can be concluded that fennel, by creating positive effects on IGF1 gene expression can be used to improve muscle structure.

Genetic analysis and physiological relationships of drought response in fennel: Interaction with mating system.

The consequences of water deficit and its interaction with pollination system (deliberate selfing compared with open-pollination) on physiological, agronomic and phytochemical traits are not understood in fennel (Foeniculum vulgare Mill.). A research was started by creating selfed (S1) and half-sib (HS) families on a fennel germplasm in 2018. Populations were studied in the field, applying a normal and a water deficit condition during two years (2019-2020). Considerable genotypic variation was observed within S1 and HS families for all of the evaluated traits, demonstrating that selection for these traits would be successful. Consequences of water deficit were manifested as declined most of the traits; and significantly increased essential oil content, harvest index, and proline content, in both populations. Mandatory selfing reduced the performance of genotypes for most of the traits confirming the existence of inbreeding depression (ID) with higher values for plant dry weight, seed yield, essential oil content, and number of umbelets per umbel. In S1 population, some of the studied traits had higher heritability estimates under normal condition and some of them showed higher heritability under water deficit. Positive relationship between GCA and STI in OP population indicated that it is possible to identify genotypes having high values of combining ability and drought tolerance. Results of the present study suggest that physiological traits cannot be used as an indicator to distinguish drought-tolerant genotypes in S1 progenies, whereas in OP progenies Chl a, Chl b, TChl, CAR, PRO, and RWC, which had significant correlations with drought tolerance, may be used for this purpose. Based on the results contrasting genotypes were identified, which can be used to develop mapping populations for genome studies of drought tolerance and physiological traits of this species in future studies.

Antibacterial Activity and Epigenetic Remodeling of Essential Oils from Calabrian Aromatic Plants.

Natural compounds have historically had a wide application in nutrition. Recently, a fundamental role has been identified for essential oils extracted from aromatic plants for their nutritional, antimicrobial, and antioxidant properties, and as food preservatives. In the present study, essential oils (EOs) from ten aromatic plants grown in Calabria (Italy), used routinely to impart aroma and taste to food, were evaluated for their antibacterial activity. This activity was investigated against Escherichia coli strain JM109, and its derived antibiotic-resistant cells selected by growing the strain at low concentrations of ampicillin, ciprofloxacin, and gentamicin by measuring the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). Although all the essential oils showed bactericidal activity, those from Clinopodium nepeta, Origanum vulgare, and Foeniculum vulgare displayed the greatest inhibitory effects on the bacterial growth of all cell lines. It is plausible that the antibacterial activity is mediated by epigenetic modifications since the tested essential oils induce methylation both at adenine and cytosine residues in the genomes of most cell lines. This study contributes to a further characterization of the properties of essential oils by shedding new light on the molecular mechanisms that mediate these properties.

Doubled Haploidy for Fennel (Foeniculum vulgare Mill.) and Dill (Anethum graveolens L.).

Doubled haploidy technology is a powerful tool to accelerate the breeding of new crop varieties. Protocols are not universal, as even species within the same family require a specific process. Here we describe methods for developing doubled haploids for fennel and dill, both Apiaceae species which are used for food, flavorings, and medicine.

The Mitochondrial Genome Assembly of Fennel (Foeniculum vulgare) Reveals Two Different atp6 Gene Sequences in Cytoplasmic Male Sterile Accessions.

Cytoplasmic male sterility (CMS) has always aroused interest among researchers and breeders, being a valuable resource widely exploited not only to breed F1 hybrid varieties but also to investigate genes that control stamen and pollen development. With the aim of identifying candidate genes for CMS in fennel, we adopted an effective strategy relying on the comparison between mitochondrial genomes (mtDNA) of both fertile and sterile genotypes. mtDNA raw reads derived from a CMS genotype were assembled in a single molecule (296,483 bp), while a draft mtDNA assembly (166,124 nucleotides, 94 contigs) was performed using male fertile sample (MF) sequences. From their annotation and alignment, two atp6-like sequences were identified. atp6-, the putative mutant copy with a 300 bp truncation at the 5'-end, was found only in the mtDNA of CMS samples, while the wild type copy (atp6+) was detected only in the MF mtDNA. Further analyses (i.e., reads mapping and Sanger sequencing), revealed an atp6+ copy also in CMS samples, probably in the nuclear DNA. However, qPCRs performed on different tissues proved that, despite its availability, atp6+ is expressed only in MF samples, while apt6- mRNA was always detected in CMS individuals. In the light of these findings, the energy deficiency model could explain the pollen deficiency observed in male sterile flower. atp6- could represent a gene whose mRNA is translated into a not-fully functional protein leading to suboptimal ATP production that guarantees essential cellular processes but not a high energy demand process such as pollen development. Our study provides novel insights into the fennel mtDNA genome and its atp6 genes, and paves the way for further studies aimed at understanding their functional roles in the determination of male sterility.

The leaf transcriptome of fennel (Foeniculum vulgare Mill.) enables characterization of the t-anethole pathway and the discovery of microsatellites and single-nucleotide variants.

Fennel is a plant species of both agronomic and pharmaceutical interest that is characterized by a shortage of genetic and molecular data. Taking advantage of NGS technology, we sequenced and annotated the first fennel leaf transcriptome using material from four different lines and two different bioinformatic approaches: de novo and genome-guided transcriptome assembly. A reference transcriptome for assembly was produced by combining these two approaches. Among the 79,263 transcripts obtained, 47,775 were annotated using BLASTX analysis performed against the NR protein database subset with 11,853 transcripts representing putative full-length CDS. Bioinformatic analyses revealed 1,011 transcripts encoding transcription factors, mainly from the BHLH, MYB-related, C2H2, MYB, and ERF families, and 6,411 EST-SSR regions. Single-nucleotide variants of SNPs and indels were identified among the 8 samples at a frequency of 0.5 and 0.04 variants per Kb, respectively. Finally, the assembled transcripts were screened to identify genes related to the biosynthesis of t-anethole, a compound well-known for its nutraceutical and medical properties. For each of the 11 genes encoding structural enzymes in the t-anethole biosynthetic pathway, we identified at least one transcript showing a significant match. Overall, our work represents a treasure trove of information exploitable both for marker-assisted breeding and for in-depth studies on thousands of genes, including those involved in t-anethole biosynthesis.

Leaf proteomics of drought-sensitive and -tolerant genotypes of fennel.

Fennel is attracted attention as a useful resource as researching medicinal plant for drought tolerance. To elucidate the response mechanism in drought-sensitive and -tolerant genotypes of fennel leaf, a gel-free/label-free proteomic technique was used. Fifty-day-old plants were subjected to drought stress for 60days. The relative water and proline contents were decreased and increased in sensitive genotypes, respectively; however, they were not a big change in tolerant genotypes. Photosynthesis was decreased in the sensitive genotypes under drought; however, it was increased in the tolerant genotype. In both drought-sensitive and -tolerant genotypes, proteins related to protein metabolism and cell organization were predominately affected under drought stress. The abundance of phosphoribulokinase and phosphoglycerate kinase enzymes were decreased and increased in drought-sensitive and -tolerant genotypes, respectively; however, the abundance of RuBisCO and glyceraldehyde-3-phosphate dehydrogenase enzymes were increased and decreased in drought-sensitive and -tolerant genotypes, respectively. Under drought stress, the abundance of glycolysis-related proteins was decreased in sensitive genotypes; however, they were increased in tolerance genotypes. Commonly changed proteins with polyethylene glycol fractionation such as cobalamin-independent methionine synthase were decreased and increased in drought-sensitive and -tolerant genotypes, respectively. These results suggest that cobalamin-independent methionine synthetase is involved in the tolerance of drought-tolerant fennel leaf under drought stress.

Seed germination of medicinal plant, fennel (Foeniculum vulgare Mill), as affected by different priming techniques.

Reduced seed germination is among the most important factors adversely affecting crop stand and subsequent plant growth. Fennel (Foeniculum vulgare Mill) is an important medicinal plant with poor seed germination rate, occasionally. It is accordingly pertinent to find methods which can enhance fennel seed germination and remove the barriers of dormancy breaking. The present experiments studied the effects of two different priming (cold moist stratification and osmopriming) and 14 dormancy breaking techniques (hormonal, osmopriming, biopriming, chemical priming, and hydropriming) on the seed germination and seedling growth of two different fennel genotypes under growth chamber conditions. In the first and second experiment, the priming techniques including the time lengths of cold moist stratification (0, 15, 30, and 45 days) and the concentrations of polyethylene glycol 6000 (PEG6000, osmopriming at -0.99, -1.35, and -2.33 MPa) were used as the main plots. However, in both experiments, the dormancy breaking techniques and fennel genotypes were factorially combined and used as the subplots. Different seed- and seedling-related parameters including germination (%), plumule, radicle and seedling length, average germination time, rate and homogeneity of germination, and seed vigor index were determined. Both priming techniques were efficient on the enhancement of seed germination and seedling growth. Among the dormancy breaking techniques, Aminol Forte (biopriming), kadostim (biopriming), benzyl adenine + kinetin (biopriming), distilled water (hydropriming), gibberellin + kinetin (hormonal priming), and benzyl adenine + kinetin + gibberellin (biopriming) were the most effective ones. The related concentrations were equal to 100 mg/l, 10(-5) M, and 0.4 %. The fennel genotypes reacted significantly different under priming conditions. It is possible to enhance seed germination and seedling growth of fennel using priming and dormancy breaking techniques, which is useful for the increased production of fennel under different conditions. The results indicate that bio and hydropriming techniques were among the most effective ones, which significantly increased seed germination and seedling growth, and removed the seed dormancy barriers.

Foeniculum vulgare Mill: a review of its botany, phytochemistry, pharmacology, contemporary application, and toxicology.

Foeniculum vulgare Mill commonly called fennel has been used in traditional medicine for a wide range of ailments related to digestive, endocrine, reproductive, and respiratory systems. Additionally, it is also used as a galactagogue agent for lactating mothers. The review aims to gather the fragmented information available in the literature regarding morphology, ethnomedicinal applications, phytochemistry, pharmacology, and toxicology of Foeniculum vulgare. It also compiles available scientific evidence for the ethnobotanical claims and to identify gaps required to be filled by future research. Findings based on their traditional uses and scientific evaluation indicates that Foeniculum vulgare remains to be the most widely used herbal plant. It has been used for more than forty types of disorders. Phytochemical studies have shown the presence of numerous valuable compounds, such as volatile compounds, flavonoids, phenolic compounds, fatty acids, and amino acids. Compiled data indicate their efficacy in several in vitro and in vivo pharmacological properties such as antimicrobial, antiviral, anti-inflammatory, antimutagenic, antinociceptive, antipyretic, antispasmodic, antithrombotic, apoptotic, cardiovascular, chemomodulatory, antitumor, hepatoprotective, hypoglycemic, hypolipidemic, and memory enhancing property. Foeniculum vulgare has emerged as a good source of traditional medicine and it provides a noteworthy basis in pharmaceutical biology for the development/formulation of new drugs and future clinical uses.