A combination of Citrus aurantifolia fruit rind and Theobroma cacao seed extracts supplementation enhances metabolic rates in overweight subjects: a randomized, placebo-controlled, cross-over study.

Background and objective: LN19183 is a proprietary, synergistic combination of Citrus aurantifolia fruit rind and Theobroma cacao seed extracts that increased resting energy expenditure (REE) in high-fat diet (HFD)-fed obese rats. The objective of this study was to validate the thermogenic potential of LN19183 in obese Sprague Dawley (SD) rats and to assess its clinical efficacy in a proof-of-concept, randomized, placebo-controlled, cross-over human trial. Methods: In the rat study, HFD-fed obese rats were supplemented with either HFD alone or with 45, 90, or 180 mg LN19183 per kg body weight (BW) for 28 days. In the human study, 60 overweight adults (male and female, aged 20-39 years) were randomized. Subjects took LN19183 (450 mg) or a matched placebo capsule on two consecutive days in phases one and two of the study, separated by a 10-day washout period. In each phase, on day 1, REE at pre-dose, 60-, 120-, and 180-min post-dose, and on day 2, metabolic rates at pre-dose and post-dose during and 20 min after exercise were measured using indirect calorimetry. Results: In rats, LN19183 significantly increased REE, reduced BW gain and fat masses, and increased fat and carbohydrate metabolism marker proteins including beta 3 adrenergic receptor (ß3-AR), phospho-AMP-activated protein kinase (AMPK), glucagon-like peptide-1 receptor (GLP-1R) in the liver, and serum adiponectin levels. Furthermore, LN19183-supplemented human volunteers increased (P < 0.05, vs. placebo) the metabolic rates at rest and with exercise; their fat oxidation was increased (P < 0.05, vs. placebo) at rest and 20 min post-exercise. The groups' systolic and diastolic blood pressure (BP), heart rates (HR), and safety parameters were comparable. Conclusion: These observations suggest that LN19183 is a thermogenic botanical composition with no stimulatory effects on BP and HR.

Investigation the effect of a herbal composition based on blackseed on patients with primary hypothyroidism: A randomized controlled trial.

Objective: Hypothyroidism is characterized by insufficient production of thyroxine by the thyroid gland. Levothyroxine may not fully alleviate patients' symptoms. This study aimed to assess the impact of a herbal product on weight, body mass index (BMI), thyroid hormones, lipid profile, fasting blood sugar (FBS), depression, and quality-of-life scores in patients. Materials and Methods: 72 patients with primary hypothyroidism, aged between 20 and 65 years old, participated in the trial and they were randomly allocated into two groups. The intervention group received the herbal powder containing Trachyspermum ammi L., Nigella sativa L., and Citrus aurantifolia L. while the control group received Avicel for 8 weeks. Results: Treatment with the herbal product resulted in statistically significant reductions in anthropometric variables such as BMI (p=0.03), hip circumference (HC) (p=0.008), waist circumference (WC) (p<0.001), and waist-to-hip circumference ratio (WHR) (p=0.003) in the intervention group in comparison between intervention and control groups. However, the decrease in weight was not statistically significant (p=0.08) in the intervention group compared the control group. In comparison between two groups, the depression score exhibited a statistically significant decrease in the intervention (p=0.001) and control groups (p=0.01), while there was a statistically significant increase in the quality-of-life score only in the intervention group (p<0.001) in comparison between intervention and control groups. Conclusion: The results indicate the potential beneficial effects of the herbal product on anthropometric variables in patients. Furthermore, the intervention yielded significant improvements in depression symptoms and quality-of-life scores among the patients.

Elucidation of the composition, antioxidant, and antimicrobial properties of essential oil and extract from Citrus aurantifolia (Christm.) Swingle peel.

The most effective methodologies for generating Musa spp. explants involve the utilization of plant tissue culture micropropagation techniques. However, the pervasive challenge of microbial contamination significantly impedes the successful micropropagation of Musa spp. This study examined the antioxidant and antibacterial characteristics of the essential oil (LPO) and extract (LPE) obtained from the peel of Citrus aurantifolia. Additionally, we explored their mechanisms against common microbial contaminants in Musa spp. micropropagation. Using gas chromatography-mass spectrometry, we identified 28 components in LPO, with δ-limonene, ß-pinene, citral, trans-citral, ß-bisabolene, geranyl acetate, and α-pinene as the primary constituents. Meanwhile, liquid chromatography-mass spectrometry detected 17 components in LPE, highlighting nobiletin, tangeretin, scoparone, sinensetin, tetramethylscutellarein, 5-demethylnobiletin, and pyropheophorbide A as the predominant compounds. Evaluation using the DPPH and ABTS methods revealed the IC50 values for LPE at 0.66 ± 0.009 and 0.92 ± 0.012 mg/mL, respectively, indicating higher antioxidant activity compared to LPO, with IC50 values of 3.03 ± 0.019 and 4.27 ± 0.023 mg/mL using the same methods. Both LPO and LPE exhibited antimicrobial activities against all tested contaminant microorganisms through in vitro assays. Mechanistic investigations employing time-kill analysis, assessment of cell membrane integrity, and scanning electron microscopy (SEM) revealed changes in the morphological characteristics of the tested microbial contaminants, intensifying with increased concentration and exposure duration of LPO and LPE. These alterations led to substantial damage, including cell wall lysis, leakage of intracellular components, and subsequent cell death. Consequently, LPO and LPE emerge as promising alternatives for addressing microbial contamination in banana tissue cultures.

Sensory Impact of Novel Dihydrocoumarins in Native Lime Oils.

Citrus fruits have been known and valued for their aroma in food and perfume ever since humans have maintained written records. Often described as the "champagne" of citrus oils, especially cold pressed lime peel oils have raised attention. Particularly peel oils of Citrus latifolia exhibit a pleasant coumarinic, sweet, and balsamic aroma in comparison to its close relative, the Citrus aurantifolia. Those coumarinic notes have not been completely understood until today. Thus, this study aimed to identify the responsible substances and elucidate their contribution and impact on the aroma of cold-pressed lime oil. By combining distillation, fractionation, olfactory detection, and structure elucidation, the responsible key aroma components were identified. A combination of coumarins and their corresponding saturated analogs have been identified to significantly contribute to the typical coumarinic-like aroma, including three new flavor compounds that have not yet been described in the literature as lime oil constituents: 7-methoxy-2-chromanone (3,4-dihydro-7-methoxy-2H-1-benzopyran-2-one; CAS 20921-02-2), 5,7-dimethoxy-2-chromanone (3,4-dihydro-5,7-dimethoxy-2H-1-benzopyran-2-one; CAS 82243-01-4) and 5,6-dihydrobergaptene (5,6-dihydro-4-methoxy-7H-furo[3,2-g][1]benzopyran-7-one; CAS 29050-61-1). The sensorial evaluation of the impact of these components on the lime aroma profile has shown flavor-modulating effects and the ability to enhance aldehydic-peely, juicy, and fruity notes as well as their importance in reproducing the authentic, typical coumarin-like notes.

Potassium Spraying Preharvest and Nanocoating Postharvest Improve the Quality and Extend the Storage Period for Acid Lime (Citrus aurantifolia Swingle) Fruits.

Citrus fruits are one of the most abundant crops globally in more than 140 countries throughout the world. Acid lime (Citrus aurantifolia swingle) is one of the citrus fruits which popularly has rich nutritional and therapeutic features. The storage period is the important factor that affects the economic and quality properties of this fruit. This study aims to demonstrate the enhancing effect of preharvest spraying with potassium, in addition to the postharvest dipping of fruits in some edible coatings, on the quality and storability of acid lime fruits. Preharvest spraying with organic and mineral forms of potassium, namely, potassium thiosulfate 1.75 g/L (S) and potassium tartrate 2 g/L (T), were carried out at three different times, in May, June, and July. On the other hand, postharvest treatments were carried out via dipping fruits in different types of biopolymers (carboxymethyl cellulose (E2) and gum arabic (E3)) and carboxymethyl cellulose/gum arabic composite (E4) as well as nanocoating formulation based on both biopolymers and doped zinc oxide nanoparticles (ZnONPs) (E1), which were prepared via acid lime peel waste extract. Herein, the physiochemical and morphological characterizations confirmed that the nanocoating was prepared at the nanoscale and doped with green synthesis ZnONPs, with recorded sizes of around 80 and 20 nm, respectively. Preharvest spraying with potassium tartrate enhanced fruit traits (Spraying with potassium tartrate at pre-harvest and nanocoating dipping at post-harvest (TE1), spraying with potassium tartrate at pre-harvest and carboxy methyl cellulose dipping at post-harvest (TE2), spraying with potassium tartrate at pre-harvest and gum arabic dipping at post-harvest (TE3) and spraying with potassium tartrate at pre-harvest and carboxymethyl cellulose/gum arabic composite dipping at post-harvest (TE4)), followed by potassium thiosulfate (spraying with potassium thiosulfate at pre-harvest and nanocoating dipping at post-harvest (SE1), spraying with potassium thiosulfate at pre-harvest and carboxy methyl cellulose dipping at post-harvest (SE2), spraying with potassium thiosulfate at pre-harvest and gum arabic dipping at post-harvest (SE3) and spraying with potassium thiosulfate at pre-harvest and carboxymethyl cellulose/gum arabic dipping at post-harvest (SE4)), compared to control. For postharvest treatments, E1 improved fruit quality, followed by E2, E4, and E3, respectively. The integration between pre- and postharvest treatments showed a clear superiority of TE2, followed by TE4, SE1, and SE2, respectively.

Efficient transformation and regeneration of transgenic plants in commercial cultivars of Citrus aurantifolia and Citrus sinensis.

Citrus is one of the major horticultural crops with high economic and nutraceutical value. Despite the fact that conventional research has developed numerous improved varieties, citriculture is still susceptible to various stresses and requires innovative solutions such as genetic engineering. Among all the currently available modern approaches, Agrobacterium-mediated transformation is the most efficient method for introducing desired traits in citrus. However, being a non-host for Agrobacterium, various citrus species, including Citrus aurantifolia and Citrus sinensis, are recalcitrant to this method. The available reports on Agrobacterium-mediated transformation of commercial citrus cultivars show very low transformation efficiency with poor recovery rates of whole transgenic plantlets. Here, we provide an efficient and reliable procedure of Agrobacterium-mediated transformation for both C. aurantifolia and C. sinensis. This protocol depends on providing callus-inducing treatment to explants before and during Agrobacterium co-cultivation, using optimum conditions for shoot regeneration and modifying in-vitro micrografting protocol to combat the loss of transgenic lines. As transgenic citrus shoots are difficult to root, we also developed the ideal conditions for their rooting. Using this protocol, the whole transgenic plantlets of C. aurantifolia and C. sinensis can be developed in about ~ 4 months, with transformation efficiency of 30% and 22% for the respective species.

Evaluating the genotoxicity of salinity stress and secondary products gene manipulation in lime, Citrus aurantifolia, plants.

Salinity is a significant abiotic stress that has a profound effect on growth, the content of secondary products, and the genotoxicity of cells. Lime, Citrus aurantifolia, is a popular plant belonging to the family Rutaceae. The interest in cultivating this plant is due to the importance of its volatile oil, which is included in many pharmaceutical industries, but C. aurantifolia plants are affected by the NaCl salinity levels. In the present study, a comet assay test has been applied to evaluate the genotoxic impact of salinity at 0, 50, 100, and 200 mM of NaCl on C. aurantifolia tissue-cultured plants. Furthermore, terpene gene expression was investigated using a semi-quantitative real-time polymerase chain reaction. Results from the two analyses revealed that 200 mM of NaCl stress resulted in high levels of severe damage to the C. aurantifolia plants' DNA tail 21.8%, tail length 6.56 µm, and tail moment 3.19 Unit. The relative highest expression of RtHK and TAT genes was 2.08, and 1.693, respectively, when plants were exposed to 200 mM of NaCl, whereas pv4CL2RT expressed 1.50 in plants subjected to 100 mM of NaCl. The accumulation of transcripts for the RTMYB was 0.951 when plants were treated with NaCl at 50 mM, and RtGPPS gene was significantly decreased to 0.446 during saline exposure at 100 mM. We conclude that the comet assay test offers an appropriate tool to detect DNA damage as well as RtHK, TAT, and pv4CL2RT genes having post-transcriptional regulation in C. aurantifolia plant cells under salinity stress. Future studies are needed to assess the application of gene expression and comet assay technologies using another set of genes that show vulnerability to different stresses on lime and other plants.

Bioactive Components and Their Activities from Different Parts of Citrus aurantifolia (Christm.) Swingle for Food Development.

Citrus aurantifolia is part of the Rutaceae family and belongs to the genus Citrus. It is widely used in food, the chemical industry, and pharmaceuticals because it has a unique flavor and odor. It is nutrient-rich and is beneficial as an antibacterial, anticancer, antioxidant, anti-inflammatory, and insecticide. Secondary metabolites present in C. aurantifolia are what give rise to biological action. Flavonoids, terpenoids, phenolics, limonoids, alkaloids, and essential oils are among the secondary metabolites/phytochemicals discovered in C. aurantifolia. Every portion of the plant's C. aurantifolia has a different composition of secondary metabolites. Environmental conditions such as light and temperature affect the oxidative stability of the secondary metabolites from C. aurantifolia. The oxidative stability has been increased by using microencapsulation. The advantages of microencapsulation are control of the release, solubilization, and protection of the bioactive component. Therefore, the chemical makeup and biological functions of the various plant components of C. aurantifolia must be investigated. The aim of this review is to discuss the bioactive components of C. aurantifolia such as essential oils, flavonoids, terpenoids, phenolic, limonoids, and alkaloids obtained from different parts of the plants and their biological activities such as being antibacterial, antioxidant, anticancer, an insecticide, and anti-inflammatory. In addition, various extraction techniques of the compounds out of different parts of the plant matrix as well as the microencapsulation of the bioactive components in food are also provided.

Bio-Fabrication of Silver Nanoparticles Using Citrus aurantifolia Fruit Peel Extract (CAFPE) and the Role of Plant Extract in the Synthesis.

The green synthesis of silver nanoparticles has been proposed as an eco-friendly and cost-effective substitute for chemical and physical methods. The aim of this study was to synthesize and characterize silver nanoparticles using the peel extract of Citrus aurantifolia fruit, and to determine the possible phytochemical constituents' presence in the plant extracts that might be responsible for the synthesis. Citrus aurantifolia fruit peel extraction was followed by phytochemical studies of secondary metabolites, FTIR analysis confirmation of functional groups, and GC-MS analysis. Silver nanoparticles were synthesized through bio-reduction of silver ions (Ag+) to silver nanoparticles using CAFPE and characterized using UV-Vis spectroscopy, HR-TEM, FESEM, EDX, XRD, DLS, and FTIR. The presence of plant secondary metabolites such as alkaloids, flavonoids, tannins, saponins, phenols, terpenoids, and steroids was detected. The FTIR analysis of the extract revealed the presence of functional groups like hydroxyl, carboxyl, carbonyl, amine, and phenyl, whereas the GC-MS analysis indicated presence of chemical compounds such as 1,2,4-Benzenetricarboxylic acid, Fumaric acid, nonyl pentadecyl, and 4-Methyl-2-trimethylsilyloxy-acetophenone, etc., with similar functional groups. The synthesized silver nanoparticle (AgNP) has displayed the characteristics of a surface plasmon resonance (SPR) band peak from 360-405 nm. High resolution transmission electron microscope (HR-TEM) and field emission scan electron microscope (FESEM) confirm polydisperse, spherical shaped, and smooth surface nanoparticles with an average size of 24.023 nm. Energy dispersive X-ray (EDX) analysis further revealed that silver is the most abundant element found in the micrograph of the nanoparticles, and FTIR analysis further confirmed the presence of different functional groups in the surface of the nanoparticle. The XRD analysis also confirmed that the nanoparticles synthesized are crystalline in nature. Based on the findings of this study, it is understood that the variety of natural compounds that are present in plant extracts of Citrus aurantifolia fruit peel can act as both reducing and stabilizing agents for the synthesis of silver nanoparticles. It is, therefore, concluded that Citrus aurantifolia peel extract can be potentially used for the large production of silver nanoparticles for several applications.

In vitro antioxidant, anti-inflammatory, and anticancer activities of mixture Thai medicinal plants.

BACKGROUND: The phytochemical study of medicinal plants is rapidly gaining popularity with many pharmacologic effects. This study aims to determine the antioxidant capacity as well as anticancer and antimigration activities of Clear belongs Plus extract (CBL-P) which consisted of five medicinal plants namely, Alpinia galanga, Piper nigrum, Citrus aurantifolia, Tiliacora triandra, and Cannabis sativa on human colon cancer cells SW620 and HCT116 cell lines, and human non-small cell lung cancer cells A549 and NCI-H460 cell lines. METHODS: In this study the dried-plant powder was extracted using 90% ethanol. Additionally, CBL-P was studied antioxidative activity via DPPH and ABTS assays and anti-inflammatory activities using nitric oxide assay using Griess reaction. Antiproliferation and antimigration of CBL-P were investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and scratch assay. RESULTS: The results showed that CBL-P had potent antiproliferative activity with IC50 values in a concentration- and time-dependent manners for all four cell lines. CBL-P also possessed potent antimigration activity against all studied cancer cells. CBL-P demonstrated antimigration activity on four different types of cancer cells (A549, NCI-H460, HCT116, and SW620) after 48 h of incubation, with the greatest effect seen at the highest concentration tested (15 µg/mL) in A549 cells (10.23% of wound closure) and NCI-H460 cells (9.16% of wound closure). CBL-P was also effective in reducing migration in HCT116 and SW620 cells, with a range of closure area from 10-50%. In addition, CBL-P had antioxidant activity with IC50 values of 8.549 ± 0.241 mg/mL and 2.673 ± 0.437 mg/mL for DPPH and ABTS assays, respectively. CBL-P also showed anti-inflammatory activity with the best inhibitory activity on NO production at a concentration of 40 µg/mL. CONCLUSION: In conclusion, the mixture extract possessed antioxidant and anti-inflammatory activity. Furthermore, the mixture plant extract significantly exhibited antiproliferative and antimigration activities on SW620, HCT116, A549, and NCI-H460 cells (P ≤ 0.05). Taken together, our results suggest that medicinal plants may have synergistic effects that could potentially enhance the effectiveness of cancer treatment when used as adjuvants. These findings provide a solid scientific foundation for future efforts to explore the mechanism of action.

PRpnp, a novel dual activity PNP family protein improves plant vigour and confers multiple stress tolerance in Citrus aurantifolia.

Under field conditions, plants are often simultaneously exposed to several abiotic and biotic stresses resulting in significant reductions in growth and yield; thus, developing a multi-stress tolerant variety is imperative. Previously, we reported the neofunctionalization of a novel PNP family protein, Putranjiva roxburghii purine nucleoside phosphorylase (PRpnp) to trypsin inhibitor to cater to the needs of plant defence. However, to date, no study has revealed the potential role and mechanism of either member of this protein group in plant defence. Here, we overexpressed PRpnp in Citrus aurantifolia which showed nuclear-cytoplasmic localization, where it functions in maintaining the intracellular purine reservoir. Overexpression of PRpnp significantly enhanced tolerance to salt, oxidative stress, alkaline pH, drought and two pests, Papilio demoleus and Scirtothrips citri in transgenic plants. Global gene expression studies revealed that PRpnp overexpression up-regulated differentially expressed genes (DEGs) related to ABA- and JA-biosynthesis and signalling, plant defence, growth and development. LC-MS/MS analysis validated higher endogenous ABA and JA accumulation in transgenic plants. Taken together, our results suggest that PRpnp functions by enhancing the endogenous ABA and JA, which interact synergistically and it also inhibits trypsin proteases in the insect gut. Also, like other purine salvage genes, PRpnp also regulates CK metabolism and increases the levels of CK-free bases in transgenic Mexican lime. We also suggest that PRpnp can be used as a potential candidate to develop new varieties with improved plant vigour and enhanced multiple stress resistance.

Pectin extraction from lime pomace by cold-active polygalacturonase-assisted method.

An Antarctic yeast was cultivated to produce and enzymatic extract with polygalacturonase activity, whose biochemical properties were studied. It assisted pectin extraction from lime pomace at 20 °C. The extract produced by Tausonia pullulans 8E had an optimum temperature of 40 °C and optimum pH of 5.0. At 20 °C, it displayed 54% of relative activity. A good pH-stability and thermostability were observed. Hg+2 and Co+2 inhibited its activity in 40 and 11%, respectively. Pectin was obtained from lime pomace at 20 °C, with 15% of yield after 120 min extraction. Uronic acid (46%) and neutral sugars (53%) were determined. Pectin's molecular weight was estimated in the order of 100,000 g mol-1. By anion-exchange chromatography, pectin-linked and free neutral sugars were observed. The high-methoxyl pectin was used to prepare low-calorie gels. It was demonstrated that this cold-active enzyme allows enzymatic-assisted pectin extraction at 20 °C.

Cold-pressed oil from Citrus aurantifolia inhibits the proliferation of vascular smooth muscle cells via regulation of PI3K/MAPK signaling pathways.

Vascular occlusive disease is a chronic disease with significant morbidity and mortality. Although a variety of therapies and medications have been developed, the likelihood of disease re-emergence is high and this can be life-threatening. Based on a previous screening experiment related to vascular obstructive diseases using 34 types of essential oils, cold-pressed oil (CpO) from Citrus aurantifolia (lime) has been demonstrated to have the best effect for the inhibition of vascular smooth muscle cells (VSMCs) proliferation. The aim of the present study was to evaluate the effect of lime CpO on the pathological changes of VSMCs. To determine this, the effect of lime CpO on VSMC proliferation, a major cause of vascular disease, was investigated. To determine the safe concentration interval for toxicity of CpO during VSMC culture, a dilution of 1x10-5 was determined using Cell Counting Kit-8 assay, which was confirmed to be non-toxic using a lactate dehydrogenase assay. To examine the effect of lime CpO in cellular signaling pathways, changes in phosphorylation of both the PI3K/AKT/mTOR and extracellular signal-regulated MEK/ERK signaling pathways with serum were investigated. Furthermore, lime CpO with FBS also significantly decreased the expression levels of the cell cycle regulators cyclin D1 and proliferating cell nuclear antigen. Additionally, lime CpO with FBS significantly inhibited the sprouting of VSMCs in an ex vivo culture system. These results suggested that lime CpO inhibited the abnormal proliferation of VSMCs and can be developed as a nature-based therapeutic agent for obstructive vascular disease.

In vitro pharmacological screening of antioxidant, cytotoxic and enzyme inhibitory activities of Citrus aurantifolia Linn. Dried fruit extract.

The Lime Basra (Citrus aurantifolia Linn., Rutaceae) plant also known as dried lime, and Limoo Omani, is used both as a spice in meals and as an herbal tea in the treatment of some diseases in the Middle East. It was aimed to determine the biological activity screening of the 70% methanol, ethanol extracts and infusion which were prepared from dried fruits. 1,1-diphenyl-2-picrylhydrazyl (DPPH●) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS+●) radical scavenging activities, ferric reducing activity, cytotoxicity on A 549, MCF 7 and L929 cell lines and α-amylase inhibitory effects were determined. According to the results, 70% methanol extract was more active in antioxidant activity tests and ethanol extract was more active in cytotoxicity tests. Interestingly both 70% methanol and ethanol extracts were found to have potent hypoglycemic activity. The present findings shed light on the fact that it is important to research and scientifically evaluate plants with traditional medicinal use.

Identification of active compounds in ethyl acetate, chloroform, and N-hexane extracts from peels of Citrus aurantifolia from Maribaya, West Java, Indonesia.

Lime peel contains essential oils are used as anti-oxidants or anti-cancer compounds. As a traditional medicine, lime has been widely used as a substitute for antibiotics. This study aimed to identify active compounds in peel extracts from Citrus aurantifolia that grows in Indonesia. Extraction was carried out by maceration using three different solvents: ethyl acetate, chloroform, and n-hexane. The extracts were analyzed using gas chromatography-mass spectrometry. The results showed that lime peel contained many important compounds and that 28, 27, and 24 different chemical compounds, both minor and major constituents, were extracted by ethyl acetate, chloroform, and n-hexane, respectively. Four compounds were found in all three solvent extracts, namely, D-limonene, phytol, α-tocopherol, and 5, 7-dimethoxycoumarin. Forty-seven compounds were uniquely present in one solvent, including 17 in ethyl acetate, 17 in chloroform, and 13 in n-hexane. Among the active compounds extracted, several are of biological importance, for example, stigmasterol, D-limonene, Vitamin E, and α-tocopherol. It can be concluded that a variety of distinct compounds are extracted from the same lime peel sample when different solvents are used and that some of these are bioactive compounds with anti-oxidant, anti-microbial, or anti-cancer properties.

De novo assembly and functional annotation of Citrus aurantifolia transcriptome from Candidatus Liberibacter asiaticus infected and non-infected trees.

Mexican lime (Citrus aurantifolia) belongs to the Rutaceae family and nowadays is one of the major commercial citrus crops in different countries. In Mexico, Mexican lime production is impaired by Huanglongbing (HLB) disease associated to Candidatus Liberibacter asiaticus (CLas) bacteria. To date, transcriptomic studies of CLas-Citrus interaction, have been performed mainly in sweet citrus models at symptomatic (early) stage where pleiotropic responses could mask important, pathogen-driven host modulation as well as, host antibacterial responses. Additionally, well-assembled reference transcriptomes for acid limes including C. aurantifolia are not available. The development of improved transcriptomic resources for CLas-citrus pathosystem, including both asymptomatic (early) and symptomatic (late) stages, could accelerate the understanding of the disease. Here, we provide the first transcriptomic analysis from healthy and HLB-infected C. aurantifolia leaves at both asymptomatic and symptomatic stages, using a RNA-seq approach in the Illumina NexSeq500 platform. The construction of the assembled transcriptome was conducted using the predesigned workflow Transflow and a total of 41,522 tentative transcripts (TTs) obtained. These C. aurantifolia TTs were functionally annotated using TAIR10 and UniProtKB databases. All raw reads were deposited in the NCBI SRA with accession numbers SRR10353556, SRR10353558, SRR10353560 and SRR10353562. Overall, this dataset adds new transcriptomic valuable tools for future breeding programs, will allow the design of novel diagnostic molecular markers, and will be an essential tool for studying the HLB disease.

Effects of lime (Citrus aurantifolia) peel to the expression of mRNA toll-like receptors 4 in balb/c mice-infected Salmonella typhi.

Lime peel contains metabolic compounds that have lethal effects of bacterial cells, but its effect as an antibacterial modulate innate immunity pathways, especially toll-like receptor 4 (TLR-4) signaling pathway, is unclear. This study examined the effects of lime peel extract (LPE) on the activity of TLR 4 in Balb/c mice induced by Salmonella typhi. Mice were induced intraperitoneally and then 3 days after induction, LPE was given orally on two doses (510 and 750 mg/kg BW). The number of bacterial colonization was counted using peritoneal fluid samples by the method of plate count agar. Intervention LPE for 5 days can degrade TLR-4 and the number of colonies of S. typhi. On day 3 after was induced S. typhi, TLR-4 gene expression of Balb/c mice is increased. Postintervention LPE for 5 days, the expression of TLR-4 gene decreased, significantly different at a dose of 750 mg/kg BW (P = 0.04). There was a positive correlation between the expression of TLR-4 gene by the number of bacterial colonization, decreasing gene expression of TLR-4, the number of bacterial colonization is also getting smaller (P = 0.013, r = 0.408). LPE can modulate the TLR-4 signaling pathway in host immunity so that the gene TLR-4 is expressed fewer in numbers. This mechanism causes the bacterial colony number to decrease, not even growth.

Insecticidal activities of Citrus aurantifolia essential oil against Aedes aegypti (Diptera: Culicidae).

BACKGROUND: In the recent time, global attention for the control of vectors has shifted from chemical insecticides to botanicals. In the present investigation, authors attempted to evaluate the efficacy of peel and leaf essential oil (EO) of Citrus aurantifolia against Aedes aegypti. RESULTS: The results revealed that both the oils possess more ovicidal activity (LC50 value of 5.26 ppm and 17.71 ppm for leaf and peel oil respectively at 72 h) than larvicidal activity. As larvicide, the essential oil from the peel of Citrus aurantifolia showed rapid effect with LC50 value of 128.81 ppm at 24 h which reduced to 106.77 ppm at 72 h while the leaf oil showed slow effect with LC50 value of 188.59 ppm, 107.37 ppm and 104.59 ppm at 24 h, 48 h and 72 h respectively. Again, the two essential oils did not show significant adulticidal activity. GC-MS analysis of both the oils recorded presence of different compounds. As a major constituent compound of the leaf EO of Citrus aurantifolia, citral was tested for their ovicidal, larvicidal and adulticidal activities against Aedes aegypti. The result showed highest ovicidal activities (LC50 value of 4.84 ppm at 72 h) of citral followed by larvicidal (LC50 value of 87.02 ppm at 24 h) and adulticidal (LC50 value of 103.88 ppm at 24 h) activities. CONCLUSION: From this study, it can be concluded that the essential oil extracted from the leaf and peel of Citrus aurantifolia and one of its major constituent compound citral can be included in the mosquito control programme of Aedes aegypti.

Chemical profile and in vivo toxicity evaluation of unripe Citrus aurantifolia essential oil.

Citrus aurantifolia (Christm.) Swingle (syn. C. MEDICA var. ACIDA Brandis) (family: Rutaceae) essential oil is one of the cheapest oils found in local markets. Although, it is generally accepted as non-toxic to vital organs and cells, majority of people are cynical about it usage. Herein, the present study reports the chemical composition and in vivo oral toxicity study of unripe C. aurantifolia essential oil found in Ghana. The toxicity of C. aurantifolia essential oil extract was investigated via oral administration using two methods: The acute toxicity single dose study (SDS) and the repeated dose method. The oil exhibited no acute toxicity but in the sub-chronic studies, the effects was dose and time-dependent. Chemical profile investigation of the oil showed 9 constituent of phytochemicals (Germacrene isomers (61.2%), Pineen (14%), Linalool dimmer (2.9%), Bornane (11%), Citral (2.9%), Anethole (1.5%), Anisole (1.1%), Safrole (0.3%) and Demitol (0.6%)). Histopathological studies revealed conditions such as necrosis, edema and inflammatory reaction in the liver, spleen and kidneys. Marginal upsurge of biochemical parameters above normal and elevated levels of lymphocytes (35.20-46.40 g/dL) demonstrated mild toxicity among the 100 mg/kg and 500 mg/kg dose groups at the sub-chronic stage. Low levels of hemoglobin (13.60 to 12.70 g/dL), MCV (34.20-24.0 fL), MCH (40.20-36.40 g/dL) along with high levels of liver enzymes confirmed the mild toxicity of the oil at sub-chronic stage. These results demonstrate that, despite consideration of lime essential oil as safe, it can have mild hematotoxic, nephrotoxic and hepatotoxic effects.

Hormonal and metabolic responses of Mexican lime plants to CTV infection.

Plant viral infections alter gene expression and metabolism in infected host. To study the molecular responses of Mexican lime to CTV infection, an analysis of plant metabolome in response to infection with severe (T318) or mild (T385) isolates of CTV was performed. Healthy plants and those infected with any of the two virus strains showed different metabolite profiles, at different stages of new sprout development. Proline content increased in plants infected with CTV, proportionally to the virulence of the virus strain. Abscisic acid content decreased after virus infection whereas jasmonic and salicylic acid levels increased. CTV infection had an impact on plant secondary metabolism, by stimulating the synthesis of different metabolites such as l-methylhistidine, phenylpropanoid derivatives. These metabolites are common responses of different organisms, including higher mammals, to viral diseases, and its presence in this system points to the existence of universal responses to virus infection among different kingdoms.