scholarly journals Induced expression of rolC for study of its effect on the expression of genes associated with nicotine synthesis in tobacco

2020 ◽  
Vol 18 (4) ◽  
pp. 413-422
Author(s):  
Gita Amini ◽  
Sofia V. Sokornova ◽  
Hanieh Mohajjel-Shoja ◽  
Andrey N. Stavrianidi ◽  
Igor A. Rodin ◽  
...  
Keyword(s):  
Inducible Promoter ◽  
Rol Genes ◽  
Primary Role ◽  
Rt Pcr ◽  
Rolc Gene ◽  
Nicotine Content ◽  
Expression Of Genes ◽  
Transgenic Lines ◽  
Tobacco Roots ◽  
And Control

Background. Agrobacterium rhizogenes rol genes cause not only hairy root syndrome in plants, but also affect their secondary metabolism. There are cases of increasing of nicotine content in transgenic tobacco roots expressing rolC alone or in combination with other rol genes. In this work, we evaluated the change in the expression of nicotine synthesis genes and their regulators in response to the induction of expression of rolC. Materials and methods. Plant material was represented by three Nicotiana tabacum genotypes: cv. Samsun and two transgenic lines, derived from this cultivar and containing rolC under dexamethasone inducible promoter: A. rhizogenes rolC (Pdex-A4rolC) and N. tabacum rolC (Pdex-trolC) correspondingly. Fluidigm Biomark RT-PCR was used for evaluation of expression of QPT1, QPT2, A622, ODC, ADC, PMT1, PMT2, PMT3, PMT4, MPO1, MPO2, BBL, MATE1, MATE2, ARF6, ERF168, ERF189, A4rolC, NtrolC, and reference gene gapdh. HPLC-MS / MS analysis was used to determine content of nicotine and its derivatives in plant tissues. Results. Expression of PMT genes for the synthesis of the pyrrolidine ring, as well as the genes, controlling enzyme for final stages of nicotine synthesis, was higher in transgenic lines without induction of rolC expression. Regulatory genes were activated by dexamethasone in both transgenic and control lines, indicating the inapplicability of rolC dexamethasone induction for their study. The level of expression of PMT and MPO genes increased over time in transgenic dexamethasone-induced lines. Nicotine content decreased in transgenic dexamethasone-induced plants. Conclusions. The rolC gene does not play a primary role in the regulation of nicotine synthesis genes. The mechanism of regulation of different nicotine biosynthesis genes and TFs varies.

scholarly journals Molecular epidemiology of coxsackievirus A16 circulating in children in Beijing, China from 2010 to 2019

2021 ◽  
Author(s):  
Ya-Fang Hu ◽  
Li-Ping Jia ◽  
Fang-Yuan Yu ◽  
Li-Ying Liu ◽  
Qin-Wei Song ◽  
...  
Keyword(s):  
Molecular Epidemiology ◽  
Evolutionary Rate ◽  
Foot And Mouth Disease ◽  
Recent Common Ancestor ◽  
Rt Pcr ◽  
Coxsackievirus A16 ◽  
Most Recent Common Ancestor ◽  
Etiological Agents ◽  
High Level ◽  
And Control

Abstract Background Coxsackievirus A16 (CVA16) is one of the major etiological agents of hand, foot and mouth disease (HFMD). This study aimed to investigate the molecular epidemiology and evolutionary characteristics of CVA16. Methods Throat swabs were collected from children with HFMD and suspected HFMD during 2010–2019. Enteroviruses (EVs) were detected and typed by real-time reverse transcription-polymerase chain reaction (RT-PCR) and RT-PCR. The genotype, evolutionary rate, the most recent common ancestor, population dynamics and selection pressure of CVA16 were analyzed based on viral protein gene (VP1) by bioinformatics software. Results A total of 4709 throat swabs were screened. EVs were detected in 3180 samples and 814 were CVA16 positive. More than 81% of CVA16-positive children were under 5 years old. The prevalence of CVA16 showed obvious periodic fluctuations with a high level during 2010–2012 followed by an apparent decline during 2013–2017. However, the activities of CVA16 increased gradually during 2018–2019. All the Beijing CVA16 strains belonged to sub-genotype B1, and B1b was the dominant strain. One B1c strain was detected in Beijing for the first time in 2016. The estimated mean evolutionary rate of VP1 gene was 4.49 × 10–3 substitution/site/year. Methionine gradually fixed at site-23 of VP1 since 2012. Two sites were detected under episodic positive selection, one of which (site-223) located in neutralizing linear epitope PEP71. Conclusions The dominant strains of CVA16 belonged to clade B1b and evolved in a fast evolutionary rate during 2010–2019 in Beijing. To provide more favorable data for HFMD prevention and control, it is necessary to keep attention on molecular epidemiological and evolutionary characteristics of CVA16.

scholarly journals A Synergistic Anti-Diabetic Effect by Ginsenosides Rb1 and Rg3 through Adipogenic and Insulin Signaling Pathways in 3T3-L1 Cells

2021 ◽  
Vol 11 (4) ◽  
pp. 1725
Author(s):  
Hee-Do Hong ◽  
Sun-Il Choi ◽  
Ok-Hwan Lee ◽  
Young-Cheul Kim
Keyword(s):  
Transcription Factors ◽  
Insulin Signaling ◽  
Adipocyte Differentiation ◽  
Red Ginseng ◽  
Inhibitory Effects ◽  
Rt Pcr ◽  
High Concentration ◽  
Expression Of Genes ◽  
The Individual ◽  
Maximal Expression

Although ginsenosides Rb1 and Rg3 have been identified as the significant ginsenosides found in red ginseng that confer anti-diabetic actions, it is unclear whether insulin-sensitizing effects are mediated by the individual compounds or by their combination. To determine the effect of ginsenosides Rb1 and Rg3 on adipocyte differentiation, 3T3-L1 preadipocytes were induced to differentiate the standard hormonal inducers in the absence or presence of ginsenosides Rb1 or Rg3. Additionally, we determined the effects of Rb1, Rg3, or their combination on the expression of genes related to adipocyte differentiation, adipogenic transcription factors, and the insulin signaling pathway in 3T3-L1 cells using semi-quantitative RT-PCR. Rb1 significantly increased the expression of CEBPα, PPARγ, and aP2 mRNAs. However, Rg3 exerted its maximal stimulatory effect on these genes at 1 μM concentration, while a high concentration (50 μM) showed inhibitory effects. Similarly, treatment with Rb1 and Rg3 (1 μM) increased the expression of IRS-1, Akt, PI3K, GLUT4, and adiponectin. Importantly, co-treatment of Rb1 and Rg3 (9:1) induced the maximal expression levels of these mRNAs. Our data indicate that the anti-diabetic activity of red ginseng is, in part, mediated by synergistic actions of Rb1 and Rg3, further supporting the significance of minor Rg3.

ESTIMATION OF SARS-COV-2 SPECIFIC ANTIBODIES SEROPREVALENCE IN HEALTHCARE WORKERS IN DISTRICT UDAIPUR, INDIA

2021 ◽  
pp. 61-63
Author(s):  
Shelesh Kumar Swami ◽  
Nitesh Kumar Chauhan ◽  
Shuchi Goyal ◽  
A.K. Verma ◽  
Shweta Biyani
Keyword(s):  
Healthcare Workers ◽  
Clinical Symptoms ◽  
Control Measures ◽  
Rt Pcr ◽  
High Seroprevalence ◽  
Know How ◽  
Novel Coronavirus ◽  
And Control ◽  
Infection Eradication ◽  
Cumulative Prevalence

Background:Current pandemic caused by Novel coronavirus (COVID-19) causes clinical symptoms from fever to acute respiratory distress syndrome but may remain mild or asymptomatic. To evaluate the cumulative prevalence of SARSCoV-2 infection in a community and know how immune response develops in the population, reliable assay alongwith RT-PCR for detection of SARS-CoV 2 antibodies is needed. Healthcare workers (HCWs) represent a high-risk populat - ion for infection with SARS-CoV-2. Methods: We evaluated total antibodies recognizing the SARS CoV 2 receptor binding domain (S1-RBD) - - - or the Spike protein over a period of six months in a total of 310 healthcare workers engaged in hospital using SARS-CoV-2 Total antibody assay kit. Findings: The overall seroprevalence found in our analysis was 41.93%. In case of males the percentage positive was found to be signicantly higher at 43.91%, compared to females at 36.25%. Seroprevalence was signicantly higher in 50 years above age group in comparison to 20-50 years old aged healthcare workers. The seroprevalence was higher in doctors, nursing staff and lab technicians than other healthcare professionals as 44.6%. Conclusions: This study showed high seroprevalence of SARS-CoV-2 in healthcare workers which means remaining proportion of the healthcare workers are still susceptible to the infection. Good compliance to infection eradication and control measures, adequate PPEs, and early detection and isolation of healthcare workers infected with SARS-CoV-2 are mandatory to reduce the risk of SARS-CoV-2 infection.

scholarly journals Effects of Dietary Supplementation with Different Polyunsaturated Fatty Acids on Expression of Genes Related to Somatotropic Axis Function in the Liver, Selected Blood Indicators, Milk Yield and Milk Fatty Acids Profile in Dairy Cows

2016 ◽  
Vol 16 (4) ◽  
pp. 1045-1058 ◽  
Author(s):  
Essa Dirandeh ◽  
Armin Towhidi ◽  
Zarbakht Ansari ◽  
Saeeid Zeinoaldini ◽  
Mehdi Ganjkhanlou
Keyword(s):  
Fatty Acids ◽  
Plasma Concentrations ◽  
Dietary Supplementation ◽  
Omega 3 ◽  
Milk Fatty Acids ◽  
Somatotropic Axis ◽  
Fatty Acids Profile ◽  
Expression Of Genes ◽  
Omega 6 ◽  
And Control

Abstract The objective of this study was to investigate whether dietary supplementation with different polyunsaturated fatty acids (PUFA s) affects expression of genes related to somatotropic axis and the plasma concentrations of insulin, glucose, non-esterified fatty acids (NEFA), beta hydroxyl butyrate acids (BHBA) and insulin-like growth factor 1 (IGF1) and milk fatty acids profile. Right after calving, Holstein cows (n=45) were randomly assigned to one of three diets supplemented with roasted whole soybean as a source of omega-6 PUFA (omega-6, n=15), linseed as a source of omega-3 PUFA (omega-3, n=15) or palm oil (control, n=15). Each cow was in the study over a period of 70 days. Blood samples were collected every two weeks from day 1 to 70 of lactation and plasma concentrations of insulin, glucose, NEFA, BHBA and IGF1 were determined. Liver samples were taken from a subset of 18 cows (6 per diet) at day 70 postpartum and hepatic mRNA level of total growth hormone-receptor 1A (GHR1A), insulin receptor (INSR), IGF1 and insulinlike growth factor binding protein (IGFBP2) was assessed. Experimental diets did not affect milk yield. Plasma glucose and insulin concentrations were greater for omega-3 treatment compared to omega-6 and control treatments. Cows fed diets enriched in omega-3 exhibited greater INSR and GHR1A mRNA expression, and a tendency for greater IGF1 mRNA expression in the liver compared to omega-6 and control cows. Plasma IGF1 concentration was significantly higher in omega-3 treatment compared with omega-6 and control treatments. Results of this study suggest that feeding omega-3 PUFA s during early postpartum couples with the somatotropic axis, leading to an increase in plasma IGF1 concentration in dairy cows.

scholarly journals Експериментальне зараження поросят вірусом епідемічної діареї свиней

2017 ◽  
Vol 19 (77) ◽  
pp. 208-213
Author(s):  
D. Masiuk ◽  
A. Sosnitskiy ◽  
A. Kokarev ◽  
S. Koliada
Keyword(s):  
Real Time ◽  
The Body ◽  
Control Group ◽  
Rt Pcr ◽  
Internal Organs ◽  
Real Time System ◽  
E Coli ◽  
Neonatal Piglets ◽  
And Control ◽  
Experimental Group

There were infected neonatal piglets in the first days of their lives PED virus suspension derived from pigs previously PED patients. Diagnosis for PED in piglets donor virus PED was inserted complex method for clinical and epizootic performance and confirmed the identification PEDV by PCR-RT using the test system «EZ-RED/TGE/PDCoV MPX 1.0 Real time RT-PCR» company Tetracore (USA) Thermocyclers CFX 96 Real-Time System company BIO RAD (USA). Homogenate small intestine of pigs PEDV donor, prepared in a blender for PCR in a thick band of 18 animal carcasses, frozen at -18 °C without cryopreservation and kept 359 days. Before infecting pigs and strip defrost by RT-PCR identified the concentration of the virus genome equivalents (GE) without establishing viable virions quantitative pathogen. For Sample 20 selected analog neonatal piglets, divided them into 3 experimental groups (group 1 – 5 piglets, group 2 – 5 piglets and group 3 – 7 piglets) and one control (3 piglets). Research pigs infected per os virus-containing suspension with a concentration PEDV 1.03×106 GE/cm3. The dose for infection first group was 6 cm3 (6.18×106 GE/cm3), for the second – 5 cm3 (5,15 × 106 GE/cm3), for the third – 4 cm3 (4.12 GE×106/cm3) homogenate. The fourth group – control (not infected). All the pigs were in identical conditions that fully meet the physiological needs of the body. Of the 17 infected pigs only 2 was infected PEDV. PED was confirmed by laboratory methods. In bacteriological examination of internal organs of pigs that came out of a research experiment and control group were diagnosed colibacteriosis. In the control group was isolated from heart and intestinal non-pathogenic for white mice E. coli. From pigs 1 and 2 research groups has been allocated to white mice nonpathogenic E. coli, is set colibacteriosis; 2 experimental group found in one pig hemolytic E. coli; 3 experimental group from the internal organs of pigs in conjunction with non-pathogenic for mice intestinal former cane isolated Klesiella spp., is diagnosed with mixed infection (E. coli, Klesiella spp.). From the intestine of experimental and control pigs do not identified beneficial microflora – aerococcus, lactobacteria, bifidobacteria and cultured putrefactive anaerobic spore facultative and non spore microflora.

scholarly journals Transcriptomics of Improved Fruit Retention by Hexanal in ‘Honeycrisp’ Reveals Hormonal Crosstalk and Reduced Cell Wall Degradation in the Fruit Abscission Zone

2021 ◽  
Vol 22 (16) ◽  
pp. 8830
Author(s):  
Karthika Sriskantharajah ◽  
Walid El Kayal ◽  
Davoud Torkamaneh ◽  
Murali M. Ayyanath ◽  
Praveen K. Saxena ◽  
...  
Keyword(s):  
Cell Wall ◽  
Ethylene Biosynthesis ◽  
Abscission Zone ◽  
Cell Wall Degrading Enzymes ◽  
Fruit Abscission ◽  
Fruit Drop ◽  
Expression Of Genes ◽  
Hormonal Crosstalk ◽  
Honeycrisp Apples ◽  
And Control

Apples (Malus domestica Borkh) are prone to preharvest fruit drop, which is more pronounced in ‘Honeycrisp’. Hexanal is known to improve fruit retention in several economically important crops. The effects of hexanal on the fruit retention of ‘Honeycrisp’ apples were assessed using physiological, biochemical, and transcriptomic approaches. Fruit retention and fruit firmness were significantly improved by hexanal, while sugars and fresh weight did not show a significant change in response to hexanal treatment. At commercial maturity, abscisic acid and melatonin levels were significantly lower in the treated fruit abscission zone (FAZ) compared to control. At this stage, a total of 726 differentially expressed genes (DEGs) were identified between treated and control FAZ. Functional classification of the DEGs showed that hexanal downregulated ethylene biosynthesis genes, such as S-adenosylmethionine synthase (SAM2) and 1-aminocyclopropane-1-carboxylic acid oxidases (ACO3, ACO4, and ACO4-like), while it upregulated the receptor genes ETR2 and ERS1. Genes related to ABA biosynthesis (FDPS and CLE25) were also downregulated. On the contrary, key genes involved in gibberellic acid biosynthesis (GA20OX-like and KO) were upregulated. Further, hexanal downregulated the expression of genes related to cell wall degrading enzymes, such as polygalacturonase (PG1), glucanases (endo-β-1,4-glucanase), and expansins (EXPA1-like, EXPA6, EXPA8, EXPA10-like, EXPA16-like). Our findings reveal that hexanal reduced the sensitivity of FAZ cells to ethylene and ABA. Simultaneously, hexanal maintained the cell wall integrity of FAZ cells by regulating genes involved in cell wall modifications. Thus, delayed fruit abscission by hexanal is most likely achieved by minimizing ABA through an ethylene-dependent mechanism.

scholarly journals Strain-specific differences in age-related changes in rat susceptibility to experimental autoimmune encephalomyelitis and dendritic cell cytokine gene expression

Genetika ◽  
2014 ◽  
Vol 46 (1) ◽  
pp. 287-301 ◽  
Author(s):  
Biljana Bufan ◽  
Jasmina Djikic ◽  
Mirjana Nacka-Aleksic ◽  
Zorica Stojic-Vukanic ◽  
Mirjana Dimitrijevic ◽  
...  
Keyword(s):  
Dendritic Cell ◽  
Rt Pcr ◽  
Autoimmune Encephalomyelitis ◽  
Expression Of Genes ◽  
Age Related ◽  
Genes Encoding ◽  
Immunosuppressive Cytokines ◽  
Age Related Changes ◽  
Experimental Autoimmune

Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis, a prototype of Th1/Th17-mediated organ-specific autoimmune disease. In the rat, susceptibility to development of these diseases is shown to be strain-and age-dependent. In adult rats of distinct strains, it correlates with splenic dendritic cell (DC) subset composition, which also exhibit age-related changes. The aim of this study was to examine influence of aging on: i) Albino Oxford (relatively resistant to EAE) and Dark Agouti (susceptible to EAE) rat development of EAE and ii) their splenic conventional (OX62+) DC population in respect to its subset composition and expression of mRNAs for proinflammatory and immunosuppressive cytokines. We used 3month-old (young) and 26-month-old (aged) rats of AO and DA strain. The rats were immunized for EAE with rat spinal cord homogenate in complete Freund?s adjuvant and clinical course of the disease was followed. Fresh OX62+DCs were examined for the expression of CD4 (using flow cytometry) and genes encoding cytokines influencing DC activation/maturation (TNF-? and IL-6) using RT-PCR. Additionally, in vitro lipopolysaccharide (LPS) activated/matured DCs were examined for the expression of genes encoding cytokines controlling Th1/Th17 cell polarization using RT-PCR. With aging, AO rats became more susceptible, whereas DA rats largely lose their susceptibility to the induction of EAE. In AO rats aging shifted CD4+:CD4DC ratio towards CD4-cells, producing large amount of proinflammatory cytokines, whereas in DA rats CD4+:CD4-DC ratio remained stable with aging. In fresh DCs from rats of both the strains the expression of TNF-? mRNA increased with aging, whereas that of IL-6 mRNA decreased and increased in DCs from AO and DA rats, respectively. Following in vitro LPS stimulation OX62+ DCs from aged AO rats up-regulated the expression of mRNA for IL-23p19 (specific subunit of IL-23; crucial for sustained IL-17 production) and IL-1? (positive IL-17 regulator), whereas down-regulated the expression of IL-10 (negative IL-17 regulator) when compared with young strain-matched rats. In DA rats aging incresed IL-23p19 mRNA expression in LPS-stimulated DCs, whereas exerted the opposing effects on the expression of mRNAs for IL-10 and IL-1? compared to AO rats. Irrespective of the rat strain, aging did not influence mRNA expression for IL-12p35 (driving Th1 polarization) in DCs. Overall, results suggest role of changes in the expression of genes encoding proinflammatory and immunosuppressive cytokines in development of age-related alterations in rat susceptibility to EAE induction.

scholarly journals Epidemiology and Control of Congo Fever in Sacrificial Animals of Pakistan

2019 ◽  
Vol 1 (2) ◽  
Author(s):  
Hafiz Muhammad Rizwan ◽  
Muhammad Sohail Sajid ◽  
Haider Abbas ◽  
Muhammad Fiaz Qamar ◽  
Qaiser Akram
Keyword(s):  
Tick Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Many Body ◽  
Rt Pcr ◽  
Body Tissues ◽  
Crimean Congo Haemorrhagic Fever ◽  
Polymerase Chain ◽  
And Control ◽  
Successful Prevention ◽  
Venereal Transmission

The cases and deaths due to Crimean-Congo haemorrhagic fever (CCHF) [49] virus commonly known as Congo virus (fatality rate 15%) have been reported throughout Pakistan from the last five years especially during religious occasion, Eid-ul-Azha. The annual increase in death rates due to CCHF demonstrate the importance of awareness of Congo fever at academia as well as public level. The symptoms of Congo fever which appear one to nine days after tick bite, include sudden high fever, muscle aches, abdominal pain, headache, dizziness, sore eyes, jaundice, mood swings, confusion, aggression, and sensitivity to light. The other signs include sore throat, joint pain, vomiting, diarrhea, hemorrhages, and bleeding from skin and large intestine. The Infection has been reported in many species of wild as well as domestic animals including hares, cattle, sheep, goats, dogs, mice and hedgehogs. At least 31 species of Hyalomma, Boophilus, Rhipicephalus, Dermacentor (Ixodidae: hard ticks) act as vector of CCHF in which transovarial, transstadial and venereal transmission occurs. The virus attacks the immune system of the host and influences the immune cells. The Congo fever virus can be isolated from blood, plasma and many body tissues (kidneys, liver, spleen, lungs, brain and bone marrow). Mice inoculation, enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR) can be used for detection of the infection. Furthermore, IgM and IgG antibodies against CCHFV can also be detected and quantified. Education of general public, tick control with acaricides, use of anti-CCHFV immunoglobulin, usage of approved repellents to prevent tick bites, wearing neutral-coloured garments, application of a permethrin spray to the clothing, avoiding tall grasses and shrubs, applying sunscreen, avoiding direct contact with the blood or tissues of animals are the factors for successful prevention of the infection.

scholarly journals Recombinase-mediated integration of a multigene cassette in rice leads to stable expression and inheritance of the stacked locus

2020 ◽  
Author(s):  
Bhuvan Pathak ◽  
Vibha Srivastava
Keyword(s):  
Metabolic Engineering ◽  
Rice Genome ◽  
Inducible Promoter ◽  
High Rate ◽  
Site Specific ◽  
Multigene Transformation ◽  
Site Specific Integration ◽  
Targeted Integration ◽  
Transgenic Lines ◽  
Inducible Genes

SummaryEfficient methods for multigene transformation are important for developing novel crop varieties. Methods based on random integrations of multiple genes have been successfully used for metabolic engineering in plants. However, efficiency of co-integration and co-expression of the genes could present a bottleneck. Recombinase-mediated integration into the engineered target sites is arguably a more efficient method of targeted integration that leads to the generation of stable transgenic lines at a high rate. This method has the potential to streamline multigene transformation for metabolic engineering and trait stacking in plants. Therefore, empirical testing of transgene(s) stability from the multigene site-specific integration locus is needed. Here, the recombinase technology based on Cre-lox recombination was evaluated for developing multigenic lines harboring constitutively-expressed and inducible genes. Targeted integration of a 5 genes cassette in the rice genome generated a precise full-length integration of the cassette at a high rate, and the resulting multigenic lines expressed each gene reliably as defined by their promoter activity. The stable constitutive or inducible expression was faithfully transmitted to the progeny, indicating inheritance-stability of the multigene locus. Co-localization of two distinctly inducible genes by heat or cold with the strongly constitutive genes did not appear to interfere with each other’s expression pattern. In summary, high rate of co-integration and co-expression of the multigene cassette installed by the recombinase technology in rice shows that this approach is appropriate for multigene transformation and introduction of co-segregating traits.Significance StatementRecombinase-mediated site-specific integration approach was found to be highly efficacious in multigene transformation of rice showing proper regulation of each gene driven by constitutive or inducible promoter. This approach holds promise for streamlining gene stacking in crops and expressing complex multigenic traits.

scholarly journals Antibacterial, Antihemolytic, Cytotoxic, Anticancer, and Antileishmanial Effects of Ajuga bracteosa Transgenic Plants

Plants ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1894
Author(s):  
Samina Rubnawaz ◽  
Mohammad K. Okla ◽  
Nosheen Akhtar ◽  
Imdad Ullah Khan ◽  
Muhammad Zeeshan Bhatti ◽  
...  
Keyword(s):  
Neglected Tropical Diseases ◽  
Human Cancer ◽  
Conventional Polymerase Chain Reaction ◽  
Rol Genes ◽  
Human Cancer Cell Lines ◽  
Traditional Medicines ◽  
Synthetic Drugs ◽  
Rolb Gene ◽  
Transgenic Lines

Herbal and traditional medicines can play a pivotal role in combating cancer and neglected tropical diseases. Ajuga bracteosa, family Lamiaceae, is an important medicinal plant. The genetic transformation of A. bracteosa with rol genes of Agrobacterium rhizogenes further enhances its metabolic content. This study aimed at undertaking the molecular, phytochemical, and in vitro biological analysis of A. bracteosa extracts. We transformed the A. bracteosa plant with rol genes and raised the regenerants from the hairy roots. Transgenic integration and expression of rolB were confirmed by conventional polymerase chain reaction (PCR) and qPCR analysis. The methanol: chloroform crude extracts of wild-type plants and transgenic regenerants were screened for in vitro antibacterial, antihemolytic, cytotoxic, anticancer, and leishmanial activity. Among all plants, transgenic line 3 (ABRL3) showed the highest expression of the rolB gene. Fourier transform infra-red (FTIR) analysis confirmed the enhanced number of functional groups of active compounds in all transgenic lines. Moreover, ABRL3 exhibited the highest antibacterial activity, minimum hemolytic activity (CC50 = 7293.05 ± 7 μg/mL) and maximum antileishmanial activity (IC50 of 56.16 ± 2 μg/mL). ABRL1 demonstrated the most prominent brine shrimp cytotoxicity (LD5039.6 ± 4 μg/mL). ABRL3 was most effective against various human cancer cell lines with an IC50 of 57.1 ± 2.2 μg/mL, 46.2 ± 1.1 μg/mL, 72.4 ± 1.3 μg/mL, 73.3 ± 2.1 μg/mL, 98.7 ± 1.6 μg/mL, and 97.1 ± 2.5 μg/mL against HepG2, LM3, A549, HT29, MCF-7, and MDA-MB-231, respectively. Overall, these transgenic extracts may offer a cheaper therapeutic source than the more expensive synthetic drugs.

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