RESUMO
Orobanche cumana Wallr. (Orobanche cernua Loefl.) causes severe yield losses of confectionary sunflower in China. While germination of O. cumana is stimulated by sesquiterpene lactones (STLs) from host sunflower (Helianthus annuus L.). Dehydrocostus lactone and costunolide isolated from sunflower root exudates are known as STLs to specifically induce O. cumana germination. Two major confectionary sunflower cultivars, SH363 (highly susceptible to O. cumana) and TH33 (resistant to O. cumana), were planted in China. However, STLs in these two sunflower cultivars has remained unknown. To identify STLs from root and exudates of sunflower for better understanding the role of stimulants in parasitic interaction of sunflower and O. cumana, we tested dehydrocostus lactone (DCL) and costunolide (CL) in root and root exudates of susceptible and resistant sunflower cultivars. The stimulant activity of sunflower root exudate and root extract to germination of O. cumana were also determined. Dehydrocostus lactone and costunolide were identified through ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS). Both DCL and CL were found in root extracts and root exudates in the whole tested time point from two sunflower cultivars. The concentration of dehydrocostus lactone was higher than that of costunolide at the same tested growth stage of each sunflower cultivar. It was observed that higher quantity of dehydrocostus lactone in susceptible cultivar than resistant cultivar of root and root exudates at later tested developmental stages. However, the amount of CL was no significant difference between SH363 and TH33 at all tested stages. The release amount of DCL from susceptible cultivar is 3.7 folds that of resistant cultivar at 28 DAT. These findings suggested that DCL was the one of the major signal compound in these two sunflower cultivars, and lower dehydrocostus lactone might contribute to the resistance of sunflower TH33 to O. cumana.
Assuntos
Helianthus , Orobanche , Sesquiterpenos , Cromatografia Líquida , Exsudatos e Transudatos , Germinação , Lactonas/química , Lactonas/farmacologia , Extratos Vegetais , Raízes de Plantas , Espectrometria de Massas em TandemRESUMO
Glyphosate and Acetyl-coenzyme A Carboxylase (ACCase) inhibitors are popular herbicides that control goosegrass. However, some populations are difficult to control due to resistance resulting from the increasing selection pressure. The objectives of this research were to detect the multiple resistance levels, resistance mechanisms, and fitness costs of two goosegrass populations collected in China. The resistance indices of two resistant populations (denominated as R1 and R2) to glyphosate were 3.8 and 2.3, respectively; and it was 18.0 and 14.2 to quizalofop-p-ethyl, respectively. Shikimate accumulation in R1 and R2 populations was only 8% of that of the susceptible population after glyphosate treatment. A Pro-106-Ala mutation in EPSPS and an Asp-2078-Gly mutation in ACCase were present in both resistant populations. Both the expression level of EPSPS and ACCase in resistant populations were similar to that of susceptible populations. The leaf area of the individuals in wild-type populations was more than three times of the leaf area in the resistant populations. Similarly, resistant plants were 45-49% shorter, had 70-76% less fresh shoot weight, and 67-69% fewer seeds than wild-type plants. Goosegrass populations have evolved multiple resistance to glyphosate and the ACCase inhibitor quizalofop-p-ethyl in China. The Pro-106-Ala mutation in the EPSPS and the Asp-2078-Gly mutation in the ACCase were responsible for this resistance. In addition, a fitness cost exists in the resistant populations, and more work should conduct to clear which mutation is responsible for the fitness penalty.
Assuntos
Eleusine , Herbicidas , 3-Fosfoshikimato 1-Carboxiviniltransferase/genética , Acetil-CoA Carboxilase/genética , China , Eleusine/genética , Eleusine/metabolismo , Regulação da Expressão Gênica de Plantas , Glicina/análogos & derivados , Resistência a Herbicidas/genética , Herbicidas/toxicidade , Mutação , Propionatos , Quinoxalinas , GlifosatoRESUMO
BACKGROUND AND AIM: Classic daily-ingestion single-film protocol using radiopaque markers for colonic transit time (CTT) is unsuitable for Chinese patients because of rapid colonic motility. A new modified method needs to be established. METHODS: The triple-phase study was performed. In Phase I, the classic protocol was assessed to evaluate its feasibility for Chinese subjects. In Phase II, a modified protocol was performed in two centers on 180 healthy subjects and 90 constipated patients to determine optimal conditions. In Phase III, the simplified protocol was validated on 90 constipated patients. RESULTS: All the subjects of the Phase I expelled more than 95% of the markers during the examination period, which means that the classic protocol is unsuitable for Chinese patients. The 20.9-h mean total CTT for healthy Chinese subjects was much faster than that seen in Western countries. As shown by Phase II, the numbers of subjects went beyond the upper limit were 22 in P1TCTT and 10 in P2TCTT (8.14% vs 3.70%, P = 0.029). The percentage of values fall outside of the measurement range of excretion ratio was around half of our study subjects (45-70%), whereas this percentage was only 3.70% using P2TCTT. The simplified protocol had a diagnostic accuracy for constipation of 0.81, with a sensitivity and specificity of 0.46 and 0.97, respectively. CONCLUSIONS: Colon movement in Chinese individuals is significantly faster than that of Western populations. The modified protocol generated in this study is appropriate for diagnosis of constipation in population with rapid colon motility.
Assuntos
Colo/fisiologia , Colo/fisiopatologia , Constipação Intestinal/diagnóstico , Constipação Intestinal/fisiopatologia , Trânsito Gastrointestinal , Voluntários Saudáveis , Povo Asiático , Feminino , Trânsito Gastrointestinal/fisiologia , Humanos , Masculino , Estudos ProspectivosRESUMO
Amaranthus retroflexus L. is one of the most troublesome weeds in autumn-crop fields in Northeast China. In recent years, field applications of fomesafen have failed to control an A. retroflexus population in Heilongjiang Province, China. Therefore, in this study, experiments were conducted to determine the resistance of A. retroflexus to fomesafen and investigate the molecular basis of herbicide resistance. Whole-plant dose-response experiments showed that the resistant (R) population exhibited 41.8-fold resistance to fomesafen compared with the susceptible (S) population. Target-gene sequence analysis revealed an Arg-128-Gly substitution in the protoporphyrinogen oxidase (PPO) in the R population. The response of PPO2 transgenic Arabidopsis thaliana to fomesafen demonstrated that the Arg-128-Gly substitution conferred high resistance to fomesafen. Cross- and multiple-resistance analyses indicated that the R population was cross-resistant to lactofen and carfentrazone-ethyl but was sensitive to imazethapyr, thifensulfuron-methyl, atrazine, and glyphosate. This study indicated that the Arg-128-Gly substitution is the main reason for A. retroflexus resistance to fomesafen. To our knowledge, this is the first report of a target-site based mechanism for the resistance to a PPO-inhibiting herbicide in A. retroflexus.
Assuntos
Amaranthus , Herbicidas , Benzamidas , China , Resistência a HerbicidasRESUMO
Goosegrass is one of the most widespread weeds in orchards and tea plantations in China, and glyphosate is a popular herbicide used to control it. However, high glyphosate selection pressure has led to some populations becoming resistant. The objectives of this research were to determine resistance levels and possible resistance mechanisms of goosegrass populations from several tea plantations in Zhejiang Province in China. The resistance indexes in four goosegrass populations (SH, SY, CA and CX) ranged from 4.9 to 13.4, and lower shikimate accumulation in these populations compared with a glyphosate-susceptible (GS) population confirmed their resistance to glyphosate. No mutations in the target gene EPSPS were found in populations SH and SY, however, the expression of EPSPS in these two populations was 9.3 and 29.7 times higher than that in the GS population, respectively. In the CX population, a P106S mutation in EPSPS was found in 6.7% of the individuals and another 80.0% of individuals had EPSPS amplification. In population CA, all the individuals had a P106A mutation and 86.7% of them had amplification in EPSPS. The EPSPS copy numbers ranged from 5.2 to 62.3 in these four resistant populations. There was a positive correlation between signal intensities of primary anti-EPSPS antibody and the copy number of the EPSPS protein, as indicated by immunoblot analysis. In population CA, with high-level resistance to glyphosate, both P106A mutation and amplification in EPSPS evolved in the same individuals in this population.
Assuntos
Eleusine , Herbicidas , 3-Fosfoshikimato 1-Carboxiviniltransferase , China , Regulação da Expressão Gênica de Plantas , Glicina/análogos & derivados , Resistência a Herbicidas , Mutação , GlifosatoRESUMO
Convolvulus arvensis is a troublesome weed that is naturally tolerant to glyphosate. This weed tolerates glyphosate at a rate 5.1 times higher than that of glyphosate-susceptible Calystegia hederacea. Glyphosate-treated C. arvensis plants accumulated less shikimic acid than C. hederacea plants. The overexpression of EPSPS genes from the two species in transgenic Arabidopsis thaliana resulted in similar glyphosate tolerance levels. qPCR of genomic DNA revealed that the EPSPS copy number in C. arvensis was approximately 2 times higher than that in C. hederacea. Moreover, glyphosate treatment caused a marked increase in EPSPS mRNA in C. arvensis compared to C. hederacea. GUS activity analysis showed that the promoter of CaEPSPS (CaEPSPS-P) highly improved GUS expression after glyphosate treatment, while no obvious differential GUS expression was observed in ChEPSPS-P transgenic A. thaliana in the presence or absence of glyphosate. Based on the obtained results, two coexisting mechanisms may explain the natural glyphosate tolerance in C. arvensis: (i) high EPSPS copy number and (ii) specific promoter-mediated overexpression of EPSPS after glyphosate treatment.
Assuntos
3-Fosfoshikimato 1-Carboxiviniltransferase/genética , Arabidopsis/efeitos dos fármacos , Calystegia/efeitos dos fármacos , Convolvulus/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas , Glicina/análogos & derivados , 3-Fosfoshikimato 1-Carboxiviniltransferase/biossíntese , Arabidopsis/enzimologia , Bioensaio , Calystegia/enzimologia , Convolvulus/enzimologia , Bases de Dados Genéticas , Relação Dose-Resposta a Droga , Tolerância a Medicamentos , Glicina/química , Resistência a Herbicidas/genética , Herbicidas/química , Plantas Geneticamente Modificadas , Reação em Cadeia da Polimerase , Pós , Regiões Promotoras Genéticas , Ácido Chiquímico/metabolismo , GlifosatoRESUMO
Buffalobur (Solanum rostratum Dunal), which belongs to the Solanaceae family, is a worldwide noxious invasive weed and is listed as one of the top 10 alien invasive species in China. It is harmful to humans and livestock because the entire plant is covered with spines containing toxins. Many studies have analysed the gene expression in this weed species under different stress conditions using quantitative real-time PCR (qPCR). However, until now, there has been no report on suitable reference genes in buffalobur. Herein, 14 candidate reference genes were selected and evaluated for their expression stability in buffalobur in different tissues, at different developmental stages, and in response to several stress conditions using the geNorm, NormFinder, BestKeeper and RefFinder statistical algorithms. The results showed that EF1α, ACT and SAND are suitable reference genes across all samples tested. We recommend the normalization of target gene expression under different experimental conditions using these three genes together. Validation of selected reference genes was achieved by assessing the relative expression levels of P5CS and GI. This work identified the appropriate reference genes for transcript normalization in buffalobur, which will be helpful in future genetic studies of this invasive weed.
Assuntos
Regulação da Expressão Gênica de Plantas , Genes de Plantas , Reação em Cadeia da Polimerase em Tempo Real , Padrões de Referência , Seleção Genética , Solanum/genética , Alelos , Perfilação da Expressão Gênica , Estabilidade de RNA , Reprodutibilidade dos Testes , Estresse Fisiológico , TranscriptomaRESUMO
Tausch's goatgrass (Aegilops tauschii Coss.) is one of the most troublesome weeds in winter wheat-growing regions of China. In recent years, the recommended field rate of mesosulfuron-methyl failed to control the Tausch's goatgrass population in Shanxi province (SX), China. Experiments were conducted to characterize the herbicide resistance level and investigate the basis of mesosulfuron-methyl resistance in Tausch's goatgrass. Whole-plant dose-response tests showed that the SX population exhibited 11.42-fold resistance to mesosulfuron-methyl than the susceptible HN population, and the resistance level in the SX population could be significantly reduced by malathion, a cytochrome P450 inhibitor. The SX population also exhibited cross-resistance to imazethapyr, pyroxsulam and bispyribacsodium. Acetohydroxyacid synthase (AHAS) sequencing and enzyme activity assays demonstrated that the mesosulfuron-methyl resistance was not conferred by target-site substitution. A sensitive AHAS, together with the malathion revisable resistance, suggested that herbicide metabolism likely plays a main role in the mechanism of mesosulfuron-methyl resistance in the SX population. To our knowledge, this is the first report elucidating the mesosulfuron-methyl resistance in Tausch's goatgrass.
Assuntos
Aegilops/efeitos dos fármacos , Herbicidas/farmacologia , Compostos de Sulfonilureia/farmacologia , Acetolactato Sintase/metabolismo , Aegilops/metabolismo , Benzoatos/farmacologia , Ácidos Nicotínicos/farmacologia , Pirimidinas/farmacologiaRESUMO
The evolution of weed-resistant species threatens the sustainable use of glyphosate, which is the most important herbicide widely used in agriculture worldwide. Moreover, the high glyphosate resistance (>180-fold based on LD50) of Eleusine indica found in Malaysia, which carries a double mutation in its 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), made the control of this species more difficult. By contrast, the same species carrying the same double mutation in EPSPS (T102I+P106S) but found in China only shows a resistance level of not more than 14-fold based on GR50. The resistance level of this population is four times higher than that of the population carrying a single mutation (P106L). Although the members of this population survive under a high glyphosate dosage of 10,080gaeha-1, their growth was significantly inhibited by glyphosate under the recommend dose (840gaeha-1), where in the fresh weight was 85.4% of the control. EPSPS expression, relative copy number, and EPSPS activity in this population were similar to those of the susceptible population. In addition, the expression of two glutathione transferase (GST) genes (GST-U8 and GST-23) and the enzyme activity of the GST in this population did not significantly differ from those of the susceptible population. This finding is important in elucidating the resistance of the naturally evolved glyphosate-resistant (GR) weed species carrying a double mutation in EPSPS to glyphosate.
Assuntos
3-Fosfoshikimato 1-Carboxiviniltransferase/genética , Eleusine/genética , Resistência a Herbicidas/genética , Eleusine/metabolismo , Amplificação de Genes , Regulação da Expressão Gênica de Plantas , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Glicina/análogos & derivados , Glicina/farmacologia , Herbicidas/farmacologia , Mutação , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , GlifosatoRESUMO
Goosegrass (Eleusine indica) is one of the most serious annual grassy weeds worldwide, and its evolved herbicide-resistant populations are more difficult to control. Quantitative real-time PCR (qPCR) is a common technique for investigating the resistance mechanism; however, there is as yet no report on the systematic selection of stable reference genes for goosegrass. This study proposed to test the expression stability of 9 candidate reference genes in goosegrass in different tissues and developmental stages and under stress from three types of herbicide. The results show that for different developmental stages and organs (control), eukaryotic initiation factor 4 A (eIF-4) is the most stable reference gene. Chloroplast acetolactate synthase (ALS) is the most stable reference gene under glyphosate stress. Under glufosinate stress, eIF-4 is the best reference gene. Ubiquitin-conjugating enzyme (UCE) is the most stable reference gene under quizalofop-p-ethyl stress. The gene eIF-4 is the recommended reference gene for goosegrass under the stress of all three herbicides. Moreover, pairwise analysis showed that seven reference genes were sufficient to normalize the gene expression data under three herbicides treatment. This study provides a list of reliable reference genes for transcript normalization in goosegrass, which will facilitate resistance mechanism studies in this weed species.
Assuntos
Eleusine/genética , Glicina/análogos & derivados , Resistência a Herbicidas/genética , Herbicidas/farmacologia , Seleção Genética , Estresse Fisiológico/genética , Acetolactato Sintase/genética , Proteínas de Cloroplastos/genética , Eleusine/enzimologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Glicina/farmacologia , Estresse Fisiológico/efeitos dos fármacos , GlifosatoRESUMO
One new flavonoid, 5,7,8,4'-tetrahydroxy-3-methoxyflavone-8-O-ß-d-xylopyranoside (2), along with other four known flavones (1, 3-5) were isolated from the aerial parts of Solanum rostratum. 8-hydroxyflavonoid was isolated from series Androceras for the first time. The structure of the new compound 2 was determined on the basis of spectroscopic techniques, including IR, NMR and HRESI-MS. The chemotaxonomic significance of these compounds was summarised.
Assuntos
Flavonas/química , Flavonoides/química , Solanum/química , Xilose/análogos & derivados , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Solanum/classificação , Espectrometria de Massas por Ionização por Electrospray , Xilose/químicaRESUMO
Glyphosate is an important non-selective herbicide that is in common use worldwide. However, evolved glyphosate-resistant (GR) weeds significantly affect crop yields. Unfortunately, the mechanisms underlying resistance in GR weeds, such as goosegrass (Eleusine indica (L.) Gaertn.), an annual weed found worldwide, have not been fully elucidated. In this study, transcriptome analysis was conducted to further assess the potential mechanisms of glyphosate resistance in goosegrass. The RNA sequencing libraries generated 24 597 462 clean reads. De novo assembly analysis produced 48 852 UniGenes with an average length of 847 bp. All UniGenes were annotated using seven databases. Sixteen candidate differentially expressed genes selected by digital gene expression analysis were validated by quantitative real-time PCR (qRT-PCR). Among these UniGenes, the EPSPS and PFK genes were constitutively up-regulated in resistant (R) individuals and showed a higher copy number than that in susceptible (S) individuals. The expressions of four UniGenes relevant to photosynthesis were inhibited by glyphosate in S individuals, and this toxic response was confirmed by gas exchange analysis. Two UniGenes annotated as glutathione transferase (GST) were constitutively up-regulated in R individuals, and were induced by glyphosate both in R and S. In addition, the GST activities in R individuals were higher than in S. Our research confirmed that two UniGenes (PFK, EPSPS) were strongly associated with target resistance, and two GST-annotated UniGenes may play a role in metabolic glyphosate resistance in goosegrass.
Assuntos
Eleusine/efeitos dos fármacos , Eleusine/genética , Glicina/análogos & derivados , Resistência a Herbicidas/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Glicina/farmacologia , Herbicidas/farmacologia , Anotação de Sequência Molecular , Proteínas de Plantas/genética , Reação em Cadeia da Polimerase em Tempo Real , GlifosatoRESUMO
OBJECTIVE: To investigate the interaction of polymorphisms of PPAR-γ2 gene -C34G and NADPH oxidase subunit p22phox gene -C242T with helicobacter pylori (H. pylori) infection in esophageal squamous cell carcinoma (ESCC) . METHODS: A total of 200 cases of LSCC of Broder grade I, 200 of Broder grade II and of grade III were enrolled in this study with 200 healthy individuals as the control group. The genetic polymorphisms of PPAR-γ2 gene -C34G and NADPH oxidase subunit p22phox gene -C242T were analyzed using PCR-RFLP in peripheral blood leukocytes. 14C-urea breath test (14C-UBT) was used to test 14C disntegration per minute (DPM) for evaluating the infection status of H. pylori. An unconditional logistic regression model was used to analyze the interaction of nucleotide polymorphisms and H. pylori infection. RESULTS: The risk of ESCC significantly increased in subjects with -C34G (CG), -C34G(GG), -C242T (CT), and -C242T (TT) genotypes. Combined analysis of the polymorphisms showed that the subjects carrying -C34G (GG)/ -C242T (TT) had a high risk of ESCC, and a positive interaction was found between -C34G (GG) and -C242T (TT) in increasing the risk of ESCC. Positive interactions in the pathogenesis of ESCC were also found between -C34G (CG) and -C242T (TT), between -C34G (CG) and -C242T (CT), and between -C34G (GG) and -C242T (CT) (γ>1). The risk of ESCC significantly increased in subjects with H. pylori infection, which showed positive interactions with -C34G (CG), -C34G (GG), -C242T (CT) and -C242T (TT) in increasing the risk of ESCC (γ>1). CONCLUSION: Individuals carrying -C34G(CG), -C34G(GG), -C242T (CT) and -C242T (TT) genotypes have a high risk of developing ESCC, and these genotypes interact with H. pylori infection in the pathogenesis of LSCC, suggesting the importance of eradicating H. pylori for prevention of ESCC.
Assuntos
Carcinoma de Células Escamosas/genética , Neoplasias Esofágicas/genética , Helicobacter pylori , NADPH Oxidases , PPAR gama/genética , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/microbiologia , Estudos de Casos e Controles , Neoplasias Esofágicas/metabolismo , Neoplasias Esofágicas/microbiologia , Carcinoma de Células Escamosas do Esôfago , Genótipo , Infecções por Helicobacter , Modelos Logísticos , NADPH Oxidases/genética , Reação em Cadeia da Polimerase , Polimorfismo Genético , Polimorfismo de Fragmento de RestriçãoRESUMO
OBJECTIVE: To investigate the correlation between Helicobacter pylori (H. Pylori) infection and polymorphism of adiponectin gene promoter -11391G/A, extracellular superoxide dismutase (EC-SOD) gene in nonalcoholic fatty liver disease (NAFLD).⩠METHODS: From June, 2010 to July, 2014, a hospital-based 1:1 matched case-control study was carried out, with 600 cases of NAFLD and 600 healthy people in the First Affiliated Hospital of Xinxiang Medical University. The genetic polymorphisms of adiponectin gene promoter -11391G/A and EC-SOD were analyzed by polymorphism-polymerase chain reaction (PCR) technique in peripheral blood leukocytes of the subjects. 14C-urea breath test (14C-UBT) was used to test 14C disntegration per minute (DPM) for evaluating the infections status of H. Pylori. The synergistic effect between the two mutants and the gene-environment interaction of the genotypes with H. Pylori infection were analyzed.⩠RESULTS: The frequencies of -11391G/A (AA) and EC-SOD (CG+GG) were 50.67% and 50.33% in NAFLD cases, 23.83% and 24.17% in healthy controls, respectively. Statistical tests showed significantly higher frequencies of -11391G/A (AA) and EC-SOD (CG+GG) in the NAFLD group (-11391G/A: P=0.0051; EC-SOD: P=0.0057). The risk of NAFLD with -11391G/A (AA) was significantly higher than those with -11391G/A(GG+GA) (OR=3.2822, 95% CI 1.9170 to 5.2039). The individuals who carried EC-SOD (CG+GG) had a high risk of NAFLD (OR=3.1800, 95% CI 1.7974 to 5.2391). Combined analysis of the polymorphisms showed that percentage of -11391G/A (AA)/EC-SOD (CG+GG) in the NAFLD group was significantly higher than that in the control groups (25.50% vs 5.83%, P=0.0039). The people who carried with -11391G/A (AA)/EC-SOD (CG+GG) had a high risk of NAFLD (OR=10.3190, 95% CI 8.1869 to 20.5102). The H. Pylori infection rate in the NAFLD group was significantly higher than that in the control group (OR=3.1667, 95% CI 1.9139 to 5.7443, P=0.0062), and statistical analysis suggested a positive correlation between H. Pylori infection and NAFLD with -11391G/A (AA) and EC-SOD (CG+GG) (-11391G/A: γ=1.8532; EC-SOD: γ=1.7899).⩠CONCLUSION: These carriers of -11391G/A(AA) and EC-SOD (CG+GG) genotypes may have a high risk of NAFLD, and the gene genotypes can interact with H. Pylori infection in the pathogenesis of NAFLD. Therefore, effective prevention measures for NAFLD should consider eradicating H. Pylori or regulating gene expression.
Assuntos
Adiponectina/genética , Infecções por Helicobacter/genética , Hepatopatia Gordurosa não Alcoólica/genética , Superóxido Dismutase/genética , Estudos de Casos e Controles , Interação Gene-Ambiente , Genótipo , Infecções por Helicobacter/complicações , Helicobacter pylori , Humanos , Hepatopatia Gordurosa não Alcoólica/complicações , Reação em Cadeia da Polimerase , Polimorfismo Genético , Regiões Promotoras Genéticas , Fatores de RiscoRESUMO
OBJECTIVE: To investigate the interaction between polymorphism of Toll-like receptor 4 (TLR4) gene G11367C in 3' untranslated region (UTR) and inhibitor of nuclear factor kappaB (IκB)-α Hae III in acute pancreatitis (AP) and the degree of severity.â© METHODS: A total of 450 patients with confirmed AP (AP group), who came from the First Affiliated Hospital of Xinxiang Medical College from May 2013 to June 2015, were divided into a mild AP subgroup (MAP subgroup), a moderately severe AP (MSAP subgroup), and a severe acute AP (SAP subgroup) (n=150 in each group). One hundred fifty healthy persons were served as a control group. There was no significant difference in age, gender, ethnicity and birthplace among all groups. The genetic polymorphisms of TLR4 gene G11367C in 3' untranslated region and IκB-α Hae III were analyzed by polymerase chain reaction (PCR). Eligible participants were personally interviewed by a questionnaire. Unconditional logistic regression model and single factor analysis were performed to calculate the adjusted odds ratios (OR) and 95% confidence intervals (95% CI) of G11367C and IκB-α Hae III polymorphisms, respectively. The interaction of nucleotide polymorphisms was analyzed.â© RESULTS: The frequencies of G11367C (GC), IκB-α Hae III (AG) and IκB-α Hae III (GG) were 69.56%, 33.78% and 36.22% in the AP group; 49.33%, 24.67% and 26.00% in the MAP subgroup; 70.67%, 34.67% and 36.67% in the MSAP subgroup; 88.67%, 42.00% and 46.00% in the SAP subgroup and 26.67%, 14.00% and 14.67% in the control group, respectively. There was significant difference in the frequencies betweenc the AP group and the control group, or among each AP subgroup (all P<0.01). The risk of AP was significantly increased in the subjects with G11367C (GC) genotype (ORAP=6.2828, ORMAP=2.6776, ORMSAP=6.6250, ORSAP=21.5147), which was also increased in those with IκB-α Hae III (AG) genotype (ORAP=5.7369, ORMAP=2.5277, ORMSAP=6.1824, ORSAP=17.8572) and in those with IκB-α Hae III (GG) genotype (ORAP=5.8724, ORMAP=2.5902, ORMSAP=6.4027, ORSAP=18.9022). The combined analysis of the polymorphisms showed that the percentage of G11367C (GC)/ IκB-α Hae III (GG) in the AP group, the MAP subgroup, the MSAP subgroup, the SAP subgroup and the control groups was 26.44%, 12.67%, 26.00%, 40.67% and 4.00%, respectively, with significant difference in the frequency among all groups (all P<0.01). The people who carried with Pro12Ala (AA)/Pro198Leu (LL) had a high risk of AP (ORAP=30.1314, ORMAP=6.7612, ORMSAP=39.5000, ORSAP=401.5833), and the statistical analysis suggested a positive interaction between Pro12Ala (AA) and Pro198Leu (LL) in increasing the risk of AP (All γ>1). Similarly, there were also positive interactions in the pathogenesis of AP between G11367C (GC) and IκB-α Hae III (AG) (All γ>1). â© CONCLUSION: These carriers of G11367C(GC), IκB-α Hae III(AG) and IκB-α Hae III (GG) genotypes may have a high risk of AP occurency, and there are significant interactions between genetic polymorphisms of G11367C and IκB-α Hae III, which increaes the risk of the occurrence and development of AP.
Assuntos
Regiões 3' não Traduzidas , Pancreatite , Polimorfismo de Nucleotídeo Único , Doença Aguda , Desoxirribonucleases de Sítio Específico do Tipo II , Etnicidade , Predisposição Genética para Doença , Genótipo , Humanos , Quinase I-kappa B , Modelos Logísticos , Inibidor de NF-kappaB alfa , Razão de Chances , Reação em Cadeia da Polimerase , Regiões Promotoras Genéticas , Receptor 4 Toll-LikeRESUMO
Experiments were conducted to confirm imazethapyr resistance in redroot amaranth (Amaranthus retroflexus L.) and study the target-site based mechanism for the resistance. Whole-plant response experiments revealed that the resistant (R) population exhibited 19.16 fold resistance to imazethapyr compared with the susceptible (S) population. In vitro ALS activity assay demonstrated that the imazethapyr I50 value of the R population was 21.33 times greater than that of the S population. However, qRT-PCR analysis revealed that there is no difference in ALS gene expression between the R and S populations. Sequence analysis revealed an Asp-376-Glu substitution in ALS in the R population. In order to verify that the imazethapyr resistance was conferred by Asp-376-Glu mutation, the ALS-R and ALS-S genes were fused to the CaMV 35S promoter and introduced into Arabidopsis respectively. The expression of ALS-R in transgenic Arabidopsis plants exhibited 13.79 fold resistance to imazethapyr compared to ALS-S transgenic Arabidopsis.
Assuntos
Amaranthus/efeitos dos fármacos , Ácidos Nicotínicos/farmacologia , Amaranthus/genética , Relação Dose-Resposta a Droga , Expressão Gênica , Plantas Geneticamente ModificadasRESUMO
BACKGROUND: The number of studies on adiponectin, GPx-1 gene polymorphisms, and nonalcoholic fatty liver disease (NAFLD) susceptibility is increasing, but none have investigated the effect of cigarette smoking in combination with the gene polymorphisms on the susceptibility to NAFLD. In order to understand the distribution of adiponectin and GPx-1 in the local population, to explore the possible association of cigarette smoking with adiponectin and GPx-1 gene polymorphisms in the pathogenesis of NAFLD, we conducted this research, examining the distribution of polymorphisms of adiponectin and GPx-1 in NAFLD patients and healthy controls, analyzing the association between these polymorphisms and cigarette smoking. METHODS: Two hundred nonalcoholic simple fatty liver (NAFL), 200 nonalcoholic steatohepatitis (NASH), and 200 nonalcoholic fatty hepatic cirrhosis (NAFHC) cases from the First Affiliated Hospital of Xinxiang Medical College in China from February 2011 to November 2014 were selected for this study, and 200 healthy individuals as a control group. No significant difference among the four groups in age, sex, ethnicity, and birthplace was observed. The genetic polymorphisms of adiponectin gene promoter-11377C/G and GPx-1 gene C594T were analyzed using polymerase chain reaction-restriction fragment length polymorphisms in peripheral blood leukocytes of the above-mentioned cases. The interaction between the two mutants and the gene-environment association of the genotypes with cigarette smoking were analyzed. RESULTS: The frequencies of adiponectin gene promoter-11377C/G(CG), -11377C/G (GG), GPx-1 gene C594T (CT) and C594T (TT) were 24.50%, 26.00%, 24.00%, and 25.50% in the NAFL group, 34.50%, 37.00%, 35.00%, and 36.00% in the NASH group, 42.00%, 46.00%, 43.50%, and 45.50% in the NAFHC group, and 14.00%, 14.50%, 13.00%, and 14.00% in the control group, respectively. Statistical tests showed a significant difference in the frequencies among each group (p < 0.01). The risk of NAFLD significantly increased in patients with adiponectin gene promoter-11377C/G (CG) genotype [odds ratio (OR)NAFL = 2.5278; ORNASH = 6.1823; ORNAFHC = 17.8570), in those with -11377C/G (GG) genotype (ORNAFL = 2.5900; ORNASH = 6.4017; ORNAFHC = 18.9023), in those with GPx-1 gene C594T (CT) genotype (ORNAFL = 2.6687; ORNASH = 6.7772; ORNAFHC = 22.2063), and in those with C594T (TT) genotype (ORNAFL = 2.6330; ORNASH = 6.4729; ORNAFHC = 21.5682). Combined analysis of the polymorphisms showed that percentages of adiponectin gene promoter -11377C/G (GG)/GPx-1 gene C594T (TT) in the NAFL, the NASH, NAFHC, and control groups was 7.00%, 13.50%, 21.00%, and 2.00%, respectively (p < 0.01). The people who carried the adiponectin gene promoter -11377C/G (GG)/GPx-1 gene C594T (TT) had a high risk of NAFLD (ORNAFL = 7.2800; ORNASH = 41.2941; ORNAFHC = 363.9724), and statistical analysis suggested a positive association between -11377C/G (GG) and C594T (TT) in increasing the risk of NAFLD (γ2NAFL = 2.2071, γ4 NAFL = 2.0773; γ2 NASH = 2.1084; γ4NASH = 2.0543; γ2 NAFHC = 2.1387; γ4NAFHC = 2.0004). Likewise, there were also positive association in the pathogenesis of NAFLD between -11377C/G (CG) and C594T (TT), -11377C/G (CG) and C594T (CT), -11377C/G (GG), and C594T (TT) (CT).The frequencies of smoking index (SI) ≤ 400 and SI > 400 were 22.50% and 26.50% in the NAFL group, 29.00% and 40.50% in the NASH group, 34.00% and 51.50% in the NAFHC group, and 15.50% and 12.00% in the control group, respectively. Statistical tests showed a significant difference in the frequencies among each group (all p < 0.01). The risk of NAFLD significantly increased in patients with SI ≤ 400 (ORNAFL = 2.0636; ORNASH = 4.4474; ORNAFH C = 10.9677) and in those with SI > 400 (ORNAFL = 3.1393; ORNASH = 8.0225; ORNAFHC = 21.4583), and statistical analysis suggested a positive association between cigarette smoking and -11377C/G (CG), -11377C/G (GG), C594T (CT), and C594T (TT) in increasing the risk of NAFLD (all γ > 1). CONCLUSION: Adiponectin gene promoter -11377C/G (CG), -11377C/G (GG), GPx-1 gene C594T (CT), C594T (TT), and cigarette smoking are risk factors in NAFLD, and the significant association between genetic polymorphisms of -11377C/G, C594T, and cigarette smoking amplify the risk of NAFLD.
Assuntos
Adiponectina/genética , Glutationa Peroxidase/genética , Hepatopatia Gordurosa não Alcoólica/etiologia , Polimorfismo Genético , Regiões Promotoras Genéticas , Fumar/efeitos adversos , Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica/genética , Glutationa Peroxidase GPX1RESUMO
OBJECTIVE: To investigate the interaction of single nucleotide polymorphisms of macrophage migration inhibitory factor (MIF) gene -173G/C and glutathione peroxidase 1(GPX1) gene 594C/T polymorphisms and high-fat diet in ulcerative colitis (UC). METHODS: The genetic polymorphisms of MIF -173G/C and GPX1 594C/T were determined with a polymorphism-polymerase chain reaction (PCR)-endonuclease method in peripheral blood leukocytes derived from 1500 UC cases and 1500 healthy controls. RESULTS: The frequencies of MIF -173CC and GPX1 594TT were 55.60% and 55.73% in the UC cases and 16.67% and 16.47% in the healthy controls, respectively. Statistical tests also showed a significant difference in the frequencies between the two groups (P<0.01; P<0.01, respectively). Individuals carrying MIF -173CC also had a significantly higher risk of UC compared with those with MIF -173GG (OR=6.8662, 95%CI: 4.5384-9.6158). Individuals carrying GPX1 594TT had a high risk of UC (OR=7.0854, 95%CI: 4.4702-10.5283). Combined analysis showed that the percentages of MIF -173CC/GPX1 594TT in the UC and control groups were 31.00% and 2.73%, respectively (P<0.01). Individuals carrying MIF -173CC/GPX1 594TT had a high risk of UC (OR=49.0113, 95%CI: 31.7364-61.8205). The high-fat diet rate of the case group was significantly higher than that of the control group (OR=3.3248, 95%CI: 1.9461-5.0193, P<0.01), and statistic analysis suggested an interaction between high-fat diet and MIF -173CC and GPX1 594TT which increase risk of UC (γ =6.9293; γ =6.9942). CONCLUSION: MIF -173CC and GPX1 594TT and high-fat diet are the risk factors for UC, and the significant interactions between genetic polymorphisms of MIF -173G/C, GPX1 594C/T and high-fat diet may increase the risk for UC.
Assuntos
Colite Ulcerativa/genética , Colite Ulcerativa/metabolismo , Gorduras na Dieta/metabolismo , Glutationa Peroxidase/genética , Oxirredutases Intramoleculares/genética , Fatores Inibidores da Migração de Macrófagos/genética , Polimorfismo de Nucleotídeo Único , Estudos de Casos e Controles , Colite Ulcerativa/enzimologia , Colite Ulcerativa/psicologia , Dieta Hiperlipídica/efeitos adversos , Comportamento Alimentar , Feminino , Interação Gene-Ambiente , Predisposição Genética para Doença , Humanos , Masculino , Fatores de Risco , Glutationa Peroxidase GPX1RESUMO
OBJECTIVE: To investigate the interaction of polymorphisms of monocyte chemoattractant protein-I (MCP-1) receptor CCR2 gene 190A/G, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunit p22phox gene C242T and cigarette smoking in nonalcoholic fatty liver disease (NAFLD ). METHODS: The genetic polymorphisms of MCP-1 receptor CCR2 gene 190A/G and NADPH oxidase subunit p22phox gene C242T were analyzed by the technique of polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) in peripheral blood leukocytes of 600 NAFLD cases and 600 healthy persons. RESULTS: The frequencies of 190A/G (GG) and C242T (TT) were 50. 17% and 50. 00% in NAFLD cases and 23. 83% and 24. 17% in healthy controls, respectively. There were significant differences in the frequencies between the two groups (χ2 = 88. 8462, P = 0. 0031, χ2 = 85. 8100, P = 0. 0039). The risk of NAFLD with 190A/G (GG) was significantly higher than those with 190A/G (AA + AG) (OR = 3. 2171, 95% CI 1. 9351 - 5. 2184). The individuals who carried with C242T (TT) had a high risk of NAFLD (OR = 3. 1379, 95% CI 1. 7973 - 5. 2362). Combined analysis of the polymorphisms showed that percentage of 190A/G (GG)/C242T (TT) in NAFLD and control groups was 39. 67% and 13. 00%, respectively (χ2 = 118. 3021, P =0. 0017). The people who carried with 190A/G (GG)/C242T (TT) had a high risk of NAFLD (OR =5. 0211, 95% CI 3. 1853 -7. 7926). The cigarette smoking rate of the case group was significantly higher than that in the control group (χ2 = 92. 2234, P = 0. 0025), smokers have a higher risk of lung cancer than non-smokers (OR = 3. 3032, 95% CI 1. 9147 -5. 7413 ), and statistic analysis suggested an interaction between cigarette smoking and 190A/G (GG) and C242T (TT) which increase risk of NAFLD (r = 3. 9983, r = 3. 8553 ). CONCLUSION: 190A/G (GG), C242T (TT) and cigarette smoking are the risk factors in NAFLD, and the significant interactions between genetic polymorphisms of 190A/G (GG), C242T (TT) and cigarette smoking added the risk of NAFLD.
Assuntos
Quimiocina CCL2/genética , NADPH Oxidases/genética , Hepatopatia Gordurosa não Alcoólica/genética , Polimorfismo Genético , Receptores CCR2/genética , Fumar/efeitos adversos , Genótipo , Humanos , NADP , NADPH Oxidases/metabolismo , Hepatopatia Gordurosa não Alcoólica/sangue , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Risco , Fatores de Risco , Fumar/genética , NicotianaRESUMO
Three putative resistant Amaranthus retroflexus L. populations were collected in Heilongjiang province in China. Whole plant bioassays indicated high resistance (RI > 10) to imazethapyr in the three populations. In vitro acetolactate synthase (ALS) assays revealed that ALS from populations H3, H17 and H39 was less sensitive to imazethapyr inhibition compared to the susceptible population H76. The half-maximal inhibitory concentration (I50) values for H3, H17 and H39 were 14.83, 15.27 and 268 times greater, respectively, than that of the susceptible population H76. Three nucleotide mutations resulted in three known resistance-endowing amino acid substitutions, Ala-205-Val, Trp-574-Leu and Ser-653-Thr in the three resistant populations respectively. Therefore, ALS target-site mutations in resistant A. retroflexus could be responsible for imazethapyr resistance.