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Development of a Fluorescence Quantitative PCR Method for Detection of Marteilia refringens in Shellfish 认领 引用
1
作者 谢丽基 谢芝勋 +3 位作者 庞耀珊 刘加波 邓显文 谢志勤 《Agricultural Science & Technology》 CAS 2012年第7期1591-1594,共4页
[Objective] This paper was to develop a fluorescence quantitative PCR method for detection of M. refringens in shellfish. [Method] A pair of primers and a TaqMan probe were designed and synthesized according to the co... [Objective] This paper was to develop a fluorescence quantitative PCR method for detection of M. refringens in shellfish. [Method] A pair of primers and a TaqMan probe were designed and synthesized according to the conserved gene sequences of M. refringens in GenBank, so as to develop a fluorescence quantitative PCR method for detection of M. refringens. The developed fluorescence quantitative PCR method was compared with conventional PCR detection. [Result] The fluorescence quantitative PCR could detect 40 template copies of plasmid DNA, and its sensitivity was 100 times higher than the conventional PCR. The detection results of Perkinsus sp, Haplosporidium sp, Aeromonas hydrophila, Pseudomonas fluorescens, Vibrio parahaemolyticu, Vibrio alginolyticu, Vibrio rluvialis and Vibrio mimicus were negtive. [Conclusion] The fluorescence quantitative PCR method for M. refringens established in this paper is specific, sensitive, rapid and quantitative with good repeatability, which can be used for clinical detection of M. refringens infection. 展开更多
关键词 Shellfish Marteilia refringens Fluorescence quantitative PCR
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Method for Solving Non-specific Amplification Interference of Fluorescence Quantitative PCR in Gene Detection 认领 引用
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作者 Jinku Zhang Jirui Sun +2 位作者 Haizhi Qiao Lu Han Yunjia Liu 《Proceedings of Anticancer Research》 2021年第1期49-52,共4页
Objective:To explore a method to solve the issue of interference in fluorescence quantitative PCR non-specific amplification for gene detection.Method:A three-step method was used for amplification,and the quantitativ... Objective:To explore a method to solve the issue of interference in fluorescence quantitative PCR non-specific amplification for gene detection.Method:A three-step method was used for amplification,and the quantitative fluorescence signal collection process was set in the extension stage.Results:Three-step amplification has the advantages of wide application range;improved accuracy;and reduced primer design requirements.Conclusion:The interference of non-specific amplification signals was effectively avoided,the melting curve plotting process was omitted,the reaction time was shortened,and the detection accuracy was improved. 展开更多
关键词 Fluorescence quantitative PCR Specific amplification Gene detection
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Development and Preliminary Application of SYBR Green I Real-Time Fluorescence Quantitative PCR Method for Detecting Porcine Parvovirus Virus 认领 引用
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作者 SHEN Zhi-qiang WANG Jin-liang +3 位作者 GUO Xian-po WANG Xiao-hu WANG Ming ZHAO De-ming 《Animal Husbandry and Feed Science》 CAS 2009年第11期42-46,共5页
According to VP2 gene sequence of the porcine parvovirus virus strain NADL-2 （NC001718） available in GenBank （NC_001718）, a pair of specific primer was designed, and the target fragment of 431 bp was obtained by P... According to VP2 gene sequence of the porcine parvovirus virus strain NADL-2 （NC001718） available in GenBank （NC_001718）, a pair of specific primer was designed, and the target fragment of 431 bp was obtained by PCR amplification. The products were ligated with pMD18- T vector and then transformed into bacteria DH5α for recombinant plasmid extraction. After PCR identification and sequencing, recombinant plasmid was used as a standard template to establish the standard curve of SYBR Green I fluorescence quantitative PCR. Sensitivity test, specificity test and repeatability test were also determined. The results indicated that there was a good linear relationship between threshold cycle of the standard curve and template concentration, R2 =0.997 6. Tm ranged from 82.3 to 82.9 ℃, while the sensitivity was 72.1 copies/μl with good specificity and repeatability. The developed SYBR Green I real-time quantitative PCR method to detect PPV VP2 gene laid the basis for further studies on patho- oenesis, early clinical diaonosis of this virus and quantitative analysis of PPV infection. 展开更多
关键词 Porcine parvovirus virus Real-time fluorescence quantitative PCR Detection
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Detection of Ratoon Stunting Disease in Virus-free Seedcane via Real-time Fluorescence Quantitative PCR 认领 引用 被引量：1
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作者 Ming DAN Song LI +3 位作者 Kunxing YU Limin LIU Hongjian LIU Manman LU 《Agricultural Biotechnology》 2012年第5期24-26,共3页
This study was to develop the real-time fluorescence quantitative PCR technique for detecting the ratoon stunting disease （RSD） in virus-free seedcane seedlings. Healthy tissue culture seedlings were obtained from s... This study was to develop the real-time fluorescence quantitative PCR technique for detecting the ratoon stunting disease （RSD） in virus-free seedcane seedlings. Healthy tissue culture seedlings were obtained from six plants of sugarcane ROC22, which had been confirmed RSD-positive by detecting the sugarcane juice, by employing the sugarcane seedlings production protocol. Real-time fluorescence quantitative PCR was used to detect RSD pathogens in tissue culture sam- pies. The results showed that target fragment of RSD pathogens was not found in all 10 samples in real-time fluorescence quantitative PCR, with the Ct values of 37 - 39. The healthy tissue culture sugarcane seedlings do not carry RSD pathogens, indicating that adopting healthy seedcane seedlings production technique could thoroughly get rid of RSD pathogens. 展开更多
关键词 Sugarcane Virus-free seedcane Ratoon stunting disease Real-time fluorescence quantitative PCR
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Establishment and Application of the TaqMan Real-Time Fluorescence Quantitative PCR Detection Assay for Koi Herpes Virus 认领 引用
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作者 Qingfeng MENG Haibin LI +2 位作者 Xiaofeng SHAN Weili WANG Aidong QIAN 《Agricultural Biotechnology》 2013年第3期36-38,42,共3页
[ Objective ] This study aimed to establish a rapid and effective quarantine method of Koi herpes virus. [ Method] Primers and corresponding TaqMan probe were designed based on the conserved sequence of Koi herpes vir... [ Objective ] This study aimed to establish a rapid and effective quarantine method of Koi herpes virus. [ Method] Primers and corresponding TaqMan probe were designed based on the conserved sequence of Koi herpes virus （KHV） pol-ymerase gene （Sph） to establish a rapid and effective fluorescence quantitative PCR method for Koi herpes virus detection. The cell cultures were detected by using the established fluorescence quantitative PCR assay, and the results were com- pared with that of conventional PCR. [ Result] The sensitivity of fluorescence quantitative PCR was higher than that of conventional PCR. The minimum copy num- ber that could be detected was 1.6 - 102 copies/p.1. The established method was adopted for sample detection, and a reliable diagnostic result could be obtained within 4 h. [Conclusion] The established method is rapid, sensitive, specific and repeatable, which is conducive to the rapid detection of Koi herpes virus. Key words Koi herpes virus; Fluorescence quantitative PCR; Detection 展开更多
关键词 Koi herpes virus Fluorescence quantitative PCR Detection
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Detection on hepatitis c virus of blood samples with fluorescence quantitative PCR 认领 引用
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《中国输血杂志》 CAS 2001年第S1期
关键词 Detection on hepatitis c virus of blood samples with fluorescence quantitative PCR
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Application of Real-time Fluorescent Quantitative PCR in Plant 认领 引用 被引量：1
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作者 崔颖 贾晋 +2 位作者 莎娜 李俊芳 王国泽 《Agricultural Science & Technology》 CAS 2016年第2期273-278,共6页
Real-time fluorescent quantitative PCR （RQ-PCR） is a detection method by adding fluorescent dye or fluorescent probe into the PCR reaction system, using fluorescent signal accumulation to monitor amplification react... Real-time fluorescent quantitative PCR （RQ-PCR） is a detection method by adding fluorescent dye or fluorescent probe into the PCR reaction system, using fluorescent signal accumulation to monitor amplification reactions of PCR reaction process, and finally the unknown template can be quantitatively analyzed through the standard curve. So the detection level of PCR has improved from the qualitative to the quantitative. In order to provide a theoretical reference for further application, the principle, classification, advantages and disadvantages of RQ-PCR were intro- duced, and its application and progress in plants in recent years were reviewed. 展开更多
关键词 Real-time fluorescent quantitative PCR （RQ-PCR） Principle Reference gene Stress resistance of plant Transgenic product
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Establishment and Application of a Real-time Fluorescent Quantitative PCR Method for Detection of Porcine Circovirus Type 2 认领 引用
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作者 Dong Lin Wei Feng +2 位作者 Guan Yu Liu Zengshan Shen Zhiqiang 《Animal Husbandry and Feed Science》 CAS 2015年第4期249-252,256,共4页
[ Objective ] To establish a real-time fluorescent quantitative polymerase chain reaction （PCR） method with SYBR Green I for the detection of porcine circovirus type 2 （PCV2）. [Methods] Specific primers were desig... [ Objective ] To establish a real-time fluorescent quantitative polymerase chain reaction （PCR） method with SYBR Green I for the detection of porcine circovirus type 2 （PCV2）. [Methods] Specific primers were designed to amplify the conserved gene segments of PCV2 with a size of 177 bp by PCR. The ampli- fied gene was cloned into the vector of pMD 18-T and transformed into DHSct to screen positive clones. After being extracted and purified, the recombinant plasraids pMD 18-T-177 were taken as the standard DNA templates to establish the fluorescence quantitative PCR method for the detection of PCV2, and the PCR re- action conditions were optimized. [ Results] Ct value of the established PCR method showed a good linear relationship with the standard DNA templates within a viral load of 3.21 × 100 -4.16 × 108 copies/μL , the correlation coefficient was O. 998 8 and the slope was - 3.286. The method did not show any cress-reactions with the genomes of PRRSV, PCV1, CSFV, PRV, PPV and Escherichia coli. Sensitivity of this method was proved to be 3.21 × 10 copies/μL, which was 1 000 times higher as conventional PCR method. Variation coefficients of the repeated trims among same batch or different batches were both less than 3.00%. Positive rate of clinical samples detected by the established PCR method was 58.94%, which was significantly higher than the detection rate by conventional PCR. [ Conclusions ] A reM-time fluorescent quantitative PCR method with SYBR Green I for the detection of PCV2 was established, which was better for conducting the quan- titative analysis and the early diagnosis of PCV2 infection. 展开更多
关键词 Porcine circovirus type 2 Fluorescent quantitative PCR SYBR Green I
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Detection of the covalently closed circular DNA of duck hepatitis B virus by Taq-Man fluorescent quantitative PCR assay 认领 引用
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作者 MEI LI FU QING LIN +3 位作者 XIAO PENG LIU SHUI LAN SHI DONG LIANG LI ZI RONG CHEN 《Journal of Microbiology and Immunology》 2007年第1期35-39,共5页
To develop a fluorescent quantitative PCR assay based on Taq-Man chemistry to detect the covalenfly closed circular DNA （eccDNA） of duck hepatitis B virus （DHBV）, a pair of primers was designed from both sides of ... To develop a fluorescent quantitative PCR assay based on Taq-Man chemistry to detect the covalenfly closed circular DNA （eccDNA） of duck hepatitis B virus （DHBV）, a pair of primers was designed from both sides of the nick in the minus strand of DHBV and a Taq-Man probes between the primers, modified with 6-Fam at 5＇ end and Tamra at its 3＇ end was designed to detect the PCR products during PCR cycles. The DHBV DNA fragment was cloned into vector PUCm-T, and the recombinant plasmid was purified and subsequently qualified as the HBV DNA standard. The experimental conditions and reagents used in PCR assay for amplification were sophisticatedly optimized in order to yield a perfect amplification efficacy and reduce the possibility to produce non-specific amplification. It was demonstrated that the detect limit of assay was 10^3 copies/ml, and a linear standard curve was obtained between 10^5 -10^9 copies/ml [ C1 =-2.8361 ln（x） ＋ 41.45, r =-0.9985]. The coefficient of variation was 0.2%-3.14% and 2.22%-4.43% for intra- and inter-assay respectively. After a dynamic survey on the contents of DHBV DNA in serum of ducks, it was found that its peak value appeared at the second week of birth in ducks. It is evident that this method of Taq-Man fluorescent quantitative PCR assay appears to be simple, sensitive and specific. 展开更多
关键词 Duck Hepatitis B virus Covalently closed circular DNA（cccDNA） Fluorescence quantitative PCR
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Application of Real-time Fluorescent Quantitative PCR in Studies on Plants 认领 引用 被引量：3
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作者 Yueping MA Silan DAI Yanrong MA 《Agricultural Biotechnology》 2012年第1期1-7,共7页
Real-Lime fluorescent quantitative PCR is a method for quantitative analysis of gene expression developed in recent years, which has been widely used in various fields such as basic scientific research, clinical diagn... Real-Lime fluorescent quantitative PCR is a method for quantitative analysis of gene expression developed in recent years, which has been widely used in various fields such as basic scientific research, clinical diagnosis, disease study, drug research and development since its appearance. It starts relatively late in study on plants, but has already been used for analysis of gene expression in plants and gene identification of exogenous genes. The principles or advantages and dis- advantages of real-time fluorescent quantitative PCR, or its potential problems and condition optimizations in tests were introduced in this study, and then the appli- cation and prospect of real-time fluorescent quantitative PCR in study on plants were also been discussed. 展开更多
关键词 Real-time fluorescent quantitative PCR （FQ-PCR） Plant C, ene expression
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Detection of Lactobacillus acidophilus in Fermented Material by Real-time Fluorescent Quantitative PCR 认领 引用 被引量：4
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作者 Guo Zihao Fang Hua +4 位作者 Xia Zhisheng Zhu Xiaoshi Sun Zhongchao Yu Hanli Xia Jiaji 《Animal Husbandry and Feed Science》 CAS 2016年第1期54-57,共4页
The species distinctive PCR primer of Lactobacillus acidophilus （ L. acidophilus） was designed according to 16S rRNA gene sequences of conunon Lac- tobacillus species in fermented material. Bacterial genome DNA of s... The species distinctive PCR primer of Lactobacillus acidophilus （ L. acidophilus） was designed according to 16S rRNA gene sequences of conunon Lac- tobacillus species in fermented material. Bacterial genome DNA of separated L. acidophilus in fermented sample was taken as template, and L. acidophilus in fer- mented material was conducted the quantitative determination by real-time quantitative PCR （RT-PCR）. Analysis on RT-PCR results shown that contents of L. aci- dophilus in the test sample reached 1.5 billion CFU / g. Test results shown that contents of L. acidophilus in fermented material could be detected accurately by the established RT-PCR method in the test. indicating that the established RT-PCR method could be aookued to the detection of L. acidophilus in fermented material. 展开更多
关键词 Real-time fluorescent quantitative PCR Lactobacillus acidophilus Quantitative analysis Fermented material
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Establishment of a Real-time Fluorescent Quantitative PCR Assay for Detection of Genetically Modified Maize Line MON88017 认领 引用
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作者 Jun SONG Dong WANG 《Agricultural Biotechnology》 2017年第1期15-19,22,共5页
In order to improve the standardized technical systems of quantitative analyses for genetically modified organisms （GMOs） and products, ensure bio-safety and reduce ecological risk in China, a real-time fluorescent ... In order to improve the standardized technical systems of quantitative analyses for genetically modified organisms （GMOs） and products, ensure bio-safety and reduce ecological risk in China, a real-time fluorescent quantitative PCR assay was established for detection of genetically modified maize line MON88017. The established method was evaluated based on the specificity, sensitivity, accuracy and measurement uncertainty. The results showed that the established method had strong specificity in detection of genetically modified maize line MON88017. 1.50% MON88017 sample was detected with 29 replica- tions. The average measured value （ 1. 541% ） was close to the actual value （ 1.50% ） and the relative deviation was 2.70%. The variation coefficient of the measured value was 0.110 g ; the recovery was 100.00% and the measurement uncertainty was 0. 096. The limit of detection for genetically modified maize line MON88017 with the established method was 5 copies at the 97.5% confidence level. Thus, the real-time fluorescent quantitative PCR assay established in this study exhibited high specificity, accuracy and sensitivity, which could provide technical support for the safety supervision of genetically modified organ- isms and products in China. 展开更多
关键词 Genetically modified maize Real-time fluorescent quantitative PCR Specificity Sensitivity Accuracy Measurement uncertainty
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Development of a real-time PCR method for Thalassiosira rotula rapid detection 认领 引用 被引量：5
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作者 HE Shanying YU Zhigang MI Tiezhu 《Acta Oceanologica Sinica》 SCIE CAS 2007年第2期133-139,共7页
Gene specific primers and DNA probe were designed based on the sequence of 18S rDNA cloned from the red tide alga Thalassiosira rotula. A real-time fluorescent quantitative PCR （RFQ - PCR） method was developed for q... Gene specific primers and DNA probe were designed based on the sequence of 18S rDNA cloned from the red tide alga Thalassiosira rotula. A real-time fluorescent quantitative PCR （RFQ - PCR） method was developed for quantitative detection of T. rotula. The RFQ - PCR assay data showed that the results obtained with the RFQ - PCR quite good agreement with those with the light microscope （LM） counting method, which suggested that the RFQ - PCR could be a useful method for red tide alga detection. 展开更多
关键词 red tide Thalassiosira rotula fluorescent quantitative PCR 18S rDNA
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Development of real-time PCR method for rapid detection and quantification of Heterosigma akashiwo 认领 引用 被引量：2
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作者 何闪英 于志刚 米铁柱 《Journal of Harbin Institute of Technology(New Series)》 EI CAS 2008年第1期118-123,共6页
To rapidly detect the harmful algae H.akashiwo qualitatively and quantitatively, sequences of the 18S rDNA deduced from H.akashiwo were used for designing species-specific primers, and a RFQ-PCR (Real-time Fluorescent... To rapidly detect the harmful algae H.akashiwo qualitatively and quantitatively, sequences of the 18S rDNA deduced from H.akashiwo were used for designing species-specific primers, and a RFQ-PCR (Real-time Fluorescent Quantitative Polymerase Chain Reaction) method was developed for quantitative detection of H.akashiwo. Primer H.akashiwo and TaqMan probe were designed, and the specificity of primer was checked with PCR. A calibration curve was constructed with cycle threshold value against visual counted cell number. And the value of the curve was tested with other H.akashiwo samples, which were assayed with both the RFQ-PCR method and visual count under microscope. 展开更多
关键词 Heterosigma akashiwo fluorescent quantitative PCR molecular probe real-time detection
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Taqman PCR-based Methods for Rapid Detection of Salmonella in Pet Food 认领 引用 被引量：1
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作者 JIA Jun-tao CUI He +3 位作者 MA Yun ZENG Jing JIANG Ying-hui LI Zheng-yi 《Chinese Food Science》 2012年第3期32-34,共3页
[ Objective] To establish a Taqman real-time PCR for detection of Salmonella in pet food. [Method] A pair of primers and a probe were designed based on published nucleotide sequence of invA gene encoding the invasion ... [ Objective] To establish a Taqman real-time PCR for detection of Salmonella in pet food. [Method] A pair of primers and a probe were designed based on published nucleotide sequence of invA gene encoding the invasion protein of Salmonella enterica. [ Result] The assay detects Salmonella specifically. The detection limit of the real-time PCR was 17 CFUest （25 uLest） for the positive strain. This method was effective to detect artificially contaminated pet food. [ Conclusion] The results showed that Taqman PCR assay was rapid and accurate for detection of Salmonella from infected pet food. 展开更多
关键词 Fluorescence quantitative PCR Salmonella Taqman probe Detection China
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Primary application of a real-time quantitative polymerase chain reaction for the detection of human breast cancer related novel gene-Metadherin expression 认领 引用 被引量：6
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作者 Bing Li Zhaozhe Liu Xiaodong Xie Yakun Wang 《The Chinese-German Journal of Clinical Oncology》 2010年第6期316-320,共5页
Objective:The aim of this study was to detect the expression level of Metadherin (MTDH) in peripheral blood of the breast cancer patients by real-time fluorescence quantitative polymerase chain reaction (PCR),and to e... Objective:The aim of this study was to detect the expression level of Metadherin (MTDH) in peripheral blood of the breast cancer patients by real-time fluorescence quantitative polymerase chain reaction (PCR),and to explore the relationship between expression of Metadherin gene in the patients peripheral blood and the clinic-pathological features in breast cancer. Methods:Real-time fluorescence quantitative polymerase chain reaction was employed to determine the expression level of Metadherin gene in 80 peripheral blood samples of breast cancer patients and healthy donors. Results:The expression of Metadherin gene in breast cancer patients peripheral blood were positive,in which 34 breast cancer patients were highly expressed,accounting for 55.7%,while the expression of Metadherin gene in normal females peripheral blood were negative,there was statistical significance (Ratio = 2.02±0.81,P < 0.05); Ratio of the Metadherin expression in breast cancer patients peripheral blood and the glyceraldehyde-3-phosphate dehydrogenase expression was 1.15 ± 0.36. REST software analysis showed that the expression of Metadherin gene was significantly up-regulated in breast cancer. Conclusion:The SYBR Green I quantitative real-time polymerase chain reaction method can successfully detect the expression level of Metadherin gene. Expression level of Metadherin gene in breast cancer patients peripheral blood is closely related to survival,and it maybe involved in the development of breast cancer and used as an indicator of prognosis. 展开更多
关键词 breast cancer Metadherin （MTDH） real-time fluorescence quantitative polymerase chain reaction （PCR）
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Temperature Stress at Grain Filling Stage Mediates Expression of Three Isoform Genes Encoding Starch Branching Enzymes in Rice Endosperm 认领 引用 被引量：5
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作者 WEI Ke-su CHENG Fang-min +1 位作者 ZHANG Qi-fang Liu Kui-gang 《Rice science》 2009年第3期187-193,共7页
An early-maturity indica rice variety Zhefu 49,whose grain quality and starch structure are sensitive to environmental temperature,was subjected to different temperatures(32℃for high temperature and 22℃for optimum t... An early-maturity indica rice variety Zhefu 49,whose grain quality and starch structure are sensitive to environmental temperature,was subjected to different temperatures(32℃for high temperature and 22℃for optimum temperature)at the grain filling stage in plant growth chambers,and the different expressions of three isoform genes(SBEI,SBEIII and SBE/V)encoding starch branching enzyme(SBE)in the endosperms were studied by the real-time fluorescence quantitative PCR(FQ-PCR)method.Effects of high temperature on the SBE expression in developing rice endosperrns were isoform-dependent.High temperature significantly down-regulated the expressions of SBEI and SBEIII,while up-regulated the expression of SBEIV.Compared with SBEIV and SBEIII,the expression of SBEI gene in Zhefu 49 rice endosperms was more sensitive to temperature variation at the grain filling stage.This study indicates that changes in weather/climate conditions especially temperature stress influence rice grain formation and its quality as evidenced by isoform expression. 展开更多
关键词 rice high temperature starch branching enzyme isoform gene expression real-time fluorescence quantitative PCR rice quality
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Transformation of ammonium nitrogen and response characteristics of nitrifying functional genes in tannery sludge contaminated soil 认领 引用 被引量：3
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作者 Xiang-ke Kong Zi-xuan Zhang +4 位作者 Ping Wang Yan-yan Wang Zhao-ji Zhang Zhan-tao Han Li-sha Ma 《Journal of Groundwater Science and Engineering》 2022年第3期223-232,共10页
High concentrations of ammonium nitrogen released from tannery sludge during storage in open air may cause nitrogen pollution to soil and groundwater.To study the transformation mechanism of NH4+-N by nitrifying... High concentrations of ammonium nitrogen released from tannery sludge during storage in open air may cause nitrogen pollution to soil and groundwater.To study the transformation mechanism of NH4+-N by nitrifying functional bacteria in tannery sludge contaminated soils,a series of contaminated soil culture experiments were conducted in this study.The contents of ammonium nitrogen(as NH4+-N),nitrite nitrogen(as NO2−-N)and nitrate nitrogen(as NO3−-N)were analyzed during the culture period under different conditions of pollution load,soil particle and redox environment.Sigmodial equation was used to interpret the change of NO3−-N with time in contaminated soils.The abundance variations of nitrifying functional genes(amoA and nxrA)were also detected using the real-time quantitative fluorescence PCR method.The results show that the nitrification of NH4+-N was aggravated in the contaminated silt soil and fine sand under the condition of lower pollution load,finer particle size and more oxidizing environment.The sigmodial equation well fitted the dynamic accumulation curve of the NO3−-N content in the tannery sludge contaminated soils.The Cr(III)content increased with increasing pollution load,which inhibited the reproduction and activity of nitrifying bacteria in the soils,especially in coarse-grained soil.The accumulation of NO2−-N contents became more obvious with the increase of pollution load in the fine sand,and only 41.5%of the NH4+-N was transformed to NO3−-N.The redox environment was the main factor affecting nitrification process in the soil.Compared to the aerobic soil environment,the transformation of NH4+-N was significantly inhibited under anaerobic incubation condition,and the NO3−-N contents decreased by 37.2%,61.9%and 91.9%under low,medium and high pollution loads,respectively.Nitrification was stronger in the silt soil since its copy number of amoA and nxrA genes was two times larger than that of fine sand.Moreover,the copy numbers of amoA and nxrA genes in the silt soil under the aerobic environment were 2.7 times and 2.2 times larger than those in the anaerobic environment.The abundance changes of the amoA and nxrA functional genes have a positive correlation with the nitrification intensity in the tannery sludge-contaminated soil. 展开更多
关键词 Tannery sludge Transformation of ammonium nitrogen Cr(III)aging Fluorescence quantitative PCR Functional gene
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Analysis of Seed-specificity of Silencing fad_2 Gene Expression in Transgenic Rapeseed Line W-4(Brassica napus L.) 认领 引用 被引量：3
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作者 陈松 彭琦 +5 位作者 周晓婴 高建芹 张维 张洁夫 浦惠明 戚存扣 《Agricultural Science & Technology》 CAS 2014年第8期1308-1311,1316,共4页
This study was to investigate the efficiency and specificity of RNAi silencing on the expression of endogenous fad2 gene in transgenic line W-4. [Method] The relative expression of fad2 gene in seeds at different deve... This study was to investigate the efficiency and specificity of RNAi silencing on the expression of endogenous fad2 gene in transgenic line W-4. [Method] The relative expression of fad2 gene in seeds at different developmental stages of 7th, 14th, 21st and 28th day after flowering （DAF） as wel as the root, stem, leaf at winter seedling stages of both the transgenic line W-4 and non-transgenic control Westar by real-time fluorescence quantitative PCR. [Results] The results showed the relative expression of fad2 gene was gradual y increasing with the days after flowering in the seeds of the control Westar, while it was found decreasing significantly since the 21st DAF in the seeds of the line W-4. The decline was up to 60% in comparison with the control Westar. However, no significant difference in the relative expression of fad2 gene in other organs like root, stem and leaf was observed between transgenic line W-4 and non-transgenic control Westar. Fatty acid composition analysis showed the oleic acid desaturation parameter（ODP） in seeds of the line W-4 was 0.07 in average, decreased by nearly 75% than control Westar which was 0.24 in average, while no significant difference in the seedling root, stem and leaf was measured between transgenic rapeseed and control. [Conclusion] The results above validated that RNA interference in transgenic rapeseed W-4 is at a seed-specific manner, not interfering with fad2 gene expression in organs such as the root, stem and leaf. The study also found that the period of fad2 gene expres-sion decline was wel coincided with the expression of napin gene, both appeared at the 21st DAF, indicating that the expression of dsRNA of fad2 gene is precisely control ed by the napin promoter. 展开更多
关键词 Transgenic rapeseed Real-time fluorescence quantitative PCR fad2gene Specific expression
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Analysis of Gene Expression of Seven Isoforms of ADP-glucose Pyrophosphorylase in Rice Endosperm under Different Temperature Conditions 认领 引用 被引量：3
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作者 袁定阳 孙志忠 +1 位作者 谭炎宁 段美娟 《Agricultural Science & Technology》 CAS 2012年第6期1226-1229,1233,共4页
[Objective] This study aimed to analyze the effects of temperature on the expression of AGPase isoform genes in rice endosperm during milk stage. [Method] Different temperature treatments （33 and 25 ℃ of daily mean ... [Objective] This study aimed to analyze the effects of temperature on the expression of AGPase isoform genes in rice endosperm during milk stage. [Method] Different temperature treatments （33 and 25 ℃ of daily mean temperature for high and normal temperature treatments, respectively） and the real-time fluorescence quantitative PCR （ FQPCR） were used to analyze the expression patterns of seven isoforms （AGPS1, AGPS2a, AGPS2b, AGPL1, AGPL2, AGPL3 and AGPL4） of ADPglucose pyrophosphorylase （AGPase） which was the key enzyme in starch synthesis and metabolism in rice endosperm of two rice varieties Teqing and Thai Fragrant Rice. [Result] The AGPase isoforms AGPS2b, AGPL2 and AGPL3 had much higher expression than the other four isoforms, thus they were thought to be the main expression patterns of AGPase in rice endosperm. The relative expressions of AGPL2 was the highest among all the isoforms. The relative expressions of AGPS2b, AGPL2 and AGPL3 were higher in the normal temperature treatment than in the high temperature treatment in both rice varieties. The relative expression of the three enzyme genes in milk stages in Teqing was higher than those in Thai Fragrant Rice under different temperature treatments. [Conclusion] This study provides a theoretical basis for further use of molecular biology techniques to cultivate stable high-quality rice varieties. 展开更多
关键词 Rice ADP-glucose pyrophosphorylase （AGPase） isoforms Gene expression characteristics Real-time fluorescence quantitative PCR （FQ-PCR）
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