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Natural variations in MdBPM2/MdRGLG3-MdNAC83 network controlling the quantitative segregation of apple fruit storability
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作者 Bei Wu Fei Shen +5 位作者 Ziying Zhou Wenhui Ren Yi Wang Ting Wu Zhenhai Han Xinzhong Zhang 《Journal of Integrative Plant Biology》 2026年第1期169-190,共22页
Dissecting quantitative traits into Mendelian factors is a great challenge in genetics.Apple fruit storability is a complex trait controlled by multi-genes with unequal effects.We previously identified62 quantitative ... Dissecting quantitative traits into Mendelian factors is a great challenge in genetics.Apple fruit storability is a complex trait controlled by multi-genes with unequal effects.We previously identified62 quantitative trait loci(QTLs)associated with apple fruit storability and genomics-assisted prediction(GAP)models were trained using 56 QTLbased markers.Here,three candidate genes,Md NAC83,Md BPM2,and Md RGLG3,were screened from the regions of QTLs with large G'value and large genetic effects.Both a 216-bp deletion and an SNP934 T/C at the promoter of Md NAC83 were associated with higher Md NAC83expression but an SNP388 G/A at the coding region significantly reduced the activity to activate the expression of the target genes Md ACO1,Md MANA3,and Md XTH28.Md BPM2 and Md RGLG3 participated in the ubiquitination of Md NAC83.SNP657 T/A of Md BPM2 and SNP167C/G of Md RGLG3 caused a reduction in the activity to ubiquitinate Md NAC83.By the addition of functional markers to the Geno Baits SNP array,the prediction accuracy of the updated GAP models increased to 0.7723/0.6231 and 0.5639/0.5345 for flesh firmness/crispness at harvest and flesh firmness/crispness retainability,respectively.The variation network involving eight simple Mendelian variations in six genes helps to gain insight into the molecular quantitative genetics,to improve breeding strategy,and to provide targets for future genome editing. 展开更多
关键词 fruit storability genetic variation Malus domestica Borkh protein ubiquitination quantitative trait
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Quantitative indices for train running performance and post-seismic traffic capacity assessment of high-speed railway bridges under random site conditions
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作者 Wangbao ZHOU Jieyu XU +4 位作者 Yu HOU Lizhong JIANG Hao LIU Yuhao WU Donghang PENG 《Science China(Technological Sciences)》 2026年第3期348-365,共18页
Under earthquake action, different site conditions have a notable impact on the dynamic response of high-speed railway bridges after earthquakes, which in turn poses a threat to the running stability of trains in the ... Under earthquake action, different site conditions have a notable impact on the dynamic response of high-speed railway bridges after earthquakes, which in turn poses a threat to the running stability of trains in the post-earthquake period. Therefore, establishing a calculation method for the post-earthquake train speed threshold that considers the influence of different site characteristics is of great engineering significance. Taking the CRTS Ⅲ slab track as the research object, this study is based on the track irregularity root mean square rate(TRR), which the authors proposed earlier to quantify the track regularity level. Using the nonlinear least squares fitting method, the mapping relationship between the TRR and the postearthquake train running performance indicators on bridges is established. Furthermore, the influence of laws governing site categories and train speeds on post-earthquake train running performance on bridges is analyzed, and a train speed threshold for bridges based on running performance under random site conditions is proposed. The research results indicate that all train running performance indicators increase significantly with the increase of train operating speed;different site categories have a significant impact on post-earthquake track residual deformation and train running stability. The greater the amplitude of postearthquake track alignment residual deformation, the lower the threshold for the stable running speed of trains after the earthquake, with the speed threshold decreasing by up to 20%. The research outcomes can provide technical references for the post-earthquake safe operation and maintenance of high-speed railway bridges under complex site conditions, as well as the formulation of targeted train speed control schemes. 展开更多
关键词 high-speed railway track regularity quantitative indicators site conditions train running performance
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A NIR and ratiometric fluorescent probe for quantitative detection of SO_(2) derivatives in Chinese medicinal materials and bioimaging in vivo
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作者 Meitong Wu Ke Wu +7 位作者 Shumin Feng Li Xu Mi Lei Jianmei Chen Shuang Li Mian Qin Dahui Liu Guoqiang Feng 《Chinese Chemical Letters》 2026年第1期434-439,共6页
Sulfur dioxide(SO_(2)) and its derivatives have been recognized as harmful environmental pollutants.However,they are often produced during the processing of traditional Chinese medicines,potentially compromising the q... Sulfur dioxide(SO_(2)) and its derivatives have been recognized as harmful environmental pollutants.However,they are often produced during the processing of traditional Chinese medicines,potentially compromising the quality of these medicinal materials and contributing to various health issues.Due to a lack of effective monitoring and imaging tools,the physiological effects of excessive SO_(2) residues in traditional Chinese medicine remain unclear.Therefore,developing a rapid and effective tool for detecting SO_(2) is crucial for understanding its metabolic pathways and effects in vivo.In this study,we developed a near infrared(NIR) and ratiometric fluorescent probe,NIR-RS,which exhibits high sensitivity,selectivity,and rapid response for SO_(2) detection.Notably,NIR-RS accurately quantifies SO_(2) contents in Pinelliae rhizoma(P.rhizoma) samples,with recovery rates from 98.46 % to 102.40 %,and relative standard deviations(RSDs)< 5.0 %.For bioimaging applications,NIR-RS has low cytotoxicity and good mitochondrial-targeting ability,making it suitable for imaging exogenous and endogenous SO_(2) in mitochondria.Additionally,NIR-RS was successfully applied to image SO_(2) content of P.rhizoma samples within cells,revealing that high SO_(2) residue elevated mitochondria adenosine triphosphate(ATP) content,these findings reveal that P.rhizoma with excessive SO_(2) can affect the organism's growth mechanisms through alterations in ATP pathways.In vivo,SO_(2) was found to predominantly accumulate in the liver following gavage with P.rhizoma solution,with accumulation levels increasing in proportion to SO_(2) residue concentration.High SO_(2) concentrations in P.rhizoma can cause pulmonary fibrosis and gastric mucosal damage.This work provides a valuable tool for regulating SO_(2) content in P.rhizoma and may help researcher better understand the metabolism of SO_(2) derivatives and explore their physiological roles in biological systems. 展开更多
关键词 Fluorescent probe SO_(2)derivatives metabolism quantitative analysis Pinelliae rhizoma Fluorescent imaging
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Genetic interaction network of quantitative trait genes for heading date in rice
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作者 Mengjiao Chen Yifeng Hong +6 位作者 Jiongjiong Fan Dengyi Cao Chong Yin Anjie Yu Jie Qiu Xuehui Huang Xin Wei 《Journal of Genetics and Genomics》 2025年第6期747-760,共14页
Several quantitative trait genes(QTGs)related to rice heading date,a key factor for crop development and yield,have been identified,along with complex interactions among genes.However,a comprehensive genetic interacti... Several quantitative trait genes(QTGs)related to rice heading date,a key factor for crop development and yield,have been identified,along with complex interactions among genes.However,a comprehensive genetic interaction network for these QTGs has not yet been established.In this study,we use 18K-rice lines to identify QTGs and their epistatic interactions affecting rice heading date.We identify 264 pairs of interacting quantitative trait loci(QTL)and construct a comprehensive genetic network of these QTL.On average,the epistatic effects of QTL pairs are estimated to be approximately 12.5%of additive effects of identified QTL.Importantly,epistasis varies among different alleles of several heading date genes.Additionally,57 pairs of interacting QTGs are also significant in their epistatic effects on 12 other agronomic traits.The identified QTL genetic interactions are further validated using near-isogenic lines,yeast two-hybrid,and split-luciferase complementation assays.Overall,this study provides a genetic network of rice heading date genes,which plays a crucial role in regulating rice heading date and influencing multiple related agronomic traits.This network serves as a foundation for understanding the genetic mechanisms of rice quantitative traits and for advancing rice molecular breeding. 展开更多
关键词 genetic network EPISTASIS Epistatic effect quantitative trait gene RICE Heading date
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QTG-LGBM:A method of prioritizing causal genes in quantitative trait loci in maize
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作者 Chuang Wang Shenshen Wu +3 位作者 Zhou Yao En Luo Junli Deng Jianxiao Liu 《The Crop Journal》 2025年第3期873-886,共14页
Efficient and accurate identification of candidate causal genes within quantitative trait loci(QTL)is a significant challenge in genetic research.Conventional linkage analysis methods often require substantial time an... Efficient and accurate identification of candidate causal genes within quantitative trait loci(QTL)is a significant challenge in genetic research.Conventional linkage analysis methods often require substantial time and resources to identify causal genes.This paper proposes a QTG-LGBM method for prioritizing causal genes in maize based on the Light GBM algorithm.QTG-LGBM dynamically adjusts gene weights and sample proportions during training to mitigate the effects of class imbalance.The method prevents overfitting in datasets with small samples by introducing a regularization term.Experimental results on maize traits,including plant height(PH),flowering time(FT),and tassel branch number(TBN),demonstrated that QTG-LGBM outperforms the commonly used methods QTG-Finder,GBDT,XGBoost,Bernoulli NB,SVM,CNN,and ensemble learning.We validated the generalization of QTG-LGBM using Arabidopsis,rice,Setaria,and sorghum.We also applied QTG-LGBM using reported QTL that affect traits of maize PH,FT and TBN,and FT in Arabidopsis,rice,and sorghum,as well as known causal genes within the QTL.When examining the top 20%of ranked genes,QTG-LGBM demonstrated a significantly higher recall rate of causal genes compared to random selection methods.We identified key gene features affecting phenotypes through feature importance analysis.QTG-LGBM is available at http://www.deepcba.com/QTG-LGBM. 展开更多
关键词 genetic analysis quantitative trait loci Light GBM Zea mays
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Quantitative Assessment of Generative Large Language Models on Design Pattern Application
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作者 Dae-Kyoo Kim 《Computers, Materials & Continua》 2025年第3期3843-3872,共30页
Design patterns offer reusable solutions for common software issues,enhancing quality.The advent of generative large language models(LLMs)marks progress in software development,but their efficacy in applying design pa... Design patterns offer reusable solutions for common software issues,enhancing quality.The advent of generative large language models(LLMs)marks progress in software development,but their efficacy in applying design patterns is not fully assessed.The recent introduction of generative large language models(LLMs)like ChatGPT and CoPilot has demonstrated significant promise in software development.They assist with a variety of tasks including code generation,modeling,bug fixing,and testing,leading to enhanced efficiency and productivity.Although initial uses of these LLMs have had a positive effect on software development,their potential influence on the application of design patterns remains unexplored.This study introduces a method to quantify LLMs’ability to implement design patterns,using Role-Based Metamodeling Language(RBML)for a rigorous specification of the pattern’s problem,solution,and transformation rules.The method evaluates the pattern applicability of a software application using the pattern’s problem specification.If deemed applicable,the application is input to the LLM for pattern application.The resulting application is assessed for conformance to the pattern’s solution specification and for completeness against the pattern’s transformation rules.Evaluating the method with ChatGPT 4 across three applications reveals ChatGPT’s high proficiency,achieving averages of 98%in conformance and 87%in completeness,thereby demonstrating the effectiveness of the method.Using RBML,this study confirms that LLMs,specifically ChatGPT 4,have great potential in effective and efficient application of design patterns with high conformance and completeness.This opens avenues for further integrating LLMs into complex software engineering processes. 展开更多
关键词 Design patterns large language models pattern application pattern-based refactoring quantitative assessment
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Detection of Exogenous Gene Copies in Transgenic Soybean by Taqman Quantitative PCR Technique 被引量:1
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作者 仇有文 张明辉 +5 位作者 高学军 曲波 敖金霞 袁肖寒 刘营 霍楠 《Agricultural Science & Technology》 CAS 2011年第4期497-499,553,共4页
[Objective] Taqman Quantitative PCR technique was adopted to detect the copies of exogenous nos terminator in transgenic hybrid soybean.[Method] With soybean Lectin as the endogenous reference gene,and gene complex DN... [Objective] Taqman Quantitative PCR technique was adopted to detect the copies of exogenous nos terminator in transgenic hybrid soybean.[Method] With soybean Lectin as the endogenous reference gene,and gene complex DNA in non-GMO soybeans as the endogenous reference standard,the method of gradient dilution was used for separately calculate Ct value of endogenous reference gene and plasmid DNA and correlation standard curve equation of logarithm of copies,and then to calculate the copies of samples through substituting thus-obtained Ct into the standard curve equation.[Result] The standard curve equation of endogenous reference gene is y=-3.422x+35.201,R2=0.998;and the standard curve equation of exogenous gene is y=-3.348x+34.890,R2=0.999.Nos terminator and its lower boundary sequences in transgenic soybean is of single copy.[Conclusion] The study has provided a theoretical basis for determining exogenous gene copies in transgenic soybean. 展开更多
关键词 Real-time PCR Transgenic hybrid soybean Copies LECTIN Boundary sequence of nos terminator gene
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Determining the Copy Number of Exogenous Gene in Transgenic Plant by SYBR Green Real-time Quantitative PCR
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作者 裘劼人 许颖 喻富根 《Agricultural Science & Technology》 CAS 2011年第6期829-831,835,共4页
[Objective] To explore the feasibility of using SYBR Green real-time quantitative PCR technique to estimate the copy numbers of exogenous gene in a transgenic plant.[Methods] Using SYBR Green real-time quantitative PC... [Objective] To explore the feasibility of using SYBR Green real-time quantitative PCR technique to estimate the copy numbers of exogenous gene in a transgenic plant.[Methods] Using SYBR Green real-time quantitative PCR technique,we have determined the copy numbers of the exogenous CYCD3;1 in transgenic Arabidopsis by comparing an endogenous single copy reference gene with CYCD3;1 copy numbers in transgenic plant,meanwhile comparing CYCD3;1 copy numbers between wild plant and transgenic plant.[Results]The exogenous CYCD3;1 copy numbers calculated by this method is identical with results of traditional Southern blot analysis which is highly accurate.[Conclusion]This method is simple,effective and safe for estimating transgene copy numbers. 展开更多
关键词 Transgenic Arabidopsis SYBR Green real-time quantitative PCR gene copy number
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Brain expression quantitative trait locus mapping informs genetic studies of psychiatric diseases
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作者 刘春宇 《Neuroscience Bulletin》 SCIE CAS CSCD 2011年第2期123-133,共11页
Genome-wide association study(GWAS) can be used to identify genes that increase the risk of psychiatric diseases.However,much of the disease heritability is still unexplained,suggesting that there are genes to be di... Genome-wide association study(GWAS) can be used to identify genes that increase the risk of psychiatric diseases.However,much of the disease heritability is still unexplained,suggesting that there are genes to be discovered.Functional annotation of the genetic variants may increase the power of GWAS to identify disease genes,by providing prior information that can be used in Bayesian analysis or in reducing the number of tests.Expression quantitative trait loci(eQTLs) are genomic loci that regulate gene expression.Genetic mapping of eQTLs can help reveal novel functional effects of thousands of single nucleotide polymorphisms(SNPs).The present review mainly focused on the current knowledge on brain eQTL mapping,and discussed some major methodological issues and their possible solutions.The frequently ignored problems of batch effects,covariates,and multiple testing were emphasized,since they can lead to false positives and false negatives.The future application of eQTL data in GWAS analysis was also discussed. 展开更多
关键词 genome-wide association study BRAIN psychiatric diseases expression quantitative trait loci geneTICS single nucleotide polymorphism
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Validation of housekeeping genes as internal controls for studying the gene expression in Pyropia haitanensis(Bangiales, Rhodophyta) by quantitative real-time PCR 被引量:6
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作者 LI Bing CHEN Changsheng +2 位作者 XU Yan JI Dehua XIE Chaotian 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2014年第9期152-159,共8页
Pyropia haitanensis is an economically important mariculture crop in China and has a high research value for several life phenomena, for example environmental tolerance. To explore the mechanisms underlying these char... Pyropia haitanensis is an economically important mariculture crop in China and has a high research value for several life phenomena, for example environmental tolerance. To explore the mechanisms underlying these characteristics, gene expression has been investigated at the whole transcriptome level. Gene expression studies using quantitative real-time PCR should start by selecting an appropriate internal control gene; therefore, the absolute expression abundance of six housekeeping genes (18S rRNA (18S), ubiquitin-conju-ating enzyme (UBC), actin (ACT), β-tubulin (TUB), elongation factors 2 (EF2), and glyceraldehyde-3-phos- phate dehydrogenase (GAPDH) examined by the quantitative real-time PCR in samples corresponding to different strains, life-cycle stages and abiotic stress treatments. Their expression stabilities were assessed by the comparative cycle threshold (Ct) method and by two different software packages: geNorm and NormFinder. The most stable housekeeping gene is UBC and the least stable housekeeping is GADPH. Thus, it is proposed that the most appropriate internal control gene for expression analyses in P. haitanensis is UBC. The results pave the way for further gene expression analyses of different aspects of P. haitanensis biology including different strains, life-history stages and abiotic stress responses. 展开更多
关键词 Pyropia haitanensis quantitative real-time PCR internal control genes gene expression
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Quantitative Trait Loci for Heading Date and Their Relationship with Genetic Control of Yield Traits in Rice (Oryza sativa) 被引量:4
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作者 GUO Liang ZHANG Zhen-hua ZHUANG Jie-yun 《Rice science》 SCIE 2013年第1期1-12,共12页
Grain yield and heading date are key factors determining the commercial potential of a rice variety. Mapping of quantitative trait loci (QTLs) in rice has been advanced from primary mapping to gene cloning, and head... Grain yield and heading date are key factors determining the commercial potential of a rice variety. Mapping of quantitative trait loci (QTLs) in rice has been advanced from primary mapping to gene cloning, and heading date and yield traits have always attracted the greatest attention. In this review, genomic distribution of QTLs for heading date detected in populations derived from intra-specific crosses of Asian cultivated rice (Oryza sativa) was summarized, and their relationship with the genetic control of yield traits was analyzed. The information could be useful in the identification of QTLs for heading date and yield traits that are promising for the improvement of rice varieties. 展开更多
关键词 RICE heading date yield traits quantitative trait locus gene
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Evaluation of reference genes for quantitative real-time PCR analysis of gene expression during early development processes of the tongue sole(Cynoglossus semilaevis) 被引量:3
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作者 MA Qian ZHUANG Zhimeng +2 位作者 FENG Wenrong LIU Shufang TANG Qisheng 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2015年第10期90-97,共8页
Differential expression of genes is crucial to growth and development of fish. To select the appropriate genes for gene normalization during Cynoglossus semilaevis early developmental process, eight candidate referenc... Differential expression of genes is crucial to growth and development of fish. To select the appropriate genes for gene normalization during Cynoglossus semilaevis early developmental process, eight candidate reference genes (ACTB, B2M, EF1A, GADPH, RPL7, TUBA, UBCE and 18S) were tested for their adequacy by using quantitative real-time PCR. The results showed that the expression of all the examined genes exhibited tissue dependent variations in the mature C. semilaevis. EFIA was listed as the most stable reference among the 14 tissues by RefFinder. Furthermore, the recommended comprehensive ranking of the stability determined by RefFinder showed that 18S was the most stable gene during the early developmental stages (from oosphere to 90 days old) in this study. However, when divided the Ct value data of the above mentioned early developmental stages into two separate periods (embryo and post-hatching periods), TUBA and 18S represented the most stable references of these two developmental periods, respectively. Consequently, the reference gene should be carefully and accurately chosen even for studies of the same species at various developmental processes. The relevant data may help in selecting appropriate reference genes for mRNA expression analysis, and is of great value in the studies of fish growth and development. 展开更多
关键词 quantitative real-time PCR reference gene early development Cynoglossus semilaevis
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Joint Analysis Method for Major Genes Controlling Multiple Correlated Quantitative Traits 被引量:5
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作者 XIAO Jing WANG Xue-feng HU Zhi-qiu TANG Zai-xiang SUI Jiong-ming LI Xin XU Chen-wu 《Agricultural Sciences in China》 CAS CSCD 2006年第3期179-187,共9页
Based on the major gene and polygene mixed inheritance model for multiple correlated quantitative traits, the authors proposed a new joint segregation analysis method of major gene controlling multiple correlated quan... Based on the major gene and polygene mixed inheritance model for multiple correlated quantitative traits, the authors proposed a new joint segregation analysis method of major gene controlling multiple correlated quantitative traits, which include major gene detection and its effect and variation estimation. The effect and variation of major gene are estimated by the maximum likelihood method implemented via expectation-maximization (EM) algorithm. Major gene is tested with the likelihood ratio (LR) test statistic. Extensive simulation studies showed that joint analysis not only increases the statistical power of major gene detection but also improves the precision and accuracy of major gene effect estimates. An example of the plant height and the number of tiller of F2 population in rice cross Duonieai x Zhonghua 11 was used in the illustration. The results indicated that the genetic difference of these two traits in this cross refers to only one pleiotropic major gene. The additive effect and dominance effect of the major gene are estimated as -21.3 and 40.6 cm on plant height, and 22.7 and -25.3 on number of tiller, respectively. The major gene shows overdominance for plant height and close to complete dominance for number of tillers. 展开更多
关键词 multiple correlated quantitative traits major gene joint segregation analysis maximum likelihood estimation EM algorithm
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Selection of Reference Genes for Gene Expression Analysis in Nilaparvata lugens with Different Levels of Virulence on Rice by Quantitative Real-Time PCR 被引量:2
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作者 WANG Wei-xia LAI Feng-xiang +1 位作者 LI Kai-long FU Qiang 《Rice science》 SCIE 2014年第6期305-311,共7页
The brown planthopper Nilaparvata lugens Stal (Homoptera: Delphacidae) can cause hopperburn by feeding on rice and also can transmit the grassy stunt disease. Resistant rice varieties have been developed, but sever... The brown planthopper Nilaparvata lugens Stal (Homoptera: Delphacidae) can cause hopperburn by feeding on rice and also can transmit the grassy stunt disease. Resistant rice varieties have been developed, but several N. lugens strains can recover their virulence to these resistant rice varieties. In the present study, reference genes with stable expression levels in N. lugens populations showed different levels of virulence to susceptible and resistant rice varieties. The expression of six candidate reference genes in N. lugens feeding on susceptible and resistant rice varieties was analyzed. These genes were evaluated for their potential use in the analysis of differential gene expression. Polymerase chain reaction data was generated from N. lugens, including two different treatments (resistant or susceptible rice) and three virulent N. lugens populations. Three software programs (BestKeeper, Normfinder and geNorm) were used to assess the candidate reference genes. Both geNorm and Normfinder identified the genes 18S, E-ACT, E-TUB and a-TUB as the most stable reference genes. BestKeeper identified ETIF1 as the optimal reference gene with the least overall variation, whereas 18S and a-TUB were the second and third most stably expressed genes, respectively. Therefore, we concluded that the genes 18S and a-TUB were the most suitable reference genes in N. lugens. These results will facilitate future transcript profiling studies on N. lugens populations that show variation in virulence levels on different rice varieties. 展开更多
关键词 reference gene Nilaparvata lugens quantitative real-time PCR gene expression RICE
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Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in Manila Clam Ruditapes philippinarum Under Hypoxic Stress 被引量:2
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作者 JING Hao ZHOU Liqing +4 位作者 GONG Miao TU Kang LIU Zhihong WU Biao SUN Xiujun 《Journal of Ocean University of China》 SCIE CAS CSCD 2023年第4期1059-1067,共9页
Quantitative real-time PCR(qRT-PCR)has been widely used for gene expression analysis,and selection of reference genes is a key point to obtain accurate results.To find out optimal reference genes for qRT-PCR in Manila... Quantitative real-time PCR(qRT-PCR)has been widely used for gene expression analysis,and selection of reference genes is a key point to obtain accurate results.To find out optimal reference genes for qRT-PCR in Manila clam Ruditapes philippinarum in response to hypoxia,different tissues were used and compared to evaluate the stability of candidate reference genes under low oxygen stress(DO 0.5mgL^(−1) and DO 2.0mgL^(−1))and normal condition(DO 7.5mgL^(−1)).Seven candidate reference genes were selected to evaluate the stability of their expression levels.The reference genes were evaluated by Delta Ct,BestKeeper,NormFinder and geNorm,and then screened by RefFinder calculation.Under hypoxic stress of 0.5mgL^(−1),the most suitable reference gene for gill and hepatopancreas was RPL31,and the optimal reference genes for axe foot and adductor muscle were TUB and HIS,respectively.For hypoxic stress of 2.0mgL^(−1),the most stable reference gene for gill and hepatopancreas was RPL31,and the optimal reference genes for axe foot and adductor muscle were RPS23 and EF1A,respectively.At the normal condition,HIS and EF1A were identified as the optimal internal reference genes in gill and hepatopancreas respectively,and GFRP2 was the best internal reference gene for axe foot and adductor muscle.The present findings will provide important basis for the selection of reference genes for qRT-PCR analysis of gene expression level in bivalves under hypoxic stress,which might be helpful for the analysis of other molluscs too. 展开更多
关键词 CLAM reference gene HYPOXIA quantitative real-time PCR
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Validation of Reference Genes for Quantitative Real-Time PCR in Laodelphax striatellus 被引量:3
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作者 HE Xiu-ting LIU Cheng-cheng +4 位作者 LI Zhao-qun ZHANG Zan LI Guo-qing LI Fei DONG Shuang-lin 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2014年第4期811-818,共8页
The normalization of quantitative real-time PCR (qPCR) is important to obtain accurate gene expression data, and the most common method for qPCR normalization is to use reference genes. However, reference genes can ... The normalization of quantitative real-time PCR (qPCR) is important to obtain accurate gene expression data, and the most common method for qPCR normalization is to use reference genes. However, reference genes can be regulated under different conditions, qPCR has recently been used for gene expression study in Laodelphax striatellus, but there is no study on validation of the reference genes. In this study, five new housekeeping genes (LstrTUB1, LstrTUB2, LstrTUB3, LstrARF and LstrRPL9) in L. striatellus were cloned and deposited in the GenBank with accession numbers of JF728809, JF728810, JF728811, JF728807 and JF728806, respectively. Furthermore, mRNA expressions of the five genes and β-actin were measured by qPCR with insect samples of different instar at nymph stage, and the expression stabilities were determined by the software geNorm and NormFinder. As a result, ARF and RPL9 were consistently more stable than β-actin, while three TUB genes were less stable than β-actin. To determine the optimal number of reference genes used in qPCR, a pairwise variations analysis by geNorm indicated that two references ARF and RPL9 were required to obtain the accurate quantification. These results were fiarther confirmed by the validation qPCR experiment with chitinase gene as the target gene, in which the standard error of the mRNA quantification by using binary reference ARF-RPL9 was much lower than those by ARF, RPL9 or β-actin alone. Taken together, our study suggested that the combination of ARF-RPL9 could replace β-actin as the reference genes for qPCR in L. striatellus. 展开更多
关键词 cDNA cloning housekeeping gene qPCR reference gene small brown planthopper
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Evaluation of bacterial pathogen diversity,abundance and health risks in urban recreational water by amplicon next-generation sequencing and quantitative PCR 被引量:7
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作者 Qijia Cui Tingting Fang +2 位作者 Yong Huang Peiyan Dong Hui Wang 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2017年第7期137-149,共13页
The microbial quality of urban recreational water is of great concern to public health.The monitoring of indicator organisms and several pathogens alone is not sufficient to accurately and comprehensively identify mic... The microbial quality of urban recreational water is of great concern to public health.The monitoring of indicator organisms and several pathogens alone is not sufficient to accurately and comprehensively identify microbial risks.To assess the levels of bacterial pathogens and health risks in urban recreational water,we analyzed pathogen diversity and quantified four pathogens in 46 water samples collected from waterbodies in Beijing Olympic Forest Park in one year.The pathogen diversity revealed by 16 S r RNA gene targeted next-generation sequencing(NGS) showed that 16 of 40 genera and 13 of 76 reference species were present.The most abundant species were Acinetobacter johnsonii,Mycobacterium avium and Aeromonas spp.Quantitative polymerase chain reaction(q PCR) of Escherichia coli(uid A),Aeromonas(aer A),M.avium(16S r RNA),Pseudomonas aeruginosa(oaa) and Salmonella(inv A) showed that the aer A genes were the most abundant,occurring in all samples with concentrations of 10^(4–6) genome copies/100 m L,followed by oaa,inv A and M.avium.In total,34.8% of the samples harbored all genes,indicating the prevalence of these pathogens in this recreational waterbody.Based on the q PCR results,a quantitative microbial risk assessment(QMRA) showed that the annual infection risks of Salmonella,M.avium and P.aeruginosa in five activities were mostly greater than the U.S.EPA risk limit for recreational contacts,and children playing with water may be exposed to the greatest infection risk.Our findings provide a comprehensive understanding of bacterial pathogen diversity and pathogen abundance in urban recreational water by applying both NGS and q PCR. 展开更多
关键词 Human pathogens Urban water Recreational activities 16S rRNA gene targeted NGS qPCR quantitative microbial risk assessment
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A Comparison Between Northern Blotting and Quantitative Real-Time PCR as a Means of Detecting the Nutritional Regulation of Genes Expressed in Roots of Arabidopsis thaliana 被引量:4
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作者 GAN Yin-bo ZHOU Zhong-jing +2 位作者 AN Li-jun BAO Sheng-jie Brian G Forde 《Agricultural Sciences in China》 CAS CSCD 2011年第3期335-342,共8页
Quantitative real-time PCR (qRT-PCR) has become a routine and robust technique for measuring the expression of genes of interest, validating microarray experiments and monitoring biomarkers. However, concerns have b... Quantitative real-time PCR (qRT-PCR) has become a routine and robust technique for measuring the expression of genes of interest, validating microarray experiments and monitoring biomarkers. However, concerns have been raised over the accuracy of qRT-PCR in China as well as in the rest of the world. We have previously used qRT-PCR to study the response of ANR1 and other root-expressed MADS-box genes to fluctuations in the supply of nitrate, phosphate and sulphate under hydroponic growth conditions. In this study, we have used both Northern blotting and qRT-PCR analyses to confirm the nutritional regulation of MADS-box genes in Arabidopsis thaliana and test whether both technologies produce the same results. The information obtained indicated that the qRT-PCR results are consistent with those obtained by Northern blotting hybridization for all the tested root-expressed MADS-box genes, in response to different nitrate, phosphate and sulphate growth conditions. Furthermore, our novel results showed that the expressions of AGL12, AGL18, and AGL19 were all down regulated in response to S and P re-supply in both qRT-PCR and Northern blotting analyses. 展开更多
关键词 Arabidopsis thaliana MADS-BOX nutrient regulation Northern blotting quantitative real-time PCR
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Heterogeneous Fenton degradation of azodyes catalyzed by modified polyacrylonitrile fiber Fe complexes: QSPR (quantitative structure peorperty relationship) study 被引量:4
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作者 Bing Li Yongchun Dong Zhizhong Ding 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2013年第7期1469-1476,共8页
The amidoximated polyacrylonitrile (PAN) fiber Fe complexeswere prepared and used as the heterogeneous Fenton catalysts for thedegradation of28 anionicwater soluble azodyes inwater under visible irradiation. The mul... The amidoximated polyacrylonitrile (PAN) fiber Fe complexeswere prepared and used as the heterogeneous Fenton catalysts for thedegradation of28 anionicwater soluble azodyes inwater under visible irradiation. The multiple linear regression (MLR) methodwas employed todevelop the quantitative structure property relationship (QSPR) model equations for thedecoloration and mineralization of azodyes. Moreover, the predictive ability of the QSPR model equationswas assessed using Leave-one-out (LOO) and cross-validation (CV) methods. Additionally, the effect of Fe content of catalyst and the sodium chloride inwater on QSPR model equationswere also investigated. The results indicated that the heterogeneous photo-Fentondegradation of the azodyeswithdifferent structureswas conducted in the presence of the amidoximated PAN fiber Fe complex. The QSPR model equations for thedyedecoloration and mineralizationwere successfullydeveloped using MLR technique. MW/S (molecularweightdivided by the number of sulphonate groups) and N N=N (the number of azo linkage) are considered as the most importantdetermining factor for thedyedegradation and mineralization, and there is a significant negative correlation between MW/S or N N=N anddegradation percentage or total organic carbon (TOC) removal. Moreover, LOO and CV analysis suggested that the obtained QSPR model equations have the better prediction ability. The variation in Fe content of catalyst and the addition of sodium chloridedid not alter the nature of the QSPR model equations. 展开更多
关键词 AZODYE heterogeneous Fentondegradation quantitative structure property relationship polyacrylonitrile fiber Fe complex
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Reference genes for quantitative real-time PCR analysis and quantitative expression of P5CS in Agropyron mongolicum under drought stress 被引量:6
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作者 TIAN Qing-song WANG Shu-yan +3 位作者 DU Jian-cai WU Zhi-juan LI Xiao-quan HAN Bing 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2016年第9期2097-2104,共8页
Reference genes, stably expressing in different tissues and cells, are commonly used as the references in expression analysis. Selecting the optimum reference gene is crucial to the success of experiments. In this stu... Reference genes, stably expressing in different tissues and cells, are commonly used as the references in expression analysis. Selecting the optimum reference gene is crucial to the success of experiments. In this study, the expression stabilities of nine common reference genes, including ACT2, 18 S r RNA, APRT, EF-1α, RNA POL II, TUBα, TUBβ, GAPDH and TLF of Agropyron mongolicum, were studied under drought condition. Among them, 18 S r RNA was found to be the most optimum reference gene under drought stress by the analyzing of ge Norm and Norm Finder software. Quantitative expression levels of P5 CS using 18 S r RNA as the reference gene, and proline contents under drought stress in A. mongolicum were further operated, and we found the expression level of P5 CS gene and proline content had a significantly positive relationship(R^2=0.7763, P〈0.05). This study established and validated 18 S r RNA as the reference genes in A. mongolicum under drought stress, providing a powerful tool for the quantitative expression analysis of drought genes in A. mongolicum. 展开更多
关键词 reference genes quantitative real-time PCR drought stress proline pyrroline-5-carboxylic acid synthetase Agropyron mongolicum
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