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| 1 | Field evaluation of effects of transgenic cry1Ab/cry1Ac,cry1C and cry2A rice on Cnaphalocrocis medinalis and its arthropod predators显示文摘The impacts of transgenic Bt rice on target pests and their predators need to be clarified prior to the commercialization of Bt rice.In this study,the percentages of folded leaves of three transgenic Bt rice lines and non-transgenic parental rice line caused by Cnaphalocrocis medinalis were studied over two successive growing seasons.In addition,the population densities,relative abundance and population dynamics of C.medinalis and four species of its natural arthropod predators were investigated at three sites in China.The results showed that rice line significantly affected the percentages of folded leaves and population densities of C.medinalis larvae.Significantly higher percentages of folded leaves were observed on the non-transgenic rice compared with the three transgenic Bt rice on most sampling dates.Significantly higher densities of C.medinalis larvae and higher relative abundance of C.medinalis within phytophages were found on non-transgenic rice compared with three transgenic Bt rice at different sites across the study period.The population dynamics of C.medinalis larvae were significantly affected by rice line,rice line×sampling date,rice line×year,rice line×sampling date×year.However,there was little,if any,significant difference in the relative abundance,population density and population dynamics of the four arthropod predators between the three Bt rice lines and non-transgenic rice.The results of this study indicate that the Bt toxin in transgenic Bt rice can effectively suppress the occurrence of C.medinalis,but has no significant effects on the occurrence of the four predatory arthropod species. | XU XueLiang HAN Yu WU Gang CAI WanLun YUAN BenQi WANG Hui LIU FangZhou WANG ManQun HUA HongXia | 2011 | Science China(Life Sciences)2011，54，11： | 14 |
| 2 | The past, present, and future of population geography in China:Progress, challenges and opportunities显示文摘人口地理(PopGeo ) 人的地理的一个学科的分支,应该很好考虑到它的国家人口在中国被开发了尺寸和唯一的人口统计的问题,地区性的社会经济的发展,和生物物理的差别。然而,它典型地落后于它的父母学科,特别人口统计学和地理的发展。明确地,在中国高等教育的 PopGeo 以纪律开发的三个主要方面仍然在底层:为高等教育,学术会议,和杂志的学术学生。在中国的 PopGeo 的研究内容在中央空间、宏空间的规模集中了于人口的生长,作文,变化,分发,和带的能力。作为在世界上的大多数占据土地,在人口安定并且移居到的地方,关于人口怎么并且为什么,并且怎么指导人口物质的健全开发到社会和经济变化询问,它的最大的带能力,总是是在在中国的 PopGeo 研究的重要话题。相反,一些新人口现象象人的时空行为那样(变换,汇款,和朋友相互作用) ,在在国外的科学界流行,充分没被调查在微水平。一会儿,因为富饶政策和国家新都市化计划的实现的调整,在中国的 PopGeo 可以在国家水平面对挑战和机会。是这个场合自从 1980 年代,要求 PopGeo 的开发的最先进的评论,越来越多的 PopGeo 的转弯的点在中国学习。我们试图在中国揭示 PopGeo 的当前的地位到世界,并且使它的进一步的学习清楚些。 | FENG Zhiming LI Fangzhou YANG Yanzhao LI Peng | 2017 | Journal of Geographical Sciences2017，27，8： | 8 |
| 3 | Ultrasmall Fe-doped carbon dots nanozymes for photoenhanced antibacterial therapy and wound healing显示文摘Pathogenic bacteria pose a devastating threat to public health.However,because of the growing bacterial antibiotic resistance,there is an urgent need to develop alternative antibacterial strategies to the established antibiotics.Herein,iron-doped carbon dots(Fe-CDs,~3 nm)nanozymes with excellent photothermal conversion and photoenhanced enzyme-like properties are developed through a facile one-pot pyrolysis approach for synergistic efficient antibacterial therapy and wound healing.In particular,Fe doping endows CDs with photoenhanced peroxidase(POD)-like activity,which lead to the generation of heat and reactive oxygen species(ROS)for Gram-positive and Gram-negative bacteria killing.This study demonstrates Fe-CDs have significant wound healing efficiency of Fe-CDs by preventing infection,promoting fibroblast proliferation,angiogenesis,and collagen deposition.Furthermore,the ultrasmall size of Fe-CDs possesses good biocompatibility favoring clinical translation.We believe that the nanozyme-mediated therapeutic platform presented here is expected to show promising applications in antibacterial. | Yunhang Liu Bolong Xu Mingzhu Lu Shanshan Li Juan Guo Fangzhou Chen Xiaolu Xiong Zhe Yin Huiyu Liu Dongsheng Zhou | 2022 | Bioactive Materials2022，7，6： | 7 |
| 4 | No expenditure difference among patients with liver cancer at stageⅠ-ⅣV:Findings from a multicenter cross-sectional study in China显示文摘Objective:The number of liver cancer patients in China accounts for more than half of the world.However,China currently lacks national,multicenter economic burden data,and meanwhile,measuring the differences among different subgroups will be informative to formulate corresponding policies in liver cancer control.Thus,the aim of the study was to measure the economic burden of liver cancer by various subgroups.Methods:A hospital-based,multicenter and cross-sectional survey was conducted during 2012・2014,covering 39 hospitals and 21 project sites in 13 provinces across China.The questionnaire covers clinical information,sociology,expenditure,and related variables.All expenditure data were reported in Chinese Yuan(CNY)using 2014 values.Results:A total of 2,223 liver cancer patients were enrolled,of whom 59.61%were late-stage cases(III-IV),and 53.8%were hepatocellular carcinoma.The average total expenditure per liver cancer patient was estimated as 53,220 CNY,including 48,612 CNY of medical expenditures(91.3%)and 4,608 CNY of non-medical expenditures(8.7%).The average total expenditures in stage I,H,m and stage IV were 52,817 CNY,50,877 CNY,50,678 CNY and 54,089 CNY(P>0.05),respectively.Non-medical expenditures including additional meals,additional nutrition care,transportation,accommodation and hired informal nursing were 1,453 CNY,839 CNY,946 CNY,679 CNY and 200 CNY,respectively.The one-year out-of-pocket expenditure of a newly diagnosed patient was 24,953 CNY,and 77.2%of the patients suffered an unmanageable financial burden.Multivariate analysis showed that overall expenditure differed in almost all subgroups(P<0.05),except for sex,clinical stage,and pathologic type.Conclusions:There was no difference in treatment expenditure for liver cancer patients at different clinical stages,which suggests that maintaining efforts on treatment efficacy improvement is important but not enough.To fiirtherly reduce the overall economic burden from liver cancer,more effort should be given to primary and secondary prevention strategies. | Haike Lei Lin Lei Jufang Shi Yongzhong Wu Ling Liang Huiyao Huang Mei He Fangzhou Bai Maomao Cao Hui Qiu Yuting Wang Chengcheng Liu Jia Du Hong Wang Yan Zhang Mengdi Cao Ji Peng Ni Li Chunfeng Qu Min Dai Wanqing Chen Jie He | 2020 | Chinese Journal of Cancer Research2020，32，4： | 7 |
| 5 | Epigallocatechin-3-gallate inhibits proliferation and migration of human colon cancer SW620 cells in vitro显示文摘目的: Epigallocatechin-3-gallate (EGCG ) 是在绿茶的主要 polyphenolic 成分。这研究的目的是在人的结肠癌 SW620 房间的增长和移植上调查 EGCG 的效果。 | FangZhou HongZhou TingWang YuanMu BiaoWu Dong-linGuo Xian-meiZhang YingWu | 2012 | Acta Pharmacologica Sinica2012，33，1： | 7 |
| 6 | Establishment and application of distant hybridization technology in fish显示文摘Hybridization is widely used. However, for a long time, systematic theories and technologies related to hybridization in fish have been lacking. In this study, through long-term systematic research, we investigated and obtained the main rules regarding inheritance and reproduction related to fish distant hybridization. Furthermore, we established one-step and multistep breeding technologies that were suitable for interspecific hybridization and intraspecific hybridization. Simultaneously, we used these two breeding technologies to produce a batch of diploid fish lineages and tetraploid fish lineages and improved fishes. In addition, we widely discuss the methods, technologies and results of hybridization breeding, referring to the domestic and foreign literature on fish hybridization. We hope that this paper will be beneficial for the research and application of fish hybrid breeding. | Shi Wang Chenchen Tang Min Tao Qinbo Qin Chun Zhang Kaikun Luo Rurong Zhao Jing Wang Li Ren Jun Xiao Fangzhou Hu Rong Zhou Wei Duan Shaojun Liu | 2019 | Science China(Life Sciences)2019，62，1： | 6 |
| 7 | The chimeric genes in the hybrid lineage of Carassius auratus cuvieri(♀)×Carassius auratus red var.(♂)显示文摘Hybridization can combine the genomes of different strains or species, which leads to changes of genotype and phenotype in the hybrids. In this study, we aimed to investigate the genetic variations of hybrids(WR-F1 and WR-F2) derived from the intraspecific hybridization of white crucian carp(Carassius auratus cuvieri, WCC, ♀) and red crucian carp(Carassius auratus red var., RCC, ♂). Here, we compared the orthologous genes in the liver transcriptomes of hybrids with those of WCC and RCC, and classified the orthologous genes into eight gene patterns within three categories(chimera, mutant, and biparental origin genes).The results revealed 19.04%, 4.17% chimeric genes and 6.90%, 5.05% mutations of orthologous genes in WR-F1 and WR-F2 respectively. Seventeen of twenty-three characterized genes(77%) were confirmed to be the chimeras at the genomic DNA level.The GO classification discovered that some chimeric and mutant genes were related to metabolic process, immune system and developmental process in WR-F1. Our results provide the new evidence that hybridization can combine the parental genomes,leading to changes in the genotype of the resultant hybrids. This is the first report on the formation of chimeric genes from fish intraspecific hybridization, which is potentially interesting from the context of both evolution and the genetic breeding of fish. | Qingfeng Liu Yanhua Qi Qiuli Liang Xiujuan Xu Fangzhou Hu Jing Wang Jun Xiao Shi Wang Wuhui Li Min Tao Qinbo Qin Rurong Zhao Zhanzhou Yao Shaojun Liu | 2018 | Science China(Life Sciences)2018，61，9： | 6 |
| 8 | Silencing of PRR11 suppresses cell proliferation and induces autophagy in NSCLC cells显示文摘Our previous studies have demonstrated that proline-rich protein 11(PRR11)is a novel tumor-related gene and implicates in regulating the proliferation in lung cancer.However,its precise role in cell cycle progression remains unclear.Our recent evidences show that PRR11 silencing has an effect on autophagy in non-small-cell lung cancer(NSCLC)cells.Two human NSCLC cell lines,H1299 and A549 were transiently transfected with against PRR11 siRNA.The Cell Counting Kit-8 and plate clone formation assay showed that downregulation of PRR11 inhibited the cell proliferation associated with cell cycle related genes reduced.And our data suggested that PRR11 depletion expression enhanced the autophagosomes formation,followed with downregulation of P62 and upregulation of LC3-II protein.Furthermore,the immunoblotting results indicated that silencing of PRR11 inactivated the Akt/mTOR signaling pathway.Collectively,these results demonstrated PRR11 had an effective role in autophagy in NSCLC cells through Akt/mTOR autophagy signaling pathways.Therefore,it is helpful to clarify the function of PRR11 in tumorigenesis of NSCLC. | Lian Zhang Yunlong Lei Ying Zhang Yi Li Youquan Bu Fangzhou Song Chundong Zhang | 2018 | Genes & Diseases2018，5，2： | 6 |
| 9 | Magnetic evidence for heavy metal pollution of topsoil in Shanghai, China显示文摘这研究介绍从土壤侧面的集中大小在 Baoshan 区域从可耕地和城市的公园收集了的磁性的危险性和重金属(Cu, Zn, Pb,和 Cr ) 获得的结果,上海的一个工业区域,中国。学习在土壤侧面在磁性的危险性和重金属集中并且在在磁性的危险性和重金属集中之间的关联上集中于垂直变化的调查。结果证明在土壤侧面的表面层的那磁性的改进与增加的重金属污染被联系。丰富因素(EF ) 和 Tomlinson 污染负担索引(PLI-EF ) 为在学习估计土壤侧面的重金属污染的水平被计算。在重金属内容,丰富因素(EF ) , Tomlinson 污染负担索引(PLI-CF ) ,修改 Tomlinson 污染负担索引(PLI-EF ) ,和磁性的危险性(c) 之间的重要积极关联重金属那么显示在学习区域的污染被连接到燃烧导出的微粒排出物。结果证实联合磁性的测量和重金属集中分析能为在城市的环境监视的土壤提供有用信息。然而,磁性的技术的使用应该被进一步的 geochemical 分析证实在这个学习区域的土壤侧面定位重金属污染边界。 | Guan WANG Yuan LIU Jiao CHEN Feifan REN Yuying CHEN Fangzhou YE Weiguo ZHANG | 2018 | Frontiers of Earth Science2018，12，1： | 5 |
| 10 | Ultrabithorax is a key regulator for the dimorphism of wings,a main cause for the outbreak of planthoppers in rice显示文摘Rice planthoppers,the most devastating rice pests,occur in two wing forms:the short-wing form for rapid population growth and long-wing form for long-distance migration,which together create the mechanism for outbreak.Here we show that Ultrabithorax(Ubx)is a key regulator for switching between the long-and short-wing forms of rice planthoppers.Ubx is expressed in both forewing and hindwing pads,which is different from the canonical model of Ubx expression.In brown planthoppers,expression of Ubx(Nl Ubx)is regulated by nutritional status of the rice host.High-quality young plants induce Nl Ubx expression leading to the short-wing form;low-quality ripe plants reduce Nl Ubx expression resulting in long-wing form.We also showed that Nl Ubx is regulated by the insulin receptors Nl In R1 and Nl In R2.The default expression of Nl In R1 inhibits Nl Ubx resulting in long-wings,while high-quality hosts induce Nl In R2 expression,which represses Nl In R1 thus promoting Nl Ubx expression to produce short-wings. | Fangzhou Liu Xiang Li Muhua Zhao Mengjian Guo Kehong Han Xinxin Dong Jing Zhao Wanlun Cai Qifa Zhang Hongxia Hua | 2020 | National Science Review2020，7，7： | 5 |
| 11 | Gut microbial balance and liver transplantation: alteration, management, and prediction显示文摘肝移植是为终端阶段肝疾病的常规治疗。然而,几复杂并发症仍然妨碍幸存率。肠的障碍破坏广泛地在收到肝的病人之中被观察移植并且因为在肠和肝之间的关系,受不了 ischemiareperfusion 或拒绝损害,两个在解剖和功能。重要地,内脏 microbiota 的产生改变在这个过程期间加重接枝机能障碍。这篇文章为内脏考察研究进步微生物引起的改变和肝移植。特别,这个工作也在内脏的管理上评估研究微生物引起的改变和在肝移植期间利用微生物引起的改变的可能的损害的预言。另外,我们为关于的研究建议可能的方向在肝移植期间毁坏微生物引起的改变并且在肝移植在微生物引起的改变的利用上提供一个假设。目的是不仅预言 perioperative 损害而且作为治疗的一个方法工作或甚至禁止肝移植的拒绝。 | Xinyao Tian Zhe Yang Fangzhou Luo Shusen Zheng | 2018 | Frontiers of Medicine2018，12，2： | 5 |
| 12 | Ultrasonic vibration assisted tungsten inert gas welding of dissimilar magnesium alloys显示文摘The effects of ultrasonic vibration assisted(UVA) treatment on the microstructures and mechanical properties of MB3/AZ31 dissimilar magnesium(Mg) alloy joints were studied by microstructural characterization, micro-hardness testing and tensile testing. Results indicate that the welding pores are eliminated and coarse a-Mg grains of fusion zone are refined to 26 μm, owing to the acoustic streaming effect and cavitation effect induced by the UVA treatment with an optimal ultrasonic power of 1.0 kW.In addition, Mg_(17)Al_(12) precipitation phases are fine and uniformly distributed in the whole fusion zone of weldment. Micro-hardness of fusion zone of the Mg alloy joints increases to 53.5 HV after UVA process,and the maximum tensile strength with optimized UVA treatment increases to 263 MPa, which leads to fracture occurrence in the Mg alloy base plate. Eventually, it is experimentally demonstrated that robust MB3/AZ31 Mg alloy joints can be obtained by UVA process. | Fangzhou Yang Jie Zhou Rongrong Ding | 2018 | Journal of Materials Science & Technology2018，34，12： | 5 |
| 13 | Ultra-high-Q resonances in terahertz all-silicon metasurfaces based on bound states in the continuum显示文摘High-Q metasurfaces have important applications in high-sensitivity sensing,low-threshold lasers,and nonlinear optics due to the strong local electromagnetic field enhancements.Although ultra-high-Q resonances of bound states in the continuum(BIC)metasurfaces have been rapidly developed in the optical regime,it is still a challenging task in the terahertz band for long years because of absorption loss of dielectric materials,design,and fabrication of nanostructures,and the need for high-signal-to-noise ratio and high-resolution spectral measurements.Here,a polarization-insensitive quasi-BIC resonance with a high-Q factor of 1049 in a terahertz all-silicon metasurface is experimentally achieved,exceeding the current highest record by 3 times of magnitude.And by using this ultra-high-Q metasurface,a terahertz intensity modulation with very low optical pump power is demonstrated.The proposed all-silicon metasurface can pave the way for the research and development of high-Q terahertz metasurfaces. | PENGFEI WANG FENGYAN HE JIANJUN LIU FANGZHOU SHU BIN FANG TINGTING LANG XUFENG JING ZHI HONG | 2022 | Photonics Research2022，10，12： | 4 |
| 14 | A survey of network anomaly visualization显示文摘Network anomaly analysis is an emerging subtopic of network security. Network anomaly refers to the unusual behavior of network devices or suspicious network status. A number of intelligent visual tools are developed to enhance the ability of network security analysts in understanding the original data, ultimately solving network security problems. This paper surveys current progress and trends in network anomaly visualization. By providing an overview of network anomaly data, visualization tasks, and applications, we further elaborate on existing methods to depict various data features of network alerts, anomalous traffic, and attack patterns data. Directions for future studies are outlined at the end of this paper. | Tianye ZHANG Xumeng WANG Zongzhuang LI Fangzhou GUO Yuxin MA Wei CHEN | 2017 | Science China(Information Sciences)2017，60，12： | 3 |
| 15 | The influence of pore size distribution on thermal conductivity,permeability, and phase change behavior of hierarchical porous materials显示文摘Porous foams have been widely employed to improve heat storage rate and prevent leakage of phase change materials(PCMs).Actual porous foams have non-uniform or hierarchical pore size distribution, which is usually neglected in most researches.Here, we establish hierarchical porous models considering different pore size distributions by using Voronoi tessellations. Effects of pore size distribution on thermal conductivity, permeability, and phase change behavior of hierarchical porous foams/PCMs composites are investigated. Uneven pore size distributions are found to decrease the thermal conductivity of porous foams to some extent. On the other hand, the permeability can be reduced by 27.6%, and the heat storage rate can be improved by 7.7% by introducing moderate hierarchy without changing the total porosity. This work opens a new route to enhance heat storage performance of PCMs via leveraging hierarchy of pore size distribution of porous foams. | LIU XiangLei SONG FangZhou XU Qiao LUO QingYang TIAN Yang LI JiaWei YANG XinMeng JIN Yi | 2021 | Science China(Technological Sciences)2021，64，11： | 3 |
| 16 | A comparison of summer precipitation structures over the South China Sea and the East China Sea based on tropical rainfall measurement mission显示文摘The three-dimensional structures of summer precipitation over the South China Sea(SCS)and the East China Sea(ECS)are investigated based on tropical rainfall measurement mission(TRMM).The primary results are as follows.First,both the convective and stratiform precipitation rates in the SCS are much higher than those of the ECS.The contribution of the convective cloud precipitation to the surface precipitation is primarily over the SCS and the ECS with a proportion of about 70%,but the contribution of convective cloud precipitation is slightly larger in the SCS than the ECS.The contribution of stratus precipitation is slightly larger in the ECS than that in the SCS.Second,the content of cloud particles and precipitation particles in the ECS in June was greater than that in the SCS,while in July and August,the content of cloud and precipitation particles in the ECS was less than that in the SCS.Third,the latent heat profile of the ECS is quite different from that of the SCS.In June,the peak values of evaporation and condensation latent heating rates in the ECS are greater than those in the SCS.In July and August,however,the peak values of evaporation and condensation latent heating rates in the ECS are about 0.05/h less than those in the SCS. | LI Jiangnan YANG Chaofeng LI Fangzhou HE Qihua LI Weibiao | 2013 | Acta Oceanologica Sinica2013，32，11： | 3 |
| 17 | Preparation and photocatalytic properties of porous C and N co-doped TiO_2 deposited on brick by a fast, one-step microwave irradiation method显示文摘A one-step microwave irradiation method was used to deposit carbon and nitrogen co-doped TiO_2((C, N)-TiO_2) on commercial brick((C, N)-TiO_2/brick). The as-prepared samples were characterized by X-ray diffraction, ultraviolet–visible(UV–vis) diffuse reflectance spectroscopy, photoluminescence spectroscopy, X-ray photoelectron spectroscopy, and scanning electron microscopy(SEM). A selective technique was also used to investigate the concentration of hydroxyl radicals during UV–vis irradiation of the Methyl Orange solution with the as-prepared samples. The C and N dopants enhanced visible light absorption and provided a longer lifetime for the photo-generated electron–hole pairs. The SEM images showed that the as-prepared sample is porous. The dark adsorption and photodegradation test for(C, N)-TiO_2/brick showed good photodegradation and good recyclability. The best photodegradation rate was 94% after 2 hr. The maximum degradation rate was maintained even after the 6th cycle. The good photocatalytic properties are attributed to the enhanced visible light absorption, enhanced pollutant adsorption arising from the porous structure of the(C, N)-TiO_2 thin film, and longer lifetime of the photo-generated electron–hole pairs.(C, N)-TiO_2/brick should have potential commercial applications in photodegradation processes because of its low cost, good photodegradation, and excellent recyclability. | Fangzhou Li Jiangshan Zhou Chun Du Wei Li Yinzhen Wang Guannan He Qinyu He | 2017 | Journal of Environmental Sciences2017，29，10： | 2 |
| 18 | 肺癌细胞中沉默PRR11诱发自噬并抑制细胞增殖显示文摘PRR11是近期研究发现的一个肿瘤相关新基因,在肺癌、胃癌及胆管癌等多种恶性肿瘤患者中呈现高表达,并与病人预后呈现负相关,提示PRR11基因高表达对于细胞增殖、侵袭迁移及体外成瘤等恶性表型具有重要作用。 | 张辉洁 Lian Zhang Yunlong Lei Ying Zhang Yi Li Youquan Bu Fangzhou Song Chundong Zhang | 2018 | 重庆医科大学学报2018，43，9： | 2 |
| 19 | Evidence for the paternal mitochondrial DNA in the crucian carp-like fish lineage with hybrid origin显示文摘In terms of taxonomic status,common carp(Cyprinus carpio,Cyprininae)and crucian carp(Carassius auratus,Cyprininae)are different species;however,in this study,a newborn homodiploid crucian carp-like fish(2n=100)(2nNCRC)lineage(F1–F3)was established from the interspecific hybridization of female common carp(2n=100)×male blunt snout bream(Megalobrama amblycephala,Cultrinae,2n=48).The phenotypes and genotypes of 2 n NCRC differed from those of its parents but were closely related to those of the existing diploid crucian carp.We further sequenced the whole mitochondrial(mt)genomes of the 2n NCRC lineage from F1 to F3.The paternal mt DNA fragments were stably embedded in the mt-genomes of F1–F3 generations of 2n NCRC to form chimeric DNA fragments.Along with this chimeric process,numerous base sites of F1–F3 generations of 2 n NCRC underwent mutations.Most of these mutation sites were consistent with the existing diploid crucian carp.Moreover,the mt DNA organization and nucleotide composition of 2n NCRC were more similar to those of the existing diploid crucian carp than those of the parents.The inheritable chimeric DNA fragments and mutant loci in the mt-genomes of different generations of 2nNCRC provided important evidence of the mt DNA change process in the newborn lineage derived from hybridization of different species.Our findings demonstrated for the first time that the paternal mt DNA were transmitted into the mt-genomes of homodiploid lineage,which provided new insights into the existence of paternal mt DNA in the mt DNA inheritance. | Shi Wang Ni Jiao Lu Zhao Meiwen Zhang Pei Zhou Xuexue Huang Fangzhou Hu Conghui Yang Yuqin Shu Wuhui Li Chun Zhang Min Tao Bo Chen Ming Ma Shaojun Liu | 2020 | Science China(Life Sciences)2020，63，1： | 2 |
| 20 | An optimized protocol for stepwise optimization of real-time RT-PCR analysis显示文摘Computational tool-assisted primer design for real-time reverse transcription(RT)PCR(qPCR)analysis largely ignores the sequence similarities between sequences of homologous genes in a plant genome.It can lead to false confidence in the quality of the designed primers,which sometimes results in skipping the optimization steps for qPCR.However,the optimization of qPCR parameters plays an essential role in the efficiency,specificity,and sensitivity of each gene’s primers.Here,we proposed an optimized approach to sequentially optimizing primer sequences,annealing temperatures,primer concentrations,and cDNA concentration range for each reference(and target)gene.Our approach started with a sequence-specific primer design that should be based on the single-nucleotide polymorphisms(SNPs)present in all the homologous sequences for each of the reference(and target)genes under study.By combining the efficiency calibrated and standard curve methods with the 2−ΔΔCt method,the standard cDNA concentration curve with a logarithmic scale was obtained for each primer pair for each gene.As a result,an R 2≥0.9999 and the efficiency(E)=100±5% should be achieved for the best primer pair of each gene,which serve as the prerequisite for using the 2^(−ΔΔCt) method for data analysis.We applied our newly developed approach to identify the best reference genes in different tissues and at various inflorescence developmental stages of Tripidium ravennae,an ornamental and biomass grass,and validated their utility under varying abiotic stress conditions.We also applied this approach to test the expression stability of six reference genes in soybean under biotic stress treatment with Xanthomonas axonopodis pv.glycines(Xag).Thus,these case studies demonstrated the effectiveness of our optimized protocol for qPCR analysis. | Fangzhou Zhao Nathan A.Maren Pawel Z.Kosentka Ying-Yu Liao Hongyan Lu James R.Duduit Debao Huang Hamid Ashrafi Tuanjie Zhao Alejandra I.Huerta Thomas G.Ranney Wusheng Liu | 2021 | Horticulture Research2021，8，1： | 2 |
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