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Identification of ZmCI-1B Promoter and Its Seven Deletions in Transgenic Arabidopsis thaliana Li Ye, Liu Xiaoqing, Li Suzhen, Zhou Xiaojin, Yang Wenzhu, Chen Rumei Biotechnology Bulletin    2014, 0 (5): 69-75.   Abstract （7087）      PDF（pc） （5083KB）（565）       Knowledge map    Save The expression vectors of ZmCI-1B promoter and its seven 5' truncated fragments were transformed into Agrobacterium tumefaciens strain GV3101. After identified by PCR assay, with Arabidopsis thaliana as genetic transformation, the expression vectors were transformed into Arabidopsis thaliana by floral-dip method. The transformed Arabidopsis thaliana plants were identified by PCR assay, then the seedings, flowers and siliques from positive plants were conducted to GUS histochemical staining. The results showed that the characterization of ZmCI-1B promoter in Arabidopsis thaliana is different from miaze. The ZmCI-1B promoter and its seven 5' truncated promoter-GUS constructs had different GUS staining in transformed Arabidopsis thaliana plants, revealing that different-length promoter had different promoting activity. The cis-acting elements on the promoter may contribute to this. Reference | Related Articles | Metrics

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Dectection of Vibrio parahaemolyticus by Outer Membrane Protein ompK Immunomagnetic Beads Li Yuchen, Wen Yiming, Tong Jiyu, Xiang Junjian Biotechnology Bulletin    2013, 0 (6): 209-214.   Abstract （5675）      PDF（pc） （2598KB）（1015）       Knowledge map    Save It was to construct recombinant gene cloning and express the membrane surface protein outer the membrane protein K(ompK)of Vibrio parahaemolyticus, developing a method to detect Vibrio parahaemolyticus via immunomagnetic separation combined with color plate method. Using Primer 5 to design primers of ompK gene, the ompK gene was amplified by PCR subsequently and cloned into pET28a(+)prokaryotic expression vector, optimally transferred into E. coli BL21 to express. The expression products were purified by Nickel column and immunized mouse to prepare polyclonal antibody. Western blotting analysis was applied to identify the recombinant protein and ELISA was used to analyze its immunogenicity. The titer and cross reactivity of the polyclonal antibody were determined using indirect ELISA, then the polyclonal antibodies we prepared were coupled with protein G immunomagnetic beads to develop a chromogenic plating detection of Vibrio parahaemolyticus. The sensitivity of immunomagnetic beads reached to 104 CFU/mL. We managed to clone and express Vibrio parahaemolyticus outer membrane protein ompK, in addition, we prepared mouse anti-Vibrio parahaemolyticus polyclonal antibody to create a new detection method using immunomagnetic beads, this method which combined with color plate method could save 72 hours comparing with traditional enrichment method, all process only need one-third of the time of traditional methods. When compared to the common immune detection methods, it was two magnitudes more sensitive. Reference | Related Articles | Metrics

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The Construction of Rhizobia Engineering Strain of Introduction a Tandem Gene nodD-lba into Sinorhizobium fredii Li Shanshan, Li Haiying, Yu Bing Biotechnology Bulletin    2013, 0 (8): 119-123.   Abstract （3950）      PDF（pc） （3244KB）（879）       Knowledge map    Save Abstract: In this study, homology-based cloning was used to clone hemoglobin gene of lba and nodulation gene of nodD. The two genes were ordered joined into downstream of lac promoter and connected by the DNA recombinant technique to constructed expression vector pTR-Plac-nodD-lba marked by luxAB gene. The vector was transformed into Sinorhizobium fredii with the method of triparental hybridization to construct genetic engineering strain. Western blot indicated the genes we used can express normally. Strains were inoculated soybean seedlings in order to get nitrogenase activity. The result showed engineering strains can improve the nitrogenase activity and the engineering strain with tandem genes can enhance nitrogenase activity 4% and 15.1%, respectively, by contrast with engineering strain with lba or nodD gene. Reference | Related Articles | Metrics

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Application and Prospect of KASP Marker Technology in Main Crops YANG Qing-qing, TANG Jia-qi, ZHANG Chang-quan, GAO Ji-ping, LIU Qiao-quan Biotechnology Bulletin    2022, 38 (4): 58-71.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1378 Abstract （2514）   HTML （136）    PDF（pc） （2179KB）（1684）       Knowledge map    Save With the development of gene sequencing technology,plant genome data are becoming more and more abundant. Single nucleotide polymorphism（SNP）data are widely used in the development and application of molecular markers because of their high density,high throughput and easy automated analysis. Kompetitive allele-specific PCR（KASP）is a high-throughput genotyping technology mainly based on SNP. Because of its high flux,low cost and strong operability,this technology has great application potential in the field of crop character improvement. This paper introduces the development,principle and method steps of KASP technology,summarizes the application of KASP technology in genetic breeding such as germplasm resource identification,molecular marker assisted breeding,gene mapping and seed purity identification of main crops,and discusses the advantages and disadvantages of KASP technology in order to provide reference basis for crop breeding research in the future. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in the Structural and Functional Analysis of Plant Transcription Factor AP2/ERF Protein Family YUE Man-fang, ZHANG Chun, WU Zhong-yi Biotechnology Bulletin    2022, 38 (12): 11-26.   DOI: 10.13560/j.cnki.biotech.bull.1985.2022-0432 Abstract （2084）   HTML （70）    PDF（pc） （2307KB）（618）       Knowledge map    Save The AP2/ERF transcription factor family contains 1-2 AP2/ERF domain（s）consisting of about 60 amino acids. It is divided into 5 subfamilies：AP2（APETALA2），ethylene response factor（ERF），dehydration response element-binding protein（DREB），RAV，and Soloist，according to the number of AP2/ERF domains and whether they contain other domains. The family plays an important role in plant growth and development，and in response to biotic and abiotic stress. This paper briefly summarizes the structural function characteristics of AP2/ERF transcription factors and the research progress of AP2/ERF family proteins in plant growth and development and response to adversity in recent years. It is expected to help with future research on such transcription factors. Table and Figures | Reference | Related Articles | Metrics

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Progress of the Structural and Functional Analysis of Plant Transcription Factor TIFY Protein Family YANG Rui-jia, ZHANG Zhong-bao, WU Zhong-yi Biotechnology Bulletin    2020, 36 (12): 121-128.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-0307 Abstract （1720）   HTML （67）    PDF（pc） （3436KB）（783）       Knowledge map    Save The TIFY protein is a kind of plant-specific transcription factor in many plants. Depending on its conservative domain,it can be classified into four subfamilies：TIFY,JAZ,ZML,and PPD. Here,we reviewed the research progress on the structure,distribution,and biological function of TIFY protein in regulating the growth and development of plants,responding to various stresses and different hormone signals,aiming to lay a theoretical foundation for in-depth research and utilization of TIFY protein family. Table and Figures | Reference | Related Articles | Metrics

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Spatially Resolved Metabolomics：Progress and Challenges YIN Zhi-bin, HUANG Wen-jie, WU Xin-zhou, YAN Shi-juan Biotechnology Bulletin    2021, 37 (1): 32-51.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-1374 Abstract （1250）   HTML （65）    PDF（pc） （2322KB）（1038）       Knowledge map    Save Spatially resolved metabolomics is developed by integrating mass spectrometry imaging（MSI）and metabolomics methods for accurate determination of types,contents,and spatial differentiation of endogenous and exogenous metabolites within plant and animal tissues. Benefiting from the unique features of label-free and non-specific detections,as well as high sensitivity,high chemical coverage,and simultaneous detection of elements and molecules,MSI has gained an increasing attention as a powerful tool in spatial and temporal visualization of various metabolites,peptides,and proteins. Herein,we introduce the current research status of metabolomics and MSI techniques,then focus on the emerging applications of spatially resolved metabolomics in plant and animal tissues,as well as at the single cell levels,and finally prospect the current bottlenecks and future developing trends in spatially resolved metabolomics. As an emerging molecular imaging-based omics technique,spatially resolved metabolomics is featured for its label-free and visualized detection of absorption,distribution,metabolism,and excretion of exogenous drugs in animal tissues,and provides insights into the biosynthesis,transport pathways,and accumulation of various metabolites in plant tissues. This technique facilitates further applications in the discovery of targeted drugs,understanding of pathological mechanisms,and a spatial metabolic network closely related to the growth and development of plants and animals. Table and Figures | Reference | Related Articles | Metrics

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Research Progress of Plant Prenyltransferases CHEN Yu, ZHU Pei-huang, LI Rong, ZHU Ling-zhi, JI Kong-shu Biotechnology Bulletin    2021, 37 (2): 149-161.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-0476 Abstract （1150）   HTML （51）    PDF（pc） （4288KB）（675）       Knowledge map    Save The biological role of secondary metabolites is recognized as the key to the survival and evolution of higher plants. Isoprenoids compounds,as a very rich class of secondary metabolites in plants,demonstrate various physiological functions in plant signal transduction,climate adaptation,reproduction and defense. In addition,plant isoprenoids compounds are also widely used in pharmaceutical,natural latex,perfume,organic synthesis and other industrial fields,with important economic values. Prenyltransferase（PT）is a key enzyme that connects the upstream mevalonate-independent and methyl-erythritol 4-phosphate pathway with the downstream branch points of isoprenoids biosynthesis of different structures. Therefore,prenyltransferase and their genes play an important role in the process of isoprenoids biosynthesis and regulation. This article introduces the biosynthesis pathway of isoprenoids in plants,prenyltransferase genes cloning and analysis,function identification of enzymes and systematic classification,and also introduces research progress in conifers. The aim is to provide assistance for the analysis of plant isoprenoids biosynthesis and regulatory mechanisms,and to point out some molecular research fields on producing rosin and biological adversity resistance of conifers. Table and Figures | Reference | Related Articles | Metrics

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Application and Research Progress on Transcriptomics CUI Kai, WU Wei-wei, DIAO Qi-yu Biotechnology Bulletin    2019, 35 (7): 1-9.   DOI: 10.13560/j.cnki.biotech.bull.1985.2019-0374 Abstract （1106）   HTML    PDF（pc） （1740KB）（1735）       Knowledge map    Save The transcriptome is the set of all RNA molecules in the specific tissue or cell,including message RNA,ribosome RNA,transfer RNA and non-coding RNAs. The rapid development of high-throughput sequencing technology provides a fast and reliable platform for the systematic study of transcriptome. Here we reviewed the current high-throughput sequencing technology and its application in transcriptome research,and discussed some issues worthy of attention in transcriptome data analysis,as well as the application direction of transcriptome sequencing technology in biological research. Reference | Related Articles | Metrics

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Structure of ABC Transporter and Research Progress of It in Bacterial Pathogenicity CHEN Fu-nuan, HUANG Yu, CAI Jia, WANG Zhong-liang, JIAN Ji-chang, WANG Bei Biotechnology Bulletin    2022, 38 (6): 43-52.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1175 Abstract （1083）   HTML （34）    PDF（pc） （3043KB）（637）       Knowledge map    Save Adenosine triphosphate binding cassette transporter（ATP-binding cassette transporter,ABC transporter）is known as the most numerous protein family with multiple roles. Functionally,ABC transporter mediate the cross-membrance transmission of multiple substrates by using the energy generated by ATP hydrolysis. ABC transporters exist in most organisms including bacteria,fungi,nematodes,fruit flies,plants,and mammals. Most of ABC transporters were originally discovered by studying the drug resistance（multiple drug resistance and multidrug resistance）of eukaryotes. Recently,there are broad researches on the roles of ABC transporters in bacterial pathogenicity. This paper summarized the structure and mechanism of ABC transporter and the role of it in bacterial pathogenicity,discussed the significance and issues in further studying the mechanism of ABC transporter and control of bacterial diseases. The cell surface or secretory factors associated with ABC transporters is potential targets of antimicrobial therapy or vaccine development,which provides a new idea for the prevention of bacterial diseases. Table and Figures | Reference | Related Articles | Metrics

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“Past and Present”Species Name of Bacillus velezensis LIU Yang, LIU Xiao-kun, CHEN Wen-hao Biotechnology Bulletin    2019, 35 (7): 230-232.   DOI: 10.13560/j.cnki.biotech.bull.1985.2019-0600 Abstract （1080）   HTML    PDF（pc） （1510KB）（808）       Knowledge map    Save Bacillus velezensis is a common functional bacterial species in the environment,and its taxonomic status and species name determination has always been the focus of debate among bacterial taxonomists. Until recently,the effective species name of B. velezensis was determined. In this paper,the evolution process of the species name of B. velezensis is systematically expounded in a time-smooth manner,in order to give readers a clear understanding of the origin and alternation of the species name of B. velezensis. Reference | Related Articles | Metrics

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Transcriptome and RNA-Seq Technology Zhang Chunlan, Qin Zijuan, Wang Guizhi, Ji Zhibin, Wang Jianmin Biotechnology Bulletin    2012, 0 (12): 51-56.   Abstract （1063）      PDF（pc） （1420KB）（7512）       Knowledge map    Save The transcriptome is the complete set of transcripts for certain type of cells or tissues in a specific developmental stage orphysiological condition. Transcriptome analysis can reveal the organism's gene expressing level, structural variation, discovery of new genes. Theresearch methods and platforms of transcriptome are undergoing rapid changes. And bioinformatics analysis is also improved gradually. RNASeqas a new research method, it can be more quickly and afford more accurate transcriptome’s information while using the Next-generationSequencing（NGS）technology. This article compares several main methods and platforms of transcriptome in recent years, and review theprinciple, purpose, steps, bioinformatics’ analysis and applications in related fields of RNA-Seq. This will be afford useful reference for relatedresearch. Reference | Related Articles | Metrics

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Research Progress in Single-Cell RNA-Seq of Plant LI Yi, SUN Chao Biotechnology Bulletin    2021, 37 (1): 60-66.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-1011 Abstract （1061）   HTML （76）    PDF（pc） （1418KB）（669）       Knowledge map    Save Single-cell RNA-Seq（scRNA-seq）is a technology to analyze gene expression at single cell level. Currently PCR plate-based and droplet-based protocols are 2 major popular ones used in the studies of plant. Here,we firstly outline the technical principles and data analysis procedure of plant scRNA-seq,and then introduce its research status,focusing on how scRNA-seq can be used to detect distinct plant cell types,to reveal cell evolution trajectories and to construct inter-cell regulatory networks. ScRNA-seq may provide a new perspective on plant research,and help to understand and identify critical cellular biological processes in complex tissues. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on the Mitochondrial Pyruvate Carrier（MPC） XIE Wen-ya, ZOU Shi-ying, Gao Ru-xin, HE Xiao-yun Biotechnology Bulletin    2019, 35 (7): 196-201.   DOI: 10.13560/j.cnki.biotech.bull.1985.2018-0926 Abstract （1057）   HTML    PDF（pc） （1110KB）（537）       Knowledge map    Save The mitochondrial pyruvate carrier（MPC）in mitochondrial inner membrane transports the pyruvate from the cytoplasm to the mitochondrial matrix. The pyruvate involves in the tricarboxylic acid cycle,gluconeogenesis,and the metabolic processes such as lipids and amino acids,which provides energy to the body. Therefore,MPC regulates energy metabolism by controlling the flux of pyruvate into the mitochondrial matrix. This paper summarized the biological characteristics of MPC and its relationship with metabolic diseases. It has been found that inhibition of MPC activity may lead to a decrease in hepatic gluconeogenesis,which in turn affects glucose homeostasis and reduces the incidence of typeⅡdiabetes. It also prevents the transmission of insulin signaling by affecting glucose-stimulated insulin secretion,resulting in that body is not sensitive to external glucose uptake;thus the MPC complex is essential to the glucose homeostasis. However,the activity of MPC is inhibited in most cancer tissues,specifically activating the MPC function should be conducted to enhance its aerobic metabolism,to inhibit that the cancer cells use lactic acid to supply energy and subsequently to hinder the proliferation of cancer cells. MPC is considered to be an initial point for studying energy metabolism regulation and potential therapeutic target sites,and also provides new therapeutic ideas for studying metabolic diseases such as obesity,diabetes and cancers. Reference | Related Articles | Metrics

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Development Status of Synthetic Biology in Globe and Its Enlightenment WANG Xiao-mei, YANG Xiao-wei, LI Hui-shang, HE Wei, XIN Zhu-lin Biotechnology Bulletin    2023, 39 (2): 292-302.   DOI: 10.13560/j.cnki.biotech.bull.1985.2022-0352 Abstract （1055）   HTML （68）    PDF（pc） （4302KB）（1228）       Knowledge map    Save Synthetic biology is a new interdisciplinary frontier science, demonstrating broad application prospects in the fields of life science, energy science and technology, medical health, material chemical industry and agricultural science and technology. The research on the development status of synthetic biology is of great significance for government departments, scientific and industrial communities to jointly promote its development. The current situation of synthetic biology policies, scientific research and industry in globe was systematically summarized. Combined with the development status and existing issues of synthetic biology in China, strategic suggestions for its development are put forward. The study showed followings. 1）Synthetic biology has entered a period of rapid development of global consensus, cooperation and competition. Many countries have assisted the scientific research and application innovation of synthetic biology, and produced many new technologies and applications with field characteristics through the top-down research and development system. 2）China is accelerating the improvement of the top-level design of synthetic biology, and a series of original discoveries and innovative achievements in the field has been achieved. However, there are also some shortcomings, such as lagging medium and long-term development planning, insufficient scientific research and innovation ability, the dislocation of application research and development subjects and the limitation of industrial application scenario expansion. 3）It is suggested to promote the development of synthetic biology in China from the aspects of strengthening macro policy guidance, building an efficient research system, cultivating high-quality industrial subjects and expanding the application scenarios of achievements. Table and Figures | Reference | Related Articles | Metrics

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Tetra-primer Amplification Refractory Mutation System PCR and Its Application in Fauna and Flora Genetics and Breeding Research YU Jun-jian, CHI Mei-li, JIA Yong-yi, LIU Shi-li, ZHU Jun-quan, GU Zhi-min Biotechnology Bulletin    2020, 36 (5): 32-38.   DOI: 10.13560/j.cnki.biotech.bull.1985.2019-0575 Abstract （1052）   HTML    PDF（pc） （1135KB）（764）       Knowledge map    Save Tetra-primer amplification refractory mutation system PCR（tetra-primer ARMS PCR）is a technique of typing SNP developed on the basis of common PCR. This technique combines the advantages of amplification refractory mutation system（ARMS）and four-primer PCR（tetra-primer PCR）,is an improvement of allele specific PCR. It has the characteristics of simple operation,rapid typing and low cost and has been applied more and more widely in the field of life sciences especially in the field of breeding. In this paper,the principle and advantages of tetra-primer ARMS PCR technique,detection methods of results and improvement methods of reaction system are introduced. In addition,the applications of this technique in genetics and breeding researches are reviewed. Reference | Related Articles | Metrics

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Research Advances in Plant Genome Assembly TANG Die, ZHOU Qian Biotechnology Bulletin    2021, 37 (6): 1-12.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0450 Abstract （1040）   HTML （102）    PDF（pc） （2931KB）（1454）       Knowledge map    Save The construction of reference genome containing genome-wide sequence is a prerequisite to genomic exploiting and utilizing of a species. Majority of angiosperms have undergone genome-wide duplication or polyploidization and subsequent chromosome rearrangement and loss. Many plants have also experienced large-scale expansion of repetitive sequences,resulting in a dramatic expansion of the genome size. These evolution events shape plant genomes with specific characteristics and extensive biodiversity,to a certain extent,has also led to many problems in plant genome assembly. Here,we classified plant genomes to simple,highly heterozygous,highly repetitive,polyploidy and pan-genome,summarized their corresponding assembly strategies and applications. Moreover,we prospecte the application trend of new sequencing technologies in resolving plant genomes. Table and Figures | Reference | Related Articles | Metrics

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Application Research Progress of Proteomics Yin Wen, Fu Xu, Li Ping Biotechnology Bulletin    2014, 0 (1): 32-38.   Abstract （1034）      PDF（pc） （1201KB）（3947）       Knowledge map    Save Proteomics is an emerging discipline for studying proteins composition and function in a type of cell, tissue or body fluids in a large-scale, high-throughput and systematic level. While genes determine the level of protein, but the level of gene expression can not represent the intracellular reactive protein levels. Proteomic analysis is a complement to the study of translation and modification and also an indispensable tool for a comprehensive understanding of genome expression. The development of proteomic technologies has greatly promoted the progress of proteomic research, and it has been widely used in various research fields.This paper revieweded the proteomic technologies and the applications in various fields are also briefly reviewed. Finally, some future issues are presented. Reference | Related Articles | Metrics

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Advances in Analysis Methods of Mass Spectrometry-based Metabolomics TIAN He, SHUI Guang-hou Biotechnology Bulletin    2021, 37 (1): 24-32.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-1383 Abstract （995）   HTML （41）    PDF（pc） （1555KB）（1177）       Knowledge map    Save Mass spectrometry（MS）-based metabolomics analyses are now widely applied to identify and quantify small-molecular metabolites in biological samples for uncovering endogenous alterations in cells after their exposure to exogenous stimulus. Duo to the complicated components in biological samples and a big span of polarities of metabolites ranged from hydrophilic to hydrophobic metabolites,a series of extraction protocols and the matched liquid chromatography（LC）and MS conditions are required to obtain the quantitative and qualitative information of target components. To some extent,the diversity of structure and physicochemical properties of small molecule metabolites limits the high-throughput acquirement of whole metabolic profiles from biological samples. Present review focuses on the introduction and discussion of extraction methods of varied metabolites,as well as their corresponding LC-MS conditions. Table and Figures | Reference | Related Articles | Metrics

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Overview of CLSI,EUCAST,and Susceptibility Breakpoints in China RUAN Zi-han, HUANG An-xiong, WANG Xiu-juan, HUANG Ling-li, HAO Hai-hong Biotechnology Bulletin    2022, 38 (9): 47-58.   DOI: 10.13560/j.cnki.biotech.bull.1985.2022-0695 Abstract （988）   HTML （32）    PDF（pc） （1270KB）（460）       Knowledge map    Save The significance of establishing susceptibility breakpoints is to interpret the results of drug susceptibility tests,and to select appropriate drugs and optimal dosing regimens in treating animals,which is an important means to control drug resistance. At present,institutions to establish susceptibility breakpoints have been established in many developed countries,among which the most influential and authoritative institutions are the Clinical and Laboratory Standards Institute（CLSI）and the European Committee on Antimicrobial Susceptibility Testing（EUCAST）. The research on drug resistance monitoring in China started relatively late,and mainly drew on the susceptibility breakpoints established by the CLSI. In 2017,with the support of the European Society for Clinical Microbiology and Infectious Diseases and EUCAST,the Chinese Committee on Antimicrobial Susceptibility Testing was established in China. Leading by the China Institute of Veterinary Drug Control（IVDC）in recent years,researchers from various research institutions have also made breakthroughs in susceptibility breakpoints and drug resistance monitoring,laying a solid foundation for controlling the development of antimicrobial resistance in China. This article first introduced the definition and significance of susceptibility breakpoints to provide a basic understanding for microbiological researchers. Then,it reviewed the basic situation of the CLSI and the EUCAST,and compared the differences in the establishment of susceptibility breakpoints between the two. Finally,this article analyzed and summarized the results of China's veterinary antimicrobial resistance monitoring and susceptibility breakpoints. In general,China is in a state of steady growth in the control of veterinary antimicrobial resistance,and many established standards for the determination of resistance are of great significance for controlling the development of resistance. However,there are still many veterinary antibiotics lacking corresponding susceptibility breakpoints,which requires more researchers to find more rapid,simple and accurate methods in the future,and fill and update the susceptibility breakpoints of veterinary antibiotics. Table and Figures | Reference | Related Articles | Metrics

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Optimization and Application of Double-plasmid CRISPR-Cas9 System in Escherichia coli WANG Kai-kai, WANG Xiao-lu, SU Xiao-yun, ZHANG Jie Biotechnology Bulletin    2021, 37 (12): 252-264.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0618 Abstract （983）   HTML （39）    PDF（pc） （4498KB）（558）       Knowledge map    Save In recent years,the CRISPR-Cas9（clustered regularly interspaced short palindromic repeats）system has been successfully exploited as versatile genome editing tools in various microorganisms. Because the application of CRISPR-Cas9 system is restricted only by the NGG of PAM（protospacer adjacent motif）sequence,therefore CRISPR-Cas9 system can theoretically edit any site or gene on the genome harboring NGG sequence. However,for the genes that have profound effects on cell growth and metabolism,the editing efficiency would be significantly reduced or even the mutants could not be obtained. Plenty of previous reports have provided valuable strategies to reduce the off-target effects of CRISPR-Cas9 system,but the reduction of editing efficiency is far from being solved. In this study,an efficient double- plasmid CRISPR-Cas9 system was established by using plasmids with different copy numbers to regulate the concentration of homology arms and the expression of Cas9 protein and gRNA,which would make them work more collaboratively for the gene editing purpose. The experimental results showed that the pfkA（6-phosphofructokinase isozyme 1）and pfkB（6-phosphofructokinase isozyme 2）genes in glycolysis pathway and the zwf（Glucose-6-phosphate 1-dehydrogenase）gene in pentose phosphate pathway were successfully deleted using the optimized double-plasmid CRISPR-Cas9 system,with the gene deletion efficiency of up to 100%;the nagABE gene cluster was also successfully replaced by glycerol kinase gene glpK,with the gene integration efficiency of 10%. By contrast,when using the single-plasmid CRISPR-Cas9 system,the pfkB gene deletion and glpK gene integration were successful in double-plasmid CRISPR-Cas9 system,and the efficiency of deleting pfkA and zwf increased by 31% and 63% respectively. The differences in carbon source utilization between mutant and wild-type strains further indicated that the gene editing efficiency was associated with special gene activity. Results from this study demonstrated that the sufficient target gene homology arms and the over-expression of gRNA may efficiently enhance the gene editing efficiency of CRISPR-Cas9 system. Table and Figures | Reference | Related Articles | Metrics

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Research Advance in Polyethylene Terephthalate Hydrolytic Enzymes SHI Li-xia, GAO Song-feng, ZHU Lei-lei Biotechnology Bulletin    2020, 36 (10): 226-236.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-0336 Abstract （978）   HTML （58）    PDF（pc） （3689KB）（1073）       Knowledge map    Save Polyethylene terephthalate（PET）is one of the most widely used plastics and accumulation of the discarded PET has become a global environmental issue. PET hydrolytic enzymes mainly hydrolyze PET into soluble building blocks：terephthalic acid（TPA）and ethylene glycol（EG）at moderate conditions. Enzymatic degradation of the recalcitrant plastic PET may provide a more environmentally friendly approach to solve the problem of “white pollution.” Here,we reviewed the 3D structures,catalytic mechanisms and PET degradation activity of four PET hydrolases,including cutinases,PETase,lipases and MHETase,mainly focusing on the protein engineering effort of cutinases and PETase. We also discussed the key scientific challenges on the development of high efficient enzymatic degradation of PET,aiming to provide theoretical basis for the engineering of more efficient PET hydrolytic enzymes. Table and Figures | Reference | Related Articles | Metrics

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Research Progress of Metabolomics in Plant Stress Biology ZHANG Feng, CHEN Wei Biotechnology Bulletin    2021, 37 (8): 1-11.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0861 Abstract （977）   HTML （100）    PDF（pc） （2025KB）（878）       Knowledge map    Save In recent years,with continuous and complex environmental changes,biotic and abiotic stresses frequently burst out in nature,and many stresses seriously affect plant normal growth and development,especially crop yield. The metabolic remodeling under stresses are the consequences of interactions between genotypes and surrounding environments,are the direct reflections of plant physiological phenotypes and biochemical activities,and largely reflects the plant response and defense to stresses. The rise of metabolomics provides a reliable way to study the metabolic remodeling in different tissues and under stresses in plants. Meanwhile,the integrations of metabolome with genome,transcriptome,proteome and phenome,especially metabolome-genome association analysis by the integration of genome and metabolome play an important role in revealing the genetic basis of plant response and adaptation to stresses,in improving crop yield and in developing stress-tolerant crop varieties. In this paper,the metabolome research method,the diversity of the metabolic remodeling and the genetic basis of plant metabolome under stresses are reviewed,and the application prospects and limitations of plant metabolomics in plant stress biology are also prospected. Table and Figures | Reference | Related Articles | Metrics

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Principle and Application of Microscale Thermophoresis in Studies of Biomolecular Interactions Ai Qiushi, Cao Xiangyu, Zhao Qian, Niu Yali, Song Shuishan, Biotechnology Bulletin    2015, 31 (6): 67-73.   DOI: 10.13560/j.cnki.biotech.bull.1985.2015.06.010 Abstract （968）   HTML    PDF（pc） （1689KB）（2455）       Knowledge map    Save Microscale Thermophoresis(MST)is a new technique to study biomolecular interactions in recent years. It is based on thermophoresis, i.e., the directional movements of molecules in a temperature gradient, which results in subsequent changes of molecular properties such as sizes, charges, hydration shell and conformation. MST combines the precise fluorescence detection and sensitive thermophoresis and provides a sensitive, fast and precise detection technique to analyze biomolecular interactions. Wor­king principle, detection process and application of MST in biology researches are reviewed here. Reference | Related Articles | Metrics

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Advances in the Regulation of Plant MYB Transcription Factors in Secondary Metabolism and Stress Response WEI Xin-xin, LAN Hai-yan Biotechnology Bulletin    2022, 38 (8): 12-23.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1350 Abstract （937）   HTML （65）    PDF（pc） （2460KB）（893）       Knowledge map    Save MYB is one of the largest transcription factor（TF）families in eukaryotes,its members play important roles in plant growth and development,secondary metabolism,biotic and abiotic stress responses and other physiological processes. MYB TFs affect the development and metabolism of plants by regulating the expressions of related genes in signaling pathways via the mutual recognition of specific functional domains with target genes. So far,the functions of some members of MYB TF family have been characterized,involving the function and expression regulation of MYB in plant secondary metabolism,biotic and abiotic stresses,etc. Based on the above research progress,hereby we summarized the recent results of MYB TF study,and which is expected to provide evidence for further study. Table and Figures | Reference | Related Articles | Metrics

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The Advantages and Limitations of CRISPR/Cas9-based Gene Editing Technology WU Yan, HAO Ya-qiao, WEI Xuan, SHEN Qi, LIU Ye-fei, WANG Sheng-hou, ZHAO Hong-xin Biotechnology Bulletin    2018, 34 (5): 1-8.   DOI: 10.13560/j.cnki.biotech.bull.1985.2017-1100 Abstract （933）   HTML    PDF（pc） （1929KB）（703）       Knowledge map    Save CRISPR/Cas9 is becoming a hot spot in genome editing,and it provides a more efficient way for gene targeting than zinc finger endonuclease（ZFN）and transcription activator-like effector nuclease（TALEN）. CRISPR/Cas9 system exists widely in bacteria and archaea,carrying out the adaptive immune responses specifically recognizing the invaded genetic elements from phage infection,plasmid conjugation and transformation as well as and degrading exogenous DNA. CRISPR/Cas9 is a precise editing technology by which the target sites are recognized by a segment of 20 bp small RNA. CRISPR/Cas9 has the advantages of simple design and operation,high efficiency,and broad generality,thus it is a novel and revolutionary precise gene editing technology. This review induced and summarized the aspects of the discovery of CRISPR/Cas,mechanisms,gene editing,and application limitations,aiming at providing references for understanding its mechanisms and applying precise gene editing technology. Reference | Related Articles | Metrics

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The Cancer-related Bioinformatics Databases Yang Jian, Cai Haoyang Biotechnology Bulletin    2015, 31 (11): 89-101.   DOI: 10.13560/j.cnki.biotech.bull.1985.2015.11.010 Abstract （918）   HTML    PDF（pc） （1223KB）（5218）       Knowledge map    Save Malignant tumor has become one of the major diseases that takes seriously risks to human health. In recent years, the rapid development of high-throughput detection technology has become an important means in cancer research. In this way, the cancer genomics data accumulated rapidly. These data is important for the research of mechanisms of tumor initiation and development. Massive biological data management and mining have become the foundation and an important field in cancer research. This article describes the frequently used bioinformatics databases of human tumors, including comprehensive databases, databases of genomics, transcriptomics, proteomics, epigenetics, etc. Here we sum up the status quo of database development in China and abroad, and discuss the existing problems to assist current research. Reference | Related Articles | Metrics

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Research Progress of Approaches in Single Cell RNA Sequencing Data Analysis ZHANG Miao, SUN Xiang-rui, XU Chun-ming Biotechnology Bulletin    2021, 37 (1): 52-59.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-0469 Abstract （912）   HTML （51）    PDF（pc） （1116KB）（796）       Knowledge map    Save Single cell RNA sequencing（scRNA-Seq）has been widely used in cell differentiation,tumor microenvironment and etiology of various diseases. Currently improving the accuracy of sequencing results by optimizing data analysis methods has become a research hotspot in the field of sequencing due to the characteristics of low capture rate,high noise,and high variability of scRNA-Seq. Here an overview of the mathematical methods used in the process of data analysis in recent years is summarized,and the advantages and issues in data analysis are discussed,aiming to provide a reference for the development and application of new algorithms,and to gradually improve the reliability of sequencing results. Reference | Related Articles | Metrics

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Application and Prospect of Gene Editing Technology in Crop Breeding LI Shu-lei, ZHENG Hong-yan, WANG Lei Biotechnology Bulletin    2020, 36 (11): 209-221.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-0328 Abstract （890）   HTML （50）    PDF（pc） （1153KB）（939）       Knowledge map    Save Genome editing is a group of technologies that allows the introduction of deletions,insertions or base substitutions by causing damage in targeted gene locus. This technology has now been utilized widely for editing the genome of various organisms,including bacteria,fungus,animals and plants. Compared to the Zinc-finger nucleases（ZFNs）and Transcription activator-like effectors nucleases（TALENs）editors,clustered regularly interspersed short palindromic repeats（CRISPR）/ CRISPR-associated protein（Cas）（CRISPR/Cas）system has developed as an efficient and versatile method for genome engineering. In this review,we describe the basic mechanism of ZFNs,TALENs and CRISPR/Cas system,and summarize the functions,limitation and application of CRISPR/Cas system in crop breeding. Finally,we discuss the potential challenges and opportunities of genome-edited crops in the future. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in the Inheritance and Breeding Improvement of Rice Quality LI Ran, QIAN Qian, GAO Zhen-yu Biotechnology Bulletin    2022, 38 (4): 4-19.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1598 Abstract （890）   HTML （62）    PDF（pc） （1174KB）（864）       Knowledge map    Save With the improvement of people’s living standard,high quality has become breeding aim of rice breeders and concern of consumers. Rice quality includes milling quality,appearance quality,eating and cooking quality,and nutrient quality. First,we reviewed the progress in studies on inheritance and breeding improvement of rice quality. Then we introduced the cloned genes related to rice quality or molecular function of QTLs for rice quality and their application in quality improvement. Based on this,we analyzed the issues encountered at present in rice quality inheritance and improvement prospected future research interests. Table and Figures | Reference | Related Articles | Metrics

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Research Progress on the MBW Complexes in Plant Anthocyanin Biosynthesis Pathway GAO Guo-ying, WU Xiao-fang, ZHANG Da-wei, ZHOU Ding-gang, ZHANG Kai-xuan, YAN Ming-li Biotechnology Bulletin    2020, 36 (1): 126-134.   DOI: 10.13560/j.cnki.biotech.bull.1985.2019-0738 Abstract （881）   HTML    PDF（pc） （1497KB）（832）       Knowledge map    Save Anthocyanin is one of the key substances in the plant coloration,which can not only endue plants with rich colors,but also have a wide range of biological functions,such as anti-oxidation,anti-ultraviolet,resistance to diseases and insect pests. The plant anthocyanin biosynthesis pathway is completed while synergizing by a series of structural and regulative genes. The structural genes are regulated and expressed by MYB,bHLH and WD40 transcription factors and the MBW complexes composed of them. This paper reviews the recent research progresses in functions of MYB,bHLH and WD40 transcription factors and MBW complexes in anthocyanin synthesis in plants,and summarizes various biological processes and regulatory networks in anthocyanin synthesis. Reference | Related Articles | Metrics

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Isolation,Identification of Carbon-fixing Bacteria and Determination of Their Carbon-fixing Abilities GUO Jun, FAN Fang-fang, WANG Li-ge, WU Ai-Lian, ZHENG Jun Biotechnology Bulletin    2019, 35 (1): 90-97.   DOI: 10.13560/j.cnki.biotech.bull.1985.2018-0544 Abstract （867）   HTML    PDF（pc） （3354KB）（558）       Knowledge map    Save Currently,as a biological carbon sequestration strategy,microbial recovery and fixing of CO2 gas,has become the focus of interest in dealing with the major environmental problem of greenhouse effect. The purpose of this study is to separate and screen efficient carbon-fixing microbial strains. By using the carbon-free source inorganic medium,24 kinds of strains with CO2 as carbon source were separated from facilities soil,activated sludge and biogas slurry. Then,the 8 strains of bacteria that grew faster were selected to conduct their morphological observations,physiological and biochemical responses as well as to perform cbbL gene and 16S rDNA sequencing analysis. Further,the sequencing results were compared with the data in BlAST database and the molecular characterization of the 8 carbon-fixing strains was carried out along with the detection of the strain content and the RubisCO enzyme activity. Finally,the C2-8R with the highest RubisCO activity among the 8 strains was selected for soil application test,and the carbon sequestration capacity of the C2-8R was determined through the detection of the RubisCO enzyme activity in soil. The above results revealed that carbon sequestration strains can be screened through carbon-free medium. The screened carbon-fixing bacteria are subordinate to the strains of Pseudomonas sp. and Methanotrophs sp.,and can be used to indicate the microbial carbon-fixing ability through RubisCO enzyme activity. Reference | Related Articles | Metrics

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Advances in HKT1 Study on the Mechanism of Salt Tolerance in Plants ZHANG Yong-lan, XIE Li-nan Biotechnology Bulletin    2021, 37 (6): 213-224.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-1379 Abstract （859）   HTML （47）    PDF（pc） （1973KB）（394）       Knowledge map    Save Soil salinization is an important factor affecting the reduction of crop yield. Under salt stress,plants have evolved a series of strategies to deal with salt. In these strategies,ion transporters play a pivotal role in plant response to salt stress. HKT1 transporters are a kind of ion transporters with the function of transporting Na+,which are mainly located near vascular bundles and widely found in monocotyledons and dicotyledons. In plants,they participate in the long-distance transport of Na+,regulate the concentration of Na+,and help to maintain ion balance. In this paper,the functions and corresponding regulatory processes of HKT1 proteins in different plants are discussed,and the different action models of HKT1 participating in Na+ long-distance transportation are summarized. By making a synthesis of the research on HKT1 in recent years and rational use of molecular breeding,it lays a theoretical foundation for increasing the salt stress tolerance of crops and improving the quality and yield of crops. Table and Figures | Reference | Related Articles | Metrics

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Research Progresses in the Synthetic Biology of Terpenoids SUN Li-chao, LI Shu-ying, WANG Feng-zhong, XIN Feng-jiao Biotechnology Bulletin    2017, 33 (1): 64-75.   DOI: 10.13560/j.cnki.biotech.bull.1985.2017.01.007 Abstract （848）   HTML    PDF（pc） （1825KB）（3408）       Knowledge map    Save The terpenoids represent the largest class of natural products with biological activities of antitumor and anti-allergy,thus they have been widely applied in the area of food,cosmetics and medical health,presenting huge potential and broad market prospects. Recent years,researchers applied functional genomics and metabonomics approaches to deeply study the biosynthesis pathways of terpenoids,providing tons of data for their synthetic biology. The construction of engineered yeasts using synthetic biology enabled the efficient synthesis of multi-target terpenoids,and highly improved the overall production level. Thus,the synthetic biology approach is expected to be an efficient way of producing plant-derived terpenoids. First,we introduced the concept of synthetic biology,summarized the important functions and applications of plant-derived terpenoids,briefly reviewed the biosynthesis pathways,and concluded the alternative production ways. Then,we discussed the design strategies of synthetic biology for terpenoids thoroughly. Finally,we elaborated the advances on the biosynthetic biology of varied terpenes with common terpenes as the studied cases. Reference | Related Articles | Metrics

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Teaching Examples of Applied Bioinformatics Course Luo Jingchu Biotechnology Bulletin    2015, 31 (11): 102-111.   DOI: 10.13560/j.cnki.biotech.bull.1985.2015.07.001 Abstract （845）   HTML    PDF（pc） （2527KB）（1194）       Knowledge map    Save In this article, we introduce the basic bioinformatics analysis methods and tools taking the hemoglobin as an example. The methods include: 1)protein and DNA sequence alignment；2)advanced search for UniProt and RefSeq database；3)Blast database similarity search；4)phylogenetic tree construction under MEGA；5)protein structure comparison using Swiss-PdbViewer. Reference | Related Articles | Metrics

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Research Progress on Plant AP2/ERF Transcription Factor Family ZHANG Qi, CHEN Jing, LI Li, ZHAO Ming-zhu, ZHANG Mei-ping, WANG Yi Biotechnology Bulletin    2018, 34 (8): 1-7.   DOI: 10.13560/j.cnki.biotech.bull.1985.2017-1142 Abstract （836）   HTML    PDF（pc） （1114KB）（810）       Knowledge map    Save As one of the largest transcription factor family,AP2/ERF transcription factors are common in plants,and have attracted much attention because of their important role in gene breeding. The AP2/ERF transcription factor family contains at least one AP2 conservative domain with about 60 amino acids. According to the number of domains and the sequence of recognition,AP2/ERF transcription factor family can be classified into 5 subfamilies of AP2,ERF,DREB,RAV,and Soloist;and the number of AP2/ERF transcription factor and their subfamily members are variable in different plants,such as Arabidopsis,rice,maize,tomato,etc. Through responses to the regulation of ethylene,cytokinin and auxin,AP2/ERF transcription factors are reported to participate directly or indirectly in multiple processes of plant development including seed development,flower and fruit organ formation. Except for primary metabolism,AP2/ERF transcription factors affect significantly the regulation of plant secondary metabolism,particularly in improving the accumulation of active ingredients of medicinal plants（such as artemisinin,paclitaxel and lignin）. Meanwhile,the AP2/ERF gene of Arabidopsis are also reported to have a positive regulatory function of resistance to Botrytis cinerea,while some AP2/ERFgenes play a vital role in response to drought,high salt,low temperature,hypoxia and other abiotic stresses. In addition,the AP2/ERF transcription factors also participate in the non-biological signal transduction pathways mediated by ethylene. In this study,the structure and classification characteristics of AP2/ERFtranscription factors,their quantity distribution in different plants will be introduced,and the roles of AP2/ERF transcription factors in plant development,secondary metabolism,biotic or abiotic stress and signal transduction will be expounded. Overall,this review is aimed to provide theory basis for cultivating transgenic crops with high-yielding and stress tolerances in the future. Reference | Related Articles | Metrics

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Research Progress in Biosynthesis of Secondary Metabolites of Microorganisms ZHAO Jiang-hua, FANG Huan, ZHANG Da-wei Biotechnology Bulletin    2020, 36 (11): 141-147.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-0475 Abstract （828）   HTML （37）    PDF（pc） （1082KB）（2512）       Knowledge map    Save Microorganisms can produce a variety of natural secondary metabolites with biological activity,which have broad application prospects in medical,agricultural and food fields. The development of molecular biology, bioinformatics and sequencing technology has laid a theoretical foundation for the mining of secondary metabolites and provided operational tools. In recent years,there are more and more studies on the secondary metabolites of microorganisms. It is difficult to get satisfactory results by traditional methods. It will be a new direction to operate microorganisms at the molecular level,such as the regulation of secondary metabolite biosynthesis gene cluster.The strategies of finding new secondary metabolites and increasing the production of secondary metabolites were reviewed,by using the methods of heterologous expression of secondary metabolite biosynthesis gene cluster,ribosome engineering,epigenetic regulation,metabolic regulation and regulation of transcription and translation level,the aim is to excavate new secondary metabolites,improve the production of secondary metabolites,and provide a breakthrough for the exploration of new drugs. Reference | Related Articles | Metrics

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Effects of Melatonin on Root Growth and Drought Tolerance of Maize Seedlings MA Xu-hui, CHEN Ru-mei, LIU Xiao-qing, ZHAO Jun, ZHANG Xia Biotechnology Bulletin    2021, 37 (2): 1-14.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-0575 Abstract （825）   HTML （58）    PDF（pc） （3968KB）（1067）       Knowledge map    Save Melatonin（N-acetyl-5-methoxy-tryptamine）,an indoleamine neurohormone widely existing in organism,involves in many physiological and biochemical processes of plants. Recent studies understand that melatonin application may improve plant stress tolerance at varied level;however,the specific underlying mechanisms are still not fully understood. The current study was carried out to investigate the effect of melatonin on maize root system and drought tolerance via 2 applications of melatonin. Firs,the method of hydroponic root maize and irrigating melatonin were used to analyze the root system and growth status of maize seedlings. The results showed that the application of melatonin significantly improved a variety of maize seedling root parameters,including root length,root surface area,root volume,number of lateral roots,etc. Secondly,the method of melatonin-soaked seeds in pot was used to determine the relative water content of leaves,photosynthesis,antioxidant enzyme activity,and above-ground biomass. The results showed that under drought stress,the method of melatonin-soaked seed improved plant photosynthesis rate,stomatal conductance and transpiration rate,enhanced antioxidant enzyme activity,and reduced reactive oxygen species and malondialdehyde content. Our current findings hereby confirmed the beneficial effects of melatonin application in enhancing maize drought tolerance by promoting root growth,reducing oxidative damages,eliminating photosynthetic inhibition,and improving plant water status. Table and Figures | Reference | Related Articles | Metrics

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Research Progress of ERF Transcription Factors in Regulating Biological Stress Responses SHAO Wen-jing, SHI Jie, ZHANG Pu, LANG Ming-lin Biotechnology Bulletin    2021, 37 (3): 136-143.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-0736 Abstract （815）   HTML （44）    PDF（pc） （1791KB）（569）       Knowledge map    Save ERF family is a major subfamily of plant-specific APETALA2/ethylene-responsive factor（AP2/ERF）transcription factor family. Its member structure is characterized by containing only one AP2/ERF domain composed of 58-60 amino acids. Most researches on the members of this family were focused on abiotic stresses such as cold and drought,and recent studies demonstrate that ERFs play an important role in resisting plant diseases and insect pests. These ERF subfamily members can bind to the GCC box element within the promoter region of their interactive genes,and activate or inhibit the expressions of these pathogenesis-related genes. Moreover,more researches confirmed that ERFs participate in salicylic acid（SA）,jasmonic acid（JA）,ethylene（ET）,H2O2,and other hormone signaling pathways,and effectively coordinate different hormones to resist the invasion of pathogenic bacteria,and to improve plants’ resistance to disease and insect through mutual promotion/antagonism. This paper reviews the structure features of ERF transcription factors（TF）,their regulatory mechanisms in different plants’ antibiotic stresses,and the latest progresses in improving plant resistance to diseases and insect pests by coordinating the interaction of different hormone signaling pathways. Finally,the paper prospects the application of ERF in plant disease resistance breeding. Table and Figures | Reference | Related Articles | Metrics