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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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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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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 （1057）   HTML （69）    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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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）（1457）       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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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 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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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 （938）   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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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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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 （860）   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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Gibberellin Metabolism Regulation and Green Revolution LI Yi-dan, SHAN Xiao-hui Biotechnology Bulletin    2022, 38 (2): 195-204.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0389 Abstract （812）   HTML （51）    PDF（pc） （4090KB）（665）       Knowledge map    Save Gibberellins（GAs）are very important plant hormones that control diverse aspects of plant growth and development. Some important regulatory genes involved in gibberellin biosynthesis and signaling transduction pathway positively impact plant architecture,yield and quality of cereal crops,which has been applied in agricultural production. The alleles of Rht-1 and sd-1,that confer a semi-dwarf feature to wheat and rice,respectively,underpin the‘Green Revolution’in the latter half of the twentieth century. This paper reviewed the research achievements on the GA metabolism regulation in impacting the height,yield,nutrition usage of‘Geen Revolution’varieties,and prospected how to develop and utilize GA regulation genes for breeding more‘Green Revolution’varieties in the future. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in CRISPR/Cas9 Genome Editing System in Edible and Medicinal Fungi LIU Xiao-tian, QIU Hao, TIAN Li, REN Ang, ZHAO Ming-wen Biotechnology Bulletin    2021, 37 (11): 4-13.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1055 Abstract （811）   HTML （68）    PDF（pc） （1708KB）（770）       Knowledge map    Save CRISPR/Cas9 is currently the most successful precise genome editing technology. So far,there have been relevant application reports in at least nine edible and medicinal fungi,including Pleurotus eryngii,Pleurotus ostreatus,Agaricus bisporus,Ganoderma lucidum,Schizophyllum commune,Coprinopsis cinerea,Flammulina velutipes,Cordyceps militaris and Shiraia bambusicola. This article briefly describes the mechanism and delivery strategies of the CRISPR/Cas9 system,and proposes solutions for the application in edible and medicinal fungi. Last but not least,referring to the current frontier development of CRISPR technology,such as multiplex gene editing, base editing, prime editing and transcription regulation, etc., we give certain enlightenment to the future application prospects in the field of edible and medicinal fungi. Table and Figures | Reference | Related Articles | Metrics

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Research Strategies of Natural Products Biosynthesis Pathways and Key Enzymes in Medicinal Plants ZHOU Zheng, LI Qing, CHEN Wan-sheng, ZHANG Lei Biotechnology Bulletin    2021, 37 (8): 25-34.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0949 Abstract （806）   HTML （62）    PDF（pc） （1166KB）（752）       Knowledge map    Save Natural products derived from medicinal plants are important sources of drug research and development. The traditional researches about natural products from medicinal plants mainly focus on their extraction and purification,chemical structure,biosynthesis and biological function. With the rapid development of genomics,bioinformatics,molecular biology,molecular genetics,synthetic biology and other disciplines and extensive cross integration among them,there are emerging opportunities for the natural products of medicinal plants. This paper systematically summarizes the research strategies of natural products biosynthesis and key enzymes in medicinal plants,included six aspects：speculation of natural products biosynthesis pathway,discovery and prediction of key enzymes in natural product biosynthesis,expression characteristics of enzymes,function of enzymes in vivo,catalytic characteristics of enzymes,analysis and optimization of enzyme structure and synthetic biology studies. Moreover,the trend of research strategies of natural products biosynthesis pathway and key enzymes in medicinal plants are prospected. Reference | Related Articles | Metrics

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Sugar Signaling and Its Role in Plant Response to Environmental Stress CHEN Hong-yan, LI Xiao-er, LI Zhong-guang Biotechnology Bulletin    2022, 38 (7): 80-89.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1289 Abstract （801）   HTML （31）    PDF（pc） （2639KB）（818）       Knowledge map    Save Sugar is not only carbon and energy sources as well as structural material in plant cells,but also a signaling molecule,which plays a key role in plant growth,development,and response to environmental stresses. Abiotic stresses,such as high temperature,low temperature,drought,salinity,and heavy metal,are the major stress factors limiting crop yield. The exact mechanism of sugar,as signaling molecule,in plants response to these stresses still remains unclear. Based on the current advance in sugar signaling and its role in the formation of stress tolerance in plants,the hexose kinase（HXK）-,regulator of G protein signaling 1（RGS1）-,glycolysis（EMP）-,and pentose phosphate pathway（PPP）-dependent signaling pathways were separately summarized;and the role of sugar signaling in the developing of plant stress tolerance,including heat,cold,drought,salt,and heavy metal stress tolerance was discussed. In addition,the future research direction of H2S signaling in the field of plant biology was prospected. Table and Figures | Reference | Related Articles | Metrics

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Humification Process and Microbial Driving Mechanism of Composting WANG Yu-yun, ZHAO Bing, MA Li-ting, LI Lan, DENG Ya-qin, XU Zhi Biotechnology Bulletin    2022, 38 (5): 22-28.   DOI: 10.13560/j.cnki.biotech.bull.1985.2022-0134 Abstract （793）   HTML （35）    PDF（pc） （1112KB）（480）       Knowledge map    Save Aerobic composting is a typical biochemical process of stabilizing and harmless organic waste. In this process,organic matter is decomposed by microorganisms and then polymerized to form humus（HS）. Due to its complex network structure,it is difficult for lignin to be completely degraded by microorganisms in the high temperature stage of compost. In addition,as the raw material and skeleton of HS formation,the deep degradation of lignin is of great significance to the humification process of composting. The composting cooling and humification stage are the key periods of HS formation,in which fungi and ligninase play an important role in deepening lignin degradation and enhancing humification. Temperature and pH are important environmental factors affecting humification process,and their regulation is an important means to strengthening humification process artificially. This paper reviewed the degradation mechanism of lignin by key enzymes,the interaction between precursor substances and humic acid formation,and the driving mechanism of fungi on humic acid formation. It is suggested that exploring the key genes and enzymes involved in HS anabolic pathway in composting is an important direction of composting humification in the future. Reference | Related Articles | Metrics

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Research on the Preparation and Purification of Kod DNA Polymerase YI Fang, LAI Peng-cheng, ZHENG Xi-ao, HU Shuai, GAO Yan-li Biotechnology Bulletin    2022, 38 (5): 183-190.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1210 Abstract （757）   HTML （28）    PDF（pc） （3661KB）（775）       Knowledge map    Save Kod DNA polymerase,as a common high-fidelity DNA polymerase,has strong DNA extension ability and 3'-5' exonuclease activities. This study is aimed to improve the expression efficiency and extension ability as well as fragment amplification ability by optimizing the isolation and purification conditions of recombinant Kod DNA polymerase. First,isopropyl β-D-thiogalactoside（IPTG）was used to induce pET-30a-Kod for expression,and enzymatic dissolution and ultrasonic fragmentation were to grind the cells and extract the crude enzyme solution,then Ni-NTA column was to elute and purify the Kod DNA polymerase. Finally,the high-purity Kod DNA polymerase was obtained by dialysis. PCR and Sanger sequencing were used to detect the activity and fidelity of self-purified Kod DNA polymerase. Meanwhile the key parameters such as IPTG and imidazole concentrations were optimized. The results showed that Kod DNA polymerase was highly expressed after induced by 0.1 mmol/L IPTG at 28℃ for 16-18 h,and the eluted effect was optimal while imidazole concentration was 200 mmol/L. The efficiency of the Kod DNA polymerase was the highest while Mg2+ was 1.5 mmol/L and Kod DNA polymerase was 0.061 25 μg/µL in the PCR reaction system,under which 3 194 bp target band was efficiently amplified;and there was no introduced mutation in the PCR product after sequence alignment. The enzymatic activity and fidelity of self-extracted Kod DNA polymerase after optimized preparation reached the same level of commercial high-fidelity DNA polymerase. Thus,this study lays a foundation for saving the cost of laboratory PCR and further developing and utilizing Kod DNA polymerase. Table and Figures | Reference | Related Articles | Metrics

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Research Progress of RNA Editing ZHU Lin, XIAN Feng-jun, ZHANG Qian-nan, HU Jun Biotechnology Bulletin    2022, 38 (1): 1-14.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1326 Abstract （748）   HTML （54）    PDF（pc） （1189KB）（1194）       Knowledge map    Save RNA editing is a post-transcriptional process on the target transcripts by base insertion,deletion,or replacement. This important epigenetic phenomenon is also considered as a correction mechanism for genetic restoration at the RNA level. Here,we reviewed the RNA editing systems based on PPR gene family in plants and CRISPR/Cas editing system in animal, introduced the important role of RNA editing in the plant growth and development, and prospected future application of RNA editing. In addition we also summarized the recent research on RNA editing database,which support the follow-up research of RNA editing. Table and Figures | Reference | Related Articles | Metrics

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Research Progress of Plant VIGS Technology and Its Application in Forestry Science GAO Peng-fei, XI Fei-hu, ZHANG Ze-yu, HU Kai-qiang, CHEN Kai, WEI Wen-tao, DING Jia-zhi, GU Lian-feng Biotechnology Bulletin    2021, 37 (5): 141-153.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-1452 Abstract （741）   HTML （30）    PDF（pc） （1868KB）（627）       Knowledge map    Save With the explosion of genomic sequence information,biological science has entered the era of big data. Functional annotation of genomic information has become an important research goal. Virus-induced gene silencing（VIGS）is a powerful tool in analysis of gene function for forest species that are lack of stable genetic transformation system. VIGS has been used to study multiple key genes in plant growth and development. VIGS works as such way,a fragment of target gene is inserted into a viral vector to generate small interfering RNAs（siRNAs）based on the mechanism of natural innate antiviral defense in plant,then the transcript of endogenous gene in the plant becomes the degraded target,resulting in down-regulation of target gene expressions. Here we systematically review the advantages and limitations of VIGS,as well as corresponding solutions. Meanwhile we also discuss the application of VIGS system in forest science. Finally,we explore the latest improving solutions and future prospects while VIGS as a powerful tool for assessing and characterizing gene functions in plant. Table and Figures | Reference | Related Articles | Metrics

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Structural and Functional Characterization of AOX Gene Family WANG Hui, MA Yi-wen, QIAO Zheng-hao, CHANG Yan-cai, ZHU Kun, DING Hai-ping, NIE Yong-xin, PAN Guang-tang Biotechnology Bulletin    2022, 38 (7): 160-170.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1285 Abstract （738）   HTML （32）    PDF（pc） （4593KB）（325）       Knowledge map    Save AOX（alternative oxidase,AOX）is a terminal oxidase in the respiratory pathway of plants. Through the identification and analysis of AOX genes in 8 species of Zea mays,Oryza sativa,Arabidopsis thaliana,Brachypodium distachyon,Solanum lycopersicum,Solanum tuberosum,Sorghum bicolor and Glycine max,we aim to further explore the function and regulatory mechanism of AOX gene. A total of 35 genes encoding AOX protein in these 8 species were identified,and these 35 genes were divided into 5 subfamilies by cluster analysis. The phylogenetic analysis and comparison of the chromosome position exon-intron structure and the conserved motif composition of AOX further supported the subfamily division model,and revealed that the AOX gene maintained a strong conserved nature during the evolution process. The prediction of cis-acting elements,miRNA target sites and the GO annotation analysis of AOX protein revealed that AOX gene may play an important role in plant growth and development and stress response. After stress treatment at the seedling stage of maize,the qRT-PCR analysis of 4 AOX genes in maize showed that the 4 AOX genes presented different degrees of inducible expression,suggesting that they were closely related in response to abiotic stress. These results lay a foundation for further understanding the biological function of AOX genes and their regulatory mechanism in response to abiotic stress. Table and Figures | Reference | Related Articles | Metrics

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The Strategy for Enhancing Foreign Proteins Expression by Signal Peptide in Bacillus subtilis MIAO Hua-biao, CAO Yan, YANG Meng-han, HUANG Zun-xi Biotechnology Bulletin    2021, 37 (6): 259-271.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-1255 Abstract （716）   HTML （31）    PDF（pc） （1658KB）（1550）       Knowledge map    Save As an important prokaryotic expression host strain,Bacillus subtilis has always been regarded as preferred model strain for foreign protein expression,featuring strong protein secretion,clear genetic background,no codon preference,rapid growth and non-pathogenicity. Signal peptide is a short peptide chain located in the N-terminal of the precursor protein,with the function of guiding and regulating the folding of the precursor protein. Meanwhile,it plays a very important role in the process of protein transfer and secretion. At present,there is no regularity to find the efficient secretion of different foreign proteins by using the signal peptide of B. subtilis. For this reason,the structural characteristics,classification,transport pathway and application of signal peptides from B. subtilis are reviewed here,aiming to provide certain reference for further screening the optimal signal peptides of foreign proteins in B. subtilis expression system. Table and Figures | Reference | Related Articles | Metrics

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Engineering Non-conventional Yeast Cell Factory for the Biosynthesis of Natural Products YE Min, GAO Jiao-qi, ZHOU Yong-jin Biotechnology Bulletin    2021, 37 (8): 12-24.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0815 Abstract （707）   HTML （53）    PDF（pc） （3826KB）（883）       Knowledge map    Save Natural products have been widely applied in medical,cosmetic and food industries. With the increasing of people’s demand,the green sustainable producing process is urgently needed. Recently,microbial cell factories for synthesizing natural products have been rapidly developing,and the current common-used cell factory hosts include Saccharomyces cerevisiae and Escherichia coli. Non-conventional yeasts have becoming very potential cell factory hosts due to their specific advantages,such as high-cell-density aerobic fermentation,tolerance to a wide range of temperature and pH,and wide substrate spectrum（long-chain hydrocarbons,fatty acids and methanol）. In this review,we summarized recent progresses on engineering non-conventional yeasts for the synthesis of natural products mainly terpenoids and flavonoids,and introduced common engineering methods and strategies. On this basis,we systematically discussed their strengths and weaknesses of novel non-conventional yeasts cell factories,and finally listed the challenges and opportunities while using non-conventional yeasts cell factories for the synthesis of natural products. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in the Mitigative Effects of Rhizosphere Microorganisms on Heavy Metal Stress in Plants and Their Mechanisms YANG Lu, XIN Jian-pan, TIAN Ru-nan Biotechnology Bulletin    2022, 38 (3): 213-225.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0811 Abstract （699）   HTML （28）    PDF（pc） （1154KB）（773）       Knowledge map    Save Phytoremediation technology has become an important means in green remediation of heavy metal pollution. Under heavy metal stress,rhizosphere microorganisms can effectively improve plant growth and physiological metabolism,and enhance the absorption and enrichment of heavy metals in plants. This paper first discusses the rhizosphere microbial composition and root-microbe interaction,and analyses the mitigative effects of plant rhizosphere growth-promoting bacteria and mycorrhizal fungi on plant resistance to heavy metal stress. Meanwhile,the article also discusses the mechanism of the rhizosphere microbes alleviating heavy metal toxicity from the aspects of microorganism,plant root exudates,plant ethylene synthesis,plant photosynthesis,plant antioxidant defense system,water and nutrient absorption and rhizospheric soil microenvironment. Finally,this paper combines the current research status and draws some conclusions. Such as,finding new strains of beneficial to plant growth under heavy metal stress,exploring the interaction between different microorganisms in the rhizosphere under heavy metal stress,and revealing the physiological and molecular mechanisms of plant root-microbial interaction system under heavy metal stress,aiming to provide guidance for the theoretical research and practice of microbial assisted phytoremediation of heavy metal pollution in the future. Table and Figures | Reference | Related Articles | Metrics

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Improvement Effect of Bifidobacterium lactis V9 on NAFLD Rats Induced by High-fat Diet ZHONG Ming-yue, LIU Chun-yan, YAN Yan, ZHANG Xiao-hui, YUAN Hai-sheng, XU Guo-quan, ZHANG He-ping, WANG Yu-zhen Biotechnology Bulletin    2022, 38 (3): 181-187.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0630 Abstract （695）   HTML （9）    PDF（pc） （3852KB）（221）       Knowledge map    Save This work aims to study the effect of probiotic Bifidobacterium lactis V9（V9）on lipid metabolism and intestinal injury in non-alcoholic fatty liver disease（NAFLD）rats induced by high-fat diet,and to explore its mechanism. Male Wistar rats were randomly divided into 5 groups（n=8）：blank control group（Control）,high-fat diet model group（HFD）,berberine-positive drug group（Berberine）,the probiotic B. lactis V9 treatment group（HFD/V9）,and the probiotic B. lactis V9 control group（V9）. Except Control and V9 were given ordinary maintenance feed,others were given a high-fat diet for 6 weeks to construct a NAFLD model. After 6 weeks,the high-fat diet was replaced with ordinary maintenance feed,and the rats were given probiotic B. lactis V9（1×10 9CFU/mL）or berberine（50 mg/kg）by gavage. After 4 weeks,blood,liver and colon tissues were collected for follow-up testing. Compared with the Control group,the mRNA expressions of liver non-esterified fatty acids（NEFA）,triglycerides（TG）,liver fatty acid synthase（FAS）and liver fatty acid binding protein 1（Fabp1）in the HFD group significantly increased（P<0.01）. At the same time,mRNA expressions of the intestinal cytokine interleukin 1β（IL-1β）,tumor necrosis factor α（TNF-α）,Toll-like receptor 2（TLR2）and Toll-like receptor 9（TLR9）in the HFD model group also significantly increased（P<0.01）. On the contrary,the expression of tight junction protein ZO-1 and occludin mRNA in the HFD model group significantly decreased（P<0.01）. After treatment with probiotic B. lactis V9,the contents of NEFA and TG in the liver significantly reduced（P<0.01）,the mRNA expressions of FAS and Fabp1 in the liver also significantly reduced（P<0.01）. Both probiotic B. lactis V9 and berberine treatment reduced the transcription levels of intestinal IL-1β,TNF-α,TLR2 and TLR9（P<0.01）,which increased the expressions of intestinal tight junction proteins ZO-1 and occludin mRNA. HE staining showed that there was no significant difference in colonic pathology among the above groups. Probiotic Probiotics B. lactis V9 can improve NAFLD induced by high-fat diet by alleviating lipid metabolism disorders,reducing intestinal inflammation and improving mulosal barrier. Table and Figures | Reference | Related Articles | Metrics

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Advances in Plant Flavonoids UDP-glycosyltransferase YAO Yu, GU Jia-jun, SUN Chao, SHEN Guo-an, GUO Bao-lin Biotechnology Bulletin    2022, 38 (12): 47-57.   DOI: 10.13560/j.cnki.biotech.bull.1985.2022-0236 Abstract （693）   HTML （19）    PDF（pc） （2676KB）（1399）       Knowledge map    Save Flavonoids are important natural products in plants and usually exist in the form of glycosides. Uridine diphosphate glycosyltransferases（UGTs）can modify flavonoids to form a variety of flavonoid glycosides, which are the active substances of flavonoid medicines in many medical plants. In recent years, as more and more plant genomes have been elucidated, a large number of glycosyltransferases involved in flavonoid synthesis have been identified. In this paper, we firstly introduced the structural features and family classification of plant UGTs, and then reviewed the advances in the study of flavonoid UGTs, had a complete summary on the selectivity of modification sites of plant flavonoid UGT in varied families, as well as the specificity of donors and receptors. It is aimed to lay a foundation for investigating the correlation between structure and function of plant flavonoid UGTs as well as the mining and identification of novel flavonoid UGTs. Table and Figures | Reference | Related Articles | Metrics

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A Rapid and Accurate Method for Tth DNA Polymerase Activity Assay CHEN Xiao-yu, ZHANG Jian, ZHANG Xin-ya, TANG Yu-ting, SHAO Yu-chen, LUO Zhi-dan, LU Chen Biotechnology Bulletin    2021, 37 (5): 281-286.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-0953 Abstract （686）   HTML （25）    PDF（pc） （2604KB）（856）       Knowledge map    Save Tth DNA polymerase is a thermostable enzyme with both DNA polymerase and reverse transcriptase activities. Traditional methods for activity assay are discontinuous,time consuming,and laborious. We developed a new method for activity assay of Tth DNA polymerase using a highly sensitive double-stranded nucleic acid specific dye PicoGreen,and a specially designed hairpin probe. A good linearity of the standard curve can be established by this method,with the coefficient of determination R2 > 0.99. This method was of high sensitivity and accuracy,and the results by it were consistent with the activity of commercial products. The verification from fluorescence quantitative PCR confirmed that this method may reveal the enzyme activity in real environment. The method can also be extended to other DNA polymerase activity assay,which will provide an effective tool for the production and transformation of DNA polymerase. Table and Figures | Reference | Related Articles | Metrics

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Effects of Endophytes on Biosynthesis of Secondary Metabolites and Stress Tolerance in Plants LIANG Zhen-ting, TANG Ting Biotechnology Bulletin    2021, 37 (8): 35-45.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0735 Abstract （677）   HTML （27）    PDF（pc） （1117KB）（621）       Knowledge map    Save Plant secondary metabolites have broad pharmacological activities and various biological functions. Biosynthesis of secondary metabolites has attracted much attention due to its low natural yield in plants and increasing demand of medicine. The plant and endophytes are long-tern mutual beneficial and symbiotic,and affects many metabolic processes and physiological activities of plants,which provides important source to solve the shortage of secondary metabolites. Here this paper reviews the biosynthesis of secondary metabolites,such as terpenoids,flavonoids and alkaloids,promoted by endophytes,as well as the regulation of secondary metabolites by endophytes to enhance plant resistance to biotic and abiotic stresses. The paper also discusses the production path and application prospect of endophytic associated plant secondary metabolites in modern biological field,aiming at providing ideal microbial resources for the biosynthesis of medicinal secondary metabolites and the improvement of crop stress tolerance. Table and Figures | Reference | Related Articles | Metrics

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Application of Single-cell Transcriptome Sequencing in Animals XIONG He-li, SHA Qian, LIU Shao-na, XIANG De-cai, ZHANG Bin, ZHAO Zhi-yong Biotechnology Bulletin    2022, 38 (3): 226-233.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0523 Abstract （674）   HTML （32）    PDF（pc） （1109KB）（633）       Knowledge map    Save Cell is the basic structural component and functional unit of organism. Cell type and function are determined by its whole transcriptional expression profile. Single-cell transcriptome sequencing can be used to obtain the transcriptional expression profile of a single cell,so as to identify cell types,cell states and rare types with high-precision resolution,thus to analyze the dynamic changes of cells and the relationship between cells at the single cell level,and further to decipher the molecular cellular mechanisms behind cell changes and cell abnormalities. With the stability and throughout improvement of single-cell sequencing,as well as the reduction of sequencing cost,single-cell transcriptome sequencing has been widely used in the fields of developmental biology,tumor,immunity and disease. However,these researches are mainly focused on human and model organisms,there are a few researches on animals. Therefore,the objective of this paper is to introduce the single-cell transcriptome sequencing and its biological application,and review some of its pioneering research in animals,aiming to provide a method reference for better application of single-cell transcriptome sequencing in animals. Reference | Related Articles | Metrics

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Advances in Compost Regulation of Rhizospheric Microbiome to Suppress Plant Diseases WANG Ning, LI Hui-xiu, LI Ji, DING Guo-chun Biotechnology Bulletin    2022, 38 (5): 4-12.   DOI: 10.13560/j.cnki.biotech.bull.1985.2022-0136 Abstract （674）   HTML （44）    PDF（pc） （1978KB）（464）       Knowledge map    Save In China,huge amount of organic wastes is produced annually and it contains large quantity of nutrients and might serve as an important resource to enhance ecological services in agroecosystem. Aerobic fermentation is a widely used method for the treatment of organic waste and resource transformation,and composting may inhibit many plant diseases. In this review,we summarized recent progress in the mechanism of the disease suppressive of composts,focusing on its regulatory effects on the structures and functions of rhizosphere microbiome as well as potential regulatory pathways. We discussed the differences between composting microbiome and soil rhizosphere microbiome,effects of composting on soil biological and physicochemical properties,and biological and abiotic environmental factors on rhizosphere microbiome. This relevant summary might provide some primary understanding of the interaction system among compost,soil and plant rhizosphere microbiome. Table and Figures | Reference | Related Articles | Metrics

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Sequencing and Analysis of Full-length Transcriptome from Medicago truncatula SHANG Xiao-yao, ZHOU Ling-fang, YIN Qian-qian, CHAO Yue-hui Biotechnology Bulletin    2021, 37 (8): 131-140.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0191 Abstract （664）   HTML （23）    PDF（pc） （5751KB）（776）       Knowledge map    Save For deep analysis and exploring of the complete structure of mRNA from Medicago truncatula,a model plant in Leguminosae,the single-molecule long-read sequencing technology（SMRT）was used to sequence and analyze the full-length transcriptome of M. truncatula. A total of 7 728 183 subreads and 509 014 full-length non-chimeric reads（FLNC）were obtained. By comparing the reference genomes of R108 and A17,94.36% and 93.01% isoforms were identified as mapped reads,respectively. A total of 8 406 alternative splicing events was identified and the majority was intron retention（RI）. The 23 926 genes were detected,of 12 049 genes had 295 545 transcripts,in which at least there was one poly（A）site. In addition,3 223 transcription factors,6 595 long non-coding RNAs（lncRNA）and 479 fusion transcripts were identified. The results showed the feasibility of deep sequencing M. truncatula transcriptome by SMRT,which provides data supplement for better utilizing the genome resource of M. truncatula. Table and Figures | Reference | Related Articles | Metrics

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Effects of High-dose Tannic Acid on the Intestinal Barrier Function and Gut Microbiota in Mice HE Ya-lun, ZENG Li-rong, LIU Xiong, ZHANG Ling, WANG Qiong Biotechnology Bulletin    2022, 38 (4): 278-287.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0860 Abstract （662）   HTML （16）    PDF（pc） （7293KB）（140）       Knowledge map    Save The objective of this study is to analyze the effects of high-dose tannic acid on the intestinal barrier and gut microbiota in normal-diet mice and high-fat diet-induced obese model mice. H&E staining,RT-qPCR,16S rRNA sequencing and other methods were used for detection and analysis. It was found that the intervention of high-dose tannic acid（400 mg/kg）reduced the body weights and food intakes of the mice,and increased the contents of various intestinal segments in the mice,among which the colon content significantly increased. In addition,high-dose tannic acid damaged the intestinal function and intestinal barrier,such as reduced the number of goblet cell and the length of crypt,and decreased intestinal tight junction protein（ZO-1,Occludin,and Claudin）expression. Moreover,oral administration of high-dose tannic acid changed the diversity of gut microbiota in the colon,and increased the abundance of SCFAs-producing bacteria Alistipes,Ruminococcus and Blautia,as well as obesity negatively related bacteria Alistipes and Oscillibacter. In conclusion,the results showed that the damage of intestinal mucosal barrier caused by high-dose tannic acid affected the digestion and absorption of food in mice,which might be the main potential reason for the rapid weight loss in the mice after the intervention of high-dose tannic acid. On the other hand,the changes of gut microbiota caused by high-dose tannic acid also had an effect on the body weights of the mice. Table and Figures | Reference | Related Articles | Metrics

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Rapid Crude Extraction of Genomic DNA from Solanum lycopersicum for PCR SHEN Heng, LIU Si-hui, LI yue, LI Jing-tao, LIANG Wen-xing Biotechnology Bulletin    2022, 38 (6): 74-80.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1123 Abstract （651）   HTML （39）    PDF（pc） （4414KB）（719）       Knowledge map    Save It is necessary to extract plant genomic DNA and perform PCR cloning when conducting the studies of molecular biology on plants. Currently,the CTAB（Cetyltrimethyl Ammonium Bromide）is commonly used to extract plant DNA in laboratories,but the steps are relatively cumbersome and require multiple organic solvents. Specifically,it takes a long time when there are many samples. The DNAs from multi-species were extracted by NaOH,and this study introduces a method of extracting plant genome DNA,which is faster,economical,safer and more efficient. The major steps include the followings：Quickly grinding the plant tissues using liquid nitrogen;quickly extracting DNA by alkaline lysis（0.5 mol/L NaOH）;neutralizing the extracted DNA sample with TE buffer;and performing PCR with neutralized DNA as template. Using this simple DNA extraction method and combining PCR detection,the rapid preliminary identification of transgenic S. lycopersicum was completed. Furthermore,it was confirmed that this NaOH method could be used for extracting DNA from the different tissues of S. lycopersicum. The entire extraction process was simple and of short-time. Thereby,DNA extraction for a large number of plant samples could be completed in a short time without using toxic regents such as chloroform and isopropanol etc. The completion time of DNA extraction is about 1/5 of the time while using CTAB method. In addition,the extracted DNA from different tissues are of good integrity and high quality,which meet regular PCR detection,which can be used for gene cloning and for transgenic tomato screening. This method demonstrates potential utilization value and prospects. Table and Figures | Reference | Related Articles | Metrics

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Recent Advances in CRISPR/Cas-based DNA Base Editing LAI Xin-tong, WANG Ke-lan, YOU Yu-xin, TAN Jun-jie Biotechnology Bulletin    2022, 38 (6): 1-12.   DOI: 10.13560/j.cnki.biotech.bull.1985.2022-0311 Abstract （650）   HTML （49）    PDF（pc） （1977KB）（777）       Knowledge map    Save CRISPR/Cas-based genome editing has revolutionized biological technology,which is achieved by introducing DSB（double strand break）in target DNA sites followed by cellular DNA repair via non-homologous end joining（NHEJ）or homology-directed repair（HDR）. NHEJ is the main repair pathway in most of cells,it characterized as repair is simple,efficiency is high and it is extremely easy to have errors,and thus presents randomly and unpredictable insertions and deletions（indels）. Point mutations represent the most prevalent form of genetic mutations,which determine the many important agronomic traits in crop plants and cause most of known human genetic diseases. Base editing enables targeted single base substitutions of high efficiency and predicted editing outcomes with neither inducing DSBs in the genomes nor requiring donor templates. Based on its great applicable potentials in the gene therapy,crop breeding as well as basic research,base editing rapidly gains much attentions with numerous applications in agricultural as well as biological fields and considerable progresses having been made to broaden its capabilities. In this review,we summarize the current advances in DNA base editing,focus on improvements in editing efficiency,precision,specificity as well as editing scope,and discuss current limitations and future directions. Table and Figures | Reference | Related Articles | Metrics

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Distribution and Function of Kazachstania Yeast in the Fermentation of Strong Flavor Baijiu YOU Ling, ZHOU Rong-qing, TAN Yi, WANG Tao, QIAO Zong-wei, ZHAO Dong Biotechnology Bulletin    2021, 37 (6): 108-116.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-1159 Abstract （650）   HTML （11）    PDF（pc） （3196KB）（1009）       Knowledge map    Save This work aims to investigate the growth characteristics and function of Kazachstania yeast in fermentation of strong flavor Baijiu. The 52 OTUs belonging to Kazachstania turicensis and 7 OTUs belonging to Kazachstania humilis were detected by high-throughput sequencing in the fermentation process of strong flavor Baijiu “Wuliangye”,and 63 K. turicensis strains and 30 K. humilis strains were isolated and identified for the study of their characteristics in solid state fermentation. It was revealed that K. turicensis and K. humilis were mainly active in the main fermentation period of 0-24 d and K. turicensis was the major dominant yeast in the main fermentation stage of strong flavor Baijiu “Wuliangye. K. turicensis produced more organic acid than K. humilis;while the difference in ethanol production and sugar utilization between K. turicensis and K. humilis was at the strain level. The average ethanol yield by K. turicensis strains was 4.94 mL/100 g and that by K. humilis strains was 5.67 mL/100 g,1 strain of K. humilis produced 11.11 mL/100 g. K. turicensis and K. humilis mainly produced the favor substances of isoamyl alcohol,isoamyl acetate,furfuryl alcohol,β-phenylethanol,and higher fatty acid ethyl ester in the fermentation of grains. The yield of isoamyl alcohol,furfuryl alcohol and β-phenylethanol by K. humilis strains was significantly higher than that by K. turiciensis strains,while the yield of acetic acid and ethyl ester of higher fatty acids above C10 were significantly lower than that by K. turiensis. The results indicated that Kazachstania yeast could be regarded as one of the landmark species of micro environment in the fermentation of strong flavor Baijiu,and have important impacts on the flavor of strong flavor Baijiu. Table and Figures | Reference | Related Articles | Metrics

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Role of WRKY Transcription Factor in Plant Response to Stresses ZHANG Tong, LI Zhi-qiang, WU Guo-qiang Biotechnology Bulletin    2021, 37 (10): 203-215.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-1481 Abstract （649）   HTML （36）    PDF（pc） （1136KB）（637）       Knowledge map    Save WRKY transcription factors（TFs）are one of the most important transcription factor families in plants. It contains a conserved WRKY domain composed of 60 amino acids. The core sequence WRKYGQK of the domain can specifically bind to W-box[（T）（T）TGAC（C/T）]in the promoter region of target genes,thus regulating the expressions of regulatory genes and/or functional genes containing W-box elements in the promoters. When plants are exposed to stresses,WRKY transcription factors are involved in the regulation of the related transcription reprogramming,and play an important role in plant growth and environmental adaptation. This paper reviews the discovery,structural characteristics,classification,regulation,and expression patterns of WRKY transcription factors,as well as the research progress in plant response to abiotic（such as drought,high temperature,low temperature,high salinity,nutrient deficiency and so on）and biological（such as disease and pest）stresses in recent years,and prospects the future research direction. Table and Figures | Reference | Related Articles | Metrics

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Identification of Key Transcription Factors in Response to Salt Stress in Rice ZHANG Bin, YANG Xin-xia Biotechnology Bulletin    2022, 38 (3): 9-15.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0802 Abstract （648）   HTML （51）    PDF（pc） （5148KB）（507）       Knowledge map    Save Salt stress is one of the most important abiotic stresses affecting rice production. In this study,the transcriptome data of rice under salt stress were used to analyze the expression changes of transcription factors at the transcriptome level by HTSeq and DESeq software. The main results were as follows：there were 26 identical transcription factors differentially expressed in rice at 1,3 and 6 h of salt stress,and an important module containing 14 genes was screened by protein interaction. GO analysis of important modules showed that 14 genes were mainly enriched in ATP binding,stress response and cytoplasmic membrane-bounded vesicle. KEGG analysis showed that it was mainly enriched in endoplasmic reticulum protein processing and endocytosis pathway. Meanwhile,the key heat shock transcription factor gene Os03g074500 and Os06g0553100 induced by salt stress were identified. It is speculated that these two genes may play a significant role in the process of rice salt stress by regulating the transcription and expression of HSP70 gene and affecting protein folding and assembly. This study provides candidate genes for subsequent genetic improvement of rice and provides scientific enlightenment for further understanding of salt tolerance mechanism. Table and Figures | Reference | Related Articles | Metrics

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Recent Advances on the Mechanism of Beneficial Microbial Fertilizers in Crops WU Qi-man, ZHANG Jin-mei, LI Yue-ying, ZHANG Ying Biotechnology Bulletin    2021, 37 (5): 221-230.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-0846 Abstract （640）   HTML （49）    PDF（pc） （1171KB）（782）       Knowledge map    Save Microbial fertilizer is a new type of environmentally friendly biological fertilizer,and it may promote plant growth,increase fruit quality,and improve soil quality. By building a fine connection between plants and microorganisms,the environment can be developed sustainably and “Circular economy” may be formed,which thereby effectively promotes the overall development of organic agriculture. This article reviews the roles of beneficial microbial fertilizers in improving soil physical and chemical properties,promoting plant growth and improving fruit quality,as well as increasing plant stress resistance and disease resistance. It also briefly introduces the molecular mechanisms of the interactions between microorganisms and plants. Meanwhile it prospects the application prospects in organic agriculture. Table and Figures | Reference | Related Articles | Metrics

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Establishing Tobacco Rattle Virus-mediated Gene Silencing System for Primula forbesii FU Si-tong, SI Wei-jia, LIU Ying, CHENG Tang-ren, WANG Jia, ZHANG Qi-xiang, PAN Hui-tang Biotechnology Bulletin    2022, 38 (4): 295-302.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-0924 Abstract （639）   HTML （23）    PDF（pc） （4986KB）（247）       Knowledge map    Save Primula forbesii is a biennial herbaceous flower,which has high ornamental value as a landscape plant. It is also an ideal material to study heterostyly. In this study,a rapid and efficient verification technology of gene function is established for P. forbesii laying a foundation for studying the function of genes in P. forbesii. The optimal infection object,infection solution formula,bacterial solution concentration and infection mode of tobacco rattle virus（TRV）vector in P. forbesii were explored,and the VIGS system suitable for P. forbesii was established. The results showed that the OD600 value of bacterial solution containing TRV1 and PfPDS-TRV2 was adjusted to 1.0 in the infection solution of 200 μmol/L AS,10 mmol/L MgCl2 and 10 mmol/L MES. After mixing,P. forbesii was infected by abaxial leaf injection. The treated plant leaves were used in PCR with primers on TRV virus vector,and the virus vectors of TRV1 and TRV2 were detected in the plants with phenotypic changes and the no-load group. The expression of PfPDS in albino plants was significantly lower than that in the no-load group and the control group. The infection efficiency of the established VIGS system was 60%,the silencing phenotype lasted for 12 months,and could play a silencing role from leaf to sepal. Due to the long duration of silencing effect,all genes can be verified by this method. Table and Figures | Reference | Related Articles | Metrics

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On the Function of Plant Cysteine Protease in Plant Growth and Development MO Li-jie, LIU Xia-tong, LI Hui, LU Hai Biotechnology Bulletin    2021, 37 (6): 202-212.   DOI: 10.13560/j.cnki.biotech.bull.1985.2020-1220 Abstract （629）   HTML （25）    PDF（pc） （1149KB）（402）       Knowledge map    Save Cysteine proteases are an important class of proteolytic enzymes and play an important role in regulating plant growth and development. Based on the expression patterns of different cysteine proteases in different tissues and organs,this article reviews the functional research and progress of cysteine proteases involved in seed germination,root development,leaf senescence,programmed cell death of tracheary elements in stems and of tapetum in anthers,aiming to provide references for further studying the function of cysteine protease. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in Plant Flavanone-3-hydroxylase Gene DUAN Yue-tong, WANG Peng-nian, ZHANG Chun-bao, LIN Chun-jing Biotechnology Bulletin    2022, 38 (6): 27-33.   DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1253 Abstract （626）   HTML （37）    PDF（pc） （1201KB）（341）       Knowledge map    Save Flavanone 3-hydroxylase（F3H）,as a central enzyme for plants to enter the branch of different flavonoid metabolites,can generate dihydroflavonol,a common precursor substance for the synthesis of anthocyanins and flavonols,and it plays a very important regulatory role in the synthesis of flavonoids. From the discovery,structure,function,and expression regulation of the F3H gene,this article reviews the research progress and regulation network of F3H gene in regulating the synthesis of anthocyanins and flavonols,and prospects the future direction. It is aimed to provide a reference for understanding the regulation mechanism of F3H gene on the metabolic synthesis pathway of plant flavonoids,and also help researchers to use F3H with genetic engineering method to obtain the new plant germplasm via directive breeding. Reference | Related Articles | Metrics